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2020 Abstracts

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CRISPR-based identification of Salmonella in local waterways

December 30, 0020 12:00 AM
Hirschi-Forster, Jeanallie; Mendoza, Matthew; Van Oene; Nicholas ; Payton, Jullian (Weber State University)
Faculty Advisor: Clark, Daniel (Science, Microbiology)

The purpose of this research is to obtain quantitative data about possible sources for Salmonella contamination including tributaries to the Great Salt Lake, namely, the Jordan River, Weber River, and Bear River in Utah. We will also analyze specific water and soil sources near poultry farms for possible contamination. In recent studies, there is a greater number of produce items that have been found to contribute to Salmonella outbreaks. Contaminated water used for irrigation of these crops has been implicated as the causative agent for food contamination.

Bacteria found in these waterways are enriched using selective and differential media. This means, the media provides Salmonella species with required nutrients to grow effectively while differential media inhibits the growth of non-Salmonella species. The enrichment media that is used during this process is 3 X Tryptic Soy Broth and Gram-Negative broth. Gram negative broth is used as an enrichment step, but also selective in that it inhibits growth of other organisms. The two types of differential media would be XLT4 and MSRV. Salmonella is a motile bacterium and thus branches out from its original location of inoculation. This creates a halo-like growth pattern that makes it possible to differentiate Salmonella on MSRV plates. Once Salmonella is confirmed through the MSRV and XLT4 media, sequencing of its two CRISPR loci is completed. These two chromosomal regions have been shown to be distinct in different serovars, and as such, they can be used to distinct what subspecies is present in the sample.
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Effects of Grape Seed Extract Metabolites on ß-cell Proliferation and Function

December 30, 0020 12:00 AM
Beales, Joseph; Lloyd, Trevor; Krueger, Emily; Barlow, Andrew (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutritional, Dietetics, and Food Science)

Worldwide, an estimated 415 million people suffer from diabetes.1 Diabetes is characterized by chronic dysfunction of the pancreatic ß-cell, which leads to unregulated insulin secretion and abnormal blood glucose levels. Therefore, methods which increase the number of ß-cells or improve their function have potential for complementary treatment of type 2 diabetes. Compounds such as antioxidants and their gut metabolites have received attention in literature as having potential ß-cell-regulating properties.2,3 Therefore, we hypothesize that supplementation of grape seed extract (GSE), which is rich in antioxidants, will enhance ß-cell proliferation and insulin secretion. Accordingly, we obtained metabolites, derived from rats on either a control or grape seed extract diet, to measure the metabolites' impact on ß-cell function through in vitro assays such as glucose stimulated insulin secretion (GSIS) and 3H-thymidine incorporation. Discoveries regarding GSE metabolites' effects on ß-cell function could be fundamental to understanding ß-cell regulation and potential pharmaceutical or dietary treatments for diabetes.

1 Ogurtsova, K., et al. "IDF Diabetes Atlas: Global Estimates for the Prevalence of Diabetes for 2015 and 2040." Diabetes Research and Clinical Practice, Elsevier, 31 Mar. 2017, www.sciencedirect.com/science/article/pii/S0168822717303753?via%3Dihub.

2 Bajaj, Sarita, and Afreen Khan. "Antioxidants and diabetes." Indian journal of endocrinology and metabolism vol. 16,Suppl 2 (2012): S267-71. Doi:10.4103/2230-8210.104057

3 Tsuda, Takanori. "Recent Progress in Anti-Obesity and Anti-Diabetes Effect of Berries." MDPI, Multidisciplinary Digital Publishing Institute, 6 Apr. 2016, www.mdpi.com/2076-3921/5/2/13.
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Drosophila melanogaster Determines Dietary Preference Through Volatile Detection

December 30, 0020 12:00 AM
Walker, Carson; Burke, Tyler; Tanner, Call; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Brigham Young University, Plant and Wildlife Sciences)

Host-microbe interactions can dramatically influence Drosophila melanogaster phenotypes, but few studies have explained how these microbes are recognized by the host. For example, fruit flies from one area, Maine, prefer to consume diets inoculated with Lactic Acid Bacteria (LABs) over diets containing Acetic acid bacteria (AABs); whereas flies from another area, Florida, show no preference for either LAB or AAB. However, the bacterial mechanisms responsible for this preference are unknown. My follow-up analyses further suggest the hypothesis that Maine flies avoid AAB, rather than are attracted to LAB. Therefore, I propose a forward genetic approach to define the AAB processes that shape this fly preference. I will do this by comparing the feeding preferences of Maine and Florida flies to diets incorporated with LAB versus specific bacterial molecules, such as acetic acid, peptidoglycan, and lipopolysaccharides. Alternatively, if these molecules do not mediate the effects I will perform a metagenome wide association assay (MGWA) to identify genes linked to this preference. Together, these approaches will help to reveal the bacterial factors that influence fly feeding preferences.
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Spatial variation in mercury concentrations of flying insects at Antelope Island

December 30, 0020 12:00 AM
Stoneham, Lisa; Brasso, Dr. Rebecka (Weber State University)
Faculty Advisor: Brasso, Rebecka (Weber State University, Zoology)

Mercury is a toxic heavy metal that poses significant health threats to people and wildlife. The organic form of mercury, methylmercury, is converted from its inorganic form via microbial methylation. Methylmercury is dangerous because it attaches to proteins in the blood, muscle, and other tissues and can cross the blood-brain and placental barriers. Microbial methylation is enhanced in anoxic environments such as wetlands, which are increasingly being classified as mercury hotspots where animals accumulate elevated concentrations relative to those in terrestrial systems. This is concerning for the wetlands of the Great Salt Lake due to its history of anthropogenic inputs of pollutants and its importance as a breeding ground and rest stop for migrating avian species. Previous research has shown significant mercury methylation occurring within the Deep Brine Layer of the GSL. The purpose of this project was to investigate potential spatial variation in mercury concentration in different portions of the GSL. With a focus on invertebrates, we collected insects including brine flies, midges, damselflies, and crane flies from three sites of varying salinity around Antelope Island State Park: Farmington Bay, White Rock Bay, and the Antelope Island Marina. Mercury concentrations in insects were determined using a Nippon MA-3000 Direct Mercury Analyzer. Our results will provide a preliminary assessment of mercury concentrations in flying insects from different habitats around the island. This will help in determining differential risk to insectivorous songbirds, waterfowl, and shorebirds foraging on these common prey species in the GSL.
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The Beta Cell Struggle: How CDKIs and Age Affect Cell Proliferation in Type 1 Diabetes

December 30, 0020 12:00 AM
Jensen, Daelin; Baxter, Melanie (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Nutrition, Dietetics, and Food Science; Life Sciences)

Approximately 1.25 million people are currently living with type 1 diabetes. By 2050, 5 million people are expected to be diagnosed with the disease1. The insulin secreting pancreatic beta cells are essential to control proper glucose absorption and storage in insulin sensitive peripheral tissue. Both type 1 and type 2 diabetes are characterized by decreased functional beta cell mass and, consequently, decreased insulin production. One potential intervention is the use of beta cell transplantation from cadaveric donors. A major impediment to greater application of this treatment is the scarcity of transplant ready beta cells. Increasing the quantity of functional beta cells for transplantation will lead to increased insulin production and better management of the disease. Various genes have been defined that can induce beta cell replication. A major caveat of these findings, however, is that these factors induce replication in young beta cells but not in aged beta cells. Age-dependent morphological changes in the beta cell are poorly understood, despite its relevance to type 1 diabetes: here, we show that insulin-positive tissue area changes with age. Given that the majority of beta cells that will be used for transplant will come from aged donors, it is imperative to understand why aged beta cells are refractory to the aforementioned proliferative mechanisms. The cell cycle is tightly regulated by cyclin-dependent kinases. Cyclin-dependent kinase inhibitors (CDKI's) bind to cyclin dependent kinases, inhibiting cell proliferation. We hypothesized that these CDKIs are responsible for the observed lack of proliferation in aged animals. We demonstrate the expression of the Ink4 and Cip/Kip family of CDKI's by mRNA, protein and histological expression in 5 week and 5 month old primary rat beta cells. In addition, we show how size-related expression differences of CDKIs relate to beta cell proliferation.
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Taller seedlings in about half the time: the effect of coyote ingestion on netleaf hackberry (Celtis reticulata) seeds

December 30, 0020 12:00 AM
Hannah A. Veltkamp, Sydney Houghton, Michael T. Stevens (Utah Valley University)
Faculty Advisor: Stevens, Micheal (Utah Valley University, Biology)

Netleaf hackberry (Celtis reticulata) is a deciduous shrub native to the southwestern United States and northern Mexico. Individual shrubs can be long-lived, but newly established stands of hackberry are rare. The lack of juvenile hackberry in the wild could be due to low germination rates reported in both laboratory and field settings. The seeds of hackberry are embedded in drupes that are an important source of food for birds and small mammals. Animals likely play an important role in seed dispersal, and passing through a digestive tract could increase the germination rates of hackberry seeds. Passage through the digestive tract of a mammal can increase the germination rates for some plant species, but not for others. We hypothesized that passage through the digestive tract of a coyote would increase the germination rates of C. reticulata. To test this hypothesis, we collected 17 coyote scats containing visible hackberry fruits from along the Bonneville Shoreline Trail east of Provo, Utah, using latex gloves. Each scat location was recorded using a GPS unit. After collecting each scat, we found the closest hackberry shrub and picked a sample of fresh hackberry fruits from it. All samples were cleaned and cold stratified and then planted into cone-tainers containing a potting soil mix and placed in the Utah Valley University greenhouse. We sowed 20 seeds from each of the 17 coyote scats and

20 seeds from each of the neighboring hackberry bushes for a total of 680 seeds. The 680 cone-tainers were labeled with plastic stakes and randomly positioned into trays that were randomly distributed on a bench in the greenhouse. The seeds, and later seedlings, were watered as needed (typically three days/week). On watering days, we checked for newly-germinated hackberry seedlings and recorded their date of emergence. Near the end of the experiment, we measured the height of each seedling. The germination rate of hackberry seeds that had passed through the

digestive tract of a coyote did not differ from the germination rate of seeds from fresh-picked fruit (42.7% vs. 46.5%, respectively; _ 2 = 0.558, df = 1, p = 0.455). However, on average, the coyote-treatment seeds took just over half as many days to germinate as did the seeds from fresh-picked (undigested) fruit (35 days vs. 69 days, respectively; p < 0.001). The seedlings from coyote-treatment seeds were 9.5% taller than were the seedlings derived from seeds from undigested fruit (6.4 cm vs. 5.8 cm, respectively; p < 0.001). Our results show that consumption by coyotes can benefit hackberries by enabling their seeds to germinate earlier in the year when

conditions for establishment are good. The earlier start on germination that coyote-ingested hackberries get translates to increased height and likely a higher rate of survival in the field.
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The Effects of Glucolipotoxicity on Nkx6.1 Expression and Insulin Secretion in the Beta Cell

December 30, 0020 12:00 AM
Elison, Weston; Bauchle, Casey; Bunker, Libby; Stephens, Samuel; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics and Food Science)

Type 2 Diabetes (T2D) effects hundreds of millions of people worldwide, with that number increasing rapidly. It is characterized by increased insulin resistance and dysfunctional insulin secretion. The beta cell of the pancreas is the primary insulin secreting tissue, found in the endocrine tissue of the pancreas called islets of Langerhans. In T2D beta cells become glucose intolerant and disease progression is characterized by loss of functional beta cell mass. Previous studies have shown that the transcription factor Nkx6.1 is vital for beta cell differentiation, identity, and insulin secretion. Research has indicated that Nkx6.1 expression and protein levels decrease in pancreatic islets from human donors with T2D. Our data indicates that glucolipotoxicity, a common model for obesity and diabetes in cell culture, leads to decreased Nkx6.1 mRNA expression, protein levels and nuclear localization in Ins-1 832/13 cells. Nkx6.1 regulates genes in the nucleus , and its loss inhibits proper insulin secretion. We propose that reactive oxygen species created by metabolism of excess fuel decreases Nkx6.1 expression and Nkx6.1 target gene expression, as measured by quantitative polymerase chain reaction (qPCR). Also, increased glucose concentrations causes increased Nkx6.1 protein degradation and translocation out of the nucleus. Protein levels will be measured by western blot and localization by confocal microscopy. In order to understand how these changes effect beta cell function, we will measure glucose stimulated insulin secretion by sandwich Enzyme Linked Immunosorbent Assay (ELISA). We further propose that Nkx6.1 overexpression will restore beta cell function. These results will assist in unraveling the cause of beta cell dysfunction in T2D.
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Stress Sensitivity to Temperature in Plateau Side-blotched Lizards (Uta stansburiana uniformis): Implications for Immune Function

December 30, 0020 12:00 AM
Lidgard, Audrey; French, Susannah; Hudson, Spencer (Utah State University)
Faculty Advisor: Lidgard, Susannah (College of Science, Biology Department)

Ectothermic organisms, such as reptiles, rely on the external environment for regulating internal temperatures necessary for vital physiological processes. When faced with environmental challenges, temperature may differentially affect how allostatic mediators (e.g., glucocorticoid hormones) are released to mediate energy allocation for handling stressors. Subsequent differences in energy mobilization and circulating metabolites during a stress response may ultimately influence self-maintenance processes such as immunity. The aims of this research were to determine how stress sensitivity varies with diurnal temperatures in the Plateau Side-blotched Lizard (Uta stansburiana uniformis) and to assess the potential implications for immune function. Both baseline and stress-induced levels of glucocorticoids (corticosterone) and energy metabolites (glucose) were compared to body temperature and the thermal environment. Variation in innate immune function (bactericidal ability) was then compared to both temperature and physiological parameters at baseline and stress-induced levels. Stress reactivity via glucocorticoid release positively corresponded with body and environmental temperatures, although glucose release did not. Bactericidal ability subsequent to a stressor negatively corresponded with body temperature and glucocorticoid release. Such findings provide further insight on how stress sensitivity and self-maintenance can vary across the thermal environment, posing potential fitness consequences for an ectothermic organism.
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The Effects Of Invasive Common Carp On Invertebrate Food Sources For Diving Ducks In Great Salt Lake Wetlands

December 30, 0020 12:00 AM
Karin, Kettenring; Robison, Talin; Leonard, Emily (Utah State University)
Faculty Advisor: Kettering, Karin (S.J. & Jessie E. Quinney College of Natural Resource, Watershed Sciences Department)

The Great Salt Lake (GSL) and its wetlands are important habitat for migrating birds. The GSL wetlands provide crucial habitat for nesting, food, and areas to recover from migration. Common carp are a threat to GSL wetlands. Carp disturb sediments in the water, blocking some of the sunlight from entering the water, which is utilized by aquatic macrophytes and algae. Carp also may be affecting invertebrate populations, which are critical food resources for migrating birds, but these effects have not been well-documented. My research addressed the question: what are the effects of invasive common carp on invertebrate food sources for diving ducks in the Great Salt Lake wetlands? I answered my research question by addressing the following objectives: (1) to identify the benthic, epiphytic, and water-column dwelling invertebrates in Farmington Bay Waterfowl Management Area (WMA), and (2) to determine if common carp are having an impact on the overall density, diversity, and abundance of the invertebrate communities fed on by diving ducks. I compared invertebrate communities (diversity and abundance) between carp-excluded boxes and control boxes. I constructed my carp exclosures of wire mesh and t-posts to prohibit carp from entering while still allowing invertebrates and water to freely move in and out of the exclosure. The control boxes were constructed of t-posts and allowed carp to freely enter and exit the box. I used dipnet and substrate core samples to determine what invertebrates are living in the water column and substrates at Farmington Bay wetlands. Although sample processing is on-going, early results indicate that carp reduce water column invertebrate abundance while effects on invertebrate diversity are thus far inconclusive. Given the importance of GSL wetlands and their invertebrate food sources to migrating diving, my research findings underscore the importance of aggressive carp management.
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The Role of Bacterial Genotype in Persistence of the Microbiota of Drosophila melanogaster

December 30, 0020 12:00 AM
Gottfredson, Sarah; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)

The microbiome of Drosophila melanogaster can have significant effects on the host, and many of these have been studied. However, the reason why the bacterial species associate with and persist in D. melanogaster has not been studied in depth. Here we define persistence as how long a microbe associates with a host. The early assumption has been that the D. melanogaster gut microbiome is established solely through diet, but recent work suggests that other factors may be at play in the microbiome establishment. This experiment aims to study the correlation between bacterial genotype and persistence in the D. melanogaster microbiome. In this study, a metagenome wide association (MGWAS) was done using 40 different strains of bacteria to find distinct bacterial genes that are significantly correlated with persistence. To do this, each strain was mono-associated with twenty-four individual flies. The flies were reared for fourteen days, transferred onto new food three times a day for two days, homogenized, and plated. Using the significant genes found through the MGWAS, the same experiment protocol will be used to test mutants of these genes for their effect on persistence. These data will provide us with distinct genes that are necessary for effective bacterial persistence.
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Temperature Effects on D. melanogaster Microbiota Content

December 30, 0020 12:00 AM
Not yet published (Brigham Young University)
Faculty Advisor: Chaston, John (Brigham young University, Life Sciences)

Within an organism's gut are many strains of bacteria that are constantly interacting with their host. Microbiota composition has been shown to impact many aspects of host health such as metabolism, fat-storage, starvation resistance, and reproduction. Certain behaviors and outcomes have been correlated with certain microbial taxa present in the host gut.

D. melanogaster serves as a useful tool for studying this relationship because its microbiota contains relatively few bacterial strains and is both widely studied and largely understood. Previous research within our lab involving D. melanogaster has found trends in many life-history strategies (ie. reproduction, fecundity, lifespan) that correlate with the presence of certain gut bacteria. While there are many aspects of health that microbiota composition affects, there are also a variety of factors that impact microbiota composition thus leading to these end results.

This experiment seeks to further understand the role that environment has in determining microbiota composition. By rearing gnotobiotic flies in environments that differ in temperature, we can then analyze microbiota content to see if any fluctuations occur due to environmental temperature. If temperature is found to have an effect on the taxa present in fully developed D. melanogaster, we can then seek to determine whether or not there are evolutions taking place in host genotype that yield differing microbiota phenotypically.
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The search for Lactobacillus wasatchensis.

December 30, 0020 12:00 AM
Thornton, Sherie; Cardona,Rebecca (Weber State University)
Faculty Advisor: Culumber, Michele (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology)

Lactobacillus wasatchensis was initially isolated from cheese produced at Utah State University and was found to be a Non-Starter Lactic Acid Bacteria (NSLAB) that causes late-gas production in cheese that can damage packaging and produce defects in the cheese. The goal of this project was to locate an environmental reservoir for Lactobacillus wasatchensis. Five samples of silage that were in different stages of fermentation and content and raw milk samples were obtained at the Utah State University dairy. Samples were serially diluted, plated on de Man, Rogosa and Sharpe agar supplemented with 1% D-Ribose (NRS-R) and incubated anaerobically for 5 days. Colonies that looked like potential Lb. wasatchensis were selected and regrown for isolation. All isolates were gram-positive rods. The isolates were further grown in broth for DNA extraction, sequencing, and analysis with API 50 carbohydrate panel (API 50CH). The API 50CH results were significantly different from Lb. wasatchensis, which only demonstrates use of ribose in this assay. Sequencing of the 16S rRNA gene, however, produced a match to three isolates from two different silage samples that had 99% sequence identity to Lb. wasatchensis. Further analysis of the isolates is being done to confirm this finding and describe the organism isolated from the soil. We hypothesize that these organisms are very closely related to Lb. wasatchensis and that silage could be an environmental source of contamination.
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The effect of IL-1β on Nf-_B and ICAM-1 mechanism

December 30, 0020 12:00 AM
Hendricks, Kyle; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)

Over 30 million Americans suffer from type 1 (T1D) or type 2 diabetes (T2D), the seventh leading cause of death in the US. T1D and T2D is caused by a significant decrease in pancreatic β-cell mass, resulting in the body's inability to regulate blood glucose. Specifically, T1D is classified as an autoimmune disease due to pancreatic β-cell death by the body's T cells. Nf-κB is required for T cell mediated β-cell destruction. Nf-κB interacts with ICAM-1 on the T cell and acts in conjunction with IL-1β which acts as a T cell activator. This pathway is part of the mechanism that contributes to T cell mediated cell destruction. Here we hypothesize that IL-1β is involved in the mechanism that contributes to Nf-κB and ICAM-1 binding. We will begin with an electrophoretic mobility shift assay to identify the interactions between the ICAM-1 site on IL-1β treated cells and the Nf-κB binding complex. A better understanding of this pathology can, in the future, lead to a treatment that could regulate T cell mediated death of β-cells.
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The role of Fibroblast Growth Factor 21 (FGF21) in Mitochondrial Disorders (MDs)

December 30, 0020 12:00 AM
Almaw, Naredos; Chaudhuri, Dipayan (University of Utah)
Faculty Advisor: Chaudhuri, Dipayan (School of Medicine, Internal Medicine)

Fibroblast Growth Factor 21 (FGF21), a regulator of metabolism that is typically expressed in the liver, has recently been shown to be induced by other tissues in the body as a response to mitochondrial stress. Elevated levels of serum FGF21 was exhibited in children with mitochondrial mutation-induced mitochondrial dysfunctions. Similarly, in dilated cardiomyopathy, a common type of heart failure (HF) mitochondrial dysfunction is associated with mitochondrial DNA damage. This study aims to determine the signaling pathway that leads to the production and effects of FGF21 during mitochondrial dysfunction associated HF. We hypothesize that in left ventricular failure, cardiomyocytes experience oxidative stress, which initiates signaling pathways that leads to the production of FGF21 by other organs.

To test this hypothesis, HF was induced in four mice models via Transverse Aortic Constriction (TAC), and tissue samples were collected. Messenger RNA (mRNA) was extracted, and quantitative Polymerase Chain Reaction (qPCR) was performed to examine the FGF21 gene expression in control and experiment mice models. The qPCR data showed an upregulation of FGF21 in the heart, liver, and pancreas of experiment mice. qPCR results were confirmed through FGF21 protein expression via western blot. Our preliminary results appear to support our hypothesis that during heart failure, the heart sends stress signals to other organs to produce FGF21. Understanding the origin of FGF21 production could help better understand the critical role it plays in preventing disease progression in HF patients.
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The Effects of the Ketogenic Diet on Learning and Memory

December 30, 0020 12:00 AM
Edwards, Jeffery; Saito, Erin; Blaylock, Tanner; Brantley, Adam; Winzenried, Eric (Brigham Young University)
Faculty Advisor: Edwards, Jeffrey (Life Sciences, Physiology and Developmental Biology)

The ketogenic diet initially began as a significant treatment to prevent epilepsy. More recently it has seen a rise in popularity again, with many attributing positive physiological and cognitive benefits. The purpose of this study is to assess the validity of those claims in an animal model in order to examine this at the cellular level as well as identify possible molecular mechanisms for the changes observed. To quantify this, mice will be fed a diet high in fats and low in carbohydrates. A Morris water maze, radial arm maze, and novel object recognition will then be used to assess the diets effect on behavioral memory. Field electrophysiology will then be performed in the CA1 region of the hippocampus, the region of the brain responsible for mediating memory, to measure two types of synaptic plasticity: long-term potentiation and long-term depression. It has been previously hypothesized that changes in BDNF concentration are a possible explanation for physiological changes caused by the keto diet. To assess this, ANA-12, a TrkB antagonist, will be used to block the effects caused by BDNF. Preliminary data gathered from bathed brain slices of both male and female animals have shown an enhancement of LTP, the cellular equivalent of learning and memory. These data lead us to our hypothesis that the ketogenic diet will cause significant changes in behavioral memory and CA1 synaptic plasticity through altered BDNF levels.
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Substrate specificity in variants of an aldehyde oxidoreductase

December 30, 0020 12:00 AM
Carter, Riley; Hertig, Jess; Durrant, Doran (Southern Utah University)
Faculty Advisor: Pierce, Elizabeth (Science and Engineering, Physical Science)

Aldehyde oxidoreductases (AOR) are enzymes used to catalyze the conversion between aldehydes and carboxylic acids. Certain bacteria use these enzymes as a source of metabolism or to detoxify aldehydes to less toxic carboxylic acids: Desulfovibrio gigas uses a highly efficient enzyme (DgAOR) to oxidize benzaldehyde in metabolism while E. coli uses a periplasmic AOR (PaoABC) to detoxify aldehydes. These AORs are members of the xanthine oxidase family, but they don't metabolize many of the normal substrates characteristic of this enzyme family, namely purines. Moreover, the active sites of these enzymes have very different environments. Correia, et al (2014) characterized the kinetics and structure of DgAOR with several substrates and found that the Phe425 and Tyr535 residues at the active site likely stabilize aromatic aldehydes by pi stacking. This active site was also buried away from solvent. The active site of PaoABC lacked any significant aromatic residues and was positioned at the surface of the protein. The substrate stabilizing elements at this active site are Leu246 and Pro352. We are interested in why these active sites both are unreactive towards purines given their different chemical and location compared to the solvent. We propose that by mutating PaoABC to have smaller, nonpolar residues at the 246 and 352 position, we may be able to change the specificity of PaoABC to include purines. We also will mutate these residues to aromatic groups to probe at the chemical environment of the active site and its similarities to DgAOR.
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Tbx2 Pigment Study by CRISPR/CAS9 Mutation

December 30, 0020 12:00 AM
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.).
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The effect of Nr4a3 gene deletion on body weight, blood glucose levels, and glucose tolerance in mice

December 30, 0020 12:00 AM
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.
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