2020 Abstracts

Bell, McKenzie; Porter, Tyrel; Naylor, Emily; Domyan, Eric (Utah Valley University)
Faculty Advisor: Domyan, Eric (Utah Valley University, Biology)

The domestic rock pigeon has been the subject of selective breeding for over a hundred years and so displays an immense variety of phenotypes. This variety provides opportunities to further understand the genetic basis of phenotypic evolution. Pigmentation of pigeon feathers is controlled by multiple alleles at different loci, which influences the type and amount of melanin deposited in the feathers. A specific phenotype, known as "recessive red", consists of distinctly red plumage and is caused by a mutation that greatly reduces the expression of the gene SOX10. This gene encodes a transcription factor, known to play a key role in melanocyte maturation and proliferation. SOX10 likely regulates the transcription of multiple downstream genes but the identities of these genes are largely unknown. To identify downstream targets of SOX10, we compared the transcriptomes of regenerating feathers from wild-type and recessive red birds to identify genes that had different expression levels between the two groups. We identified 46 genes that are expressed at different levels between wild-type and recessive red birds, and thus potential targets of SOX101. Of the 46 genes, Tbx2 was selected as a starter because it is one of the only transcription factors regulated by Sox10 in melanocytes. This mechanism makes it a plausible candidate given the critical role proteins play in phenotypic expression ("TBX2 T-box transcription factor 2—Gene—NCBI," n.d.).

Yang, Haokun; Herring, Jacob; Elison, Weston; Wynn, Adam; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)

Nearly 1 in 10 Americans have type 2 diabetes (T2D), a disease that is characterized by a loss of functional β-cell mass, resulting in decreased insulin secretion and glucose utilization. The pancreatic β-cell is responsible for producing and secreting insulin and monitoring blood glucose levels, and it is crucial to the understanding of T2D. The orphan nuclear receptor Nr4a3 (Nor1) has well-defined roles throughout the body, specifically with fuel utilization in the liver, muscle, and adipose tissues. Here we present data demonstrating that Nr4a3 KO mice have increased body weight, blood glucose levels (fasting and non-fasting), and impaired glucose tolerance when fed a standard diet. Respiration from adipose tissue is significantly impaired in male and female Nr4a3 KO animals. These data demonstrate that Nr4a3 is necessary for whole-body homeostasis. We believe that these data serve as a step toward understanding the pathway of T2D progression and finding a cure.

Garcia, Jessica; Hanks, Hana; Kist, Taylor; Suman, Tanner (Dixie State University)
Faculty Advisor: Meyer, Jennifer (Dixie State University, Physical Sciences)

Antioxidants in the human body regulate reactive oxygen species (ROS). If ROS are increased within the body it can potentially lead to oxidative stress and cell injury. Glucose-6-phosphate dehydrogenase (G6PD) is an enzyme found in the pentose phosphate pathway that assists in reducing the amount of ROS in the bloodstream. Increased concentrations of glucose, commonly found in patients that suffer from type 1 and type 2 diabetes mellitus have been shown to impair G6PD activity, thereby causing damage to erythrocytes (RBCs)2. It has been shown that hyperglycemic conditions decrease the activity of G6PD in RBCs, which is improved by the addition of pterostilbene, a potent antioxidant1. Glutathione, another antioxidant found in the pentose phosphate pathway, is decreased in RBCs within hyperglycemic conditions. The addition of pterostilbene is predicted to increase levels of glutathione under high glucose conditions.

References:

1. Richins, M., & Meyer, J. (2018). Pterostilbene Ameliorates Lipid Peroxidation and Increases Glucose-6-Phosphate Dehydrogenase Activity in Erythrocytes Subjected to High Glucose Conditions. American Heart Association Journals, 138.

2. Zang, Z., Apse, K., Pang, J., & Stanton, R. C. (2000). High glucose inhibits glucose-6-phosphate dehydrogenase via cAMP in aortic endothelial cells. The Journal of Biological Chemistry, 275(51), 40042-40047. Doi: 10.1074/jbc.M007505200

Brown, Nathan; Herring, Jacob; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham young University; Nutrition, Dietetics, and Food Science)

Diabetes is a global epidemic affecting millions of people. The total estimated cost of diabetes in the U.S. during 2017 was 327 billion dollars. Diabetes is characterized by the loss of pancreatic β-cell function which is caused by an autoimmune disorder in Type 1 diabetes or insulin resistance and β-cell exhaustion in Type 2 (T2D) diabetes.
It is shown that β-cell mitochondrial respiration is dependent on the nuclear receptor Nr4a1. Respiration rates of cells lacking Nr4a1 in the presence of 16 mM glucose resulted in a significant decrease in glucose-stimulated insulin secretion by impeding the production of ATP. It was also found that knockdown of Nr4a1 results in decreased expression of mitochondrial dehydrogenase subunits Idh3g and Sdhb. Thus, the orphan nuclear receptor Nr4a1 is critical for β-cell mitochondrial function and insulin secretion.
In subsequent studies it was shown that dihydroergotamine (DHE) induces Nr4a1 expression via recruitment of the super elongation complex to enable elongation of Nr4a1 promoter paused RNA polymerase II. While these experiments have been shown in cancer cells, I hypothesize that DHE will up-regulate Nr4a1 and other downstream targets. To test this I will use an in-vitro model to culture INS-1 832/3 rat insulinoma cell lines as a useful model for insulin secretion regulation and pancreatic islet beta-cell function studies. This study will shed further light on the regulation of the Nr4a1 nuclear receptor in pancreatic β-cells.

Berges, Bradford; Wienclaw, Trevor; Ball, Ashley; Richmond, Bradley (Brigham Young University)
Faculty Advisor: Berges, Bradford (Life Sciences, Microbiology and Molecular Biology)

Staphylococcus Aureus (SA) biofilms are serious impediments to immune defenses and antibiotics, making them a major factor in SA infection. Such infections can be highly lethal even using current treatments, representing a major challenge to the healthcare industry. Previous genetic screenings of SA have revealed several genes that may be associated with biofilm formation. While the roles of many of these genes have been studied, little research has been done on how mutations of these genes impact biofilm composition. As several therapeutic options for treating mature SA biofilms require understanding of biofilm composition, a better understanding of how genes influence that composition is critical to improving current treatments and developing new ones.
In this project, we will study the biofilm phenotypes of SA with mutations in common biofilm-associated genes. By comparing the biofilm mass and composition of the wild-type (wt) Je2 strain to strains containing mutated biofilm-associated genes, we hope to uncover the impact that each mutation has on the composition of the biofilm matrix. We will utilize crystal violet assays as well as extracellular DNA and protein quantifying procedures to determine biofilm composition, after which meaningful comparisons can be made between mutant biofilms and wt biofilms.

Murley, Jordan; Stockard, Alyssa; Payne, Andrew; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)

Brain-derived neurotrophic factor (BDNF) is implicated in varied physiological processes. Its main receptor in the central nervous system is tyrosine receptor kinase B (TrkB), and the main ligand for TrkB in the central nervous system is BDNF. It has been shown previously that activating TrkB can cause a downregulation of the chloride-exporting potassium chloride cotransporter 2 (KCC2), presumably resulting in a decreased chloride gradient. It is hypothesized that this down-regulation, caused by an increase in BDNF levels, creates hyperexcitable GABA neurons in the ventral tegmental area (VTA) due to a reduced efficacy of inhibitory currents. Here we investigate this mechanism of adaptation in the context of chronic alcohol exposure. We demonstrate that BDNF levels are elevated in the VTA during withdrawal from chronic alcohol exposure. We also observe that blocking TrkB activity decreases alcohol seeking behavior. Further, we investigate the expression patterns of KCC2 in connection with chronic alcohol administration. Additional work is underway to validate this mechanism and further elucidate its putative role in alcohol dependence.

Diaz, Paula; Baptista, Gabriella; Obray, J Daniel; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)

The prevailing view is that enhancement of dopamine (DA) transmission in the mesolimbic DA system underlies the rewarding properties of alcohol. The aim of the present study was to help reconcile the relative insensitivity of DA neurons to ethanol in vitro (EC50 = 96 mM) with the sensitivity of DA neurons in vivo (EC50 = 3 mM). To do this we investigated the role of peripheral dopamine 2 receptor (D2R) mediated neuroimmune responses in ethanol enhancement of DA release in the NAc and ethanol reward. We found that systemic administration of ethanol (0.5-4.0 g/kg) markedly enhanced DA release in the NAc while pretreatment with a peripheral-only D2 receptor (D2R) antagonist blocked these effects. A place conditioning paradigm was used to test rats for ethanol preference. Administration of a peripheral D2R antagonist before ethanol conditioning trials was found to prevent acquisition of ethanol conditioned place preference. Finally, ethanol suppression of locomotor activity in rats was attenuated by domperidone pre-administration. Domperidone pre-administration did not affect ethanol impairment of motor coordination. These findings suggest that ethanol enhancement of DA release, intoxication, and ethanol reward are all mediated, at least in part, by a peripheral mechanism involving D2Rs. These results challenge the dogma regarding direct ethanol actions on mesolimbic DA transmission and potentially provides novel pharmacological targets for the treatment of alcohol use disorder.

Desdames, Chloe; Smith, Mick (Salt Lake Community College)
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)

Spider monkeys (genus Ateles) is one of the many endangered species in Costa Rica and they can now only be found in very few places due to deforestation. It is important for conservation reasons to know what forest types spider monkeys prefer. According to prior research in Surinam, Mexico, and Bolivia, Ateles prefers primary forests (undisturbed, high canopy) over secondary forests (degraded, lower canopy). Ateles are frugivores and their habitat preference depends highly on fruit availability. Fruit is more abundant in primary over secondary forests. This goal of this study was to determine whether spider monkeys in Costa Rica have a similar habitat preference. I hypothesize that spider monkeys will be found more often in high canopy primary forests compared to secondary forests. This research was conducted at La Selva Biological Station in northeastern Costa Rica. La Selva is a protected lowland tropical rainforest consisting of both primary forest (55%) and secondary forest in various stages of regeneration. I conducted a census of Ateles geoffroyi by walking established trails, with markers every 50 meters, in May 2019. Whenever a spider monkey was spotted, the trail name, nearest meter marker, and forest type were recorded. On a census of 9 trails, five Ateles groups were located, with four of the sightings being in primary forests. My hypothesis, that Ateles geoffroyi would be found in primary over secondary forests, was supported because 80% of the sightings were in primary forests. This is especially significant because, of the 20.5 kilometers censused, only 4.6 km censused (22%) were in primary forests with the remaining 15.9 km (78%) in secondary forests. This preference for primary forests agrees with prior studies on Ateles in other Neotropical forests. This highlights the importance of conserving primary forest for the well-being of spider monkeys.

MacLachlan, Aileen; Berges, Bradford (Brigham Young University)
Faculty Advisor: Berges, Bradford (Life Sciences, Microbiology and Molecular Biology)

Staphylococcus Aureus (SA) is a well-known human pathogen causing infection in hospital settings world-wide. Given that SA is becoming increasingly resistant to antibiotics, the need to discover alternative treatments is urgent. One path that SA uses to combat antibiotics is by forming biofilms. Biofilms are microbial cell communities that form on surfaces and employ a complex extracellular polysaccharide matrix to protect the bacteria. In the past, bacteriophage (phage) has been investigated as a potential alternative to treat methicillin-resistant SA (MRSA) and break down its biofilm. Recently, students from Dr. Berges' laboratory isolated 6 novel strains of phage. In a recently published paper from Dr. Berges' lab, these phages demonstrated significant reduction of planktonic strains of SA and MRSA.

In this project, we further explore the ability of these 6 phages in breaking down biofilms from hospital associated SA strains. We plan to measure the reduction of SA biofilms caused by these novel phages against a control. The reduction results will be analyzed with previous research results to detect the presence of a polysaccharide degrading enzyme for the purpose of future research. In addition, we also plan on measuring the ability of the phage in preventing biofilm formation.

Bursell, Madeline; Dikow, Rebecca; Johnson, Warren; Koepfli, Klaus-Peter; Frandsen, Paul (Brigham Young University)
Faculty Advisor: Frandsen, Paul (Life Sciences, Plant and Wildlife Sciences)

Due to decreasing costs in genome sequencing, conservation genomics is a field that has experienced immense growth over the last few years. By comparing whole genome data within threatened and endangered populations, we can estimate important elements in conservation such as levels of homozygosity and demographic histories that reveal the level of endangerment of a species. This information informs conservation priorities and captive breeding programs. In the present study, we focus on two species of clouded leopards: Neofelis nebulosa and Neofelis diardi. N. nebulosa is a species of clouded leopard that lives in mainland southeast Asia. N.diardi inhabits the islands of Sumatra and Borneo in Indonesia. While these two species were initially thought to be a single species, evidence, such as differences in fur color and sizes of cloud markings, hint that they have diverged into two species. To shed more light on the genomic differences between them, we sequenced, assembled, and annotated whole genomes from both species. With genomes, we will explore differences in demographic histories, variation in blocks of homozygosity, and generate a whole genome phylogeny with other large cat species. Using these analyses, we share insights that will inform the conservation status of the two species.

Bonnie K. Baxter (Westminster College)
Faculty Advisor: Baxter, Bonnie (Westminster College, Biology)

Gypsum is a calcium sulfate mineral in a hydrated form. NASA's Mars Exploration Rover, Opportunity, found veins of gypsum deposited by water in 2011(Figure 7), and gypsum has been detected on Mars as early as 2005 by the ESA's Mars Express Orbiter. On Earth, gypsum is formed in hypersaline environments, in minerals left behind when water evaporates and it can trap microorganisms in fluid inclusions. Gypsum obtained from Great Salt Lake was used to develop a method to extract halophilic archaea and culture it in the lab. Our studies show that the mineral was difficult to dissolve in aqueous microbiological media. We tested various methods of dissolution involving mechanical crushing and tested solvents including microbiological media. We also employed a variety of cultivation methods. We will present data on best practices for obtaining halophilic microorganisms from gypsum samples. The method obtained could be used to isolate potential microorganisms present in gypsum samples from Mars.

Brunetti, Bryce; Escarate, Ashley; Conway, Matthew; Slezak, Cyrill; Kopp, Olga (Utah Valley University)
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology); Slezak, Cyrill (Utah Valley University, Physics)

Purpose:
This study evaluates the effect of electrohydraulic shockwaves on Staphylococcus aureus biofilms. This system could be a great alternative to the use of antibiotics, and potentially life-saving technology that could save billions of dollars.

Background:
The rise of antibiotic-resistant bacteria is a global threat. Staphylococcus aureus is typically harmless, but this gram-positive species has become highly resistant and extremely pathogenic. Strains like MRSA and VRSA have the highest rate of drug resistance and are the leading cause of chronic bacterial infections via bacterial biofilms on medical devices. Biofilms are an aggregation of microbes that excrete an extracellular matrix providing an ideal environment for gene exchange and quorum sensing. Their complexity hinders the diffusion of antimicrobials. A proposed method to prevent device-associated infection is shockwave sterilization and therapy. A shockwave is a high-energy wave causing a sudden change in temperature, pressure and density in the medium. This study investigates the potential disruption of bacterial biofilms by electrohydraulic shockwaves.

Methods:
E. coli and S. aureus biofilms were grown on polystyrene plates. Biofilms were treated with shockwaves (0.19mJ/mm2, 300 pulses, 3 Hz) in a water bath and compared with those treated with Vancomycin. Cell viability was determined through XTT/menadione absorbance and specific biofilm formation through crystal violet absorbance.

Results:
Current testing has shown that electrohydraulic shockwaves have a bacteriostatic effect on biofilms. Other finding show potential for shockwaves to increase bacterial susceptibility to lower levels of antibiotics.

Conclusions:
Device-associated infections are a serious threat to patients' health. The diminishing effectiveness of antibiotics in treating and preventing infections along with evolution of mass resistance in bacteria have given rise to the term "post-antibiotic era." The better understanding of electrohydraulic shockwaves bacteriostatic effect could lead to more effective treatments for antibiotic resistant bacteria such as S. aureus.

Sirrine, Michael; Grose, Julianne (Brigham Young University)
Faculty Advisor: Grose, Julianne (Brigham Young University, Microbiology and Molecular Biology)

PAS kinase is a serine/threonine protein kinase known to regulate the pivotal switch between cellular respiration and lipid biosynthesis. One substrate of PAS kinase is Cbf1, a known transcription factor which regulates lipid biosynthesis in yeast and mammalian cells (human homolog USF1). USF1 is associated with hyperlipidemia and hypercholesterolemia in several GWAS studies. We have recently identified and characterized a role for Cbf1 in the regulation of respiration as well, making it a key player in partitioning cellular resources towards respiration versus lipid metabolism. The goal of this proposal is to use the powerful tools of yeast genetics to identify physical interactors of Cbf1 in order to characterize the molecular mechanisms of its action.

Lemmon, Skyler; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)

Over the course of the last year, I have dedicated most of my time in the lab to learning about CRISPR/Cas9 and practicing the laboratory techniques that are necessary to make genetic changes in Drosophila melanogaster. Here I aim to expand on that expertise by applying CRISPR to study a genetic question: how the microbial composition of the D. melanogaster microbiome is affected by the modification of 4 specifically selected genes in flies from Florida and Maine. For each of the selected genes, the Florida fly allele will be put into the Maine fly genome and the Maine fly allele will be put into the Florida fly genome. The microbiome composition of these two new flies will be compared against the original lines in a factorial design. Embryos will be injected with the necessary plasmids for a double-stranded cut to take place. After injection, homology dependent repair that will incorporate the new allele. Sanger sequencing will be used to screen for successful knock-in of the allele. Finally, the concentrations of each type of bacteria found in the microbiota of the flies will be measured and compared against the flies from which the allele came from.

Hess, Kavan; Herring, Jacob; Yang, Haokun; Tessem, Jeff (Brigham Young University)
Faculty Advisor: Tessem, Jeff (Brigham Young University; Department of Nutrition, Dietetics, and Food Science)

Diabetes is the seventh leading cause of death in the United States, and often accompanies other life-threating complications. There are two main types of diabetes that are both characterized by disfunction or destruction of insulin producing beta cells found in the islets of Langerhans. Islets of Langerhans are composed of endocrine hormone secreting cells, including alpha cells (glucagon), beta cells (insulin) delta cells (somatostatin), epsilon cells (ghrelin) and PP cells (pancreatic polypeptide). While alpha and beta cells make up ~90% of all the cells in the islet, delta cells comprise only ~10% and are responsible for cross talk in the islet. Delta cells regulate intra-islet cross talk through the secretion of somatostatin-14. It has been shown that Nr4a1 and Nr4a3 overexpression induces beta cell proliferation, while Nr4a1 or Nr4a3 deletion inhibits insulin secretion when challenged with glucose. Delta cells contain three times the amount of Nr4a1 mRNA than beta cells. However, no research has been done on the role of either of these transcription factors in the cross talk between the different cell types of the islet. Here we aim to show how a lack of Nr4a1 and Nr4a3 affects delta cell somatostatin release when challenged with glucose.

Domyan, Eric; Godoy, Daniela; Gardiner, Kylan (Utah Valley University)
Faculty Advisor: Domyan, Eric (Science, Biology)

Pigmentation is one of the main traits we notice when we look at something, whether it be a flower, an animal, or another human. Variation in pigmentation arises when random mutations affect the function of a gene involved in pigmentation production. In this research our goal is to understand a specific mutation that happens in Pigeons which involves the TYRP1 gene.
The TYRP1 gene instructs the making of the tyrosinase-related protein. This enzyme is located in melanocytes, which are cells that produce melanin. Studies suggest that this enzyme may help stabilize tyrosinase, which is responsible for the first step in melanin production.
TYRP1 has a signal peptide which directs the protein to the ER (endoplasmic reticulum) where the signal peptide is removed before the mature protein is trafficked to melanosomes to perform its normal function. The Ash-red mutation, however, prevents removal of the signal peptide, which somehow results in pheomelanin synthesis (red) instead of eumelanin synthesis (dark blue). These findings suggest that the Ash-red mutation is causing the TYRP1 protein to perform a new function. The goal of this project is to better understand the synthesis and trafficking of TYRP1 throughout the cell organelles.
To study this, we plan to use transgenesis to express normal, or Ash-red versions of TYRP1 protein in melanocytes, label the different intracellular compartments using an immunostain, and determine whether normal or Ash-red TYRP1 proteins are being sent to the same, or different compartments of the cell.

Wilson, Seth; Taylor Adam; Bursell, Madeline; Frandsen, Paul; Stewart, Russell; Steeneck, Amy (Brigham Young University)
Faculty Advisor: Frandsen, Paul (Brigham Young University, Plant and Wildlife Sciences)

Caddisflies (Insecta: Trichoptera) have evolved to produce silk with adhesive and elastic properties in aqueous environments. The silk is used in several ways by different species within the order such as case making, retreat making and using the silk as an anchor in the stream. Previous research on caddisfly silk has focused on understanding the evolutionary changes in the H-fibroin gene, the main protein found in caddisfly silk, which underlies the structural transformation behind these phenotypic properties that allow for diverse usage of the silk across the order (Ashton et al. 2013). Understanding the genetic foundation of the silk is crucial to understanding the phenotypic interactions that determine the unique qualities of caddisfly silk. An accurate assembly of the caddisfly genome will allow us to resolve the H-fibroin gene that plays an integral role in the formation of the caddisfly silk. Next-generation sequencing, Oxford Nanopore, and PacBio will allow us to sequence long reads that can span repetitive regions of the genome. These regions have made it difficult to resolve the H-fibroin gene as there are many repetitive motifs found in the gene. We will combine this next-generation sequencing with second-generation sequencing, Illumina and Sanger Sequencing to optimize the assembly. In this study, we used a combination of next-generation sequencing technologies to assemble the complex H-Fibroin gene in order to look at the underlying genetic structure of the silk protein. We identified unique repetitive motifs in the gene that contribute to the silk's adhesive strength and elasticity when in aqueous environments.

Escarate, Ashley; Brunetti, Bryce; Conway, Matthew (Utah Valley University)
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology); Slezak, Cyrill (Utah Valley University, Physics)

Medical device-associated infections can lead to serious complications affecting the health of patients. Electrohydraulic shockwave treatments have shown bactericidal activity in some microorganisms. Biofilms are structures formed by microorganisms enclosed in an extracellular matrix. They form on a variety of surfaces protecting the microorganisms from antibiotics and facilitating their growth. This can result in a high rate of drug resistance and in many cases result in chronic bacterial infections.
Previously determined MIC50 concentrations of vancomycin had little effect on biofilms at twelve hours of treatment when not paired with shockwave therapy. This research evaluates the synergistic effect of different concentrations of vancomycin and shockwaves after twelve and twenty four hours of treatment given that vancomycin has shown time-dependent activity. Biofilms were grown in 96 well plates and the corresponding treatments were applied. XTT and Crystal Violet assays were used to quantify and qualify the presence of the biofilm and the antibiosis effect. The results of this experiment will be discussed in detail. A better understanding of the synergistic effects of antibiotics and shockwave therapy may lead to a more effective treatment of biofilm and device-associated infections.