Abstracts (sorted by last name of presenter)
Nichole Abrego, St. Edward’s University – 11:15-11:30 in Boax Session
Determining if extracts of Eysenhardtia polystachya, Pinus maritima, and Zingiber officinale can combat antimicrobial resistance.
Nichole Abrego and Patricia Baynham.
Antimicrobial resistance (AMR) causes more than 35,000 deaths annually in the United States, so new treatments are needed. In this study, parts of three different plants were examined for their antibacterial activity: the bark of Eysenhardtia Polystachya (palo azul) and Pinus maritima (pine) ,and the root of Zingiber officinale (ginger). The barks were extracted and were then tested for antimicrobial activity using a Kirby-Bauer disk diffusion assay against E. coli lptD4213-. While the ginger extract displayed no zone of inhibition, the average zone of inhibition was 13mm for pine bark and 11mm for palo azul. A minimum inhibitory concentration assay (MIC) was performed, to better quantify the antimicrobial activity and showed that pine bark had an MIC of 640ug/ml while the MIC of palo azul was 600ug/ml.. To investigate the mechanism of action (MOA) of the extracts, bacterial cytological profiling (BCP) was performed. BCP involves treating the bacterium with an antimicrobial substance and then visualizing the changes in shape that are different based on which part of the cell is targeted. Visualization was via confocal microscopy using FM4-64 to show the membranes and DAPI to see the DNA. The treated cells changed shape but not all cells were visualized so this will require further analysis. In the future, the cell shapes can be quantified and compared with the known MOA of known antibiotics. New treatments are crucial when AMR infections are increasing. Plant substances can be characterized and developed into possible alternative treatments to AMR microbes.
Research Advisor: Dr. Patricia Baynham
Ryan Agyemang and Altherr Joseph Alcuitas, Southwestern Oklahoma State University – 10:45-11:00 in Boax Session
The effect of female mate choice on offspring sex ratios in a freshwater amphipod species.
Ryan Agyemang and Altherr (Joseph) Alcuitas.
This experiment highlights the effect of female mate choice on offspring sex ratios in a freshwater amphipod species. There are several determinants of sex ratios in nature all falling under two main categories being genetic or environmental. The expectation from theory is that the sexes will be produced in a 1:1 ratio. However, biased sex ratios are common in nature and can be the result of both genetic and environmental factors. We tested whether females bias the sex ratio of broods in favor of sons when mating with a high quality male. We collected mating pairs of amphipods (Hyalella sp.) from two pools in the same natural spring system. Half of the females were separated and repaired with their original mate—i.e. they were allowed to choose their mate. The other half of the females were separated and assigned a new male at random. Each pair was placed in their own 50-ml jar and placed in an environmental chamber. We are checking the pairs three times a week and recording offspring sex ratios. We predict that females with a choice in their mate will pick higher quality males and in turn produce more male biased broods and females assigned a mate at random will produce more of an even brood sex ratio. We also predict that the choice pairings will have less variation in offspring sex ratios as mates would be all of high quality. On the contrary, random pairings will have a much higher variation in offspring ratios as some males assigned will be higher quality and some males assigned will be poor quality.
Research Advisor: Dr. Ricky Cothran
Sara Ambrocio Paque, University of the Ozarks – 9:45-10:00 in Boax Session
Comparison of eukaryotic soil algae in Warren Prairie Natural Area sodic saline slicks.
Sara Ambrocio Paque, Marvin W. Fawley, and Karen P. Fawley.
Warren Prairie Natural Area (WPNA) in Bradley and Drew Counties, Arkansas, is a mosaic area of saline slicks that form flat, crusty depressions in a central area with a zone of lichens and a few rare angiosperms, and an outer zone of cyanobacterial mats. The edges of the saline slicks are home to the rare, diminutive vascular plant, Geocarpon minimum Mackenzie (Caryophyllaceae), which is a federally protected threatened species. Because the Warren Prairie slicks are home to many rare and unusual vascular plants, we hypothesized that the soil algae community will also comprise many unusual species. The main objective of this study was to characterize and compare the soil crust eukaryotic algal communities in Site 1 (saline slick) and Site 2 (soil not within the sodic saline slick) in WPNA using morphological and molecular techniques. We focused on the 10 species of the green algal genera Coelastrella, Cylindrocystis, Myrmecia, and Diplosphaera that were previously isolated from a saline slick in WPNA. We used light microscopy for morphological characterization of each strain and DNA sequences (barcodes) from loci appropriate for species-level identification (tufA; ITS; rbcL). Coelastrella strains from Site 1 were identical to a strain previously described from a slick in WPNA. In contrast, Site 2 had a unique strain similar to Coelastrella yingshanensis. Cylindrocystis strains were present in the Site 1, but were not present in Site 2. Myrmecia was not found from Site 1 or Site 2, whereas Diplosphaera strains were found in both sites. Our results show that most of the species of green algal genera examined were different between the soil from saline slicks and soil outside the slick.
Research Advisor: Dr. Karen P. Fawley
Bryler Atchley, Southwestern Oklahoma State University – 1:30-3:30 in Serpent Session
Syringafactin Increases Membrane Permeability in Gram-Negative Bacteria.
Bryler Atchley and Regina McGrane.
Pseudomonas syringae is a gram-negative plant pathogen that produces a biosurfactant called syringafactin. Syringafactin, a lipo-octapeptide, has been shown to act as an antagonist molecule against competing gram-negative bacteria such as Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa. The primary antagonistic properties include induction of negative chemotaxis and cell death in competing bacteria. We hypothesized that cell death in competing cells is the result of increased membrane permeability. This work aimed to quantify the impact of syringafactin on the membrane permeability of E. coli, S. typhimurium, and P. aeruginosa. Fluorescence was detected using DiS-C2(5), a fluorescent dye sensitive to fluctuations in membrane polarity. Data was collected using a multi-mode plate reader for four hours following exposure to increasing concentrations of crude syringafactin extract. Additionally, extract free samples and samples containing crude extracts from a mutant lacking syringafactin production were tested. Samples showed increasing fluorescence correlating to increasing concentrations of syringafactin, and extract free samples lacked fluorescence. Samples exposed to extracts from a mutant lacking syringafactin production exhibited some fluorescence, but the fluorescence was significantly lower than samples exposed to syringafactin extracts. To confirm that the major difference between the two types of crude extracts was the presence or absence of syringafactin, purified samples were analyzed using mass spectroscopy and liquid chromatography. These procedures evaluate the peptide and lipid composition of the extract. Analysis demonstrated that the six lipo-octopeptides previously characterized as syringafactin were present in high concentrations in extracts from wild-type bacteria, but they were absent or in very low concentrations in mutants lacking syringafactin production. Collectively, these studies suggest that syringafactin in crude extracts is responsible for reducing cell viability by increasing membrane permeability. Identifying the mechanism responsible for syringafactin induced cell death leads to a deeper understanding of the potential effectiveness of syringafactin as an antibacterial agent.
Research Advisor: Dr. Regina McGrane
Nicholas Bauer, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
High Resolution Crystal Structure of Ro0101, a Dodecameric Dps-like Protein from Rhodococcus jostii.
Nicholas Bauer, Abigail Trejo, Leonard Thomas, Rakhi Rajan and Vijayakumar Somalinga.
Dps (DNA binding protein from starved cells) and related ferritin proteins are ubiquitous stress proteins expressed in eukaryotes and prokaryotes. Dps proteins are integral in stress tolerance and for DNA binding and protection. Furthermore, Dps and related ferritin proteins have been tested in bionanotechnological applications including nanocages, biocontrast agents, and in drug delivery. We have identified a Dps homolog, Ro00101 referred as Dps1, from Rhodococcus jostii, a soil-dwelling bacteria with unparalleled metabolic potential. The dps1 gene encodes a 19.4 kDa protein with conserved domains typical of DNA-binding proteins with ferroxidase activity. The goal of this study is to characterize Rhodococcal Dps protein using biochemical and structural techniques. Cloning and over-expression were done using standard techniques. Protein purification was done using immobilized metal affinity chromatography. The purified protein was concentrated to 17 mg/ml before crystallization trials. Preliminary 96-well sitting-drop vapor diffusion crystallization revealed crystals in drops containing 0.1 M sodium acetate trihydrate pH 4.5. and 2M ammonium sulfate pH 4.5. A single, rhomboidal crystal obtained using hanging-drop method was used for data collection and diffracted at 1.85 Angstrom. The structure for Dps1 was solved by molecular replacement using Mycobacterium smegmatis Dps1 (PDB ID: 1VEI) as a model. The structure of Dps1 revealed a dodecameric protein with conserved ferritin-like and Dps-like trimeric interfaces involved in iron channeling and protein folding respectively. The dimeric interface that harbors the iron-binding active site is also conserved. The active site of Dps1 reveals twin histidine residues from one monomer and glutamine/aspartic acid residues from the second monomer contributing to the two ferroxidase sites found in the dimeric interface. Interestingly, Dps1 crystallized without the bound ligand. Studies are underway to obtain ligand-bound structures and to determine the ferroxidase and DNA-binding properties of Dps1.
Research Advisor: Dr. Vijay Somalinga
Benjamin Berggren, Austin College – 10:00-10:15 in Boax Session
Detection of Chytrid Fungus via Environmental DNA in North Texas.
Benjamin Berggren, Loriann Garcia, Frank Goodavish, and Keegan Nichols.
EDNA, or environmental DNA, is DNA shed by organisms in their environment. Ecologists are beginning to use this DNA shed into the environment to detect elusive species, track disease progression, or perform general biodiversity surveys. EDNA detection has been used in many different projects, and the methods can be adjusted to fit the needs of the study. In the summer of 2021, we extracted eDNA from soil and water samples from two sites in Grayson, Co. TX to detect the presence of Blanchard’s Cricket frog and the Texas Smallmouth salamander. We successfully detected Blanchard’s Cricket frog. This fall and spring, I used eDNA to attempt to detect the presence or absence of the amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd). I sampled two sites in Grayson Co. which border counties in Oklahoma where chytrid has been detected. Detecting this fungus as early as possible will allow ecologists to mitigate the effects on larger amphibian populations. Our tests so far have not detected Bd from our samples, but our test results show an error in our methods. To rectify this, we will update our methods, continue to sample, and test, and send our current isolated eDNA to be sequenced.
Research Advisor: Dr. Loriann Garcia
Sebastian Calderon, Houston Baptist University – 9:15-9:30 in Boax Session
Determining the Presence of E. coli 0157:H7 in Bovine Population.
Sebastian Calderon.
Shiga toxin producing E. coli are derived from the guts of most cattle. The 0157:H7 strain of E. coli is the most prominent strain that infects humans, and it can cause severe damage to the lining of one's intestines and kidneys. The ideal situation is to attack it at the source. Vaccines that reduce the amount of bacterial shedding exist, but the beef industry has no incentive to use them. Thus, the research aims to determine the prevalence of E. coli 0157 in bovine feces. Through the process of isolating the E. coli, extracting genomic DNA, and PCR there was data determining that there is a relatively large amount of this strain of E. coli within cattle.
Research Advisor: Dr. Jacqueline Peltier Horn
Eduardo Carrillo, St. Edward’s University – 9:30-9:45 in Balanos Session
Defects in autophagy flux related to microtubule associated protein UNC-33 mediate changes in basal and starvation-induced autophagy.
Eduardo Carrillo, Alexa Ott, Emily Holechek, Arianna Vacio, and Andrea Holgado.
Autophagy is the process in which cellular components are degraded and recycled. Regulation of autophagy is essential for axonal development and maintenance of synapses. UNC-33/CRMP-2 is a microtubule associated protein (MAP) that contributes to neuronal development and transport of autophagosomes. UNC-33 is part of a ternary complex that anchors microtubule bundles to the cortex and inhibits microtubule sliding. Problems with microtubule sliding, and cargo transport and degradation in unc-33 mutants may contribute to the development of neurodegenerative diseases. Our laboratory studies autophagy via cleavage of the double fluorescently tagged protein CERULEAN-VENUS LGG-1(dFP) in C. elegans unc-33 mutants. We predict that lack of UNC-33 will result in defects in autophagy. To test this hypothesis we monitored autophagosome maturation and pathway flux using western blots and the levels of dFP(MW 75) versus the cleaved monomeric fluorescent protein (mFP)(25 kDa). In these experiments, we induced autophagy via starvation and compared the results against basal autophagy. Analysis of western blot results show that unc-33 mutants undergoing induced autophagy have an accumulation of the cleaved mFP. Conversely in basal autophagy, the accumulation of mFP was decreased.These results suggest that UNC-33 may contribute to autophagy flux and clearance of cargo. Microtubule sliding, a phenomenon seen in unc-33 mutants, may affect autophagosome transport and cargo degradation. These findings support a role of UNC-33 in autophagy in neurons.
Research Advisor: Dr. Andrea Holgado
Bethany Chapman, Angelo State University – 1:30-3:30 in Bird Session
An Observational Study of Mast Cells in the Mephitidae Family.
Bethany Chapman and Laurel Fohn.
Mast cells are important effector cells in the immune system of vertebrates. While mast cells, or mastocytes, are primarily associated with allergic reactions and anaphylaxis, they are also involved in defense against pathogens and immune tolerance. These cells are found in connective tissues, especially that of the skin, lungs, and intestines. The dysregulation of these cells can lead to mast cell tumors, a common type of tumor in household pets. Though these cells and tumors are commonly observed in cats and dogs, they have yet to be studied in many other mammals. The observation of mastocytes in other mammals may give us clues as to how they function in wild animals as compared with domesticated animals. This study seeks to observe mast cells and their prevalence within skin tissue samples of the skunk family, Mephitidae. Tissues will be formalin fixed and paraffin embedded before being stained with hematoxylin and eosin as well as toluidine blue for better identification of mast cells.
Research Advisor: Dr. Laurel Fohn
Mary Cooperrider, Oral Roberts University – 9:30-9:45 in Boax Session
A Preliminary Analysis of Full Spectrum Hemp Oil in Triple-Negative Mammary Epithelia.
Mary Claire Cooperrider and William Ranahan.
Cancer is one of the leading causes of death in the world and there are limited options for noninvasive, therapeutic strategies. In recent years, more individuals have opted to use Cannabis sativa, hemp, to soothe their ailments ranging from pain to epilepsy. The common misconception surrounding hemp is that it is identical to marijuana. A hemp plant contains a markedly lower percentage of tetrahydrocannabinol (THC) than a marijuana plant. Hemp oils and extracts are widely available for purchase in the United States, and researchers have yet to fully document the capabilities and treatment implications of hemp extracts. It is known that hemp reduces inflammation in the body, indicating it as a potential method for targeting cancer cells because inflammation promotes the proliferation of tumors and sustainability of tumor microenvironments. Full spectrum hemp extracts contain various compounds with possible therapeutic capabilities to be used in targeting tumors. Researchers know of many genes that may be monitored for changes in expression in order to observe efficacy of treatment. The current study aimed to analyze the variations in gene expression in a triple-negative breast cancer cell line, MDA-MB-468 cells, following the topical application of full spectrum hemp oil. The gene expression levels of MDA-MB-468 cells that were treated with the drug were compared to expression levels of untreated MDA-MB-468 cells using a PrimePCR Assay. The data suggested that treatment with full spectrum hemp extract yielded upregulation in genes SUMO1 and DKK1 and downregulation in genes BMP2 and MCM2. This preliminary data suggests that multiple pathways involved in tumor survival and progression are affected by components of full spectrum hemp extract. Further studies will include repeated analysis of gene expression and cell viability, as well as gene-specific analysis to observe individual alterations in cell mechanisms of cancer cells.
Research Advisor: Dr. William Ranahan
Larry Cossey, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
Detecting Variations in Microbial Concentrations in Oklahoma Lakes.
Larry Cossey and Regina McGrane.
The microbial composition of water can be the difference between life or death for many organisms. Microbes in lakes can reproduce and spread to other parts of the water cycle. Ice nucleating Pseudomonads are microbes that have been shown to be in all parts of the water cycle and damage plants. These Pseudomonads conduct ice nucleation by inducing freezing of supercooled water molecules. Through evaporation and rainfall, Pseudomonads can disperse anywhere. When bacterial ice nucleation occurs around plant tissue, ice crystals form within the tissue and damages the plant. To examine microbial composition, two Oklahoma lakes: the urban lake---Lake Hefner and the rural lake--Crowder Lake, were examined. In each lake, water samples were collected from three specific locations in summer and winter months. Samples were inoculated on 10% tryptic soy agar to determine total bacterial load and King’s B agar supplemented with boric acid, cycloheximide, and cephalexin to isolate Pseudomonads. At Lake Hefner, the eastern shore had significantly larger bacterial concentration in both seasons compared to other locations. At Crowder Lake, the center of the lake had significantly lower bacterial concentration in both seasons compared to other locations. Seasonally, there was lower bacterial concentration in the winter at Lake Hefner compared to the summer, while Crowder Lake had similar bacterial concentration year-round. Lake Hefner had higher bacterial concentration compared to Crowder Lake. Pseudomonads isolated from both seasons and lakes were tested for ice nucleation activity by inoculating in super cooled water and detecting ice formation. Twenty-two ice nucleators were detected at Crowder Lake and three ice nucleators were detected at Lake Hefner in the summer, and none were detected in the winter. The seasonal differences in the concentration of ice nucleating bacteria were surprising because other studies have shown that cooler environments select for increased ice nucleating bacteria.
Research Advisor: Dr. Regina McGrane
Alejandra Cristancho, St. Edward’s University – 1:30-3:30 in Serpent Session
Does the extension of lifespan equate to improved quality of life?
Alejandra Cristancho, Alexia Samaro, Ruby Valtierra, Skye Beck, Maria Miranda, and Andrea Holgado.
Collapsin response mediator protein-2 (CRMP-2) in humans (UNC-33 in C. elegans) mediates axonal outgrowth, maintenance, and degeneration. UNC-33/CRMP-2 has been hypothesized as a potential drug target for the treatment of Alzheimer’s and other neurodegenerative diseases. In aging, CRMP-2 becomes hyperphosphorylated, destabilizing the cellular skeleton and leading to neurodegeneration. In C. elegans, aging can be slowed by entering dauer diapause; a non-aging developmental stage turned on when the DAF-7/TGFβ signaling pathway is silenced in response to environmental stress. In our laboratory, we discovered that unc-33 mutants cannot form dauers in response to environmental stress. This study investigates whether a mutation in the daf-7 gene can rescue phenotype characteristic of unc-33 mutants. To this end, we created unc-33; daf-7 double mutants and quantified dauer formation, axonal elongation, and locomotion after exposure to unfavorable conditions. Our analysis of unc-33; daf-7 double mutants showed that introducing the daf-7 mutation into an unc-33 mutant rescued dauer formation. Studies of the rescue of axonal elongation and locomotion are currently underway, and results will be presented at the symposium. So far, these findings suggest that unc-33 mutants may have a problem suppressing DAF-7 signaling, leading to the continuation of reproductive programs even under unfavorable conditions. Furthermore, these results point out a role of CRMP2/UNC-33 and DAF-7 in the extension of the lifespan.
Research Advisor: Dr. Andrea Holgado
Lily Ellzey, Angelo State University – 1:30-3:30 in Bird Session
Detection and Identification of Coronavirus Strains in Myotis velifer.
Lily Ellzey and Laurel Fohn.
SARS-CoV-2 is a member of the family of viruses known as the coronaviruses. This family of viruses includes two subfamilies of viruses. Of these, the subfamily Coronavirinae contains four genera of viruses, two of which are known to infect humans. Bats are a natural reservoir for many kinds of coronaviruses, but bats collected in West Texas have not yet been analyzed to determine which strains of coronaviruses these species of bats carry. RNA was extracted from the Angelo State Natural History Collection gastrointestinal tissue samples of bats from West Texas and subjected to RT-PCR to assess for the presence or absence of coronavirus. If present, amplicons will be submitted for sequencing and coronavirus strain identification.
Research Advisor: Dr. Laurel Fohn
Payden Farnsley, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
Development of Isolation Protocols to Study the Distribution of Pseudomonas syringae in Western Oklahoma.
Payden Farnsley and Regina McGrane.
Pseudomonas syringae is a plant pathogen that causes significant economic losses due to reduced crop yield caused by disease. Studies have shown that P. syringae is globally distributed and is linked to the water cycle due to its ability to induce ice nucleation, the crystallization of supercooled water. Our goal is to better understand how this plant pathogen affects western Oklahoma by studying its distribution in non-agricultural systems. If P. syringae is present in samples, it will help us predict outbreaks that might affect Oklahoma crops. To study the distribution of P. syringae on wild plants, we are currently developing protocols for isolation. To evaluate the validity of these protocols, we collected leaf samples at a local state park from two different species of wild plant in two different areas of the park. Leaf samples were suspended in washing buffer and sonicated the sample to break up bacterial aggregates. Samples were then inoculated on 10% tryptic soy agar to determine total microbial load and King B’s agar supplemented with boric acid, cycloheximide, and cephalexin, which is a medium selective for P. syringae. Potential P. syringae isolates were then subcultured on tryptic soy agar for storage. Isolates were then evaluated for oxidases, which P. syringae does not produces. To test isolates for ice nucleation isolates were inoculated in super cooled water and observed for induction of freezing. We have stored thirteen isolates, which each tested negative for oxidase activity and positive for ice nucleation activity because they are likely P. syringae strains. This bacterium produces a biosurfactant – syringafactin – and we plan to test for production of this biosurfactant to further characterize their identity. We also plan to perform 16s rRNA sequencing to confirm their identity.
Research Advisor: Dr. Regina McGrane
Emily Foss, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
Identification and Characterization of Bacterial Carbonic Anhydrases for Enzyme Accelerated Carbon Capture and Sequestration.
Emily Foss, Amy Grunden, and Vijayakumar Somalinga.
Carbon capture and sequestration (CCS) is one of the numerous mitigation efforts that are being implemented to minimize the amount CO2 being dumped into earth’s atmosphere. Current CCS technologies aimed at removing CO2 from industrial gases rely on energy intensive scrubbing or amine-based CCS processes. Enzyme based CCS technologies have recently been used to improve industrial CCS, although enzyme stability in solvents and at high temperature has impeded the large-scale implementation of this technology. Carbonic anhydrases (CA) are ubiquitous enzymes that catalyze the interconversion of CO2 to bicarbonate. The catalytic activity of CA’s can be exploited in mineralization of CO2 in the form of carbonates, a well-known and a safe CO2 disposal method. We have previously identified and characterized the structure of an a-CA from Photobacterium profundum and recently identified several bacterial CA’s including an active b-CA from Streptococcus sanguinis. The goal of this study is to elucidate the biochemical characteristics of unique CA’s for use in CCS technologies. Using in-gel carbonic anhydrase activity assay (protonography) we have previously shown that a-CA is capable of CO2 hydration activity and in this study, using protonography, we confirmed that b-CA from S. sanguinis is also capable of CO2 hydration activity. Further biochemical analysis of a-CA revealed a bimodal pH maxima with peak activity observed at acidic (pH 5.0) and alkaline (pH 11) pH. a-CA enzyme activity dramatically declined at temperatures above 30oC, while maximal activity was observed at lower temperatures (4 to 30oC). A unique redox dependent activity was observed for a-CA which has not been previously reported in a-CA’s. Finally, biomineralization assay revealed that a-CA accelerates CO2 sequestration into carbonates under alkaline condition. Our preliminary biochemical characterization indicates that a-CA possess several unique biochemical characteristics and can be exploited for use in CCS technologies.
Research Advisor: Dr. Vijay Somalinga
Ella Fotinos, St. Edward’s University – 9:45-10:00 in Balanos Session
Using lipopolysaccharides to evaluate the effect of energetic stress on Italian honeybee immune response.
Ella Fotinos.
Colony collapse disorder (CCD) is a phenomenon in which colonies of honeybees rapidly lose all of their workers and consequently become nonfunctional. Due to honeybee use in agricultural and ecological pollination services, our ability to combat CCD has real world consequences. And, because CCD has been linked to pathogens, it is essential to better understand how honeybees respond immunologically to pathogenic threats to help inform best practices for minimizing the impact of CCD. Our study focused on the immune response of Italian honeybees to the introduction of lipopolysaccharides (LPS) to emulate a pathogenic threat. We examined bees from one hive containing frames with a plastic foundation and another containing no foundation in order to evaluate if energetic expenditure associated with honeycomb production has an effect on immune responses. These two forms of colony structure are common in beekeeping operations to either maximize honey versus honeycomb production respectively. After feeding specimens a sugar water solution containing 1mg/ml of LPS (Escherichia coli R515), we evaluated their immune response using a several colorimetric immunoassays at staggered feeding timepoints (0, 1, or 2 hours after ingestion). Bees with no foundation had significantly higher protein concentration in their hemolymph than those with a foundation, and there was trend of protein concentration spiking in the no foundation treatment two hours post-ingestion. Prophenoloxidase immune assays are currently ongoing. Our results indicate that honeybees used for honeycomb production may be more energetically stressed and thus increase their circulating protein concentration to preventatively prepare for combating pathogenic threats.
Research Advisor: Dr. Matt Steffenson
Annabelle Hawkins, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
Investigating impacts of syringafactin on bacterial chemotaxis.
Annabelle Hawkins and Regina McGrane.
Pseudomonas syringae is a bacterium that infects plants. Our laboratory demonstrated that syringafactin, a biosurfactant produced by P. syringae, has antimicrobial properties and repels human pathogens on semi-solid surfaces. This suggests syringafactin may be useful in controlling the growth of pathogenic bacteria. The purpose of this study was to determine whether syringafactin acts as a repellant of bacteria in liquid suspensions and identify potential Pseudomonas aeruginosa receptors involved in syringafactin detection. To study if syringafactin works as a repellant in liquid, normalized cultures of Escherichia coli, Salmonella enteritidis, and P. aeruginosa were exposed to suspended pipette tips containing syringafactin or sterile deionized water for 15 min. The contents of the pipette tips were then spread onto agar media, and colonies were counted to quantify bacteria that moved into the pipette tip. Samples from tips containing syringafactin were expected to have lower bacterial concentrations compared to samples from tips containing water. Surprisingly, our studies showed that samples with syringafactin were not significantly different from those with water. Therefore, we concluded that syringafactin does not work as a chemorepellant in liquids. To determine the chemoreceptors responsible for P. aeruginosa detection of syringafactin, we exposed P. aeruginosa mutants with disruptions in genes encoding for putative chemoreceptors to wild-type P. syringae on soft-agar media. The ability of the mutants to respond to and move away from P. syringae was observed. Mutants unable to move away from P. syringae on soft agar media likely have disruptions in syringafactin specific chemoreceptors. Collectively, this work is the first to demonstrate that syringafactin does not impact chemotaxis in liquids and attempt to identify chemoreceptors involved in detecting syringafactin.
Research Advisor: Dr. Regina McGrane
Makayla Hicks, Southwestern Oklahoma State University – 9:15-9:30 in Balanos Session
Evaluating pathogenicity of Pseudomonas aeruginosa through assessment of motility and biofilm mutants.
Makayla Hicks and Regina McGrane.
Pseudomonas aeruginosa is an opportunistic pathogen that causes nosocomial infections, particularly in the lungs of cystic fibrosis patients. Motility and biofilm formation are known to contribute to this bacterium’s ability to cause disease. These processes are modulated by many biological molecules including biosurfactants, the flagella, the pilus, and several two-component regulatory systems. In this research, we evaluated the biofilm formation and virulence of a variety of P. aeruginosa mutants with disruptions in genes encoding for biosurfactant (rhlA), pili (pilA), and flagella (fliC, motB, and motD) biosynthesis along with surface sensing (wspA). We hypothesize that deletion of these genes will cause this bacterium to be impaired in biofilm formation and pathogenicity. Biofilm production was evaluated using a crystal violet microtiter plate assay. We found that mutants lacking functional fliC, motB, motD, wspA, and pilA were all significantly impaired in biofilm formation. Virulence was tested by inoculating Galleria mellonella larvae, a wax moth that is used as a model organism for the understanding of bacterial pathogenicity, with mutant or wild type cultures. The survivability, coloration, movement, and silk production of the larvae were evaluated every day for up to seven days or until all the larvae died, whichever occurred first. Significant differences were detected between the wild type and mutants with deletions in rhlA, wspA, motB, and motD in silk cocoon formation and movement of the larvae. This suggests that these mutations alter the ability of P. aeruginosa to cause disease. It is likely that impaired virulence is directly related to the ability of mutants to produce biofilms. Understanding the role of biofilm formation in the pathogenicity of P. aeruginosa in wax moth larvae could lead to a better understanding of how this bacterium causes infection in humans and ultimately could lead to a solution for problematic infections.
Research Advisor: Dr. Regina McGrane
Taylor-Jade Higgins, Austin College – 11:00-11:15 in Balanos Session
Investigating the Role of Epithelial to Mesenchymal Transition on PA28γ Expression.
Taylor-Jade Higgins.
The 20S proteasome degrades and recycles proteins following activation by PA28γ, a ubiquitin- and ATP-independent proteasome activator. Several tumor tissues express elevated levels of PA28γ, and studies have identified that it also induces the epithelial to mesenchymal transition (EMT). With PA28γ and EMT playing a role in tumor development, the objective of this study was to identify whether PA28γ activity and expression are affected by EMT. EpH4 murine epithelial cells were treated with varying dosages and exposure times to Transforming Growth Factor β-1 (TGF-β1) to induce EMT, and the cells were characterized by being compared to normal EpH4 cells and 4T1 murine mammary carcinoma cells. RT-qPCR showed no statistically significant differences in the increased mRNA expression of EMT markers Snail and Slug, while mRNA expression of vimentin was not observed. Vimentin expression at the protein level was lowly detected by immunofluorescence, suggesting that the transformation was not completed in the TGF-β-treated EpH4 cells. Further analysis on the TGF-β treatment and the expression of PA28γ downstream targets – c-Myc, phosphorylated c-Myc (Thr58 and Ser62), and PP2A will determine what is preventing the EMT transformation from reaching its end-stage. While therapies currently exist to target EMT, revealing this blockage can help understand the relationship between PA28γ and EMT as well as to elucidate the mechanism by which PA28γ provides cancer cells the ability to acquire stemness through EMT.
Research Advisor: Dr. Lance Barton
Katie Holland, Angelo State University – 1:30-3:30 in Bird Session
Adenovirus screening in Texas bats.
Katie Holland and Loren K Ammerman.
Viruses in the family Adenoviridae (AdVs) are nonenveloped, double-stranded DNA viruses that infect a variety of hosts, including reptiles, birds, fish, and mammals. Viruses in the genus Mastadenovirus may cause respiratory, ocular, and gastrointestinal disease in mammals, including humans. It is, therefore, important to understand the distribution and transmission of adenoviruses in infected organisms. Bats have been found to serve as reservoirs in the evolution of adenoviruses due to bats’ atypical ability of harboring genetically diverse viruses within a single geographic location or host species. Adenovirus DNA has been detected at unknown prevalence in Cave Myotis (Myotis velifer) in Oklahoma, but adenoviruses have not yet been detected in Texas bats. Prevalence in European bats has been found to be around 8%, so we hypothesized the prevalence of adenoviruses in Texas bats to be low. We screened intestinal and fecal samples of M. velifer collected in 2018-2021 from Texas counties and found all samples to be negative for adenovirus DNA. The absence of detection supports the hypothesis of low presence of adenoviruses in Cave Myotis in Texas.
Research Advisor: Dr. Loren Ammerman
Saraya Hunt, Southwestern Oklahoma State University – 1:30-3:30 in Serpent Session
Microbial Analysis of Regolith-Grown Species (M.A.R.S.) on Mars.
Saraya Hunt, Lisa Castle, Emilee Adams, Molly Byers, Payden Farnsley, Lauren Hartsell, Kyla Langstraat, Chloe Mellott, Joseph Olonovich, Niki Strauch, and Rachel Uhlig.
As part of the on-going Plant Mars Challenge, SWOSU students are growing plants for a duration of ten weeks in simulated Mars regolith. While an official challenge goal is to have the most crop yield in 10 weeks, the SWOSU researchers are also investigating the interactions of plant diversity and soil microbe diversity and measuring chlorophyll content and diversity within microbial communities as dependent variables. This poster describes the experimental design and rationale behind the project, as well as potential implications of the results
Research Advisor: Dr. Regina McGrane
Sneha Jacob, Oral Roberts University – 1:30-3:30 in Serpent Session
Determining Optimal Extraction Conditions to Isolate a Peptide Containing Polysaccharide with anti-cancer properties from Ganoderma Lucidum.
Sneha Ann Jacob and William P Ranahan.
Ganoderma Lucidum, commonly known as Reishi, is a popular mushroom used in Asian countries for promotion of health and longevity. It was therefore called the “Mushroom of Immortality”. In ancient Chinese medicine, Ganoderma Lucidum has been used to treat various human diseases like bronchitis, chronic hepatitis, hyperglycemia, inflammation, and cancer. In our research, a peptide containing polysaccharide, Felix, isolated from Ganoderma Lucidum showed cytotoxicity in triple-negative breast cancer cells. However, because of the availability of Ganoderma Lucidum from different sources, the biological activity and concentration of the isolated compound (Felix) varied. To determine the optimal conditions, temperature, and time, needed to extract the compound of interest, we took extraction from Ganoderma Lucidum mycelia grown at six different conditions (varying temperature and time). It was determined that extraction at 37 degrees Celsius for 48 hours gave the highest concentration of Felix (peptide containing polysaccharide). It was also found that once the extraction is done on the mycelium plate, they are no longer alive. This means we need to have a constant supply of fresh mycelium each time for the extraction and isolation of Felix. The isolated Felix was then tested on triple-negative breast cancer cells for cytotoxicity. Further research should be done on finding the concentration of Felix needed to produce the cytotoxic effect and the mechanism pathway.
Research Advisor: Dr. Julianna Goelzer
Laura Matt, Texas Wesleyan University – 10:00-10:15 in Balanos Session
Antibacterial Properties of a Novel 1,2,4 Oxadiazole.
Laura L. Matt.
Antibiotics have revolutionized medicine for the last 100 years, but not without consequences. Over the last 100 years, bacteria have become resistant to common antibiotics, like penicillin, methicillin, and vancomycin. Oxadiazoles are a class of organic compounds that are known to possess antibiotic properties. The study of oxadiazoles is new, and not much is known about how they work as antibiotics, specifically against antibiotic resistant bacteria. In the summer of 2021, a 1,2,4-Oxadiazole compound was synthesized at Texas Wesleyan University. This compound has a guanidine functional group on carbon 3 and a 4-methoxybenzene on carbon 5, which contributes to making this compound an eligible antibiotic. Similar studies done at Texas Wesleyan University have shown that oxadiazole compounds have antibacterial properties toward gram-positive and gram-negative bacteria. Assuming this novel oxadiazole has antibacterial properties, this study is focused on quantitative results to determine the minimum inhibitory concentration of the compound. The minimum inhibitory concentration will give information about how little compound is needed to inhibit growth of the bacteria. These results are presented in growth curves. The antibacterial properties of the novel oxadiazole have been tested on Enterococcus faecalis, a gram-positive bacterium, and Escherichia coli, a gram-negative bacterium. In an ever-increasing era of antibiotic resistance, it is worth characterizing the antibacterial properties of the oxadiazole against gram-positive and gram-negative bacteria and considering their applications in modern medicine. Negative control assays have been done to observe the growth of each bacterium uninhibited by antibiotics. Positive control assays have been done to observe the growth (and lack thereof) of each bacterium under the influence of ampicillin, a cell wall biosynthesis attacking antibiotic, and chloramphenicol, a translation attacking antibiotic. The antibacterial properties are currently being characterized, but there is evidence that the novel 1,2,4-Oxadiazole does work as an antibiotic.
Research Advisor: Dr. Christopher Parker
Emily Penner, Oral Roberts University – 10:45-11:00 in Balanos Session
Bioassessment of Fred Creek and Haikey Creek, Tulsa, OK water quality using aquatic macroinvertebrate diversity.
Emily Penner.
Monitoring the quality of public water sources is important due to the many potential health risks and ecological impacts involved with contaminated and polluted water. Biological monitoring can be used to evaluate the physical, chemical, and biological condition of the body of water because the populations living in a body of water provide a fair representation of the condition of the ecosystem. Aquatic macroinvertebrate abundance and diversity are good indicators of water quality; there are some populations that are pollution sensitive and others that are pollution tolerant. The relative abundances of sensitive and tolerant taxa gives a baseline idea of the overall water quality and provides data to compare with previous years and other watersheds. This study aims to evaluate the quality of Fred Creek and Haikey Creek in Tulsa, OK by collecting and categorizing aquatic macroinvertebrates by pollution tolerance, calculating the EPT/C ratio, and performing a Bio-Assess Stream Quality Assessment to indicate the relative water integrity. Samples were taken from each location once monthly from March to May and August to November, and the organisms were organized by taxa. The data was compared between the two creeks, and the data from Fred Creek was compared with existing data from previous years, including before significant construction was done at the site. Fred Creek was expected to have better diversity than Haikey based on previous studies characterizing the impairment of both creeks based on macroinvertebrates. In this study, while the two sites had similar EPT/C ratios, Fred Creek was found to have consistently higher cumulative indices for each sample, indicating better water quality than Haikey.
Research Advisor: Dr. Julianna Goelzer
Stephenie Rogers, Houston Baptist University – 11:15-11:30 in Balanos Session
Potential for Combination Antibiotic-Phage Treatment as a Viable Therapeutic.
Stephenie Rogers.
The misuse of antibiotics in medicine and industry has contributed to an increase in bacterial resistance in every known antibiotic, including vancomycin. According to the Center for Disease Control (CDC), each year nearly 3 million people in the United States suffer from antibiotic resistant infections, while deaths associated with these illnesses exceed 50,000 annually. The production of new antibiotics, while a costly endeavor, has temporarily slowed the resistance crisis. However, the rate of bacterial mutation has shown to render these new treatments ineffective within a short time, proving the need for a more permanent solution. In the midst of the current antibiotic resistance crisis, bacteriophages have come to light as a potential alternative to antibiotic therapy because of their high binding affinity and specificity for corresponding bacterial strains. Phage therapy has previously been successful as an individual therapy, as well as in combination with antibiotic therapy. Overall, there is insufficient literature related to phage-host interactions and combination antibiotic-phage therapy. This study aims to explore the potential for phage therapy and combination antibiotic-phage therapy in the treatment of Escherichia coli infections by identifying a baseline minimum inhibitory concentration (MIC) for these treatments. Three different antibiotics and a T2 bacteriophage strain were used in varying concentrations in repeated trials to determine the mean MIC for each antibiotic and phage and combination. While isolated phage may not be the most effective treatment for mild infections, there is potential for using combination antibiotic-phage therapy in various agricultural industries as an alternative to animal antibiotic treatments and bactericides, one of the main contributors to antibiotic resistance worldwide.
Research Advisor: Dr. Curtis Henderson
Kaylee Salazar, St. Edward’s University – 11:30-11:45 in Boax Session
Evaluating the link between bacterial infections and Schizophrenia.
Kaylee Salazar, Eric Torres, Angelica Carrasco-Pena, Maria Miranda, Sarah Avant, and Andrea Holgado.
The relationship between Schizophrenia and external factors controlling genetic components has been made evident, yet it has been difficult to identify the causative molecular players involved. Yoshihara and colleagues recently hypothesized that factors inactivating CRMP-2 may participate in the pathogenesis of Schizophrenia. UNC-33 in C. elegans, CRMP-2/DPYSL2 in humans, regulates neuronal development and axonal outgrowth in healthy brains and is suppressed in the brain of schizophrenic patients. Our previous findings support the hypothesis that external stressors such as temperature play a role in regulating the activity of the promoter controlling neuronal levels of UNC-33. Herein we studied the effects of pathogens on the expression driven by the unc-33 promoter. To this end, C. elegans hermaphrodites were incubated on plates seeded with Escherichia coli OP50 or Pseudomonas aeruginosa PA14 until they reached the L3/L4 stage. Confocal microscopy was used to image the head of the C. elegans to determine GFP expression driven by the unc-33 promoter in neurons and glial cells. The analysis of the quantification of twenty-five worms per condition showed that C. elegans infected since embryo with Pseudomonas aeruginosa PA14 have decreased neuronal GFP driven by the unc-33 promoter. This study provides insights into the role of the Pseudomonas aeruginosa PA14 infection in regulating the activity of the unc-33 promoter, potentially linking bacterial infections with faulty neuronal development and Schizophrenia.
Research Advisor: Dr. Andrea Holgado
Danitra Sargent, University of the Ozarks – 1:30-3:30 in Serpent Session
Two new species in the genus Vacuoliviride (Eustigmatophyceae, Goniochloridales) revealed by light microscopy and DNA sequence analysis.
Danitra P. Sargent, Marvin W. Fawley, and Karen P. Fawley.
The Eustigmatophyceae are a group of unicellular eukaryotic algae that were recently thought to be rare, but with the aid of molecular techniques are now understood to be more diverse. The name Eustigmatophyceae is a result of the morphological feature of a unique eyespot sitting outside of the chloroplast of the zoospore. This group also has commercial importance in areas like aquaculture, agriculture, medicine and biofuel production. The recently described genus Vacuoliviride is a member of the Eustigmatophyceae with a large open vacuole, a polyhedral pyrenoid and an unusual v-shaped crystal. The objective of this study is to describe new diversity in the genus Vacuoliviride. We recently isolated new strains of eustigmatophytes that can be placed in the genus Vacuoliviride based on morphological characteristics and phylogenetic analysis of DNA sequences. Our results show that both strains are new species of Vacuoliviride. We propose a new species of Vacuoliviride for strain BogD 8/9 T-2w, isolated from a small bog in Itasca State Park, Minnesota. We also propose another species of Vacuoliviride for strain Bat 8/9-5m from Batsto Lake in the pine barrens of New Jersey.
Research Advisor: Dr. Karen P. Fawley
Kristina Simons, Texas A&M University - Galveston – 1:30-3:30 in Serpent Session
A systems toxicology analysis of the effects of oxybenzone on the metabolic physiology of embryo-larval zebrafish (Danio rerio).
K.I. Simons, L. Thibault, R. Nolen, Y. Umeki, L.H. Petersen, and D. Hala.
Oxybenzone is a chemical ingredient commonly used in sunscreen lotions, as well as other hygienic products considered to be an emerging contaminant in aquatic and marine systems frequented by tourism. There is growing concern that oxybenzone can cause adverse effects in exposed invertebrate and vertebrate organisms due to metabolic and endocrine disrupting effects. Embryo-larval fish are expected to be particularly sensitive at this early life stage by oxybenzone exposure as their developmental processes are likely to be perturbed. In this study I present results of an in vivo toxicological study with embryo-larval life stages of zebrafish (Danio rerio). Fish were exposed from 2 - 9 days post fertilization (7 days exposure) to encompass environmentally relevant concentrations of 1 and 10 µg/L Oxybenzone. The levels chosen are environmentally relevant as oxybenzone levels in surface waters have been reported at <=10 µg/L (Downs et el., 2015). Micro-respirometry measurements of metabolic rate (or oxygen consumption) in fish showed a significantly elevated respiration rate at 1 µg/L; while at 10 µg/L oxygen consumption was not significantly different from the control. Whole-organism RNA-sequencing analysis indicated widespread transcriptional effects on gene regulation DNA repair and morphological systems. Subsequent computational biology analysis using a genome-scale metabolic model of zebrafish identified glycerolipid metabolism as being highly dysregulated in the 1 µg/L treatment group vs. solvent control (0.01% dimethyl sulfoxide). This prediction agrees with a metabolomics study by Ziarrusta et al., (2018) which showed glycerolipid metabolism to also be highly dysregulated in gilthead seabream exposed to 50 µg/L oxybenzone (14-day exposure). Based on my research I hypothesize that glycerolipid metabolism is diagnostic of the increased metabolic rate measured of oxybenzone at 1 µg/L in my study.
Research Advisor: Dr. David Hala
Anastasia Smith, Oral Roberts University – 1:30-3:30 in Bird Session
Analysis of Effects of Full Spectrum CBD Extracts on Gene Expressions in Triple Negative Breast Cancer Cells.
Anastasia Smith and William Ranahan.
Contemporary cancer treatments such as radiation, chemotherapy, and surgery cause negative off-target effects in nontumorigenic tissue. Alternative remedies may offer treatment with little to no side effects. Medicinal plants have been used worldwide as adjuvant therapy and treatment for cancer. Given that initial in vitro and animal model studies investigating the effect of cannabinoids on tumors show promising results, the authors proposed to investigate a full spectrum extraction of cannabinoids from hemp (A et al. 2011; Dariš et al. 2019). The authors hypothesize that this extraction will negatively affect the viability of cancer cells and change gene expression. Cannabinoids from a hemp extract were analyzed using high pressure chromatography in the NGC chromatography machine. Full cannabinoid extractions tested on ER-/PR-/Her2- cancer cell model MDA MB-468 decreased cell viability when applied in amounts of four, eight and twelve µl in 100 µl of total suspension with 20,000 cells per well. The extractions were again tested on MDA MB-468’s, but in a six cm petri dish and successful results were used to obtain RNA. cDNA was made from RNA samples and added in a primePCR assay. The primePCR assay well’s contained preloaded primers for 91 genes and four controls. Of these genes, three were specifically examined: HMOX1, IL6, and VIM. The authors found these genes to be over expressed when compared to the control tested with olive oil. Overexpression of these genes is correlated with increased cancer aggression and poor clinical prognosis Thus, the authors hypothesis that upregulated of HMOX1, IL6, and VIM may be an epigenetic modification the MDA-MB-468’s to evade the treatment. Future studies will aim to examine other genes analyzed in the primePCR assay, test isolated cannabinoids, and examine the effects of full hemp extracts on different cell types.
Research Advisor: Dr. Julianna Goelzer
Emma Solis, Austin College – 10:15-10:30 in Boax Session
Evaluation of the relationship between a high-fat diet and gut microbiome health in thirteen-lined ground squirrels (Ictidomys tridecemlineatus).
Emma Solis and Jessica Healy.
Hibernating species typically undergo extreme cycles of fat accumulation and loss throughout the year as they maintain their normal circannual rhythm of euthermia and torpor. With feeding patterns changing drastically during the year, there have been several papers detailing subsequent seasonal differences in gut microbiota structure and diversity according to a standard hibernation cycle. However, little research has been done to evaluate if other factors such as dietary fat content or natural vs. lab diets have any direct effects on the gut health and microbial community of hibernating species. In this project we are using behavioral (food intake), morphological (body mass and composition), and molecular (qPCR) techniques to evaluate the physiological effects of a high-fat diet on our animal of study, the thirteen-lined ground squirrel (Ictidomys tridecemlineatus) to observe potential differences that emerge as wild-caught animals are transitioned to a lab diet.
Research Advisor: Dr. Jessica Healy-LaPrice
Megan Strotman, Southwestern Oklahoma State University – 1:30-3:30 in Serpent Session
Investigating Potential Circadian Oscillations in Pseudomonas syringae.
Megan Strotman and Regina McGrane.
Pseudomonas syringae is a gram-negative plant pathogen that encodes a kaiC ortholog. KaiC has been proven to regulate circadian oscillations in cyanobacteria, which control transcriptional oscillations, cell division, and photosynthesis. We hypothesize that P. syringae is capable of circadian oscillation and that kaiC contributes to regulation of stress response, biofilm development, motility, and plant colonization. To determine if P. syringae has circadian oscillations, the wild type strain or mutants lacking kaiC were spotted onto King’s B agar media supplemented with coomassie blue, which binds proteins, or congo red, which binds exopolysaccharides. The plates were then exposed to day-night cycles (12 hours light, 12 hours dark) or dark conditions for seven days. Oscillations in protein and exopolysaccharide production were then observed due to varying intensity of either blue or red rings in the colonies. While oscillations were detected, analysis of the plates indicated no significant differences in the number or pattern of the rings when comparing the mutants to the wild type or the plates exposed to day-night cycles to dark conditions. To determine if kaiC regulated P. syringae stress response, wild type or mutants lacking kaiC were exposed to day-night cycles or dark conditions as described above and inoculated in MinA media supplemented with glucose and varying concentrations of salt. Growth was then recorded by detecting absorbance every 30 minutes over a 48-hour period. Analysis revealed significant differences in the growth of the wild type exposed to day-night cycles compared to samples kept in the dark; however, these differences were not observed in mutant strains. This suggests that kaiC may modulate the ability of P. syringae to combat stressful environments. Determining the function of kaiC in P. syringae may allow for development of techniques that target this ortholog and prevent P. syringae growth on crops.
Research Advisor: Dr. Regina McGrane
Uzziah Urquiza, Southwestern Oklahoma State University – 11:00-11:15 in Boax Session
Investigation of the molecular mechanisms underlying the cytotoxicity of novel lactate dehydrogenase A inhibitors against human pancreatic cancer.
Uzziah Urquiza, Hanna Hill, Pragya Sharma, and Horrick Sharma.
Pancreatic cancer is the 3rd leading cause of cancer-related death in the United States. Chemotherapy is one of the standard treatment options for pancreatic cancer; however, the rapid development of resistance to chemotherapeutic agents represents a significant bottleneck in treatment options. The average life expectancy after diagnosis with metastatic pancreatic cancer is just three to six months reflecting an urgent unmet need to identify new targets. Cancer cells are well-known to alter their metabolic pathways to support their growth and proliferation. Unlike normal cells, cancer cells prefer aerobic glycolysis, a phenomenon described by Otto Warburg, known as the Warburg effect. This unique metabolic alteration in cancerous cells is regulated by lactate dehydrogenase A (also known as LDH5), an enzyme that is overexpressed in many tumor types, including pancreatic cancer. In collaboration, our lab has identified several novel lactate dehydrogenase inhibitors, which have shown promising anticancer activity against pancreatic cancer cell lines. The goal of the project study is to uncover the molecular mechanisms of antiproliferative activity of our lead compounds. Our preliminary data indicate the activation of the pro-apoptotic protein pathway as a possible mechanism of cytotoxicity of pancreatic cancer cells.
Research Advisor: Dr. Pragya Sharma
Arianna Vacio, St. Edward’s University – 1:30-3:30 in Serpent Session
UNC-33L partially rescues lifespan and locomotion defects in unc-33 mutants but fails to rescue dauer formation defects.
Arianna Vacio.
Aging is a process that gradually decreases an organism's physical and mental capacity, increasing the risk of disease and death. Neurodegenerative diseases, such as Alzheimer’s and Parkinson’s, result from accelerated neuronal cell death, which leads to neurophysiological decline, deterioration of the body, and reduction in the standard of living. UNC-33, homolog to CRMP2, serves as a biomarker for these neurodegenerative diseases. In healthy tissue, CRMP2 regulates neuronal development and axonal outgrowth. In disease, CRMP2 is inactivated by hyperphosphorylation, compromising the maintenance of axons. To better understand the role of CRMP2/UNC-33 in the aging process, we tested the ability of UNC-33L, one of the isoforms coded by the unc-33 gene, to rescue the shortened lifespan, defective locomotion, and the failure to make dauers, a non-aging life stage, in C. elegans unc-33 mutants. Analysis of results demonstrates that the presence of UNC-33L does not rescue the defective dauer phenotype in unc-33 mutants. However, UNC-33L significantly rescued premature death and uncoordinated locomotion in young unc-33 adults. The degree of UNC-33L-mediated rescue was less noticeable as the nematodes aged, denoting that both age and the presence of UNC-33L interact in the production of the phenotypes.
Research Advisor: Dr. Andrea Holgado
Luke Whelchel, Dallas Baptist University – 10:15-10:30 in Balanos Session
Urinary Inflammatory Biomarkers and Gut Microbiome are Weight Dependent in Adolescents.
Luke Whelchel.
Obesity has risen dramatically in recent decades. Studies have shown obesity causes inflammation through activation of pro-inflammatory cytokines. Obesity has also been linked with dysbiosis of the gut microbiome. The goal of the study was to evaluate noninvasive biomarkers of inflammation and gut microbiome and follow impacts of lifestyle modification in adolescents that were overweight and obese. Thirty-eight adolescents were enrolled: sixteen healthy (BMI 30). Anthropometric measurements were taken at enrollment and were followed overtime. HbA1c, blood pressure, and lipid profiles were taken at the time of enrollment. A lifestyle modification program with the goal of promoting healthier eating and more active lifestyles was implemented. Urine was collected from all participants at enrollment and at 3- and 6-months in the program and analyzed for TNF-α, IL-6, and ET-1 by ELISA. Stool samples were collected at enrollment and compared to healthy peers. Adolescents of the OW/OB group exhibited elevated levels of urinary inflammatory markers compared to healthy control groups at enrollment. After six months, urinary pro-inflammatory TNF-a significantly decreased. Bacterial DNA analysis indicated that fecal samples from the OW/OB group demonstrated differing microbiome profiles as well as an increased presence of pathogenic bacterial families compared to healthy controls. Adolescents that were OW/OB demonstrated increased levels of urinary inflammatory cytokines, which improved under the lifestyle modification program. Notable differences in the gut microbiome compositions between adolescents who were OW/OB and their healthy counterparts were observed.
Research Advisor: Dr. Tetyana L. Vasylyeva
Determining if extracts of Eysenhardtia polystachya, Pinus maritima, and Zingiber officinale can combat antimicrobial resistance.
Nichole Abrego and Patricia Baynham.
Antimicrobial resistance (AMR) causes more than 35,000 deaths annually in the United States, so new treatments are needed. In this study, parts of three different plants were examined for their antibacterial activity: the bark of Eysenhardtia Polystachya (palo azul) and Pinus maritima (pine) ,and the root of Zingiber officinale (ginger). The barks were extracted and were then tested for antimicrobial activity using a Kirby-Bauer disk diffusion assay against E. coli lptD4213-. While the ginger extract displayed no zone of inhibition, the average zone of inhibition was 13mm for pine bark and 11mm for palo azul. A minimum inhibitory concentration assay (MIC) was performed, to better quantify the antimicrobial activity and showed that pine bark had an MIC of 640ug/ml while the MIC of palo azul was 600ug/ml.. To investigate the mechanism of action (MOA) of the extracts, bacterial cytological profiling (BCP) was performed. BCP involves treating the bacterium with an antimicrobial substance and then visualizing the changes in shape that are different based on which part of the cell is targeted. Visualization was via confocal microscopy using FM4-64 to show the membranes and DAPI to see the DNA. The treated cells changed shape but not all cells were visualized so this will require further analysis. In the future, the cell shapes can be quantified and compared with the known MOA of known antibiotics. New treatments are crucial when AMR infections are increasing. Plant substances can be characterized and developed into possible alternative treatments to AMR microbes.
Research Advisor: Dr. Patricia Baynham
Ryan Agyemang and Altherr Joseph Alcuitas, Southwestern Oklahoma State University – 10:45-11:00 in Boax Session
The effect of female mate choice on offspring sex ratios in a freshwater amphipod species.
Ryan Agyemang and Altherr (Joseph) Alcuitas.
This experiment highlights the effect of female mate choice on offspring sex ratios in a freshwater amphipod species. There are several determinants of sex ratios in nature all falling under two main categories being genetic or environmental. The expectation from theory is that the sexes will be produced in a 1:1 ratio. However, biased sex ratios are common in nature and can be the result of both genetic and environmental factors. We tested whether females bias the sex ratio of broods in favor of sons when mating with a high quality male. We collected mating pairs of amphipods (Hyalella sp.) from two pools in the same natural spring system. Half of the females were separated and repaired with their original mate—i.e. they were allowed to choose their mate. The other half of the females were separated and assigned a new male at random. Each pair was placed in their own 50-ml jar and placed in an environmental chamber. We are checking the pairs three times a week and recording offspring sex ratios. We predict that females with a choice in their mate will pick higher quality males and in turn produce more male biased broods and females assigned a mate at random will produce more of an even brood sex ratio. We also predict that the choice pairings will have less variation in offspring sex ratios as mates would be all of high quality. On the contrary, random pairings will have a much higher variation in offspring ratios as some males assigned will be higher quality and some males assigned will be poor quality.
Research Advisor: Dr. Ricky Cothran
Sara Ambrocio Paque, University of the Ozarks – 9:45-10:00 in Boax Session
Comparison of eukaryotic soil algae in Warren Prairie Natural Area sodic saline slicks.
Sara Ambrocio Paque, Marvin W. Fawley, and Karen P. Fawley.
Warren Prairie Natural Area (WPNA) in Bradley and Drew Counties, Arkansas, is a mosaic area of saline slicks that form flat, crusty depressions in a central area with a zone of lichens and a few rare angiosperms, and an outer zone of cyanobacterial mats. The edges of the saline slicks are home to the rare, diminutive vascular plant, Geocarpon minimum Mackenzie (Caryophyllaceae), which is a federally protected threatened species. Because the Warren Prairie slicks are home to many rare and unusual vascular plants, we hypothesized that the soil algae community will also comprise many unusual species. The main objective of this study was to characterize and compare the soil crust eukaryotic algal communities in Site 1 (saline slick) and Site 2 (soil not within the sodic saline slick) in WPNA using morphological and molecular techniques. We focused on the 10 species of the green algal genera Coelastrella, Cylindrocystis, Myrmecia, and Diplosphaera that were previously isolated from a saline slick in WPNA. We used light microscopy for morphological characterization of each strain and DNA sequences (barcodes) from loci appropriate for species-level identification (tufA; ITS; rbcL). Coelastrella strains from Site 1 were identical to a strain previously described from a slick in WPNA. In contrast, Site 2 had a unique strain similar to Coelastrella yingshanensis. Cylindrocystis strains were present in the Site 1, but were not present in Site 2. Myrmecia was not found from Site 1 or Site 2, whereas Diplosphaera strains were found in both sites. Our results show that most of the species of green algal genera examined were different between the soil from saline slicks and soil outside the slick.
Research Advisor: Dr. Karen P. Fawley
Bryler Atchley, Southwestern Oklahoma State University – 1:30-3:30 in Serpent Session
Syringafactin Increases Membrane Permeability in Gram-Negative Bacteria.
Bryler Atchley and Regina McGrane.
Pseudomonas syringae is a gram-negative plant pathogen that produces a biosurfactant called syringafactin. Syringafactin, a lipo-octapeptide, has been shown to act as an antagonist molecule against competing gram-negative bacteria such as Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa. The primary antagonistic properties include induction of negative chemotaxis and cell death in competing bacteria. We hypothesized that cell death in competing cells is the result of increased membrane permeability. This work aimed to quantify the impact of syringafactin on the membrane permeability of E. coli, S. typhimurium, and P. aeruginosa. Fluorescence was detected using DiS-C2(5), a fluorescent dye sensitive to fluctuations in membrane polarity. Data was collected using a multi-mode plate reader for four hours following exposure to increasing concentrations of crude syringafactin extract. Additionally, extract free samples and samples containing crude extracts from a mutant lacking syringafactin production were tested. Samples showed increasing fluorescence correlating to increasing concentrations of syringafactin, and extract free samples lacked fluorescence. Samples exposed to extracts from a mutant lacking syringafactin production exhibited some fluorescence, but the fluorescence was significantly lower than samples exposed to syringafactin extracts. To confirm that the major difference between the two types of crude extracts was the presence or absence of syringafactin, purified samples were analyzed using mass spectroscopy and liquid chromatography. These procedures evaluate the peptide and lipid composition of the extract. Analysis demonstrated that the six lipo-octopeptides previously characterized as syringafactin were present in high concentrations in extracts from wild-type bacteria, but they were absent or in very low concentrations in mutants lacking syringafactin production. Collectively, these studies suggest that syringafactin in crude extracts is responsible for reducing cell viability by increasing membrane permeability. Identifying the mechanism responsible for syringafactin induced cell death leads to a deeper understanding of the potential effectiveness of syringafactin as an antibacterial agent.
Research Advisor: Dr. Regina McGrane
Nicholas Bauer, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
High Resolution Crystal Structure of Ro0101, a Dodecameric Dps-like Protein from Rhodococcus jostii.
Nicholas Bauer, Abigail Trejo, Leonard Thomas, Rakhi Rajan and Vijayakumar Somalinga.
Dps (DNA binding protein from starved cells) and related ferritin proteins are ubiquitous stress proteins expressed in eukaryotes and prokaryotes. Dps proteins are integral in stress tolerance and for DNA binding and protection. Furthermore, Dps and related ferritin proteins have been tested in bionanotechnological applications including nanocages, biocontrast agents, and in drug delivery. We have identified a Dps homolog, Ro00101 referred as Dps1, from Rhodococcus jostii, a soil-dwelling bacteria with unparalleled metabolic potential. The dps1 gene encodes a 19.4 kDa protein with conserved domains typical of DNA-binding proteins with ferroxidase activity. The goal of this study is to characterize Rhodococcal Dps protein using biochemical and structural techniques. Cloning and over-expression were done using standard techniques. Protein purification was done using immobilized metal affinity chromatography. The purified protein was concentrated to 17 mg/ml before crystallization trials. Preliminary 96-well sitting-drop vapor diffusion crystallization revealed crystals in drops containing 0.1 M sodium acetate trihydrate pH 4.5. and 2M ammonium sulfate pH 4.5. A single, rhomboidal crystal obtained using hanging-drop method was used for data collection and diffracted at 1.85 Angstrom. The structure for Dps1 was solved by molecular replacement using Mycobacterium smegmatis Dps1 (PDB ID: 1VEI) as a model. The structure of Dps1 revealed a dodecameric protein with conserved ferritin-like and Dps-like trimeric interfaces involved in iron channeling and protein folding respectively. The dimeric interface that harbors the iron-binding active site is also conserved. The active site of Dps1 reveals twin histidine residues from one monomer and glutamine/aspartic acid residues from the second monomer contributing to the two ferroxidase sites found in the dimeric interface. Interestingly, Dps1 crystallized without the bound ligand. Studies are underway to obtain ligand-bound structures and to determine the ferroxidase and DNA-binding properties of Dps1.
Research Advisor: Dr. Vijay Somalinga
Benjamin Berggren, Austin College – 10:00-10:15 in Boax Session
Detection of Chytrid Fungus via Environmental DNA in North Texas.
Benjamin Berggren, Loriann Garcia, Frank Goodavish, and Keegan Nichols.
EDNA, or environmental DNA, is DNA shed by organisms in their environment. Ecologists are beginning to use this DNA shed into the environment to detect elusive species, track disease progression, or perform general biodiversity surveys. EDNA detection has been used in many different projects, and the methods can be adjusted to fit the needs of the study. In the summer of 2021, we extracted eDNA from soil and water samples from two sites in Grayson, Co. TX to detect the presence of Blanchard’s Cricket frog and the Texas Smallmouth salamander. We successfully detected Blanchard’s Cricket frog. This fall and spring, I used eDNA to attempt to detect the presence or absence of the amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd). I sampled two sites in Grayson Co. which border counties in Oklahoma where chytrid has been detected. Detecting this fungus as early as possible will allow ecologists to mitigate the effects on larger amphibian populations. Our tests so far have not detected Bd from our samples, but our test results show an error in our methods. To rectify this, we will update our methods, continue to sample, and test, and send our current isolated eDNA to be sequenced.
Research Advisor: Dr. Loriann Garcia
Sebastian Calderon, Houston Baptist University – 9:15-9:30 in Boax Session
Determining the Presence of E. coli 0157:H7 in Bovine Population.
Sebastian Calderon.
Shiga toxin producing E. coli are derived from the guts of most cattle. The 0157:H7 strain of E. coli is the most prominent strain that infects humans, and it can cause severe damage to the lining of one's intestines and kidneys. The ideal situation is to attack it at the source. Vaccines that reduce the amount of bacterial shedding exist, but the beef industry has no incentive to use them. Thus, the research aims to determine the prevalence of E. coli 0157 in bovine feces. Through the process of isolating the E. coli, extracting genomic DNA, and PCR there was data determining that there is a relatively large amount of this strain of E. coli within cattle.
Research Advisor: Dr. Jacqueline Peltier Horn
Eduardo Carrillo, St. Edward’s University – 9:30-9:45 in Balanos Session
Defects in autophagy flux related to microtubule associated protein UNC-33 mediate changes in basal and starvation-induced autophagy.
Eduardo Carrillo, Alexa Ott, Emily Holechek, Arianna Vacio, and Andrea Holgado.
Autophagy is the process in which cellular components are degraded and recycled. Regulation of autophagy is essential for axonal development and maintenance of synapses. UNC-33/CRMP-2 is a microtubule associated protein (MAP) that contributes to neuronal development and transport of autophagosomes. UNC-33 is part of a ternary complex that anchors microtubule bundles to the cortex and inhibits microtubule sliding. Problems with microtubule sliding, and cargo transport and degradation in unc-33 mutants may contribute to the development of neurodegenerative diseases. Our laboratory studies autophagy via cleavage of the double fluorescently tagged protein CERULEAN-VENUS LGG-1(dFP) in C. elegans unc-33 mutants. We predict that lack of UNC-33 will result in defects in autophagy. To test this hypothesis we monitored autophagosome maturation and pathway flux using western blots and the levels of dFP(MW 75) versus the cleaved monomeric fluorescent protein (mFP)(25 kDa). In these experiments, we induced autophagy via starvation and compared the results against basal autophagy. Analysis of western blot results show that unc-33 mutants undergoing induced autophagy have an accumulation of the cleaved mFP. Conversely in basal autophagy, the accumulation of mFP was decreased.These results suggest that UNC-33 may contribute to autophagy flux and clearance of cargo. Microtubule sliding, a phenomenon seen in unc-33 mutants, may affect autophagosome transport and cargo degradation. These findings support a role of UNC-33 in autophagy in neurons.
Research Advisor: Dr. Andrea Holgado
Bethany Chapman, Angelo State University – 1:30-3:30 in Bird Session
An Observational Study of Mast Cells in the Mephitidae Family.
Bethany Chapman and Laurel Fohn.
Mast cells are important effector cells in the immune system of vertebrates. While mast cells, or mastocytes, are primarily associated with allergic reactions and anaphylaxis, they are also involved in defense against pathogens and immune tolerance. These cells are found in connective tissues, especially that of the skin, lungs, and intestines. The dysregulation of these cells can lead to mast cell tumors, a common type of tumor in household pets. Though these cells and tumors are commonly observed in cats and dogs, they have yet to be studied in many other mammals. The observation of mastocytes in other mammals may give us clues as to how they function in wild animals as compared with domesticated animals. This study seeks to observe mast cells and their prevalence within skin tissue samples of the skunk family, Mephitidae. Tissues will be formalin fixed and paraffin embedded before being stained with hematoxylin and eosin as well as toluidine blue for better identification of mast cells.
Research Advisor: Dr. Laurel Fohn
Mary Cooperrider, Oral Roberts University – 9:30-9:45 in Boax Session
A Preliminary Analysis of Full Spectrum Hemp Oil in Triple-Negative Mammary Epithelia.
Mary Claire Cooperrider and William Ranahan.
Cancer is one of the leading causes of death in the world and there are limited options for noninvasive, therapeutic strategies. In recent years, more individuals have opted to use Cannabis sativa, hemp, to soothe their ailments ranging from pain to epilepsy. The common misconception surrounding hemp is that it is identical to marijuana. A hemp plant contains a markedly lower percentage of tetrahydrocannabinol (THC) than a marijuana plant. Hemp oils and extracts are widely available for purchase in the United States, and researchers have yet to fully document the capabilities and treatment implications of hemp extracts. It is known that hemp reduces inflammation in the body, indicating it as a potential method for targeting cancer cells because inflammation promotes the proliferation of tumors and sustainability of tumor microenvironments. Full spectrum hemp extracts contain various compounds with possible therapeutic capabilities to be used in targeting tumors. Researchers know of many genes that may be monitored for changes in expression in order to observe efficacy of treatment. The current study aimed to analyze the variations in gene expression in a triple-negative breast cancer cell line, MDA-MB-468 cells, following the topical application of full spectrum hemp oil. The gene expression levels of MDA-MB-468 cells that were treated with the drug were compared to expression levels of untreated MDA-MB-468 cells using a PrimePCR Assay. The data suggested that treatment with full spectrum hemp extract yielded upregulation in genes SUMO1 and DKK1 and downregulation in genes BMP2 and MCM2. This preliminary data suggests that multiple pathways involved in tumor survival and progression are affected by components of full spectrum hemp extract. Further studies will include repeated analysis of gene expression and cell viability, as well as gene-specific analysis to observe individual alterations in cell mechanisms of cancer cells.
Research Advisor: Dr. William Ranahan
Larry Cossey, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
Detecting Variations in Microbial Concentrations in Oklahoma Lakes.
Larry Cossey and Regina McGrane.
The microbial composition of water can be the difference between life or death for many organisms. Microbes in lakes can reproduce and spread to other parts of the water cycle. Ice nucleating Pseudomonads are microbes that have been shown to be in all parts of the water cycle and damage plants. These Pseudomonads conduct ice nucleation by inducing freezing of supercooled water molecules. Through evaporation and rainfall, Pseudomonads can disperse anywhere. When bacterial ice nucleation occurs around plant tissue, ice crystals form within the tissue and damages the plant. To examine microbial composition, two Oklahoma lakes: the urban lake---Lake Hefner and the rural lake--Crowder Lake, were examined. In each lake, water samples were collected from three specific locations in summer and winter months. Samples were inoculated on 10% tryptic soy agar to determine total bacterial load and King’s B agar supplemented with boric acid, cycloheximide, and cephalexin to isolate Pseudomonads. At Lake Hefner, the eastern shore had significantly larger bacterial concentration in both seasons compared to other locations. At Crowder Lake, the center of the lake had significantly lower bacterial concentration in both seasons compared to other locations. Seasonally, there was lower bacterial concentration in the winter at Lake Hefner compared to the summer, while Crowder Lake had similar bacterial concentration year-round. Lake Hefner had higher bacterial concentration compared to Crowder Lake. Pseudomonads isolated from both seasons and lakes were tested for ice nucleation activity by inoculating in super cooled water and detecting ice formation. Twenty-two ice nucleators were detected at Crowder Lake and three ice nucleators were detected at Lake Hefner in the summer, and none were detected in the winter. The seasonal differences in the concentration of ice nucleating bacteria were surprising because other studies have shown that cooler environments select for increased ice nucleating bacteria.
Research Advisor: Dr. Regina McGrane
Alejandra Cristancho, St. Edward’s University – 1:30-3:30 in Serpent Session
Does the extension of lifespan equate to improved quality of life?
Alejandra Cristancho, Alexia Samaro, Ruby Valtierra, Skye Beck, Maria Miranda, and Andrea Holgado.
Collapsin response mediator protein-2 (CRMP-2) in humans (UNC-33 in C. elegans) mediates axonal outgrowth, maintenance, and degeneration. UNC-33/CRMP-2 has been hypothesized as a potential drug target for the treatment of Alzheimer’s and other neurodegenerative diseases. In aging, CRMP-2 becomes hyperphosphorylated, destabilizing the cellular skeleton and leading to neurodegeneration. In C. elegans, aging can be slowed by entering dauer diapause; a non-aging developmental stage turned on when the DAF-7/TGFβ signaling pathway is silenced in response to environmental stress. In our laboratory, we discovered that unc-33 mutants cannot form dauers in response to environmental stress. This study investigates whether a mutation in the daf-7 gene can rescue phenotype characteristic of unc-33 mutants. To this end, we created unc-33; daf-7 double mutants and quantified dauer formation, axonal elongation, and locomotion after exposure to unfavorable conditions. Our analysis of unc-33; daf-7 double mutants showed that introducing the daf-7 mutation into an unc-33 mutant rescued dauer formation. Studies of the rescue of axonal elongation and locomotion are currently underway, and results will be presented at the symposium. So far, these findings suggest that unc-33 mutants may have a problem suppressing DAF-7 signaling, leading to the continuation of reproductive programs even under unfavorable conditions. Furthermore, these results point out a role of CRMP2/UNC-33 and DAF-7 in the extension of the lifespan.
Research Advisor: Dr. Andrea Holgado
Lily Ellzey, Angelo State University – 1:30-3:30 in Bird Session
Detection and Identification of Coronavirus Strains in Myotis velifer.
Lily Ellzey and Laurel Fohn.
SARS-CoV-2 is a member of the family of viruses known as the coronaviruses. This family of viruses includes two subfamilies of viruses. Of these, the subfamily Coronavirinae contains four genera of viruses, two of which are known to infect humans. Bats are a natural reservoir for many kinds of coronaviruses, but bats collected in West Texas have not yet been analyzed to determine which strains of coronaviruses these species of bats carry. RNA was extracted from the Angelo State Natural History Collection gastrointestinal tissue samples of bats from West Texas and subjected to RT-PCR to assess for the presence or absence of coronavirus. If present, amplicons will be submitted for sequencing and coronavirus strain identification.
Research Advisor: Dr. Laurel Fohn
Payden Farnsley, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
Development of Isolation Protocols to Study the Distribution of Pseudomonas syringae in Western Oklahoma.
Payden Farnsley and Regina McGrane.
Pseudomonas syringae is a plant pathogen that causes significant economic losses due to reduced crop yield caused by disease. Studies have shown that P. syringae is globally distributed and is linked to the water cycle due to its ability to induce ice nucleation, the crystallization of supercooled water. Our goal is to better understand how this plant pathogen affects western Oklahoma by studying its distribution in non-agricultural systems. If P. syringae is present in samples, it will help us predict outbreaks that might affect Oklahoma crops. To study the distribution of P. syringae on wild plants, we are currently developing protocols for isolation. To evaluate the validity of these protocols, we collected leaf samples at a local state park from two different species of wild plant in two different areas of the park. Leaf samples were suspended in washing buffer and sonicated the sample to break up bacterial aggregates. Samples were then inoculated on 10% tryptic soy agar to determine total microbial load and King B’s agar supplemented with boric acid, cycloheximide, and cephalexin, which is a medium selective for P. syringae. Potential P. syringae isolates were then subcultured on tryptic soy agar for storage. Isolates were then evaluated for oxidases, which P. syringae does not produces. To test isolates for ice nucleation isolates were inoculated in super cooled water and observed for induction of freezing. We have stored thirteen isolates, which each tested negative for oxidase activity and positive for ice nucleation activity because they are likely P. syringae strains. This bacterium produces a biosurfactant – syringafactin – and we plan to test for production of this biosurfactant to further characterize their identity. We also plan to perform 16s rRNA sequencing to confirm their identity.
Research Advisor: Dr. Regina McGrane
Emily Foss, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
Identification and Characterization of Bacterial Carbonic Anhydrases for Enzyme Accelerated Carbon Capture and Sequestration.
Emily Foss, Amy Grunden, and Vijayakumar Somalinga.
Carbon capture and sequestration (CCS) is one of the numerous mitigation efforts that are being implemented to minimize the amount CO2 being dumped into earth’s atmosphere. Current CCS technologies aimed at removing CO2 from industrial gases rely on energy intensive scrubbing or amine-based CCS processes. Enzyme based CCS technologies have recently been used to improve industrial CCS, although enzyme stability in solvents and at high temperature has impeded the large-scale implementation of this technology. Carbonic anhydrases (CA) are ubiquitous enzymes that catalyze the interconversion of CO2 to bicarbonate. The catalytic activity of CA’s can be exploited in mineralization of CO2 in the form of carbonates, a well-known and a safe CO2 disposal method. We have previously identified and characterized the structure of an a-CA from Photobacterium profundum and recently identified several bacterial CA’s including an active b-CA from Streptococcus sanguinis. The goal of this study is to elucidate the biochemical characteristics of unique CA’s for use in CCS technologies. Using in-gel carbonic anhydrase activity assay (protonography) we have previously shown that a-CA is capable of CO2 hydration activity and in this study, using protonography, we confirmed that b-CA from S. sanguinis is also capable of CO2 hydration activity. Further biochemical analysis of a-CA revealed a bimodal pH maxima with peak activity observed at acidic (pH 5.0) and alkaline (pH 11) pH. a-CA enzyme activity dramatically declined at temperatures above 30oC, while maximal activity was observed at lower temperatures (4 to 30oC). A unique redox dependent activity was observed for a-CA which has not been previously reported in a-CA’s. Finally, biomineralization assay revealed that a-CA accelerates CO2 sequestration into carbonates under alkaline condition. Our preliminary biochemical characterization indicates that a-CA possess several unique biochemical characteristics and can be exploited for use in CCS technologies.
Research Advisor: Dr. Vijay Somalinga
Ella Fotinos, St. Edward’s University – 9:45-10:00 in Balanos Session
Using lipopolysaccharides to evaluate the effect of energetic stress on Italian honeybee immune response.
Ella Fotinos.
Colony collapse disorder (CCD) is a phenomenon in which colonies of honeybees rapidly lose all of their workers and consequently become nonfunctional. Due to honeybee use in agricultural and ecological pollination services, our ability to combat CCD has real world consequences. And, because CCD has been linked to pathogens, it is essential to better understand how honeybees respond immunologically to pathogenic threats to help inform best practices for minimizing the impact of CCD. Our study focused on the immune response of Italian honeybees to the introduction of lipopolysaccharides (LPS) to emulate a pathogenic threat. We examined bees from one hive containing frames with a plastic foundation and another containing no foundation in order to evaluate if energetic expenditure associated with honeycomb production has an effect on immune responses. These two forms of colony structure are common in beekeeping operations to either maximize honey versus honeycomb production respectively. After feeding specimens a sugar water solution containing 1mg/ml of LPS (Escherichia coli R515), we evaluated their immune response using a several colorimetric immunoassays at staggered feeding timepoints (0, 1, or 2 hours after ingestion). Bees with no foundation had significantly higher protein concentration in their hemolymph than those with a foundation, and there was trend of protein concentration spiking in the no foundation treatment two hours post-ingestion. Prophenoloxidase immune assays are currently ongoing. Our results indicate that honeybees used for honeycomb production may be more energetically stressed and thus increase their circulating protein concentration to preventatively prepare for combating pathogenic threats.
Research Advisor: Dr. Matt Steffenson
Annabelle Hawkins, Southwestern Oklahoma State University – 1:30-3:30 in Bird Session
Investigating impacts of syringafactin on bacterial chemotaxis.
Annabelle Hawkins and Regina McGrane.
Pseudomonas syringae is a bacterium that infects plants. Our laboratory demonstrated that syringafactin, a biosurfactant produced by P. syringae, has antimicrobial properties and repels human pathogens on semi-solid surfaces. This suggests syringafactin may be useful in controlling the growth of pathogenic bacteria. The purpose of this study was to determine whether syringafactin acts as a repellant of bacteria in liquid suspensions and identify potential Pseudomonas aeruginosa receptors involved in syringafactin detection. To study if syringafactin works as a repellant in liquid, normalized cultures of Escherichia coli, Salmonella enteritidis, and P. aeruginosa were exposed to suspended pipette tips containing syringafactin or sterile deionized water for 15 min. The contents of the pipette tips were then spread onto agar media, and colonies were counted to quantify bacteria that moved into the pipette tip. Samples from tips containing syringafactin were expected to have lower bacterial concentrations compared to samples from tips containing water. Surprisingly, our studies showed that samples with syringafactin were not significantly different from those with water. Therefore, we concluded that syringafactin does not work as a chemorepellant in liquids. To determine the chemoreceptors responsible for P. aeruginosa detection of syringafactin, we exposed P. aeruginosa mutants with disruptions in genes encoding for putative chemoreceptors to wild-type P. syringae on soft-agar media. The ability of the mutants to respond to and move away from P. syringae was observed. Mutants unable to move away from P. syringae on soft agar media likely have disruptions in syringafactin specific chemoreceptors. Collectively, this work is the first to demonstrate that syringafactin does not impact chemotaxis in liquids and attempt to identify chemoreceptors involved in detecting syringafactin.
Research Advisor: Dr. Regina McGrane
Makayla Hicks, Southwestern Oklahoma State University – 9:15-9:30 in Balanos Session
Evaluating pathogenicity of Pseudomonas aeruginosa through assessment of motility and biofilm mutants.
Makayla Hicks and Regina McGrane.
Pseudomonas aeruginosa is an opportunistic pathogen that causes nosocomial infections, particularly in the lungs of cystic fibrosis patients. Motility and biofilm formation are known to contribute to this bacterium’s ability to cause disease. These processes are modulated by many biological molecules including biosurfactants, the flagella, the pilus, and several two-component regulatory systems. In this research, we evaluated the biofilm formation and virulence of a variety of P. aeruginosa mutants with disruptions in genes encoding for biosurfactant (rhlA), pili (pilA), and flagella (fliC, motB, and motD) biosynthesis along with surface sensing (wspA). We hypothesize that deletion of these genes will cause this bacterium to be impaired in biofilm formation and pathogenicity. Biofilm production was evaluated using a crystal violet microtiter plate assay. We found that mutants lacking functional fliC, motB, motD, wspA, and pilA were all significantly impaired in biofilm formation. Virulence was tested by inoculating Galleria mellonella larvae, a wax moth that is used as a model organism for the understanding of bacterial pathogenicity, with mutant or wild type cultures. The survivability, coloration, movement, and silk production of the larvae were evaluated every day for up to seven days or until all the larvae died, whichever occurred first. Significant differences were detected between the wild type and mutants with deletions in rhlA, wspA, motB, and motD in silk cocoon formation and movement of the larvae. This suggests that these mutations alter the ability of P. aeruginosa to cause disease. It is likely that impaired virulence is directly related to the ability of mutants to produce biofilms. Understanding the role of biofilm formation in the pathogenicity of P. aeruginosa in wax moth larvae could lead to a better understanding of how this bacterium causes infection in humans and ultimately could lead to a solution for problematic infections.
Research Advisor: Dr. Regina McGrane
Taylor-Jade Higgins, Austin College – 11:00-11:15 in Balanos Session
Investigating the Role of Epithelial to Mesenchymal Transition on PA28γ Expression.
Taylor-Jade Higgins.
The 20S proteasome degrades and recycles proteins following activation by PA28γ, a ubiquitin- and ATP-independent proteasome activator. Several tumor tissues express elevated levels of PA28γ, and studies have identified that it also induces the epithelial to mesenchymal transition (EMT). With PA28γ and EMT playing a role in tumor development, the objective of this study was to identify whether PA28γ activity and expression are affected by EMT. EpH4 murine epithelial cells were treated with varying dosages and exposure times to Transforming Growth Factor β-1 (TGF-β1) to induce EMT, and the cells were characterized by being compared to normal EpH4 cells and 4T1 murine mammary carcinoma cells. RT-qPCR showed no statistically significant differences in the increased mRNA expression of EMT markers Snail and Slug, while mRNA expression of vimentin was not observed. Vimentin expression at the protein level was lowly detected by immunofluorescence, suggesting that the transformation was not completed in the TGF-β-treated EpH4 cells. Further analysis on the TGF-β treatment and the expression of PA28γ downstream targets – c-Myc, phosphorylated c-Myc (Thr58 and Ser62), and PP2A will determine what is preventing the EMT transformation from reaching its end-stage. While therapies currently exist to target EMT, revealing this blockage can help understand the relationship between PA28γ and EMT as well as to elucidate the mechanism by which PA28γ provides cancer cells the ability to acquire stemness through EMT.
Research Advisor: Dr. Lance Barton
Katie Holland, Angelo State University – 1:30-3:30 in Bird Session
Adenovirus screening in Texas bats.
Katie Holland and Loren K Ammerman.
Viruses in the family Adenoviridae (AdVs) are nonenveloped, double-stranded DNA viruses that infect a variety of hosts, including reptiles, birds, fish, and mammals. Viruses in the genus Mastadenovirus may cause respiratory, ocular, and gastrointestinal disease in mammals, including humans. It is, therefore, important to understand the distribution and transmission of adenoviruses in infected organisms. Bats have been found to serve as reservoirs in the evolution of adenoviruses due to bats’ atypical ability of harboring genetically diverse viruses within a single geographic location or host species. Adenovirus DNA has been detected at unknown prevalence in Cave Myotis (Myotis velifer) in Oklahoma, but adenoviruses have not yet been detected in Texas bats. Prevalence in European bats has been found to be around 8%, so we hypothesized the prevalence of adenoviruses in Texas bats to be low. We screened intestinal and fecal samples of M. velifer collected in 2018-2021 from Texas counties and found all samples to be negative for adenovirus DNA. The absence of detection supports the hypothesis of low presence of adenoviruses in Cave Myotis in Texas.
Research Advisor: Dr. Loren Ammerman
Saraya Hunt, Southwestern Oklahoma State University – 1:30-3:30 in Serpent Session
Microbial Analysis of Regolith-Grown Species (M.A.R.S.) on Mars.
Saraya Hunt, Lisa Castle, Emilee Adams, Molly Byers, Payden Farnsley, Lauren Hartsell, Kyla Langstraat, Chloe Mellott, Joseph Olonovich, Niki Strauch, and Rachel Uhlig.
As part of the on-going Plant Mars Challenge, SWOSU students are growing plants for a duration of ten weeks in simulated Mars regolith. While an official challenge goal is to have the most crop yield in 10 weeks, the SWOSU researchers are also investigating the interactions of plant diversity and soil microbe diversity and measuring chlorophyll content and diversity within microbial communities as dependent variables. This poster describes the experimental design and rationale behind the project, as well as potential implications of the results
Research Advisor: Dr. Regina McGrane
Sneha Jacob, Oral Roberts University – 1:30-3:30 in Serpent Session
Determining Optimal Extraction Conditions to Isolate a Peptide Containing Polysaccharide with anti-cancer properties from Ganoderma Lucidum.
Sneha Ann Jacob and William P Ranahan.
Ganoderma Lucidum, commonly known as Reishi, is a popular mushroom used in Asian countries for promotion of health and longevity. It was therefore called the “Mushroom of Immortality”. In ancient Chinese medicine, Ganoderma Lucidum has been used to treat various human diseases like bronchitis, chronic hepatitis, hyperglycemia, inflammation, and cancer. In our research, a peptide containing polysaccharide, Felix, isolated from Ganoderma Lucidum showed cytotoxicity in triple-negative breast cancer cells. However, because of the availability of Ganoderma Lucidum from different sources, the biological activity and concentration of the isolated compound (Felix) varied. To determine the optimal conditions, temperature, and time, needed to extract the compound of interest, we took extraction from Ganoderma Lucidum mycelia grown at six different conditions (varying temperature and time). It was determined that extraction at 37 degrees Celsius for 48 hours gave the highest concentration of Felix (peptide containing polysaccharide). It was also found that once the extraction is done on the mycelium plate, they are no longer alive. This means we need to have a constant supply of fresh mycelium each time for the extraction and isolation of Felix. The isolated Felix was then tested on triple-negative breast cancer cells for cytotoxicity. Further research should be done on finding the concentration of Felix needed to produce the cytotoxic effect and the mechanism pathway.
Research Advisor: Dr. Julianna Goelzer
Laura Matt, Texas Wesleyan University – 10:00-10:15 in Balanos Session
Antibacterial Properties of a Novel 1,2,4 Oxadiazole.
Laura L. Matt.
Antibiotics have revolutionized medicine for the last 100 years, but not without consequences. Over the last 100 years, bacteria have become resistant to common antibiotics, like penicillin, methicillin, and vancomycin. Oxadiazoles are a class of organic compounds that are known to possess antibiotic properties. The study of oxadiazoles is new, and not much is known about how they work as antibiotics, specifically against antibiotic resistant bacteria. In the summer of 2021, a 1,2,4-Oxadiazole compound was synthesized at Texas Wesleyan University. This compound has a guanidine functional group on carbon 3 and a 4-methoxybenzene on carbon 5, which contributes to making this compound an eligible antibiotic. Similar studies done at Texas Wesleyan University have shown that oxadiazole compounds have antibacterial properties toward gram-positive and gram-negative bacteria. Assuming this novel oxadiazole has antibacterial properties, this study is focused on quantitative results to determine the minimum inhibitory concentration of the compound. The minimum inhibitory concentration will give information about how little compound is needed to inhibit growth of the bacteria. These results are presented in growth curves. The antibacterial properties of the novel oxadiazole have been tested on Enterococcus faecalis, a gram-positive bacterium, and Escherichia coli, a gram-negative bacterium. In an ever-increasing era of antibiotic resistance, it is worth characterizing the antibacterial properties of the oxadiazole against gram-positive and gram-negative bacteria and considering their applications in modern medicine. Negative control assays have been done to observe the growth of each bacterium uninhibited by antibiotics. Positive control assays have been done to observe the growth (and lack thereof) of each bacterium under the influence of ampicillin, a cell wall biosynthesis attacking antibiotic, and chloramphenicol, a translation attacking antibiotic. The antibacterial properties are currently being characterized, but there is evidence that the novel 1,2,4-Oxadiazole does work as an antibiotic.
Research Advisor: Dr. Christopher Parker
Emily Penner, Oral Roberts University – 10:45-11:00 in Balanos Session
Bioassessment of Fred Creek and Haikey Creek, Tulsa, OK water quality using aquatic macroinvertebrate diversity.
Emily Penner.
Monitoring the quality of public water sources is important due to the many potential health risks and ecological impacts involved with contaminated and polluted water. Biological monitoring can be used to evaluate the physical, chemical, and biological condition of the body of water because the populations living in a body of water provide a fair representation of the condition of the ecosystem. Aquatic macroinvertebrate abundance and diversity are good indicators of water quality; there are some populations that are pollution sensitive and others that are pollution tolerant. The relative abundances of sensitive and tolerant taxa gives a baseline idea of the overall water quality and provides data to compare with previous years and other watersheds. This study aims to evaluate the quality of Fred Creek and Haikey Creek in Tulsa, OK by collecting and categorizing aquatic macroinvertebrates by pollution tolerance, calculating the EPT/C ratio, and performing a Bio-Assess Stream Quality Assessment to indicate the relative water integrity. Samples were taken from each location once monthly from March to May and August to November, and the organisms were organized by taxa. The data was compared between the two creeks, and the data from Fred Creek was compared with existing data from previous years, including before significant construction was done at the site. Fred Creek was expected to have better diversity than Haikey based on previous studies characterizing the impairment of both creeks based on macroinvertebrates. In this study, while the two sites had similar EPT/C ratios, Fred Creek was found to have consistently higher cumulative indices for each sample, indicating better water quality than Haikey.
Research Advisor: Dr. Julianna Goelzer
Stephenie Rogers, Houston Baptist University – 11:15-11:30 in Balanos Session
Potential for Combination Antibiotic-Phage Treatment as a Viable Therapeutic.
Stephenie Rogers.
The misuse of antibiotics in medicine and industry has contributed to an increase in bacterial resistance in every known antibiotic, including vancomycin. According to the Center for Disease Control (CDC), each year nearly 3 million people in the United States suffer from antibiotic resistant infections, while deaths associated with these illnesses exceed 50,000 annually. The production of new antibiotics, while a costly endeavor, has temporarily slowed the resistance crisis. However, the rate of bacterial mutation has shown to render these new treatments ineffective within a short time, proving the need for a more permanent solution. In the midst of the current antibiotic resistance crisis, bacteriophages have come to light as a potential alternative to antibiotic therapy because of their high binding affinity and specificity for corresponding bacterial strains. Phage therapy has previously been successful as an individual therapy, as well as in combination with antibiotic therapy. Overall, there is insufficient literature related to phage-host interactions and combination antibiotic-phage therapy. This study aims to explore the potential for phage therapy and combination antibiotic-phage therapy in the treatment of Escherichia coli infections by identifying a baseline minimum inhibitory concentration (MIC) for these treatments. Three different antibiotics and a T2 bacteriophage strain were used in varying concentrations in repeated trials to determine the mean MIC for each antibiotic and phage and combination. While isolated phage may not be the most effective treatment for mild infections, there is potential for using combination antibiotic-phage therapy in various agricultural industries as an alternative to animal antibiotic treatments and bactericides, one of the main contributors to antibiotic resistance worldwide.
Research Advisor: Dr. Curtis Henderson
Kaylee Salazar, St. Edward’s University – 11:30-11:45 in Boax Session
Evaluating the link between bacterial infections and Schizophrenia.
Kaylee Salazar, Eric Torres, Angelica Carrasco-Pena, Maria Miranda, Sarah Avant, and Andrea Holgado.
The relationship between Schizophrenia and external factors controlling genetic components has been made evident, yet it has been difficult to identify the causative molecular players involved. Yoshihara and colleagues recently hypothesized that factors inactivating CRMP-2 may participate in the pathogenesis of Schizophrenia. UNC-33 in C. elegans, CRMP-2/DPYSL2 in humans, regulates neuronal development and axonal outgrowth in healthy brains and is suppressed in the brain of schizophrenic patients. Our previous findings support the hypothesis that external stressors such as temperature play a role in regulating the activity of the promoter controlling neuronal levels of UNC-33. Herein we studied the effects of pathogens on the expression driven by the unc-33 promoter. To this end, C. elegans hermaphrodites were incubated on plates seeded with Escherichia coli OP50 or Pseudomonas aeruginosa PA14 until they reached the L3/L4 stage. Confocal microscopy was used to image the head of the C. elegans to determine GFP expression driven by the unc-33 promoter in neurons and glial cells. The analysis of the quantification of twenty-five worms per condition showed that C. elegans infected since embryo with Pseudomonas aeruginosa PA14 have decreased neuronal GFP driven by the unc-33 promoter. This study provides insights into the role of the Pseudomonas aeruginosa PA14 infection in regulating the activity of the unc-33 promoter, potentially linking bacterial infections with faulty neuronal development and Schizophrenia.
Research Advisor: Dr. Andrea Holgado
Danitra Sargent, University of the Ozarks – 1:30-3:30 in Serpent Session
Two new species in the genus Vacuoliviride (Eustigmatophyceae, Goniochloridales) revealed by light microscopy and DNA sequence analysis.
Danitra P. Sargent, Marvin W. Fawley, and Karen P. Fawley.
The Eustigmatophyceae are a group of unicellular eukaryotic algae that were recently thought to be rare, but with the aid of molecular techniques are now understood to be more diverse. The name Eustigmatophyceae is a result of the morphological feature of a unique eyespot sitting outside of the chloroplast of the zoospore. This group also has commercial importance in areas like aquaculture, agriculture, medicine and biofuel production. The recently described genus Vacuoliviride is a member of the Eustigmatophyceae with a large open vacuole, a polyhedral pyrenoid and an unusual v-shaped crystal. The objective of this study is to describe new diversity in the genus Vacuoliviride. We recently isolated new strains of eustigmatophytes that can be placed in the genus Vacuoliviride based on morphological characteristics and phylogenetic analysis of DNA sequences. Our results show that both strains are new species of Vacuoliviride. We propose a new species of Vacuoliviride for strain BogD 8/9 T-2w, isolated from a small bog in Itasca State Park, Minnesota. We also propose another species of Vacuoliviride for strain Bat 8/9-5m from Batsto Lake in the pine barrens of New Jersey.
Research Advisor: Dr. Karen P. Fawley
Kristina Simons, Texas A&M University - Galveston – 1:30-3:30 in Serpent Session
A systems toxicology analysis of the effects of oxybenzone on the metabolic physiology of embryo-larval zebrafish (Danio rerio).
K.I. Simons, L. Thibault, R. Nolen, Y. Umeki, L.H. Petersen, and D. Hala.
Oxybenzone is a chemical ingredient commonly used in sunscreen lotions, as well as other hygienic products considered to be an emerging contaminant in aquatic and marine systems frequented by tourism. There is growing concern that oxybenzone can cause adverse effects in exposed invertebrate and vertebrate organisms due to metabolic and endocrine disrupting effects. Embryo-larval fish are expected to be particularly sensitive at this early life stage by oxybenzone exposure as their developmental processes are likely to be perturbed. In this study I present results of an in vivo toxicological study with embryo-larval life stages of zebrafish (Danio rerio). Fish were exposed from 2 - 9 days post fertilization (7 days exposure) to encompass environmentally relevant concentrations of 1 and 10 µg/L Oxybenzone. The levels chosen are environmentally relevant as oxybenzone levels in surface waters have been reported at <=10 µg/L (Downs et el., 2015). Micro-respirometry measurements of metabolic rate (or oxygen consumption) in fish showed a significantly elevated respiration rate at 1 µg/L; while at 10 µg/L oxygen consumption was not significantly different from the control. Whole-organism RNA-sequencing analysis indicated widespread transcriptional effects on gene regulation DNA repair and morphological systems. Subsequent computational biology analysis using a genome-scale metabolic model of zebrafish identified glycerolipid metabolism as being highly dysregulated in the 1 µg/L treatment group vs. solvent control (0.01% dimethyl sulfoxide). This prediction agrees with a metabolomics study by Ziarrusta et al., (2018) which showed glycerolipid metabolism to also be highly dysregulated in gilthead seabream exposed to 50 µg/L oxybenzone (14-day exposure). Based on my research I hypothesize that glycerolipid metabolism is diagnostic of the increased metabolic rate measured of oxybenzone at 1 µg/L in my study.
Research Advisor: Dr. David Hala
Anastasia Smith, Oral Roberts University – 1:30-3:30 in Bird Session
Analysis of Effects of Full Spectrum CBD Extracts on Gene Expressions in Triple Negative Breast Cancer Cells.
Anastasia Smith and William Ranahan.
Contemporary cancer treatments such as radiation, chemotherapy, and surgery cause negative off-target effects in nontumorigenic tissue. Alternative remedies may offer treatment with little to no side effects. Medicinal plants have been used worldwide as adjuvant therapy and treatment for cancer. Given that initial in vitro and animal model studies investigating the effect of cannabinoids on tumors show promising results, the authors proposed to investigate a full spectrum extraction of cannabinoids from hemp (A et al. 2011; Dariš et al. 2019). The authors hypothesize that this extraction will negatively affect the viability of cancer cells and change gene expression. Cannabinoids from a hemp extract were analyzed using high pressure chromatography in the NGC chromatography machine. Full cannabinoid extractions tested on ER-/PR-/Her2- cancer cell model MDA MB-468 decreased cell viability when applied in amounts of four, eight and twelve µl in 100 µl of total suspension with 20,000 cells per well. The extractions were again tested on MDA MB-468’s, but in a six cm petri dish and successful results were used to obtain RNA. cDNA was made from RNA samples and added in a primePCR assay. The primePCR assay well’s contained preloaded primers for 91 genes and four controls. Of these genes, three were specifically examined: HMOX1, IL6, and VIM. The authors found these genes to be over expressed when compared to the control tested with olive oil. Overexpression of these genes is correlated with increased cancer aggression and poor clinical prognosis Thus, the authors hypothesis that upregulated of HMOX1, IL6, and VIM may be an epigenetic modification the MDA-MB-468’s to evade the treatment. Future studies will aim to examine other genes analyzed in the primePCR assay, test isolated cannabinoids, and examine the effects of full hemp extracts on different cell types.
Research Advisor: Dr. Julianna Goelzer
Emma Solis, Austin College – 10:15-10:30 in Boax Session
Evaluation of the relationship between a high-fat diet and gut microbiome health in thirteen-lined ground squirrels (Ictidomys tridecemlineatus).
Emma Solis and Jessica Healy.
Hibernating species typically undergo extreme cycles of fat accumulation and loss throughout the year as they maintain their normal circannual rhythm of euthermia and torpor. With feeding patterns changing drastically during the year, there have been several papers detailing subsequent seasonal differences in gut microbiota structure and diversity according to a standard hibernation cycle. However, little research has been done to evaluate if other factors such as dietary fat content or natural vs. lab diets have any direct effects on the gut health and microbial community of hibernating species. In this project we are using behavioral (food intake), morphological (body mass and composition), and molecular (qPCR) techniques to evaluate the physiological effects of a high-fat diet on our animal of study, the thirteen-lined ground squirrel (Ictidomys tridecemlineatus) to observe potential differences that emerge as wild-caught animals are transitioned to a lab diet.
Research Advisor: Dr. Jessica Healy-LaPrice
Megan Strotman, Southwestern Oklahoma State University – 1:30-3:30 in Serpent Session
Investigating Potential Circadian Oscillations in Pseudomonas syringae.
Megan Strotman and Regina McGrane.
Pseudomonas syringae is a gram-negative plant pathogen that encodes a kaiC ortholog. KaiC has been proven to regulate circadian oscillations in cyanobacteria, which control transcriptional oscillations, cell division, and photosynthesis. We hypothesize that P. syringae is capable of circadian oscillation and that kaiC contributes to regulation of stress response, biofilm development, motility, and plant colonization. To determine if P. syringae has circadian oscillations, the wild type strain or mutants lacking kaiC were spotted onto King’s B agar media supplemented with coomassie blue, which binds proteins, or congo red, which binds exopolysaccharides. The plates were then exposed to day-night cycles (12 hours light, 12 hours dark) or dark conditions for seven days. Oscillations in protein and exopolysaccharide production were then observed due to varying intensity of either blue or red rings in the colonies. While oscillations were detected, analysis of the plates indicated no significant differences in the number or pattern of the rings when comparing the mutants to the wild type or the plates exposed to day-night cycles to dark conditions. To determine if kaiC regulated P. syringae stress response, wild type or mutants lacking kaiC were exposed to day-night cycles or dark conditions as described above and inoculated in MinA media supplemented with glucose and varying concentrations of salt. Growth was then recorded by detecting absorbance every 30 minutes over a 48-hour period. Analysis revealed significant differences in the growth of the wild type exposed to day-night cycles compared to samples kept in the dark; however, these differences were not observed in mutant strains. This suggests that kaiC may modulate the ability of P. syringae to combat stressful environments. Determining the function of kaiC in P. syringae may allow for development of techniques that target this ortholog and prevent P. syringae growth on crops.
Research Advisor: Dr. Regina McGrane
Uzziah Urquiza, Southwestern Oklahoma State University – 11:00-11:15 in Boax Session
Investigation of the molecular mechanisms underlying the cytotoxicity of novel lactate dehydrogenase A inhibitors against human pancreatic cancer.
Uzziah Urquiza, Hanna Hill, Pragya Sharma, and Horrick Sharma.
Pancreatic cancer is the 3rd leading cause of cancer-related death in the United States. Chemotherapy is one of the standard treatment options for pancreatic cancer; however, the rapid development of resistance to chemotherapeutic agents represents a significant bottleneck in treatment options. The average life expectancy after diagnosis with metastatic pancreatic cancer is just three to six months reflecting an urgent unmet need to identify new targets. Cancer cells are well-known to alter their metabolic pathways to support their growth and proliferation. Unlike normal cells, cancer cells prefer aerobic glycolysis, a phenomenon described by Otto Warburg, known as the Warburg effect. This unique metabolic alteration in cancerous cells is regulated by lactate dehydrogenase A (also known as LDH5), an enzyme that is overexpressed in many tumor types, including pancreatic cancer. In collaboration, our lab has identified several novel lactate dehydrogenase inhibitors, which have shown promising anticancer activity against pancreatic cancer cell lines. The goal of the project study is to uncover the molecular mechanisms of antiproliferative activity of our lead compounds. Our preliminary data indicate the activation of the pro-apoptotic protein pathway as a possible mechanism of cytotoxicity of pancreatic cancer cells.
Research Advisor: Dr. Pragya Sharma
Arianna Vacio, St. Edward’s University – 1:30-3:30 in Serpent Session
UNC-33L partially rescues lifespan and locomotion defects in unc-33 mutants but fails to rescue dauer formation defects.
Arianna Vacio.
Aging is a process that gradually decreases an organism's physical and mental capacity, increasing the risk of disease and death. Neurodegenerative diseases, such as Alzheimer’s and Parkinson’s, result from accelerated neuronal cell death, which leads to neurophysiological decline, deterioration of the body, and reduction in the standard of living. UNC-33, homolog to CRMP2, serves as a biomarker for these neurodegenerative diseases. In healthy tissue, CRMP2 regulates neuronal development and axonal outgrowth. In disease, CRMP2 is inactivated by hyperphosphorylation, compromising the maintenance of axons. To better understand the role of CRMP2/UNC-33 in the aging process, we tested the ability of UNC-33L, one of the isoforms coded by the unc-33 gene, to rescue the shortened lifespan, defective locomotion, and the failure to make dauers, a non-aging life stage, in C. elegans unc-33 mutants. Analysis of results demonstrates that the presence of UNC-33L does not rescue the defective dauer phenotype in unc-33 mutants. However, UNC-33L significantly rescued premature death and uncoordinated locomotion in young unc-33 adults. The degree of UNC-33L-mediated rescue was less noticeable as the nematodes aged, denoting that both age and the presence of UNC-33L interact in the production of the phenotypes.
Research Advisor: Dr. Andrea Holgado
Luke Whelchel, Dallas Baptist University – 10:15-10:30 in Balanos Session
Urinary Inflammatory Biomarkers and Gut Microbiome are Weight Dependent in Adolescents.
Luke Whelchel.
Obesity has risen dramatically in recent decades. Studies have shown obesity causes inflammation through activation of pro-inflammatory cytokines. Obesity has also been linked with dysbiosis of the gut microbiome. The goal of the study was to evaluate noninvasive biomarkers of inflammation and gut microbiome and follow impacts of lifestyle modification in adolescents that were overweight and obese. Thirty-eight adolescents were enrolled: sixteen healthy (BMI 30). Anthropometric measurements were taken at enrollment and were followed overtime. HbA1c, blood pressure, and lipid profiles were taken at the time of enrollment. A lifestyle modification program with the goal of promoting healthier eating and more active lifestyles was implemented. Urine was collected from all participants at enrollment and at 3- and 6-months in the program and analyzed for TNF-α, IL-6, and ET-1 by ELISA. Stool samples were collected at enrollment and compared to healthy peers. Adolescents of the OW/OB group exhibited elevated levels of urinary inflammatory markers compared to healthy control groups at enrollment. After six months, urinary pro-inflammatory TNF-a significantly decreased. Bacterial DNA analysis indicated that fecal samples from the OW/OB group demonstrated differing microbiome profiles as well as an increased presence of pathogenic bacterial families compared to healthy controls. Adolescents that were OW/OB demonstrated increased levels of urinary inflammatory cytokines, which improved under the lifestyle modification program. Notable differences in the gut microbiome compositions between adolescents who were OW/OB and their healthy counterparts were observed.
Research Advisor: Dr. Tetyana L. Vasylyeva
