Abstracts (sorted by last name of presenter)

Emily J. Aller, Austin College – 10:30-10:45 AM in Balanos Session 
The Function of The Oncogene c-Myc Is Affected By PA28γ In Cancer.
Emily J. Aller.
Cancers are identifiable by the demonstration of ten hallmarks, such as increased proliferation and capacity for metastasis. Proteins contributing to multiple hallmarks are generally more likely to be altered by mutation in cancers making them appealing targets for the development of chemotherapies. One such protein affecting multiple hallmarks is c-Myc, a transcriptional regulator involved in growth, proliferation, metabolism, and differentiation. Meta-analysis of gene expression data indicates a correlation between cellular levels of c-Myc and the proteasome activator, PA28γ in multiple cancers. Recent studies have demonstrated contradicting roles for PA28γ in regulating c-Myc’s cellular abundance, with one study indicating that it degrades c-Myc via the ubiquitin-independent proteasomal protein system (UIPPS), and another indicating that it stabilizes c-Myc via an unknown mechanism. This project investigated the relative importance of the ubiquitin proteasome system (UPS) and the UIPPS in regulating c-Myc’s stability in cancer. Using normal and cancer cell lines expressing differing amounts of PA28γ, I identified a correlation between elevated PA28γ expression and increased c-Myc levels in breast cancer.  Increased c-Myc levels, however, are not due to alterations in UPS-mediated c-Myc degradation nor alterations in UIPPS function alone. Therefore, further analysis of PA28γ function in these cells is required to determine its viability as a chemotherapy target.
Research Advisor: Dr. Lance Barton
 
Sara Ambrocio Paque, University of the Ozarks – 1:15-1:30 PM in Boax Session 
New species of the green algal genus Coelastrella Chodat from Warren Prairie Natural Area in southeast Arkansas.
Sara Ambrocio Paque, Marvin Fawley and Karen Fawley.
Warren Prairie Natural Area 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 our study was to characterize the soil crust eukaryotic algal communities using morphological and molecular techniques. We have characterized strains isolated from soil samples collected in February, 2016 and December, 2017.  We generated 18S ribosomal RNA gene sequences for the strains and used BLAST searches of the GenBank database to determine preliminary identifications.  Several strains were consistent with the coccoid green algal genera Coelastrella Chodat and Asterarcys Comas (Chlorophyta; Chlorophyceae). Morphology analysis was made by using a light microscope, the cells had irregular spherical shape, uninucleate with presence of asexual reproduction. Phylogenetic analyses of both the ribosomal 18S and ribosomal RNA internal transcribed spacer (ITS) and tufA regions indicated that some of the strains are a new species of the genus Coelastrella.  Other strains are closely related to the recently described species, Coelastrella yingshanensis Qinghua Wang et al., but there is some evidence that these strains may also be one or more new species.
Research Advisor: Dr. Karen P. Fawley
 
Riley Andrews, Oral Roberts University – 9:00-9:15 AM in Balanos Session 
Attenuation of Angiomotin Expression via CRISPR in Triple-Negative Mammary Epithelia.
Riley Andrews and William Ranahan.
Breast cancer is the most commonly diagnosed form of cancer among women living in the United States, and it is characterized by uncontrolled cell growth in either the lobules or ducts of the breast tissue. Additionally, breast cancer accounts for the second highest number of cancer deaths, only trailing behind lung cancer. Current efforts to treat this form of cancer include various combinations of chemotherapy, mastectomy, radiation, and endocrine therapy; however, there is still a need for treatments with increased selectivity. Use of the CRISPR/Cas9 gene-editing tool is a method that could be used to directly target cancer cells. This gene-editing tool is guided to the appropriate sequence through a complementary small guide RNA (sgRNA) that can be designed for any gene of interest. Angiogenesis, which is required in order for tumor cells to survive, is the process in which new blood vessels are formed from pre-existing vessels, and angiomotin (Amot) is directly involved in this process. Additionally, various studies have found that Amot enables the proliferation of mammary epithelia through the activation of extracellular signal-regulated kinases (ERK1/2). Therefore, Amot is thought to be a putative oncogene involved in breast cancer. The current study focused on reducing Amot expression in tumorigenic MDA-MB-468 cells via CRISPR. Cancer cells were transfected with varying guide RNA (gRNA) sequences and varying concentrations of CRISPR-containing DNA constructs. qPCR data suggested that the gRNA sequence “Amot 174” was most effective at reducing Amot mRNA transcripts. Amot mRNA reduction correlated with a decrease in proliferating cell nuclear antigen (PCNA) mRNA transcripts, suggesting a decrease in cell viability. Follow up studies will include Western blotting to confirm reduction in Amot protein, cell viability assays to confirm cytotoxicity, and end-point PCR to confirm editing of the Amot gene.
Research Advisor: Dr. William Ranahan
 
Rachel S. Armfield, University of Central Oklahoma – 2:30-2:45 PM in Johnson Poster Session 
Differential Gene Expression in a Murine Model of Maternal Phenylketonuria.
Rachel S Armfield.
Maternal PKU affects embryos exposed to high concentrations of Phenylalanine (Phe) in utero due to PKU in the mother.  MPKU leads to craniofacial, cardiac and cognitive abnormalities.  The mechanism of MPKU is unknown, as are the specific genes which are differentially expressed in the presence of high Phe.  Cranial neural crest cells (O9-1 cell line) from Mus musculus (mouse) were exposed to the high Phe concentrations that occur in utero.  mRNA was then extracted and converted to cDNA.  Quantitative Real-Time PCR (qRT-PCR) was performed on this cDNA, initially for 3 ADH (alcohol dehydrogenase) and 2 ALDH (aldehyde dehydrogenase) genes, and then for a Retinoic Acid pathway array of 84 genes.  Data were analyzed using ΔΔCt method, and expression changes were given as a log2 fold change.  Thirteen genes in the Retinoic Acid pathway were found to be significantly upregulated in the presence of Phe.  Several are known to contribute to heart development in the embryo, and one is known to be necessary for neuron development in the brain.
Research Advisor: Dr. Nikki Seagraves
 
Bryler Atchley, Southwestern Oklahoma State University – 9:45-10:00 AM in Balanos Session 
Characterization of the Antimicrobial Properties of Syringafactin.
Bryler Atchley, Virginie Sjoelund, and Regina McGrane.
Pseudomonas syringae is a gram-negative plant pathogen that produces a biosurfactant called syringafactin. Syringafactin, a lipooctapeptide, 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 lipooctopeptides 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 for the potential effectiveness of syringafactin as an antibacterial agent.
Research Advisor: Dr. Regina McGrane
 
Christian Baker and Juan Gonzalez, Northwestern State University of Louisiana – 12:15-12:30 PM in Johnson Poster Session 
Helminthological survey of channel catfish (Ictalurus punctatus) in aquaculture hatchery (Natchitoches, Louisiana, USA)​.
Christian Baker and Juan Gonzalez.
Helminths are important parasitic worms that cause disease and reduced growth in fish and other vertebrates. Helminthological surveys can be used to determine the community structure within host populations and can provide information useful for the mitigation of communicable disease in aquaculture. This project aims to survey helminths in farmed Ictalurus punctatus (Natchitoches National Fish Hatchery) and compare the parasite community structure across different host age classes. Ten fish were collected from each of three age classes, for a total sample size of 30. Parasites were collected from the gastrointestinal tract, body cavity, and musculature, and stained using an aceto-carmine and alcohol dehydration protocol, then permanently mounted on microscope slides. Parasites were enumerated and identified based on morphological traits compared to taxonomic keys and published literature. In the one-year age class, 71 parasites were collected (Hysteromorpha sp. n=49, Clinostomum sp. n=21, Alloglossidium sp. n=1). In the three-year age class, 88 parasites were collected (Hysteromorpha sp. n=77, Clinostomum sp. n=2, Alloglossidium sp. n=9). In the four-year age class, 144 parasites were collected (Hysteromorpha sp. n=100, Clinostomum sp. n=16, Alloglossidium sp. n=28). Total parasitic load was analyzed via simple linear regression.​
Research Advisor: Dr. Cynthia Doffitt
 
Shawn Bounkhong, Southwestern Oklahoma State University – 12:45-1:00 PM in Johnson Poster Session 
Characterization of Biosurfactant Chemoreceptors in Pseudomonas aeruginosa.
Shawn Bounkhong and Regina McGrane.
Pseudomonas aeruginosa is a bacterium that causes nosocomial infections. Pathogenesis of P. aeruginosa involves chemotactic abilities in which bacteria swim toward environments that have sufficient nutrients to promote growth, proliferation, and biofilm formation. Chemotaxis in P. aeruginosa involves 26 chemoreceptors; however, only 4 of these chemoreceptors have been fully characterized. The primary interest of this study was to investigate P. aeruginosa chemoreception of the lipopeptide biosurfactant, syringafactin, produced by the phytopathogen, Pseudomonas syringae. Syringafactin was previously shown in our laboratory to repel P. aeruginosa via chemotaxis. To investigate which chemoreceptor is involved in detecting syringafactin, we utilized P. aeruginosa strains harboring mutations in chemoreception genes. The chemotactic abilities of mutant strains were evaluated on soft agar media by inoculating each mutant next to P. syringae. Strains with mutations in the genes PA1608, PA2573, and PA4520 displayed poor chemotaxis when inoculated near P. syringae compared to wild type P. aeruginosa, which displayed strong chemotaxis. To further characterize these mutants, chemotaxis capillary assays were performed by filling capillary tubes with purified syringafactin, placing the open capillary in cultures of chemoreceptor mutants, and comparing the concentration of bacteria in the capillary tube at the end of a 30-minute incubation to capillaries filled with water. Bacterial concentrations were determined by spread plating and counting colony forming units. If mutation in any of the three chemoreceptor genes does inhibit chemorepulsion in response to syringafactin, we expect that the mutant strain would be present in higher concentrations in the capillary tube compared to wild type P. aeruginosa. We hypothesize that bacteria that sense and respond to the presence of syringafactin are able evade it’s the antimicrobial activity. By identifying the specific chemoreceptors involved in chemotactic response to syringafactin we can gain a better understanding of how bacteria sense and respond to potential microbial antagonism.
Research Advisor: Dr. Regina McGrane
 
Caden Bowles, Southwestern Oklahoma State University – 1:00-1:15 PM in Johnson Poster Session 
Foraging behavior of fruit flies (Drosophila melanogaster) on patches of sugar: the effects of distance and food quality.
Caden Bowles, Tommy Nguyen, Raistlin Hiner, and Jimena Aracena.
Animals use searching mechanisms that optimize energy while foraging on patches, which are food sources arranged in groups. We used fruit flies (Drosophila melanogaster) to test the effect of food quality and distance between patches on their foraging behavior. The flies were allowed to choose between a patch with red 0.25 M sucrose solution and a patch with blue 0.125 M solution. We tested five distances (0 to 14 cm) between patches and allowed 100 flies to feed in the dark for one hour at each distance. The flies were then scored according to their abdomen color to assess their preference. There was no clear distance effect on the preference for the higher concentration patch. However, there was a higher proportion of visits when the patches were close together. The 0.25 M and 0.125 M concentrations were probably not different enough for the flies to show a preference and allow us to detect the decision making between food patches. We plan to change the concentration difference to 0.25 M vs. 0.0625 M, which should result in a clear preference for the higher-quality patch. This will allow us to better test the effect of distance on decision making while foraging on patches.
Research Advisor: Dr. Jimena Aracena
 
Hannah Budde, Southwestern Oklahoma State University – 9:00-9:15 AM in Boudetase Session 
Investigating the impact of phytopathogen competition on the survival of Salmonella enterica and Escherichia coli on lettuce.
Hannah Budde, Payden Farnsley, and Regina McGrane.
Human pathogen colonization of crops leads to economic losses, illness, and fatality and is of growing concern due to antimicrobial resistance. The phytopathogen Pseudomonas syringae secretes syringafactin during plant colonization. Our laboratory has shown that syringafactin repels and kills gram-negative bacteria, suggesting it is a potential solution to human pathogen colonization of plants. We hypothesize that the presence of P. syringae on leaves excludes human pathogens like Salmonella enterica and Escherichia coli via syringafactin. To test this hypothesis, P. syringae and either S. enterica or E. coli were co-inoculated on lettuce and the population dynamics were compared to plants inoculated with each species alone. We found that S. enterica reached higher concentrations when co-inoculated with P. syringae compared to when S. enterica was inoculated alone. One possible explanation is that S. enterica is profiting from the ability of syringafactin to increase water and nutrient diffusion from the plant. Another explanation is that the negative chemotaxis induced in S. enterica by syringafactin allows S. enterica to evade the antimicrobial impacts of syringafactin. To investigate this, we plan to evaluate colonization of S. enterica mutants lacking chemotaxis when co-inoculated with P. syringae. We also found that when P. syringae is co-inoculated with E. coli, the E. coli concentration is comparable to when inoculated alone. However, the concentration of P. syringae was noticeably lower when co-inoculated with E. coli compared to when P. syringae was inoculated alone. This result provides evidence that bacterial competition on leaves may be a selective pressure for phytopathogens. Although the results of this experiment where not expected, evaluation of S. enterica and E. coli chemotaxis mutants, in addition to evaluation of the impact of crude syringafactin extracts on S. enterica and E. coli plant colonization, could still support the use of syringafactin as an antimicrobial agent in agriculture.
Research Advisor: Dr. Regina McGrane
 
Larry Cossey, Southwestern Oklahoma State University – 1:15-1:30 PM in Johnson Poster Session 
Effects of Controlled Traffic on the Microbial Communities on Doorhandles.
Larry Cossey and Regina McGrane.
With the onset of COVID-19 all eyes were on the microscopic world. Increased sanitation and proper hygiene slowed the spread of the virus, and there were many other tactics used to battle COVID-19. One of these tactics was to limit the number of times people crossed paths. Southwestern Oklahoma State University (SWOSU) developed guidelines regarding the movement of students through buildings when hybrid classes began in the fall 2020. In the Old Science Building at SWOSU, doors were designated entrances or exits. By implementing this strategy, many people were forced to use the same doorhandles, which are known to harbor microbes. In this work we analyzed the impact of controlling movement through a building on the microbial communities found on doorhandles. We hypothesized that high traffic doorhandles would have higher concentrations of microorganisms compared to low traffic doorhandles. The following doorhandles were examined, high traffic or low traffic and indoor or outdoor. Three sets of doors were sampled from the first floor, and three were sampled from the second floor. On each side of door, there was either a high traffic handle or a low traffic handle. Three locations, each with an approximate size of two inches squared, were sampled on each doorhandle. Sterile cotton swabs were used to collect microbes and the microbes were inoculated on tryptic soy agar. This experiment was repeated four times. Following incubation, the average colony forming units for each condition were determined. Results were highly variable. We did not detect significant differences in the concentration of microorganisms between any of the treatments. This may be the result of increased disinfection practices.
Research Advisor: Dr. Regina McGrane
 
Ethan Cranford, University of Louisiana at Monroe – 12:30-12:45 PM in Johnson Poster Session 
Analysis of Biofilming Capabilities of Novel Rhizobium radiobacter Phages from Louisiana Soil.
Ethan Cranford, Bailey Mabou, Emmanuel Perez, and Allison Wiedemeier.
Antibiotics are not always effective methods of treatment for bacterial infections. This is especially true of antibiotic resistant strains of bacteria and organisms present in biofilms. Cells within biofilms have some characteristics that differ from their planktonic counterparts, making treatment increasingly difficult. This is the case with Rhizobium radiobacter, formerly Agrobacterium tumefaciens, the causative agent of Crown Gall disease which reduces yield in many crops. This organism can position itself on the plant surface while in a biofilm state, so that when the plant is wounded entrance into the plant interior is guaranteed. Our research aims to show the effects of bacteriophage therapy on the biofilm formation of this organism. Bacteria were cultured in a 96 well plate under conditions to that caused biofilms to form. Then each well received treatment with a bacteriophage lysate. The biofilm was then stained and solubilized to measure the amount of biofilm formed. This was then compared to a control that did not receive treatment. Preliminary research led us to hypothesize that a certain phage, JP1, would reduce biofilm mass. We are currently conducting a time course experiment to elucidate the relationship between JP1 and R. radiobacter biofilm formation.
Research Advisor: Dr. Allison M Wiedemeier
 
Aria Dang, Houston Baptist University – 11:15-11:30 AM in Balanos Session 
Using malaria to understand the proteins of Toxoplasma gondii parasitism.
Aria Dang.
Toxoplasma and Plasmodium walk into a bar to discuss sugar. Both come to conclude that the protein, RON3, will be a good way to acquire such nutrients and implement RON3 into their respective parasitization strategies...“Cat scratch fever” is caused when a cat scratches and inoculates a warm-blooded host with Toxoplasma gondii, an intracellular parasite. Typical of the apicomplexan protists, T. gondii possesses a pointed apical end, housing specialized organelles called micronemes and rhoptries. These organelles secrete proteins that aid in host invasion. Of interest are rhoptry neck proteins (RONs), which create the necessary ‘landing pad’ on which the parasite may attach and enter the cell. RONs also aid in the maintenance of a parasitophorous-vacuole, an organelle in which T. gondii may reproduce and feed on the host cell’s nutrients. Yet while several Toxoplasma RON proteins (tgRON) have been characterized as necessary components of host invasion, tgRON3's physiological impact is largely undefined. Comparisons between amino acid sequences have demonstrated that the better-characterized Plasmodium RON3 is similar to tgRON3. These similarities point towards conserved functions, such as obtaining glucose. Researching tgRON3 and its potential interactions with other proteins add to our understanding of an otherwise poorly-characterized protein, and by extension, Toxoplasma’s nutrient acquisition.
Research Advisor: Dr. Matthew Blank
 
Jireh Gillian De La Cruz, University of Dallas – 2:30-2:45 PM in Boax Session 
The role of glial cells in the nociceptive circuit of Drosophila melanogaster third instar larvae.
Jireh Gillian De La Cruz, Linh Nguyen, Philip Arlinghaus, and Drew Stenesen.
Nociception provides a model sensory circuit through which potentially harmful stimuli evoke a stereotypic behavioral response.  Several genes involved in nociception within Drosophila melanogaster have been identified; however,  most studies focus on aspects necessary to individual circuit neurons. Here we identify a role for the defining glial transcription factor, reversed polarity (repo), within the nociceptive circuit. Our data demonstrates a decreased sensitivity to pain in repo[1] mutants relative to wild type controls. These findings indicate that glial cells play an important role in support of sensory neurons transmitting information related to painful stimuli and establish nociception as a tangible circuit to study the functional dependence of glial-neuronal cell relationships.
Research Advisor: Dr. Drew Stenesen
 
Hallie Dickerson, Austin College – 2:15-2:30 PM in Boax Session 
A 50 Year Restoration Difference Increased Small Mammal Abundance, but not Diversity in Blackland Prairie.
Hallie Dickerson.
Tallgrass prairie is characterized by a grass dominated landscape with a large forb variety and very few shrubs and trees. Almost all the tallgrass prairie in North America has been lost to agriculture which produces less ecosystem services: ground water absorption, soil accumulation, carbon sequestration, etc (Rowe 2010). Prairie ecosystem benefits have led many to see the gain from prairie restoration. Prairie restoration is the attempt to convert land back into its natural ecosystem of grasses and forbs with minimal trees. I investigated the restoration progress of Sneed prairie (formerly a farm) in Sherman, TX using multiple methods not previously applied systematically at this location. Initially, I completed an in-depth terrestrial vertebrate (small mammals & herpetofauna) and invertebrate census. Then I compared the small mammal species found at Sneed to those found at the Lyndon B. Johnson National Grassland field location of National Ecological Observation Network (NEON). I used the Lyndon B. Johnson (LBJ) National Grassland small mammal trapping data as a reference to assess the restoration progress of Sneed, while considering general differences between the locations. The field treatments at Sneed (prescribed burning, grazing, and mechanical management) allowed me to compare the species present in each field treatment type and identify if a certain type of prairie management is associated with higher diversity or abundance of vertebrate populations. LBJ, with 70 years of restoration management had higher species present and abundance as compared to Sneed. However, LBJ did not have significantly higher species richness or higher diversity values. Between the treatments at Sneed there was higher species diversity in the fire, cattle, and mechanical field treatment as compared to the other two.
Research Advisor: Dr. Jessica Healy
 
Kade Ezell, Southwestern Oklahoma State University – 12:00-12:15 PM in Boax Session 
Impacts of Naturally Occurring Antimicrobials on Eukaryotic Organisms.
Kade Ezell and Regina McGrane.
Antimicrobials play a major role in the fight against bacterial growth in many areas of the modern world. Without the benefits of antimicrobials, the medical field, food industry, and overall well-being of countless lives would be significantly impacted. The gram-negative, phytopathogen Pseudomonas syringae produces the biosurfactant syringafactin while colonizing plant tissue. This biosurfactant lowers surface tension, act as a lubricant for bacterial swarming motility, increases water availability, and diffusion of plant nutrients. Our laboratory has observed that P. syringae can repel leaf colonizing bacteria and gram-negative human pathogens when inoculated in close proximity to them on swarming agar. Furthermore, we have demonstrated that syringafactin causes cell death of gram-negative human pathogens. We hypothesize that syringafactin may be important for P. syringae competition on leaves through repulsion of nearby bacteria and that this microbial antagonism could be harnessed in medicine and agriculture. For syringafactin to be a viable biocontrol agent in agriculture and/or antimicrobial agent used in medicine, it must be non-toxic to eukaryotes. The goal of this work was to investigate the impact of syringafactin on wheat seed development and wax moth larvae survival. To evaluate the impact of syringafactin on seed development, wheat seeds were soaked in varying concentrations of syringafactin and germination rate, as well as length of radicals, were recorded for seven days. To evaluate the impact of syringafactin on wax moth survivability, larvae were injected with varying concentrations of syringafactin and the proportion moving and alive were recorded for four days. Our results demonstrate that syringafactin had no significant effect on the germination rate or radical length of wheat seeds nor the survivability of caterpillar larvae. Collectively, this work supports the proposal that syringafactin is a viable antimicrobial for use in agriculture and medicine.
Research Advisor: Dr. Regina McGrane
 
Abigail Fajardo, University of St. Thomas – 1:00-1:15 PM in Boax Session 
Stress Promotes a More Aggressive Phenotype in Human Melanoma Cells.
Rebecca A. Rosero , Jerry L. R. Amomoy, Elizabeth Klettke, Vanessa Phung, Jenny M. Tran, Abigail Fajardo, Laila Barkoudeh, Cecilia Nguyen, Francisca Gutierrez, Heidi Diaz, Melody Zarghooni, Kati H. Phan, Abigail Contreras,Guillermo N. Armaiz-Peña, and Gabriel J. Villares.
Growing evidence suggests that stress plays a vital role in metastasis and tumor development by activating the sympathetic nervous system. The catecholamines released into the tumor microenvironment (TME), specifically norepinephrine (NE), results in a cascade effect leading to a variety of pro‐metastatic activities that sustain tumor growth and increase melanoma aggressiveness. Most notable are the secretion of cytokines and the stimulation of tumor‐associated macrophages (TAMs). Using a co‐culture system of melanoma cancer cells and macrophages we found that sustained exposure of NE in the microenvironment induced the release of tumor‐growth and progression‐associated cytokines from TAMs. Of the upregulated cytokines found; IL‐11, IL‐24, GRO‐a, DKK‐1, and angiopoietin‐2 are known to induce growth, migration, invasion, and tumor development in human melanoma. Our next steps include validating the cytokine array using real time PCR and using an ELISA assay to quantify protein levels of antigens associated with the cytokines secreted from the cells and macrophages.
Research Advisor: Dr. Gabriel Villares
 
Kaitlin Galassini, Southwestern University – 10:15-10:30 AM in Balanos Session 
Old School or New School? Comparing the Efficacy of Traditional eDNA Hand Sampling and the Novel ANDe™ eDNA Backpack.
Kaitlin Galassini, Esther Nyaberi, Matthew A. Barnes, and Romi L. Burks.
Ecological conservation efforts increasingly employ ways of sampling that do not require collection of the whole organism. Environmental DNA (eDNA) consists of trace genetic material that an organism releases into its habitat. For aquatic samples, eDNA sampling often requires researchers to transport water samples from the field to the laboratory. A new eDNA backpack (Smith Root ANDe™) potentially allows for improved replicability of eDNA field sampling. Although easy to use, eDNA backpack sampling costs considerably more because of the special filters used. We targeted identification of Pomacea maculata, a non-native invasive apple snail species. Sampling in a Houston nature reserve invaded by the snails, we examined the amount of eDNA recovered from samples collected by hand or using the on-site filtering provided by the ANDe™ backpack. At both sites (near egg clutches and away from egg clutches), we collected four samples by hand (500 mL) and filtered four 1.0 L samples using the ANDe™ backpack. In comparing methods, on-site filtering took considerably less time than filtering samples by hand. Hand sampling successfully filtered only one fourth of the volume as the eDNA backpack. We extracted all samples using a chloroform isopropanol protocol (CTAB) that did not fully dissolve the filters within the eDNA filtering apparatus. Thus, the possible need to alter widely accepted extraction protocols for the different filters might be a cost dependent issue in field sampling. While there were no significant relationships between sampling method or collection site (R2 = 0.992, p-value = 0.08), the observed results indicated trends that there was more eDNA closer to egg clutches, more eDNA with hand sampling, and there was less of a difference between sites with eDNA backpack.
Research Advisor: Dr. Romi L. Burks
 
Juan Gonzalez and Christian Baker, Northwestern State University of Louisiana – 12:15-12:30 PM in Johnson Poster Session 
Helminthological survey of channel catfish (Ictalurus punctatus) in aquaculture hatchery (Natchitoches, Louisiana, USA)​.
Christian Baker and Juan Gonzalez.
Helminths are important parasitic worms that cause disease and reduced growth in fish and other vertebrates. Helminthological surveys can be used to determine the community structure within host populations and can provide information useful for the mitigation of communicable disease in aquaculture. This project aims to survey helminths in farmed Ictalurus punctatus (Natchitoches National Fish Hatchery) and compare the parasite community structure across different host age classes. Ten fish were collected from each of three age classes, for a total sample size of 30. Parasites were collected from the gastrointestinal tract, body cavity, and musculature, and stained using an aceto-carmine and alcohol dehydration protocol, then permanently mounted on microscope slides. Parasites were enumerated and identified based on morphological traits compared to taxonomic keys and published literature. In the one-year age class, 71 parasites were collected (Hysteromorpha sp. n=49, Clinostomum sp. n=21, Alloglossidium sp. n=1). In the three-year age class, 88 parasites were collected (Hysteromorpha sp. n=77, Clinostomum sp. n=2, Alloglossidium sp. n=9). In the four-year age class, 144 parasites were collected (Hysteromorpha sp. n=100, Clinostomum sp. n=16, Alloglossidium sp. n=28). Total parasitic load was analyzed via simple linear regression.​
Research Advisor: Dr. Cynthia Doffitt
 
Stephany A. Gutierrez, Texas Wesleyan University – 10:45-11:00 AM in Balanos Session 
Analysis of the evolution of the widerborst gene, a critical regulator of the insulin pathway, in the genus Drosophila.
Stephany A. Gutierrez, and Chitra Chandrasekaran.
The insulin pathway is important for normal human development; failure to respond to insulin is the cause of diabetes, which is one of the leading causes of illness and death in the United States. The genes of the insulin pathway are conserved in a wide variety of organisms, including the fruit fly Drosophila melanogaster. The focus of this research is to evaluate the evolution of the widerborst gene, which is a regulator of the insulin signaling pathway, in the genus Drosophila. The widerborst gene encodes a catalytic subunit of the protein phosphatase 2A (PP2A) complex, which regulates the activity of the downstream signaling molecules of the insulin pathway.  Using a bioinformatics approach, we present the results of annotating widerborst gene sequences in 10 Drosophila species.  We determined that the widerborst-PA isoform had significant similarity in its catalytic domain region in all species evaluated, and will present analysis of the amino acid similarity of these predicted proteins.
Research Advisor: Dr. Chitra Chandrasekaran
 
Elizabeth Gwartney, Oklahoma City University – 11:00-11:15 AM in Boudetase Session 
Novel Antibiotics from Oklahoma Soil.
Elizabeth Gwartney and Greg Mullen.
According to the United States Centers for Disease Control and Prevention’s (CDC) 2019 Antibiotic Resistance Threats Report, “more than 2.8 million antibiotic-resistant infections occur in the U.S. each year, and more than 35,000 people die as a result” (CDC). My research attempts to contribute to the ongoing race to discover new antibiotics from non-traditional sources. Microbes that inhabit soil often produce antibiotic substances as a way to inhibit growth of other microorganisms that compete for resources. Thus, antibiotic production confers an evolutionary advantage. Bacteria and fungi from garden soil in northeast Oklahoma were tested for the ability to inhibit growth of various pathogenic bacteria including Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and Mycobacterium smegmatis. These bacteria were chosen based on their relationship to the pathogens identified in the CDC report as serious threats and common infection-causing organisms. Four novel microorganisms were identified that secrete soluble substances which inhibited growth of common pathogens: CRB1, CRB2, CRB3 and CRB4. Inhibition assays similar to the Kirby-Bauer assay were used to define the spectrum of pathogens inhibited by each of these isolates. The antibiotic secreted by the CRB3 strain was especially broad spectrum, inhibiting a wide range of Gram-positive and -negative organisms. Sequencing of the 16S rRNA gene was used to determine the phylogenetic relationships of these isolates to known bacterial species. The results indicated that two of these organisms are species of Bacillus and one is a species of Streptomyces. The fourth species could not be amplified using the 16S-specific primers and was thus inconclusive. This organism is most likely a fungus and I am currently working to identify alternate methods of identification. Research on the antibiotic agents secreted by these novel microorganisms could be a valuable contribution to the race to develop new antibiotics and combat antibiotic resistant bacterial pathogens.
Research Advisor: Dr. Greg Mullen
 
Tristan Henderson, Houston Baptist University – 2:00-2:15 PM in Boax Session 
The neglect of microbes (and a potentially novel species of antibiotic resistant marine bacteria isolated from blood worms).
Tristan Henderson.
Microbes have been neglected for centuries, and we are only beginning to understand the true extent of their diversity, let alone their biology. In fact, most global biodiversity is microbial, with plants, animals, and fungi comprising only a minority of total eukaryotic diversity, and eukaryotes being a minority compared to bacteria. Yet most lineages of microbes are not yet characterized, giving a skewed perspective on the history of life and the standard model of biology. For reasons I will not cover here, I’ve been hunting marine worms as potential sources of novel microbial symbionts or parasites. After significant postponement, small time frames, and worm problems, I found a potentially new species of a marine bacterium in the Vibrio genus, which was isolated from the guts of blood worms. Five separate colonies were isolated from different worms, each having a distinct morphology not found in the literature. Because the colony morphology reminded me of an aerial view of a metropolis, I termed the mystery bacteria Metro. They seemed to have originated from the gut according to experimental and theoretical evidence. After sequencing the 16S rRNA gene, Metro’s closest relative was Vibrio alginolyticus, which is most known for pathogenizing humans and producing tetrodotoxin in pufferfish. Metro was also antibiotic resistant to both ampicillin and cloxacillin. Future experiments would have been done to further characterize the Metro bacteria, but they seemed to have all mysteriously died over spring break. In this presentation, I’ll bring context to Metro and ascertain its importance to Texan ecology. Then I will ponder a new direction in the study of microbes with a eukaryotic emphasis.
Research Advisor: Dr. Hannah Wingate
 
Makayla Hicks, Southwestern Oklahoma State University – 1:30-1:45 PM in Johnson Poster Session 
Evaluating pathogenicity of Pseudomonas aeruginosa by manipulating genes associated with motility and biofilm formation.
Makayla Hicks and Regina McGrane.
Pseudomonas aeruginosa is an opportunistic pathogen and causes infections in the lungs of cystic fibrosis patients.  Motility and biofilm formation are large contributing factors to the ability of P. aeruginosa to cause infection, and these processes have been found to be modulated by multiple biological molecules including rhamnolipid, a biosurfactant produced by RhlA, WspA, a protein predicted to have a role in surface sensing, and MotB and MotD, proteins that contribute to the flagella stator complex. In this work, we evaluated the impact of the deletion genes encoding for expression of these factors on the pathogenicity of P. aeruginosa. We hypothesize that loss of these motility and biofilm factors will cause P. aeruginosa to be less pathogenic. This hypothesis was tested by inoculating Galleria mellonella larvae, a wax moth recently developed as a model for understanding bacterial pathogenicity, with either wild type or mutant cultures. The survivability, coloration, movement, and silk formation of the larvae were evaluated each day until all the larvae died or for seven days, whichever occurred first. This required optimization of bacterial concentrations to ensure that the bacteria did not kill the larvae immediately and so that differences between wild type and mutant cultures could be detected. Differences between the wild type and mutants with deletions in rhlA, wspA, motB or motD were detected in the silk cocoon formation and movement of the larvae, suggesting these mutations impair the ability of P. aeruginosa to induce disease. Understanding the role of motility and biofilm formation in the pathogenicity of P. aeruginosa in wax moth larvae could lead to a greater understanding of how this bacterium causes infection in humans and ultimately could lead to a solution for problematic infections.
Research Advisor: Dr. Regina McGrane
 
Raistlin Hiner, Southwestern Oklahoma State University – 1:45-2:00 PM in Johnson Poster Session 
The effect of acceleration of high power rockets on the foraging behavior of Drosophila melanogaster.
Raistlin Hiner, Tommy Nguyen, Caden Bowles, Jimena Aracena, and Wayne Trail.
We are exploring the effects of large, short accelerations on the foraging behavior of fruit flies through the use of high power rocketry. These accelerations, as large as 490 m/s2 or more, last up to several seconds and are dependent on the motor type and characteristics of the rocket. Three containers carrying approximately 20 fruit flies each were flown in a high powered rocket for which the acceleration, velocity, and altitude were recorded. We compared the foraging of the flies flown in the rockets to field and lab controls. There was no significant effect on their survivorship or behavior in the subsequent feeding test. This suggests that there was no damage to their nervous systems while experiencing high accelerations.
Research Advisor: Dr. Jimena Aracena
 
Katie Holland, Angelo State University – 10:00-10:15 AM in Balanos Session 
Adenovirus Screening in Texas Bats.
Katie Holland and Loren K. Ammerman.
Adenoviruses (AdVs) in the genus Mastadenovirus play an ecological role in causing respiratory, ocular, and gastrointestinal pathology in mammals. Bats have been noted as important reservoirs in the evolution of adenoviruses due to bats’ atypical ability of harboring genetically diverse viruses within a single geographic location or host species. Research relating to the evolution, distribution, and diversity of adenoviruses can be beneficial for understanding epidemiological effects on bat ecology and conservation applications. Analyzing genetic diversity of viruses may also elucidate mechanics of inter- and intraspecies viral transmission in bats. We conducted RT-qPCR on 65 samples of Myotis velifer collected in 2019 and 2020 from Texas counties. For each sample, we completed two treatments to determine whether DNA extraction of intestinal tissue or DNA extraction of a phosphate buffered saline wash of intestinal tissue caused increased viral load detection via RT-qPCR. We added positive control AdV DNA to tissue prior to DNA extraction to determine the detection limits of the RT-qPCR assay. Based on these detection limits, all samples were negative for AdV DNA presence.
Research Advisor: Dr. Loren Ammerman
 
Annet Kwagala, Oral Roberts University – 9:15-9:30 AM in Balanos Session 
Gene Annotation of Select Protein-Coding Regions of Wolbachia Endosymbiont of Culex quinquefasciatus.
Annet Kwagala and Celestino Velásquez.
Genome sequencing technology is rapidly advancing, which has led to a vast collection of genetic information of various species—from humans to even the rarest of microbes. These sequenced genomes require manual curation and analysis, which is formally known as genome annotation. Genome annotation has become a significant part of genetics research for its utility in analyzing an organism’s whole genome and predicting functions of hypothetical genes based on nucleotide and amino acid sequence alone. Various genes code for proteins with specific functions, and much gene annotation analysis today focuses on the identification of protein-coding regions within sequenced genomes. This study focused on predicting the functions of five unannotated hypothetical protein-coding genes of Wolbachia endosymbiont of Culex quinquefasciatus: WP1368, WP1383, WP1384, WP1385, and WP1387. These genes were provided by the national GENI-ACT project and were analyzed using bioinformatics tools such as BLAST, MUSCLE, T-COFFEE, and WebLogo to identify sequence similarities. To predict protein localization for these genes, databases such as TMHMM, SignalP, Phobius, and PSORTb were used. Based on the results from these publicly available tools, WP1368 was predicted to code for a ferredoxin family protein, WP1383 for transcription initiation factor IF-2, WP1384 for transcription termination and antitermination protein NusA, WP1385 for efflux RND transporter permease subunit, and WP1387 for glutamine synthetase. These bioinformatic predictions are, however, hypothetical, and laboratory work is necessary to validate and prove that these genes indeed code for their predicted functions.
Research Advisor: Dr. Celestino Velásquez
 
Eddy Leardini, Oral Roberts University – 10:00-10:15 AM in Boudetase Session 
Genome Annotation of Putative Coding Regions of Listeria monocytogenes.
Eddy Leardini and Celestino Velásquez.
The genomes of many organisms have been annotated and made freely accessible on online databases thanks to bioinformatics servers that have permitted the analysis and sequencing of genetic material. Starting from the nucleotide and primary amino acid sequences of a gene, it has become possible to predict the properties of the hypothetical gene products of any organism, such as identifying the function of a protein. The work requires tools such as BLAST, MUSCLE, T- Coffee, WebLogo, SignalP, LipoP, TMHMM, PSORTb, and Phobius, which compare the input sequence with the data already manually recorded on online databases. These tools base their predictions on the properties of proteins, conserved domains, signal peptides, transmembrane regions, structure, and localization within the cell. The research focused on four putative coding regions of the Gram-positive bacterium Listeria monocytogenes, which is a foodborne human pathogen. The unannotated genes lmo2838, lmo2842, lmo2848, and lmo2854 were predicted to code for: a sugar ABC transporter permease, a substrate-binding domain-containing protein, the L-rhamnose isomerase, and the membrane protein insertase YidC, respectively. Such information was deduced to enrich the genetic knowledge about Listeria monocytogenes, which might help to understand its pathogenicity in the human body. Eventual tests to perform in the laboratory would be necessary to confirm the bioinformatics predictions about the type of protein expressed.
Research Advisor: Dr. Celestino Velásquez
 
C. Ethan Long, Southwestern Oklahoma State University – 9:15-9:30 AM in Boudetase Session 
Photoinducible Changes in Cell Morphology and Phenol Content Related to UVB Tolerance in Isolates of Zygnema (Streptophyceae).
C. Ethan Long and Steven W. O'Neal.
Zygnema is a type of algae that typically forms floating mats that rest on the top of the water which leads to different levels of light exposure among alga cells. The purpose of this study was to determine if light levels and UVB exposure affected Zygnema phenolic content, morphology, and UVB absorbance. In response to light exposure, algae produce protective compounds called phenolics that work as a sunscreen to protect the algae from harmful rays. Because of this, the algae on the top of the mat are exposed to more light and UVB than the algae underneath the surface of the water. In this study, we wanted to test if Zygnema at conditions comparable to algae at the top of the floating mat produced more or less phenolics than the algae at the bottom of the mat. In the experiment conducted, we exposed two Zygnema isolates to different light treatments that included: high light with UVA, high light without UVA, low light with UVA, and low light without UVA.  After the algae grew for 7 days, we photographed the cells and measured their length. The algae samples were then ground up and made into a testable solution. Zygnema isolates exposed to high light produced 255% more phenolic content than the samples exposed to low light. The effect of UVA on phenolic content production provided inconclusive results. Cell length decreased 51% at high light. At low light, removing UVA produced significantly larger cells. Zygnema UVB absorbance rose 296% when exposed to high light.
Research Advisor: Dr. Steven O'Neal
 
Michelle Marshall, Tarleton State University – 2:15-2:30 PM in Johnson Poster Session 
Evaluation of Diatoms in Bone Marrow as Evidence for Length of Post-Mortem Skeletal Submersion with Applications in Forensic Sciences.
Michelle Marshall.
Aquatic environments are home to many specialized plants and algae. These environments are also common sites for criminal activities such as drownings and body disposals. Based on unique characteristics and wide diversity, diatoms (Bacillariophyceae) can be used to determine the site of crime or evidence disposal, providing evidence for forensic cases (Pollanen 1998). However, not much is known of the pathways or timeframes by which diatoms enter bone marrow, presenting limitations on the use of diatoms as trace evidence of drowning versus body disposal in water (Lunetta et al. 2013). This study uses experimental bone submersion in exposure to diatoms in order to determine the potential for diatoms to enter bones post-mortem. The results of this study will help refine the interpretation of diatom presence in bone marrow as evidence of drowning and provide additional sources of trace evidence for crimes in aquatic locations. This study conducted a controlled experiment in field conditions using cow femurs. Two container bins were filled with river water and rocks with visible diatom colonies. Three bones were placed in each bin; one bin had algaecide added to kill the diatoms and serve as a control. The bones were submerged in oxygenated, circulated water in the bins and were removed for sampling at 1 week, 3 weeks, and 2 months. Preliminary results found that diatoms will form biofilm on the exterior of bones, especially in cartilage and joint areas. Bone marrow was extracted and sampled for diatoms. Observations of diatom morphology and abundance will be recorded and compared to the timeframe of submersion. This will constrain accuracy of use of diatoms as positive indicator of drowning and may increase their use as trace evidence of length of submersion in cases with skeletal remains.
Research Advisor: Dr. Victoria Chraibi
 
Robert B. McManus, Texas Wesleyan University – 9:30-9:45 AM in Boudetase Session 
Population structure of White Rosinweed (Silphium albiflorum A. Gray: Asteraceae).
Robert B. McManus.
Silphium albiflorum is a native Texas endemic plant species distributed disparately along limestone deposits. Limited population size and an Area of Occupancy (AOO) that exceeds IUCN endangered criteria threaten this grassland species' viability. Despite this, the genetic structure of S. albiflorum remains obscure. 701,485 SNPs identified by tGBS genotyping were used to survey the genetic structure of S. albiflorum populations. The methods used to determine the genetic structure include the genetic clustering algorithm STRUCTURE, principal component analysis (PCA), and phylogeny. STRUCTURE results indicate a highly subdivided species population with K=3 having the highest probability of occurring. The principal component analysis suggests three relatively distinct clusters; however, the model at PC2 cumulatively explains only 5.7% of the variance, and at PC5, 12.6% of the cumulative variance is explained. Finally, a neighbor-joining tree of the 96 samples supports the hypothesized groups by STRUCTURE and PCA previously identified according to watersheds by Hutchinson et al., (2019).  All three analyses show that S. albiflorum from the Trinity river watershed shows minor admixture amounts, whereas populations from the Colorado and Brazos watershed tend to be more similar. These results suggest that the optimal preservation of genetic diversity should be focused on samples derived from the Brazos or Colorado as samples from these locations display the highest admixture amount. Further studies regarding the correlation between geography and genetic structure will prove beneficial to preserving S. albiflorum.
Research Advisor: Dr. Bruce Benz
 
Tommy Nguyen, Southwestern Oklahoma State University – 2:00-2:15 PM in Johnson Poster Session 
Establishing quinine as a deterrent for learning experiments in fruit flies (Drosophila melanogaster).
Tommy Nguyen, Caden Bowles, Raistlin Hiner, and Jimena Aracena.
The purpose of this study is to establish a deterrent solution for future use in classical conditioning experiments. We starved groups of 50 fruit flies for 42 +/-2 hours and allowed them to feed for one hour in the dark in an arena containing a choice of red vs blue solutions. The tests were 1) 250mM pure sucrose solution vs 250mM sucrose with 4.0mM quinine, 2) control of pure sucrose vs pure sucrose, and 3) control of sucrose with quinine vs sucrose with quinine. After feeding, the flies were counted according to the color of their abdomens to determine their choice. All flies preferred the pure sugar solution over sugar with quinine. When only sucrose with quinine was present, no flies fed and when only pure sugar was present, there was a slight preference for the red solution.  We established that sugar with quinine is clearly a deterrent and that pure sucrose can be used as a reward in subsequent learning experiments.
Research Advisor: Dr. Jimena Aracena
 
Kayvan Noori and Lauren Watkins, University of Central Oklahoma – 11:00-11:15 AM in Balanos Session 
Analysis of Cardiac Teratogenicity in Maternal PKU: Cloning for In-situ hybridization probe synthesis.
Kayvan Noori and Lauren Watkins.
Maternal Phenylketonuria (MPKU) is the result of exposure of high levels of phenylalanine (PHE) to the developing embryo of mothers with PKU. High levels of PHE can lead to cranial and cardiac developmental issues in the embryo. Previous studies in our lab revealed the Retinoid pathway as a potential mechanism for these defects. Transthyretin (TTR) is a gene of the retinoic acid pathway that is effected by an excess of PHE. The TTR gene is responsible for producing the protein called Transthyretin. This protein transports retinol in the retinoic acid pathway.  The second gene of interest in this project is PlexinA2 and is also a gene effected by an excess of PHE. PlexinA2 is responsible for neural cell guidance and growth, specifically PlexinA2 is important in neural crest cell (ncc) migration. Disruption in the migration of nccs may lead to the cranial and cardiac defects observed in MPKU. The objective of this project was to clone a fragment of TTR and PlexinA2 RNA in order to transcribe a RNA probe. Cloning was done by dissecting chicken embryos and extracting RNA. RNA was then reverse transcribed and cDNA was used in PCR amplification of the two genes. The PCR product was extracted and cloned into the pGEM-T easy vector system through ligation of the DNA insert to plasmid. One Shot TOP10 chemically competent E. coli was transformed with the plasmid containing our gene insert. X-Gal was then used to select for bacterial colonies containing the DNA insert. Clonal PCR was conducted to confirm the presence of the insert. Further experiments are underway, including sequencing of the insert, RNA probe transcription, and insitu hybridization to understand the effect of excess of PHE on gene expression.
Research Advisor: Dr. Nikki Seagraves
 
Nazka Nurbyek and Michaela Vance, University of Central Oklahoma – 9:45-10:00 AM in Boudetase Session 
Effect of Phenylalanine, Retinoic Acid, Retinal, and Citral on the Proliferation of O9-1 Mouse Cranial Neural Crest Cells.
Nazka Nurbyek, Michaela Vance, and Nikki Seagraves.
Maternal phenylketonuria [MPKU] is a syndrome that causes many different types of birth defects affecting growth, and development of heart, brain, and bones of the face. The syndrome is caused by exposure to too much Phenylalanine, an amino acid found in proteins eaten by a mother with Phenylketonuria during pregnancy. It is not known why Phe causes abnormal development of embryos. Our lab has preliminary evidence that high levels of Phe could inhibit communication between cells mediated by Retinoic Acid [RA], which effects how cells divide, migrate, and specialize. Division of the neural crest cells are important in formation of components of the heart including the outflow tract (OFT) and aortic arch arteries (AAA). We hypothesize that Phe inhibits the rate that cells divide, which may be the cause of the birth defects seen in MPKU. We cultured O9-1 mouse neural crest cells and performed an experiment to determine the effect of Phe, RA, retinal, and citral exposure on how the cells divide. Images were analyzed with ImageJ and GraphPad Prism. Results suggest that Phe exposure causes a significant decrease in division of cells. It has been shown that RA and retinal increase cell division, and that citral decreases. Phe acted similar to citral, which suggests that it may act as an inhibitor of RA, which could cause the heart defects seen in MPKU. This work is significant because no one knows how Phe causes the types of defects observed in human MPKU.
Research Advisor: Dr. Nikki Seagraves
 
Denton Parsells, Southwestern Oklahoma State University – 12:15-12:30 PM in Boax Session 
Sexual Selection in Response to Varying Levels of Eutrophication.
Denton Parsells and Rickey Cothran.
Sexually selected traits are expensive to build and maintain and thus are predicted to be dependent upon condition and useful for making decisions about potential mates. However, the condition-dependence of these traits is also expected to make them very sensitive to environmental change. We explored patterns of sexual selection in populations of amphipods in the genus Hyalella exposed to varying levels of nutrient pollution. These amphipods were collected from nine natural lakes in NW Pennsylvania that have varying nutrient levels, which are suggestive of human induced change. These varying levels are most likely due to fertilizer runoff from local farms. We measured sexually dimorphic traits and control traits to examine whether the former were more sensitive to nutrient pollution. Higher levels of phosphorous, found in lakes with higher nutrient runoff, were predicted to lead to larger sexually selected traits, the posterior gnathopod (a claw-like trait) and second antenna. These larger sexual traits in males allow them to overcome female resistance and decrease the fitness of the females. A decrease in female fitness then has a negative impact on the population as a whole. Nutrient pollution is expected to cause less variation between males and lead to weaker sexual selection which can in turn decrease the overall health of the population. This decrease is caused by females not being able to use these information-rich traits to choose among potential mates. The current trait size results show a trend that follows the expected results. Lakes with higher eutrophication have males with larger trait sizes. However, we did not find a negative correlation between trait variation and increasing eutrophication. Our results do suggest that females in eutrophic lakes will have to deal with more well-armed males, which could negatively affect population fitness.
Research Advisor: Dr. Rickey Cothran
 
Michelle Ramirez, Austin College – 11:15-11:30 AM in Boudetase Session 
Investigation of Correlation between Expression of PA28 gamma and Cell Sensitivity to Cisplatin.
Michelle Ramirez.
PA28γ is a proteasome activator that has been seen to be overexpressed in many cancers. It is associated with many characteristics that help develop cancer otherwise known as hallmarks. Some of these hallmarks include evading growth suppressor, sustaining proliferative signaling, resisting cell death, genome instability and mutation, and activating invasion and metastasis. Cancers with higher PA28γ expression are shown to be more aggressive. Aggressive cancers have also been seen to be more susceptible to drug resistance. A conventional drug used to treat cancer is cisplatin, however, cells treated with this drug can also be susceptible to it. Therefore, the goal of this project is to determine whether there is a correlation between the expression of PA28γ in certain cell lines and cisplatin resistance.
Research Advisor: Dr. Lance Barton
 
Marissa Rivas, Oral Roberts University – 9:30-9:45 AM in Balanos Session 
An Analysis of Hypothetical Protein-Coding Genes of Vibrio cholerae.
Marissa Rivas and Celestino Velásquez.
With the advancement of technology as well as the launch of the Human Genome Project in the early 2000s, understanding one’s genetic information has become valuable as well as accessible. Learning such a phenomenon allows for greater insight on the proteins a specific gene will produce, furthermore, corresponding to its function. With the millions of organisms on the Earth today, it is a daunting task to analyze an abundance of genome sequences; however, the many bioinformatic tools available allow for some exploration as well as an analysis of these unexplored genomes. Some of these tools include bioinformatics programs such as BLAST, MUSCLE, T-Coffee, WebLogo, SignalP, LipoP, TMHMM, BOMP, PSORTb, and Phobius. These programs allow one to understand and predict function from a gene’s primary amino acid sequence. These tools are able to identify various properties such as sequence similarity, conserved domains, and protein localization within the cell. With this information, the purpose of this lab is to analyze five genes belonging to the genome of bacterium Vibrio cholerae: VC0007, VC0009, VC0013, VC0015, and VC0021. These genes were predicted to code for 50S ribosomal protein L34, ABC transporter permease, DNA polymerase III beta subunit, DNA gyrase subunit B, and tRNA ligase alpha subunit, respectively. Acquiring this data will then allow for possible and appropriate functional predictions of related genes. This information can only then be supported by performing a lab experiment to test for the predicted functions.
Research Advisor: Dr. Celestino Velásquez
 
Lizbeth Robles-Fernandez, East Central University – 12:00-12:15 PM in Johnson Poster Session 
Analysis of Regulating Sugar Metabolism in Escherichia coli Under Anaerobic Conditions Using Beta-Galactosidase Tests.
Lizbeth Robles-Fernandez and April D. Nesbit.
Escherichia coli is one of the bacteria found in the gut microbiome of humans.  It plays a role in maintaining gut health, although some strains can be pathogenic.  E. coli is useful for research because it has an extensively studied genome.  However, there are still genes that we know little about in E. coli.  One such gene is yfaX, and it encodes a predicted transcription factor.  Other genes in the same operon as yfaX are suspected of playing a role in L-rhamnonate metabolism based on in vitro studies.  Thus, our hypothesis was that yfaX regulated genes in response to L-rhamnonate.  To test this hypothesis, we created promoter fusions to the lacZ reporter gene and tested the effect of deleting yfaX on gene expression of these reporter gene constructs.  When we grew E. coli under anaerobic conditions with L-rhamnonate as the sole carbon source, the cells showed little to no growth.  When we grew the cells with glucosamine and L-rhamnonate, we saw that L-rhamnonate had no effect on gene expression in the presence or absence of yfaX for the one promoter tested.  Thus, we conclude that L-rhamnonate cannot be used by E. coli as a sole carbon source under anaerobic conditions, and L-rhamnonate may not play a role in regulation by YfaX.
Research Advisor: Dr. April Nesbit
 
Jason Salvato, Oral Roberts University – 12:30-12:45 PM in Boax Session 
Gene Annotation of the Hypothetical Protein-Coding Genes of Coxiella burnetii.
Jason Salvato and Celestino Velásquez.
Genetic information of organisms and microorganisms has become readily accessible due to advances in genomic sequencing and bioinformatic technology. Despite these advances, there are numerous organisms with genome sequences that have yet to be annotated. Many of these genome sequences require manual annotation, which can uncover hypothetical protein-coding genes. Through the use of publicly available online bioinformatics tools, such as BLAST, T- COFFEE, TMHMM, SignalP, Phobius, and PSORTb, the functions of hypothetical protein- coding genes can be predicted from primary amino acid sequences. Two clusters of properties that aid in determining and predicting the hypothetical genes involve sequence similarity and protein localization. The bioinformatic programs can identify properties such as protein families, conserved domains, signal peptides, and transmembrane regions that belong to the respective clusters. This research project aims to predict the functions of five unannotated hypothetical protein-coding genes in the genome of the bacterium Coxiella burnetii. The genes  BMW92_RS10760, BMW92_RS10830, BMW92_RS10835, BMW92_RS10840, and BMW92_RS10855 were analyzed and predicted to code for the following proteins: uroporphyrinogen-III synthase, pyrroline-5-carboxylate reductase, pyridoxal phosphate- dependent enzyme, phosphoenolpyruvate carboxykinase, and aspartate carbamoyltransferase, respectively. The predicted functions of the hypothetical protein-coding genes provide insight into the proteome of C. burnetii. Ultimately, the proposed gene annotations must be validated through molecular cloning and biochemical methods to determine if these proteins are indeed expressed by C. burnetii and carry out their predicted functions.
Research Advisor: Dr. Celestino Velásquez
 
Kristina M. Sattler, Mount St. Joseph University – 1:45-2:00 PM in Boax Session 
Validation of a Therapeutic Biomarker in a Cell Model of GNE Myopathy.
Kristina M. Sattler, Kara E. Bradley, Alexa P. Adams, and Kelly E. Crowe.
GNE myopathy (GNEM) is a rare, adult-onset, autosomal recessive disease that causes severe muscle wasting. Pathology in GNEM occurs due to mutations in the GNE gene, which encodes for a protein along the biosynthetic pathway of sialic acid, a terminal sugar that resides in sugar chains on the extracellular membrane of cells. Disruptions in the GNE gene result in significant reductions in the levels of sialic acid; however, the specific quantity of sialic acid is difficult to determine. In order to assess sialylation, a robust biomarker is needed to quantify levels of sialic acid. Validation of a biomarker in vitro would allow researchers to move towards development of a gene therapy. Our lab has determined a subset of lectins which show promise of a therapeutic biomarker for GNEM; however, such lectins must also be validated when GNE gene replacement occurs via transient transfection to mimic what would happen after a gene therapy.
Research Advisor: Dr. Kelly Crowe
 
Kenneth Shimer, University of Central Oklahoma – 2:45-3:00 PM in Johnson Poster Session 
Comparison of passive detection methods in determining occupancy of coyotes in rangeland in southern Oklahoma.
K.E. Shimer, S.L Webb, M.D. Proctor, and V.L. Jackson.
Passive monitoring devices have had a long-standing influence on how wildlife surveys are conducted, given their low labor investment and cost. The primary and most used example of this is triggered camera traps, particularly in terrestrial mammal surveys. However, autonomous recording units (ARUs) have been growing in popularity for sound-producing terrestrial species. ARUs allow for a broader detection range and are historically used in surveying marine and air-borne species such as birds and bats. This increase in utilization raised the question of how successful these monitors are compared to the more traditional camera trap in terrestrial environments. To address this question, we compared 29 paired sets of un-baited passive detection devices (1 camera trap, 1 ARU per site) for detection of coyotes (Canis latrans), a highly vocal species, within two rangeland sites owned by the Noble Foundation in south-central Oklahoma. We used occupancy models to compare detection binary histories of each method in order determine detection success. Our preliminary finding suggests that ARUs offer a higher-level detection of coyotes in comparison to camera traps. This study recommends that using or supplementing ARUs within survey protocols increases the potential detection of elusive sound-producing species.
Research Advisor: Dr. Vicki Jackson
 
Anastasia Smith, Oral Roberts University – 12:45-1:00 PM in Boax Session 
Identification and Characterization of ~2DKa Peptide-Containing Polysaccharide “Felix”.
Anastasia Smith and William Ranahan.
Current cancer therapies such as radiation, chemo, and surgery induce many off-target effects in healthy tissue. Thus, the search for alternative treatments with mild or no side effects continue. Several candidates have emerged from unlikely sources. The endophytic fungi Taxomyces andreanae, has given us the widely used chemotherapy called Taxol. Furthermore, as first-world countries like China and Japan routinely prescribe fungi derived compounds (Lentinan, PSK, and Schizophyllan) as standard adjuvant therapy, the authors focused this investigation on fungi. Extracts from the mushroom Ganoderma lucidum have commonly been used with a variety of health benefits. However, rather than looking at the benefits of the mycelia itself, the authors focused on the compounds secreted from the fungus. From these compounds, the authors sought to identify fungi-derived secreted compounds with anticancer cell properties. Ganoderma lucidum was first “trained,”* and secretions were collected. These secretions from the “trained” Ganoderma lucidum yielded a 2KDa peptide-containing polysaccharide named “Felix.” The impact of cell viability was tested with the ER-/PR-/Her2- cancer cell model MDA MB-468. This cancer cell model represents a mammary breast cancer with very poor clinical outcomes. Following 48hrs treatment with Felix, the authors observed a 50% decrease in cell viability. Previous studies from this lab have shown that exposure to Felix did not result in a decrease in cell viability in the non-tumorigenic mammary control cell lines. Future studies will focus on elucidation of the mechanism and range of efficacy of Felix.  * U.S. Patent No.: 15/814,068
Research Advisor: Dr. William Ranahan
 
Zoey Stormes, Angelo State University – 1:30-1:45 PM in Boax Session 
Screening of Genetic Markers to Distinguish Morphologically Similar Speices of Cottontail Rabbit.
Zoey Stormes and Loren K Ammerman.
The Davis Mountain Cottontail, Sylvilagus robustus, and the Eastern Cottontail, Sylvilagus floridanus, are two morphologically similar but genetically distinct species of cottontail rabbit. Identification of a specimen as one species over the other is currently difficult due to the lack of a single genetic marker capable of conclusively separating the two species. This study aims to address this problem by investigating five nuclear genetic markers of interest (TG, THY, SPTBN1, PRKC1, and MGF) previously used to elucidate a phylogeny for the family Leporidae. This study utilizes both the species of interest as well as the Desert Cottontail, Sylvilagus audubonii, as an outgroup. Twelve Sylvilagus robustus, thirteen Sylvilagus floridanus, and ten Sylvilagus audubonii samples were obtained and DNA was extracted and amplified for the five markers of interest. Successfully sequenced DNA was aligned and processed using phylogenetic tree software, and the effectiveness of each gene was evaluated based on branch groupings and bootstrap values within the tree. So far, none of the five genes has shown promise in their ability to confirm a specimen as being S. robustus or S. floridanus.
Research Advisor: Dr. Loren Ammerman
 
Laura Tham, Oral Roberts University – 10:15-10:30 AM in Boudetase Session 
Identification and Characterization of a Compound, “F14,” Secreted by Inonotus obliquus.
Laura Tham and William Ranahan.
Breast cancer is one of the most commonly diagnosed cancers among women. Current cancer treatments include chemotherapy, surgery, and radiation, which produce off-target-effects on patients. Currently, there are no cancer treatments which exclusively target cancer cells. In Asia and Russia, scientists have been researching and testing medicinal mushrooms, and their effects on cancer, for decades. In these developed nations, standard adjuvant therapies include PSK, Schizophyllan, and Lentinan, all mushroom-derived drugs. In the United States an extract from the Turkey Tail mushroom is currently in FDA Stage 3 clinical trials as an adjuvant during breast cancer therapy. One well studied medical mushroom is Inonotus obliquus, also known as Chaga. Given that there are no published studies of compounds secreted from I. obliquus, the authors sought to identify compounds secreted from I. obliquus with anti-cancer properties. Collected secretions were separated and concentrated through vacuum filtration and characterized through size exclusion chromatography. The authors identified and named a peptide-containing polysaccharide “F14.” The authors observed decreased cancer cell viability following treatment with the F14 compound. To better understand the changes in gene expression induced by F14 exposure, the authors analyzed expression of BCL2 and PCNA mRNA levels. qPCR analysis revealed a decrease in expression of BCl2 and PCNA in the “triple negative” mammary cancer cell model MDA-MB-468. Further studies should include qPCR assays and cell cytotoxicity assays to confirm these results.
Research Advisor: Dr. William Ranahan
 
Michaela Vance and Nazka Nurbyek, University of Central Oklahoma – 9:45-10:00 AM in Boudetase Session 
Effect of Phenylalanine, Retinoic Acid, Retinal, and Citral on the Proliferation of O9-1 Mouse Cranial Neural Crest Cells.
Nazka Nurbyek, Michaela Vance, and Nikki Seagraves.
Maternal phenylketonuria [MPKU] is a syndrome that causes many different types of birth defects affecting growth, and development of heart, brain, and bones of the face. The syndrome is caused by exposure to too much Phenylalanine, an amino acid found in proteins eaten by a mother with Phenylketonuria during pregnancy. It is not known why Phe causes abnormal development of embryos. Our lab has preliminary evidence that high levels of Phe could inhibit communication between cells mediated by Retinoic Acid [RA], which effects how cells divide, migrate, and specialize. Division of the neural crest cells are important in formation of components of the heart including the outflow tract (OFT) and aortic arch arteries (AAA). We hypothesize that Phe inhibits the rate that cells divide, which may be the cause of the birth defects seen in MPKU. We cultured O9-1 mouse neural crest cells and performed an experiment to determine the effect of Phe, RA, retinal, and citral exposure on how the cells divide. Images were analyzed with ImageJ and GraphPad Prism. Results suggest that Phe exposure causes a significant decrease in division of cells. It has been shown that RA and retinal increase cell division, and that citral decreases. Phe acted similar to citral, which suggests that it may act as an inhibitor of RA, which could cause the heart defects seen in MPKU. This work is significant because no one knows how Phe causes the types of defects observed in human MPKU.
Research Advisor: Dr. Nikki Seagraves
 
Andrew H. Vergote, Southwestern University – 10:45-11:00 AM in Boudetase Session 
Designing Novel Bioinks for 3D Printing.
Andrew H. Vergote, Cody Crosby, and Jon Smart.
3D bioprinting aims to recreate complex biological tissues for use in transplantation, drug screening, and disease modeling. Traditional fabrication methods (e.g., casting in molds) often struggle to recapitulate complex biological structures. In contrast, our research utilizes computer aided design (CAD) models to guide the printing of physiological structures from hydrogel-based bioinks. Bioinks typically consist of alginate cross-linked with calcium chloride; the benefit of cross-linking alginate with calcium chloride is that it improves the structural integrity of the otherwise unstable hydrogel. This pre-cross-linked hydrogel was loaded into a Tissue Scribe(™) printer using a blunt-tipped syringe as the means of extrusion. The hydrogel was subsequently embedded into a gelatin slurry suspension bath that provides additional structural support while encouraging cytocompatibility. This combination of functional advantages will allow for the creation of the complex biological structures necessary for biological tissues. In this work, we have demonstrated the viability of our current setup and have developed robust procedures for material synthesis. In future work, we aim to synthesize novel bioinks for medical printing applications.
Research Advisor: Dr. Cody Crosby
 
Lauren Watkins and Kayvan Noori, University of Central Oklahoma – 11:00-11:15 AM in Balanos Session 
Analysis of Cardiac Teratogenicity in Maternal PKU: Cloning for In-situ hybridization probe synthesis.
Kayvan Noori and Lauren Watkins.
Maternal Phenylketonuria (MPKU) is the result of exposure of high levels of phenylalanine (PHE) to the developing embryo of mothers with PKU. High levels of PHE can lead to cranial and cardiac developmental issues in the embryo. Previous studies in our lab revealed the Retinoid pathway as a potential mechanism for these defects. Transthyretin (TTR) is a gene of the retinoic acid pathway that is effected by an excess of PHE. The TTR gene is responsible for producing the protein called Transthyretin. This protein transports retinol in the retinoic acid pathway.  The second gene of interest in this project is PlexinA2 and is also a gene effected by an excess of PHE. PlexinA2 is responsible for neural cell guidance and growth, specifically PlexinA2 is important in neural crest cell (ncc) migration. Disruption in the migration of nccs may lead to the cranial and cardiac defects observed in MPKU. The objective of this project was to clone a fragment of TTR and PlexinA2 RNA in order to transcribe a RNA probe. Cloning was done by dissecting chicken embryos and extracting RNA. RNA was then reverse transcribed and cDNA was used in PCR amplification of the two genes. The PCR product was extracted and cloned into the pGEM-T easy vector system through ligation of the DNA insert to plasmid. One Shot TOP10 chemically competent E. coli was transformed with the plasmid containing our gene insert. X-Gal was then used to select for bacterial colonies containing the DNA insert. Clonal PCR was conducted to confirm the presence of the insert. Further experiments are underway, including sequencing of the insert, RNA probe transcription, and insitu hybridization to understand the effect of excess of PHE on gene expression.
Research Advisor: Dr. Nikki Seagraves
 
Bethany Wilkinson, Oral Roberts University – 10:30-10:45 AM in Boudetase Session 
Gene annotation of hypothetical protein-coding genes of Yersinia pestis.
Bethany Wilkinson and Celestino Velásquez.
Technological advances within the past two decades have allowed researchers the capacity to sequence and analyze any genome. However, while many organisms have had their genomes sequenced, hardly any have been manually interpreted to propose the functions of an organism’s hypothetical protein-coding genes. The primary amino acid sequences of an organism’s genes can be used to offer functions for its proteins. Online bioinformatics programs, such as BLAST, T-COFFEE, MUSCLE, TMHMM, SignalP, Phobius, and PSORTb, identify several properties such as conserved domains, protein families, protein conformations, signal peptides, and transmembrane regions of the sequences. These elements can be examined to provide insight into likely organismal lineage, protein functions, location, or intended use of the protein by the organism. This research was conducted with the purpose of proposing functions for five unannotated hypothetical protein-coding genes within the genome of the bacterium Yersinia pestis, which was most notably responsible for the Bubonic plague. The genes A1122_RS20830, A1122_RS20870, A1122_RS21135, A1122_RS21140, and A1122_RS21150 were analyzed and predicted to code for the following proteins: DNA polymerase III subunit alpha, an intermembrane protease, a domain protein of the enzyme diguanylate cyclase, a pseudogene, and a major facilitator superfamily (MFS) transporter, respectively. These predictions allow some insight into the functions of the proteome of Yersinia pestis. However, these hypothetical gene annotations must be validated through molecular cloning and biochemical methods to determine whether Yersinia pestis expresses these proteins and whether they engage in their bioinformatically proposed functions.
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