Life Sciences

Jason Ray, Benjamin Bitner, Kyle Kener, and Brent Jackson, Brigham Young University Life Sciences Type 1 and Type 2 diabetes are increasing at an alarming rate. Both types of diabetes result in decreased functional β-cell mass, which is defined as the number of β-cells multiplied by their Glucose Stimulated Insulin Secretion rate. Decreased functional β-cell mass inhibits regulation of blood glucose levels. β-cells have an extremely low proliferation rate after adolescence, meaning the functional β-cell mass cannot naturally recover. Increasing functional β-cell mass could provide a cure for diabetes, either through pancreatic islet transplants or through enhancement of the endogenous β-cell population. Nkx6.1 has been shown to increase β-cell proliferation by inducing the nuclear receptors Nr4a.1 and Nr4a.3. We have shown that Nkx6.1 increases expression of the gene c-Fos, and that c-Fos induces expression of Nr4a1 and Nr4a3. Furthermore, we have shown that c-Fos is sufficient to induce proliferation of β-cells in the INS-1 832/3 cell line and in primary rat islets. Finally, using lenti-sh-c-Fos to create a stable c-Fos deficient stable cell line, we have demonstrated that Nkx6.1 mediated proliferation is modified by the lack of c-Fos. We propose a model by which c-Fos is a critical link between Nkx6.1 and Nr4a mediated β-cell proliferation.

Jenny Pattison, Brigham Young University Life Sciences Sensory protein kinases are essential in the phosphorylation of many protein substrates, allowing them to control several metabolic functions and maintain cellular homeostasis. PAS kinase is a sensory protein kinase that is highly conserved and plays a crucial role in glucose homeostasis, however little is known about the molecular mechanisms behind its function. UGP1 is the only well-characterized substrate of PAS kinase, and its phosphorylation diverts glucose away from storage and towards cell wall biosynthesis. We have recently discovered another key substrate of PAS kinase that affects glucose metabolism in the cell, Centromere binding factor 1 (Cbf1). Cbf1 regulates genes involved in respiration, and we have shown that the phosphorylation of Cbf1 by PAS kinase inhibits Cbf1, decreasing respiration in yeast cells. We hypothesize that this is due to a decrease in mitochondrial mass in cbf1 deficient yeast. Further characterizing the effects of PAS kinase on Cbf1 will give further insight into how cells regulate their central metabolic functions, including respiration.

Chase Barker, Utah Valley University Life Sciences Central Research Question:

Stephan Maman and Holden Wagstaff, Southern Utah University Life Sciences Most plant species produce chemical compounds called secondary metabolites that enhance fitness in a variety of ways. Many of these compounds are also physiologically active in vertebrates and have widespread medicinal uses. The most ubiquitous secondary metabolites are the terpenoids, many of which cause vasodilation of the aorta and mesenteric arteries. In this study, we examined the vasoactive effects of the essential oil of Umbellularia californica, which contains the terpenoid umbellulone. Oil obtained via steam distillation using aerial portions of U. californica was applied directly to cutaneous arterioles of frogs. Arteriole diameter was monitored both before and after oil application by video microscopy. Within seconds of application, the oil caused significant vasoconstriction that persisted until the oil was washed off. Our control, medical grade sesame oil, caused no observable effects when applied using the same protocols. These results are opposite to the vasodilatory effects of terpenoids on aortic rings and mesenteric arteries. This suggests that the vasoactive effects of umbellulone are different from other terpenoids, that the vasoactive effects of terpenoids differ depending on blood vessel type, or that application of the complete essential oil affects vasculature differently than application of the isolated terpenoid.

Michael Hinckley and Cayden Westwood, Dixie State University Life Sciences Clinical laboratory standards state that urine samples should be tested within 2 hours of collection. If testing is delayed beyond that time frame the sample needs to be refrigerated to inhibit urea conversion to ammonia and an increase in urine pH. Here medical lab science students investigated whether urine samples left at room temperature for 48 hours resulted in significant increases in urine pH with concomitant microbial growth. Five clean-catch urine specimens were obtained and promptly tested for pH levels using a sterile pH meter (EcoTestr, Oakton Instruments). Samples were then aliquoted into capped and uncapped containers that were left at room temperature. Samples were tested at 0, 8, 20, 24, 32 and 48 hours. The pH values from the uncapped and capped samples from these time periods were analyzed using a paired two tailed t-test. Results indicated one sample out of five was significant (P = .05). Initial samples were gram stained, then read to determine the presence of bacteria. Specimens were cultured on 5% Sheep Blood and MacConkey agar plates. Plates were read for bacterial growth at 24 hours incubation and growth was identified on two samples. At 48 hours, both uncapped and capped urine cultures were plated again and three samples demonstrated bacterial growth the following day. Microbial testing identified normal urogenital flora and pathogenic bacteria. Urine with pathogenic bacteria demonstrated significant increases in pH, while additional cultures with bacterial growth also increased but not with significance. Future studies could employ a larger sample size from both healthy and diseased individuals. Furthermore, identification of microbes that will thrive in acidic and alkaline pH would be of interest.

Kylynn Parker, University of Utah Life Sciences Red Butte Canyon (RBC) is a Research Natural Area administered by the US Forest Service in Salt Lake City, Utah. RBC is an undisturbed area and a haven for all types of birds. Most of the avian species found in this area are migratory, and either pass through or breed in the area. The overall aim of this project is to determine if there have been any notable changes in populations of species in the area over the past 22 years. The research question that is covered in this summary are the following: has the density and relative abundance of the top five most commonly detected avian species in Red Butte Canyon notably changed through time in Transect 1? Data was collected by Mark Leppert, PhD and Sherwood Casjens, PhD of the University of Utah. They recorded the number and species of birds that were both seen and heard in 10 different transects within RBC over the past 22 years (1991-2013) and 457 survey days. In 2013 and 2014, I compiled and entered all of the data into a database with the guidance of the researchers. For analysis, I focused on the five most commonly detected species in Transect 1. These species are Black-Capped Chickadee (Poecile atricapilla), Warbling Vireo (Vireo gilvus), Yellow Warbler (Dendroica petechia), American Robin (Turdus migratorius), and Lazuli Bunting (Passerina amoena). Detection trends were found by graphing the number of individual birds seen or heard in Transect 1 over the days since surveys began in 1991 and statistical evidence was found showing significant changes in species population size of these five most commonly detected species, especially in the case of the American Robin which exhibits a decline in detections in recent years.

Coral Gardner, Southern Utah University Life Sciences Sea spiders (pycnogonids) are a small group of exclusively marine arthropods which resemble terrestrial spiders. One family, the Colossendeidae can grow to a very large adult size, much larger than any other pycnogonid. Colossendeis colossea, the largest known species, has a leg span of up to 70 cm and is found in both very deep ocean waters and shallower water in the Antarctic. Since it has been very difficult to obtain and study live specimens of this species, very little is known of their biology and natural history. We have obtained a number of preserved specimens, all labeled C. colossea, from the United States National Museum and are asking the question, are all of these specimens actually C. colossea? To answer this question, the specimens are being compared to syntypes from the Museum of Comparative Zoology and also to the original type description (Wilson, 1881). Based on previous scientific papers on C. colossea and closely related species, the following are used to separate species within this genus: relative proportions of terminal segments of walking leg and of pedipalp; location and number of eyes and shape of eye tubercle; proboscis shape and orientation; number of spine rows on terminal segments of oviger and shape of oviger terminal claw; abdomen size and orientation. Based on these, our preliminary results demonstrate that some of the United States National Museum specimens are not C. colossea since they differ from the syntypes and type description in several of the above characteristics.

Daniel Barnett, Brigham Young University Life Sciences Approximately one-third of US adults have metabolic disease, increasing their risk for diabetes, cancers and neurodegenerative disease (www.ADA.org). At the heart of these diseases are imbalances in the cellular redox state. The cofactor NAD(P), commonly known as niacin, is required for over 300 essential reactions in the cell and is largely responsible for the cellular redox state. NAD kinase regulates the NAD to NADP ratio, an important ratio for controlling cellular redox state and central metabolism. Herein we provide evidence that PAS kinase, a nutrient sensing kinase required for glucose homeostasis, phosphorylates NAD kinase. We are currently investigating the effect of this phosphorylation on the function of NAD kinase both in vitro and in vivo by measuring NAD kinase activity and associated phenotypes. This research will increase our understanding of how cells regulate central metabolism. In addition, because PAS kinase is a nonessential protein, it may prove to be an invaluable treatment target for regulating NAD(P) levels and controlling cellular redox state. This may lead to therapeutic targets for cancer and metabolic diseases such as diabetes.

Amanda Hobson, Carrie Draney, Andrew Stratford, Thomas Becker, Danhong Lu, Michelle Arlotto,

Brent Knoblauch and Marty Larsen, Dixie State University Life Sciences Chytrid fungus (Batrachochytrium dendrobatidis) is one of the major hypothesized theories behind global amphibian decline. Canyon Tree frogs (Hyla arenicolor) found in Zion National Park have been found to contain infected individuals, however population sizes have grown despite infection with the usually deadly fungus. The present study sought to demonstrate a correlation between skin temperatures and fungal proliferation, and investigated what enabled Hyla arenicolor to survive and reproduce with chytrid present. Ten frogs were sampled with sterile swabs from each of seven different study areas found in Zion National Park. Along with each swab a temperature was taken via infrared thermometer and recorded. DNA was also extracted from the swabs, and Batrachochytrium dendrobatidis- specific primers were used in a touchdown PCR protocol to determine infection rates. Swabs from specimens with higher temperatures were expected to display lower instances of infection. This would establish for the first time a clear demonstration that high skin temperatures were destroying the fungus in infected individuals in the wild.

Bryce Anderson, Kemais Ehlers, Deborah Johnson, and Stephen Persaud, Brigham Young University Life Sciences Antigen presenting cells digest and display foreign proteins from infected cells on the major histocompatibility complex (MHC) that is recognized by T cells via their T cell receptor (TCR). LLO56 and LLO118 are CD4+ helper T cells with TCRs specific for the same Listeria monocytogenes epitope. Despite differing by only 15 amino acids, these TCRs have dramatically different primary and secondary responses to infection. TCRs have very low affinity for peptide MHC. We determined to generate high affinity T cell receptors to test if T cell activation would be improved. We reasoned that the single chain LLO118 and LLO56 TCRs (Vβ2-linker-Vα2) could be subjected to directed evolution to generate mutants that are more stable and then used as a template for engineering high affinity T cell receptors. Single chain LLO118 and LLO56 were fused to the yeast surface protein Aga-2 and error prone PCR was used to generate mutagenic libraries. Stabilized single chain TCRs (scTCRs) were selected for using biotinylated Vβ2 and Vα2 antibodies and anti-biotin beads. First generation clones with increased stability compared to wild type were isolated for both LLO118 and LLO56. A second mutagenic library using the first generation mutants as templates was produced and the most stable clones were selected after temperature denaturation, permitting isolation of clones with increased stability. We are currently engineering high affinity T cell receptors by generating affinity libraries using site directed mutagenesis of the CDR3 regions. These libraries are sorted for their ability to bind to MHC tetramers and individual clones are tested using flow cytometry. Generation of pathogen specific high affinity TCRs will increase our understanding of how T cells are activated and could also provide infection specific diagnostics and therapeutics.

Benjamin Bitner, Jason Ray, Kyle Kener, and Brent Wright, Brigham Young University Life Sciences β-cell mass is lost in both major forms of diabetes. Mature β-cell have restricted proliferative capacity. Studies aimed at increasing β-cell mass frequently have the unwanted side effects of decreased glucose stimulated insulin secretion (GSIS) or increased apoptotic rates. Without functional β-cells, the body is unable to reduce elevated blood glucose, which results in hyperglycemia induced maladies. Enhanaced GSIS could be used as a treatment for diabetes. Overexpression of the β-cell transcription factor Nkx6.1 induces β-cell proliferation, enhances GSIS and protects against apoptosis. Nkx6.1 induces expression of VGF, which is necessary for enhanced GSIS. Microarray analysis of Nkx6.1 expressing primary rat islets demonstrates upregulation of c-Fos at 24 and 48 hours after adenoviral transduction. We have shown that c-Fos upregulates expression of VGF. Finally, preliminary data suggest that c-Fos modulates GSIS as demonstrated in INS-1 β-cell line, stable INS-1 knockdown cells transduced with lenti-sh-c-Fos and in primary islets. We present a model by which c-Fos is necessary for Nkx6.1 mediated enhanced GSIS by inducing expression of VGF.

Jacob Welch, Brigham Young University Life Sciences Learning and memory occur due to adaptive brain changes in response to our environment. These changes are mediated by synaptic plasticity, particularly within the hippocampus. Plasticity can either strengthen or weaken synapses, known as long-term potentiation (LTP) or long-term depression (LTD) respectively. While many forms of plasticity are NMDA-dependent, recently endocannabinoids were identified to mediate several new forms of hippocampal synaptic plasticity through the CB1 and TRPV1 receptors. However, research has demonstrated a non-CB1/TRPV1-dependent endocannabinoid synaptic plasticity in the hippocampus. Several potential candidate receptors that bind the endocannabinoid anandamide have been identified. These are among the orphan G-protein coupled receptors (GPRs) whose distribution in the brain and/or function is less well known. GPR55 is of particular interest as it activates second messenger systems. Using quantitative RT-PCR, electrophysiological and memory behavioral tasks we examined hippocampal GPR55 expression and function. GPR55 is expressed in hippocampus of both rats and mice. Cellular expression is currently being examined and appears to be rare in interneurons and more likely expressed by pyramidal cells. Interestingly, application of the GPR55 agonist LPI (2 μM) to wild-type mice demonstrates a significant enhancement of LTP in brain slices. This LPI effect was not noted in GPR55 knock-out (KO) mice, which exhibit significantly (p < 0.05) smaller LTP (146%) than wildtype (WT) (181%). GPR55 also appears to increase release probability (Sylantyev et al., PNAS, 2013), denoting a presynaptic role. Paired-pulse ratios are now being analyzed between GPR55 KO and WT mice to confirm this finding; however we did not note a change in EPSCs in CA1 in response to 2μM LPI. These data suggest GPR55 is expressed and physiologically relevant in the hippocampus. Because enhanced LTP is usually associated with better memory performance in rodents, this provides a potential target to enhance the cellular mechanism associated with memory formation.

Bryce Taylor and Ian Kesler, Southern Utah University Life Sciences Green infrastructure is the use of natural processes to manage stormwater runoff and has many positive effects on evaporative cooling, building energy demand, and wildlife habitat. Very little green infrastructure research has been conducted in arid environments such as Southern Utah. The scope of this research project is to establish a complete water budget of the green roof using a lysimeter (an acrylic box resting on a high-resolution weighing scale). The water budget will help in determining whether the cost of irrigating the green roof is off- set by the potential benefits to biodiversity and decreased energy demands.We have constructed a working lysimeter and will collect the required data by using: a high resolution scale, a tipping bucket to measure drainage, two soil moisture and temperature sensors and weather data from the campus weather station. A collection of mixed Sedum species vegetation will be planted in the lysimeter using a special green roof media to accurately simulate a greenroof environment. Both pitfall (used to capture grounded insects) and combination traps (used to capture airborne insects) will be used to measure the biodiversity. Data collection will begin during the winter of 2014 and continue indefinitely.

Anne Thomas and Richard Gill, Brigham Young University Life Sciences Providing water for expanding urban communities in the western United States is a growing concern for city planners and governmental agencies. Landscaping can claim up to 50% of the urban water budget but also has the most potential for water conservation. Landscape water use is highly variable, however, because of species-specific differences in tree water use and because of decision-making by city planners and residents in maintaining trees. The objective of this study is to improve our ability to predict urban forest water use by identifying differences in tree basal area and diversity between neighborhoods that have arisen at different periods of development in Heber Valley, Utah. We classified neighborhoods as established, exurban (rural housing), commercial, or new tract based on age, location, and lot size. We performed a stratified random survey with twenty lots in each category and collected diameter and species data for each tree in the lot. Some of the patterns we observed were easily anticipated, such as higher basal area per hectare in the older, established neighborhoods relative to newer tract housing. Surprisingly, the number of individual trees per hectare in tract and established neighborhoods is very similar. Perhaps of more interest is the low species richness of tract housing compared to exurban communities. Because exurban communities are being replaced by tract housing there is evidence that tree diversity will be lost. Another important aspect of community structure in urban forests is the ratio of conifers to broadleaf trees because of fundamental differences in water use patterns. Conifers comprised twenty-five percent of the basal area in exurban and thirty-five percent in established neighborhoods, as opposed to five percent in tract. Our data suggest that tree diversity is likely to decrease while water demand is likely to increase with changes in urban forests in the coming decade.

Westen Archibald, Trevor Smart, and Emily Forsyth, Brigham Young University Life Sciences The Beaver Dam Wash National Conservation Area is considered an important biodiversity hotspot for the Western United States. The wash is an ecotone on the Northeastern edge of the Mojave Desert and is directly adjacent to both the Colorado Plateau and the Great Basin. Thus, this area contains various species of flora and fauna from each of the surrounding biomes. Prior research of the Beaver Dam Wash has documented high aboveground biodiversity for a desert ecosystem however no research has been conducted on belowground diversity. For this reason we chose to characterize the diversity and distribution of entomophilic nematodes, as they are commonly used as a biological indicator to the surrounding ecosystems. We hypothesized that because plant and insect biodiversity is high here, we would also find a diversity of entomophilic nematodes that is higher than the surrounding areas. To test this hypothesis we collected soil samples from 15 sample sites representative of the diversity of the different ecological communities. We tested for patterns of codistribution between entomophilic nematodes and environmental variables, such as plant cover, proximity to water, presence of organic matter, elevation, ecosystem classification, and soil chemistry. We extracted nematodes from the soil samples and sequenced the 28s rDNA region of representative individuals from each sample. Nematode diversity was low; one species was broadly distributed, X others were more patchily distributed. No correlation was found between above ground factors or soil chemical properties (ppm phosphorous, percent organic matter, and percent soil moisture). We conclude that the distribution of entomophilic nematodes in this ecological confluence is to some degree stochastic and uncoupled from aboveground diversity or belowground soil conditions.

Katherine Harris, Brady Evans and Thomas Andros, Brigham Young University Life Sciences Metabolic diseases, such as obesity and diabetes, have become endemic and the need for better treatments is rising. Mutations in PAS kinase, a recently discovered sensory kinase, have been shown to cause Maturity Onset Diabetes of the Young (MODY) in humans (Semplici et al., 2011). In addition, PAS kinase deficient mice display many phenotypes related to diabetes including resistance to weight gain, insulin insensitivity and triglyceride accumulation in response to a high-fat diet (Hao et al., 2007). Despite its importance in metabolism, little is known about the regulation of PAS kinase. PAS kinase consists of a sensory PAS domain that binds to and inhibits a protein kinase domain (Amezcua et al, 2002). We are currently engaged in several yeast genetic screens which will allow identification of regions in the full length PAS kinase that are essential for activation or for binding its substrates. The first screen is based on the finding that PAS kinase overexpression rescues a temperature-sensitive mutation in Tor2, the tor2(ts). We have isolated both point mutations and truncations in PAS kinase which alleviate the tor2(ts), suggesting they are hyperactive alleles. These mutations identify novel regions involved in PAS kinase regulation. Our second screen uses the yeast 2-hybrid to select for both point mutations and truncations that increase the ability of PAS kinase to bind its substrates. These mutations will help identify key regions of PAS kinase utilized in substrate recognition. Finally, we have identified regions of PAS kinase that are well-conserved throughout evolution and will compare these regions with the regions affected by our mutations. This study will be the first reported mutagenic analysis of PAS kinase. Analysis of these specific genetic regions will help elucidate the molecular mechanisms involved in the regulation and function of PAS kinase, a key player in the development of metabolic disease.

Scott Newton, Brigham Young University Life Sciences The ventral tegmental area (VTA) is known to controls the processing of rewarding and addictive behaviors. The VTA contains dopamine (DA) cells, which release DA to downstream targets in response to rewarding stimuli, and GABA cells, which modulate DA cell activity. Therefore, both cell types are involved in associative reward learning. Synaptic plasticity plays an important role in adaptive reward signaling within the VTA. Endocannabinoids mediate or modulate synaptic plasticity at synapses within the reward circuit. However, the source of endocannabinoids within the VTA is not well understood. Therefore, our goal was to describe the distribution of endocannabinoid biosynthetic enzyme mRNA within VTA neurons. We extracted single VTA neurons via whole cell patch clamp and used single-cell real-time quantitative PCR to identify DA and GABA neurons based on mRNA expression of cell-type specific targets. Additionally, electrophysiological properties such as action potential frequency and sag potential amplitude were examined between the two cell types. Concurrent with established observations, slower firing frequencies were observed in DAergic neurons, however overlap was identified between these two cell types. VTA neurons were then probed for endocannabinoid/ biosynthetic enzyme mRNA, such as N-acyl-phosphatidylethanolamine-specific phospholipase D (NAPE- PLD), diacylglycerol lipase α (DAGLα), and 12-lipoxygenase. We also probed for type I metabotropic glutamate receptor (mGluR) mRNA, as endocannabinoid synthesis requires mGluR activation. Our data demonstrate that endocannabinoid biosynthetic enzyme mRNA is expressed in both DAergic and GABAergic cells with concurrent expression of type I mGluRs. Next, to ensure mRNA expression was representative of protein content, slices were stained using immunohistochemistry for GAD67, DAGLα, NAPE-PLD and type I mGluRs. Positive labeling for these targets was observed in VTA neurons, supporting our RT-PCR results. Collectively, these data suggest DAergic and GABAergic cells of the VTA have the capability to produce endocannabinoids and potentially alter synaptic plasticity involved in reward and addiction.

Alex Hyer, University of Utah Life Sciences Geochemical reactions associated with the process known as serpentinization can generate copious quantities of organic carbon and chemical energy that support life, but these reactions also greatly increase the pH of serpentinization sites. High-pH environments hinder ATP production because the low proton concentrations at high pH can result in a reversed proton gradient across cell membranes. Organisms present at serpentinization sites are not well-characterized, and adaptations to their alkaliphilic lifestyle are unknown. Bacillus pseudofirmus OF4 was not isolated from serpentinization sites, but it is a well- studied alkaliphile that has several distinct adaptations for ATP production in high pH environments. Its sequence for ATP synthase, the molecular motor responsible for ATP generation in the cell, contains several distinctive mutations including an AXAXAXA motif that increases the C-ring stoichiometry, a P51XXE54XXP57 motif that creates a distinct kink in the outer helix of the C-subunit, and a V21 mutation granting rotamer freedom to E54. In this study, we search for similar mutations in metagenomic databases containing DNA sequences collected from serpentinization sites. Analysis of alignments from the programs BLAST and Exonerate indicate that V21 is present in several alignments, but P51 is found in only a single alignment from serpentinization sites. The AXAXAXA motif is non- existent in our databases, but the more common GXGXGXG motif is present with alanine replacements occurring periodically. In general, these results indicate that our DNA sequences from high-pH sites of serpentinization are more reminiscent of typical neutrophiles than of the alkaliphile B. pseudofirmus OF4. Therefore, serpentinization- adapted organisms may harbor other adaptations to high pH such as local pmf regulation. Future work will test for quantitative correlations between environmental pH measurements and the incidence of mutations in ATP synthase in order to identify novel adaptations to high pH in serpentinization-driven ecosystems.

Simranvir Kaur, University of Utah Life Sciences The placenta is the major source of fetal serotonin during pregnancy, which is essential for fetal brain development. In Utah, approximately 13% of pregnant women take Selective Serotonin Reuptake Inhibitors (SSRIs) to treat depression, the use of which has been correlated to significantly lower serotonin levels in cord blood for newborns. Studies suggest association between maternal SSRI use and adverse outcomes such as preterm birth, cognitive deficit, and disruption of serotonergic systems. However the effect of SSRIs on placental gene expression, serotonin synthesis and transport in the placenta, is not known. This study evaluates the impact of maternal SSRI use on placental gene expression and levels of serotonin in the cord blood using a nested case-control observational study model. Biological samples will be collected until 20 cases (women taking SSRIs during pregnancy) and 20 appropriately matched controls have been enrolled into the study. Participants also complete an online questionnaire to measure depression and anxiety levels as well as document any medication they have taken during pregnancy. Data and sample collection for this study is still in progress. Once enough samples have been collected for batch analysis, we will complete RT-PCR and ELISA, expected in Spring 2015. Upon complete data analysis, we expect this study will help in targeting mothers who are at risk for adverse pregnancy outcomes and further provide suggestions for intervention.

Stephen Tahan, Westminster College Life Sciences

Kyle Hansen, University of Utah Life Sciences Our brain processes physical stimuli from the world around us to shape how we perceive our environment. In the olfactory system, this processing begins with olfactory sensory neurons that relay information about distinct odorants to a region of the brain called the olfactory bulb. Olfactory sensory neurons express the same odorant receptors converge onto structures within the olfactory bulb called glomeruli. Here, we map lateral interactions between glomeruli to better understand how olfactory information from the environment is processed within the olfactory bulb. Using GCaMP6, a fluorescent calcium indicator, in awake mice we can observe optogenetically-evoked patterns of lateral connectivity between glomeruli in the olfactory bulb. The findings from this study will help clarify potential interglomerular connection schemes and provide understanding of how local circuitry in the olfactory bulb transforms the representations of sensory information before it is relayed to higher brain areas.

Brady Evans, Brigham Young University Life Sciences Nutrient-sensing kinases, such as AMPK and mTOR, play a key role in regulating cellular metabolism. They recognize nutrient levels within the cell and distribute nutrients accordingly. The failure to coordinate metabolic processes can lead to diseases such as diabetes, obesity and cancer. PAS kinase is another member of the nutrient-sensing kinase group that regulates glucose distribution in the cell. Despite its clear importance, little is known about the mechanisms regarding PAS kinase activation and its function. Studies have shown that mice without the PAS kinase gene are resistant to weight gain and maintain insulin sensitivity when placed on a high fat diet. Weight gain and insulin resistance are associated with increased rates of diabetes, cancer and other metabolic diseases. AMPK is currently the key target for the treatment of diabetes and has been found to be a cancer target as well. The yeast homologue, Snf1, is necessary for activation of yeast PAS kinase. Further studies have shown that PAS kinase is activated under the same respiratory conditions as Snf1. The aims of this project is to determine if Snf1 directly phosphorylates PAS kinase and to determine the effects of this phosphorylation. We have found that PAS kinase is activated quickly when cells are placed under Snf1-activating conditions. In addition, Snf1 is necessary and sufficient for this activation. Snf1 also copurifies with PAS kinase suggesting this activation is direct. PAS kinase purified from wild type verses Snf1-deficient cells shows phosphorylation on two key residues, S1020 and S1035. In addition, Snf1 directly phosphorylates PAS kinase in vitro. Together these findings suggest that Snf1 directly phosphorylates PAS kinase in order to regulate cellular glucose homeostasis

Sharisa Nay, Brigham Young University Life Sciences Gene therapy is a growing field of science with the potential to improve thousands of lives. With an eye toward improving the effectiveness and longevity of gene therapies, my project examines the preferential binding tendencies of the histone protein variants Htz1 and Htas1. Htz1 is the Ceanorhabditis elegans homolog of H2AZ, an important variant of the H2A histone. This protein has been shown as necessary for survival and as playing a role in the prevention of ectopic heterochromatin spread. Htas1 is another variant of H2A that plays a role in the increased transcription of sperm-producing genes. The preferred positioning of these variants on naked DNA is not yet known. Through a DNA extraction, nucleosome reconstitution, and DNA digest and sequencing, we will take these variants and examine where they are prone to localize within the N2 Bristol strain of c. elegans. This will allow us to include DNA sequences on our gene insertions possessing a high binding-affinity for these transcription-promoting histones. Thus, if we can identify the locations at which these variants will localize within DNA, we will be able to insert these preferred constructs into the genes used for gene therapy and thereby increase the effectiveness of gene therapies.

Joe Passman, University of Utah Life Sciences Neurodegenerative disorders and amyloidosis are thought to be a consequence of the misfolding of intrinsically disordered proteins (IDPs). In non-pathogenic states, IDPs are thought to manipulate their configurational ensembles (CEs) – through partially folding and/or forming secondary structure – to create binding sites for the multiple interaction partners necessary for cell-signaling, recognition, and regulation. An obstacle to accurate in vivo understanding of non-pathogenic mechanisms is that little is known about the impact of the crowded cellular environment on the global (i.e. partial folding) and local structural characteristics (i.e. secondary structure) of IDPs. It is difficult to understand why an IDP may misfold if little atomistic structural understanding exists regarding IDP functional mechanisms in non-adherent physiological states.

Shwan Javdan, University of Utah Life Sciences Each year in the United States, over 22,000 new cases of ovarian carcinoma are diagnosed and 14,000 women die from its progressive cancer stages. Anthracyclines, a class of common chemotherapy drugs, have long been the primary treatment for this and many other cancers, but they often leave patients with cardiotoxicity, hepatotoxicity, and other adverse effects. Polymer-drug conjugates using poly[N-(2-hydroxypropyl)methacrylamide] (poly-HPMA) are nanosized, water-soluble constructs that accumulate passively in solid tumors by the enhanced permeability and retention effect as well as actively by cell surface targeting methods. As such, they have exhibited reduced toxicity in the body. The goal of this study was to develop a novel, targeted HPMA copolymer-drug conjugate for the treatment of ovarian carcinoma. To accomplish this, an antibody fragment called Fab’, targeted to ovarian carcinoma cell surface antigen OA3, was bound to an HPMA copolymer-gemcitabine conjugate. Gemcitabine is a nucleoside analog used in chemotherapy that has demonstrated considerable effectiveness in recent years. The copolymer-gemcitabine conjugate was successfully developed following the synthesis of all the requisite components, including Fab’, diblock chain-transfer agent (di-CTA), N-(2-(2-pyridyldithio)ethyl)methacrylamide (PDTEMA), and polymerizable backbone-degradable gemcitabine derivative. Syntheses of di-CTA and PDTEMA were optimized and then characterized by high-performance liquid chromatography, H-NMR, and mass spectrometry. Monomers were combined by reversible addition-fragmentation chain-transfer polymerization, and then the Fab’ fragment was bound to the polymer backbone via a disulfide exchange reaction with PDTEMA. The targeted copolymer-drug conjugate was then verified by fast-protein liquid chromatography as well as ultraviolet-visible spectroscopy. A number of in vitro tests have demonstrated the properties of an efficient drug delivery system. Flow cytometry displayed active accumulation of the copolymer-gemcitabine conjugate to OVCAR3 cell surfaces. Furthermore, IC50 measurement found promising drug efficacy comparable to modern anthracyclines. Future work will involve in vivo evaluation of the conjugate’s therapeutic efficacy in nude mice bearing OVCAR3-xenografts.

Julia Hull, Weber State University Life Sciences Endophytes are fungi that live within aerial portions of plants for most or all of their life cycle without causing visible signs of disease. Gall forming aphids, Pemphigus betae, are highly competitive over gall site selection (Moran 1993), forming galls on the leaves of narrowleaf cottonwood and their hybrids. The favored gall location overlaps with areas of highest endophyte probability. I hypothesized that a negative correlation would exist between endophyte infection and aphid galling on leaves of backcross hybrid cottonwood trees.

Brianne Palmer, Utah State University Life Sciences Inversions have plagued the valleys in Utah resulting in built up pollution carpeting the cities and spreading into surrounding ecosystems. The ecological impact of these inversions is unknown. Inversion-based pollution events deposit nutrients and pollutants in the ecosystems. The purpose of this study is to determine if lichens, due to their unique physiology, preserve a record of inversion-based nitrogen deposition in order to assess the ecological impact of the inversions. In June and July of 2013 we collected 111 samples of lichens from sites exposed to the inversions, along Red Butte Creek and on an elevation gradient on Grandeur Peak in Salt Lake City. The collection focused on two species of lichens (Xanthomendoza montana and Xanthomendoza fallax) because they are easily identifiable and live in all the sampling sites. The samples were ground, run through a stable-isotope mass spectrometer and analyzed for the %N and δ15N. If lichens hold a record of inversion-based depositions there should be more nitrogen in lichens from the sampling sites closer to the pollution source (Salt Lake City). Once the data was complied, the resulting graphs showed no correlation between %N and distance from Salt Lake City between δ15N and the pollutant source. However, the isotopic variability within the samples was greater than expected and requires further research.

Preston Manwill, Southern Utah University Life Sciences Plants produce compounds, secondary metabolites, which can be harnessed for medicinal uses. Caryophyllene, a secondary metabolite produced by a variety of higher plants, has shown promise as an analgesic and anti-inflammatory. Caryophyllene synthase is the final enzyme in the biosynthetic pathway that produces caryophyllene. The gene and enzyme responsible for caryophyllene production have been observed in crop plants and model organisms, but few studies have investigated caryophyllene production in indigenous plants. We investigated the production of caryophyllene in Ericameria linearifolia (Narrowleaf goldenbush) because it is a shrub native to southwest North America. The first portion of this research examined sequence similarities between caryophyllene synthase gene orthologs in E. linearifolia and other plant genera. Genomic DNA was isolated from E. linearifolia leaves collected from plants growing in southern Utah. Literature reviews and database inquiries have provided DNA sequences for similar synthases, from which primers will be designed to copy and sequence the E. linearifolia caryophyllene synthase gene, confirming the orthologs presence. Following confirmation of the gene, mRNA will be extracted from leaf material and oligo(dT) primers used to synthesize cDNA. Subsequent cDNA amplification and cloning of products into a vector will allow for transformation into a protein-producing bacterium. These final steps make the downstream procedures of protein production, structure determination, and functional characterization of the protein possible. Comparison with enzymes that produce similar organic compounds may result as well, and would provide insight that could improve the biosynthesis of medicinal compounds.

Preston Manwill, Sourthern Utah University Life Sciences Plants produce myriad secondary metabolites (essential oil) that play a role in ecological interactions. Species evolve unique mixtures of organic compounds due to strong selective pressures that act on metabolic pathways. Few studies have investigated the antimicrobial properties of secondary metabolites that evolved against soil microbes associated with native plants. The essential oil of Mojave goldenbush (Ericameria linearifolia), a shrub native to the Intermountain West, was characterized and its bacteriostatic abilities against root associated and non-root associated bacteria morphs were examined in this study. We hypothesized E. linearifolia oil would act as a greater growth inhibitor against native soil bacteria with which it has evolved than against bacteria from non-associated soils. Secondary metabolites were extracted using steam distillation and analyzed with gas chromatography mass spectroscopy. A Kirby-Bauer assay tested presence, size and retention of inhibition zones (IZ) produced against soil bacteria. The essential oil of E. linearifolia contained twenty secondary metabolites, two of which, limonene and sabinene, constituted greater than fifty percent of the oil. Presence and retention of IZ produced by E. linearifolia metabolites varied significantly by community association and bacteria morph. The hypothesis was supported by IZ that were largest against bacteria cultures isolated from soil that surrounded Mojave goldenbush roots and least effective against cultures isolated from non-root associated soil. Our findings suggest that secondary metabolites of E. linearifolia have evolved to specifically prevent negative interactions with bacteria most associated with the plant.

Cameron Sargent, Brigham Young University Life Sciences Although viruses present a potential solution for treating and studying many harmful diseases, they have yet to be utilized successfully, largely due to an overall lack of understanding of the interactions between viruses and their hosts. This study, however, demonstrates the efficacious usage of comparative genomics in further elucidating viral reproduction mechanisms, a step in overcoming this hurdle. Following the sequencing and annotation of genomes of the B4 subcluster of mycobacteriophage, a previously unstudied selection of viruses, comparative genomics analysis identified regions of proteins from these viruses that are highly conserved, or highly similar. One of these B4 phage proteins was then compared on the basis of its predicted folded structure and its amino acid composition to a protein with the same function from another phage, whose structure and DNA binding mechanism was determined via NMR spectrometry. Gene sequence alignments identified the region of highest conservation in the B4 phage. Protein folding then revealed that this region created a structure that was analogous to the functional region of the protein studied elsewhere, indicating that it too recognizes, binds to, and modifies DNA in the same manner. Furthermore, another DNA-binding motif with high conservation among B4 phage was observed in both the folded B4 proteins and the protein analyzed via NMR spectrometry. This comparison procedure not only further elucidated the function of an important gene in B4 phage but also displayed that comparative genomics, a resource-effective and computer-based approach, can successfully identify the critical functional regions of a protein. This study therefore provides a novel procedure for further understanding the mechanisms of virus-host interactions, which in turn facilitates the successful application of viruses in treating and studying diseases.

Eliza Warren, Brigham Young University Life Sciences We have previously demonstrated that ethanol inhibition of medium spiny neurons (MSNs) in the nucleus accumbens (NAc), and γ-aminobutyric acid (GABA) neurons in other ethanol-sensitive brain areas, is mediated by GABA(B) receptors. The aim of this study was to evaluate the involvement of GABA and glutamate (GLU), and in particular GABA(A), GABA(B) and NMDA receptors, in mediating ethanol inhibition of dopamine (DA) release in the NAc. Using fast scan cyclic voltammetry (FSCV), we evaluated the effects of ethanol on DA release in the NAc core of C57/BL6 and CD-1 mice. In the slice preparation, local stimulation evoked robust, frequency-dependent DA release in the NAc, with maximal release at 20 Hz. Ethanol decreased DA release with an IC50 of 60 mM in C57Bl6 mice. In anesthetized C57BL6 mice, ethanol decreased DA release with an IC50 of 2.0 g/kg. Superfusion of the GABA(B) receptor antagonist CGP55845 enhanced DA release 85%, while neither the GABA(A) receptor antagonist bicuculline, the GABA(A) receptor agonist muscimol, the GABA(B) receptor agonist baclofen, nor the NMDA GLU antagonist APV significantly affected DA release. CGP55845 blocked ethanol inhibition of DA release, while neither bicuculline nor APV altered ethanol’s effects. GABA inhibited DA release at 0.5-10 uM but not at 0.1-1 mM. Compared to ethanol effects on DA release in C57BL6 mice, CD-1 GAD GFP knock-in mice, which only express GAD at 50% levels, were significantly less sensitive to ethanol. As DA release was sensitive to low concentrations of GABA, CGP55845 may act as a GABA(A) rho receptor antagonist, and neither bicuculline nor baclofen had any effect on DA release, ethanol may be acting on extrasynaptic GABA(A) rho receptors on DA terminals to inhibit DA release in the NAc.

Ryan Watkins, Univeristy of Utah Life Sciences The lymphatic system is responsible for controlling systemic fluid buildup. Lymphangiogenesis is a dynamic process involving sprouting and maintaining new lymphatic vasculature. Vascular endothelial growth factor C (VEGF-C) is known as a key growth factor through VEGF receptor 2 and 3 (VEGFR2 and VEGFR3). Interestingly, the cornea expresses VEGF-C but is alymphatic. We found that the soluble isoform of VEGFR3, which lacks tyrosine kinase domains, is responsible for the alymphatic nature of the cornea by sequestering endogenous VEGF-C. Although soluble VEGFR3 or soluble VEGFR2 can be useful for inhibition of VEGF-C derived lymphangiogenesis, they also bind VEGF-A. Inhibition of VEGF-A suppresses blood vessel formation, damaging tissue and creating additional side effects. The development of a new anti-lymphangiogenic drug, that only blocks VEGF-C, has many implications: preventing tumor metastasis and reducing rejection rates of tissue and organ transplants. VEGF-C mainly binds to VEGFR3 domain 2 and VEGFR2 domain 3. To develop an anti-lymphangiogenic drug that specifically binds VEGF-C the binding domains were inserted into a vector that produces a recombinant protein (VEGF-C trap) that sequesters VEGF-C, suppressing lymphangiogenesis. Mice in a cornea transplantation model were treated with VEGF-C trap. After 8 weeks, 60% of the treated cornea survived (no rejection) compared to 10% in the empty vector control group. A 60% transplant survival rate is one of the highest rates compared to other single treatment methods. Blood and lymph vessel area was calculated and showed a decrease in lymph vasculature but not blood vessel. Suggesting only VEGF-C activity was affected. In a tumor metastasis model, nude mice will be injected subcutaneously with cultured MCF-7 cells which have been transfected with VEGF-C trap or an empty vector. We expect to see a decrease of lymphatic vasculature in the mice injected with MCF-7 cells containing the VEGF-C trap and ultimately less metastasis.

Kevin Nay, Southern Utah University Life Sciences Leech taxonomy has traditionally been based on morphological characters, but with new developments in DNA technology many taxonomists are starting to use genetic information in descriptions of new species. Leeches in southern Utah are poorly inventoried with respect to many other aquatic animals. There have been few morphological inventories of leeches and even fewer descriptions of the genetic diversity within leeches. Landscape genetics is a powerful tool used to understand geographic patterns of genetic diversity. Southern Utah has many naturally isolated bodies of water due to the climate and the dramatic changes in elevation in this part of the country. The landscape genetic study of leeches in southern Utah will provide us with a better understanding of genetic differentiation within southern Utah leeches. The mitochondrial DNA (CO I region) will be used to estimate genetic diversity and examine the relationships among individuals in two populations of leeches. I hypothesize that leeches in southern Utah will have greater genetic diversity then historically recognized from morphological studies suggesting a new species of leech. The study will lead to better understanding of the taxonomy and identification of southern Utah leeches.

Preston Manwill, Southern Utah University Life Sciences Members of Anthoxanthum (sweetgrass) have a history of ceremonial and medicinal use. Coumarin, a secondary metabolite produced by the grass, is an anticoagulant and antimicrobial agent. Antimicrobial properties of commercially available European A. odoratum metabolites have been nominally investigated, but no North American species have been studied. Additionally, European sweetgrass is purported to be available in both diploid and polyploid strains. Our research objectives were to: 1) determine if ploidy level strains do exist, and 2) investigate secondary metabolite production and evolution in commercial strains and A. hirtum, a Great Basin native. Using epidermal casts and fuschin staining, differences in guard cells and nuclei were surveyed. Secondary metabolites from the A. odoratum strains and A. hirtum were obtained through steam distillation and a vacufugation protocol that concentrated hydrosols. GC/MS analyses characterized and quantified secondary metabolites. The secondary metabolites coumarin, dihydrobenzofuron, and dihydroactinidiolide were identified, with the first and latter greatest in A. hirtum. Significant guard cell differences between strains, as well as species, were observed. Two ploidy strains of A. odoratum were suggested. A Kirby-Bauer assay tested presence, size and retention of inhibition zones (IZ) produced against soil bacteria. Gram-staining initially characterized bacterial morphs. Secondary metabolites from the diploid strain were most effective against all bacterial morphs, but polyploid metabolites also generated and retained IZ against diploid root associated and non-root associated morphs. The native sweetgrass produced IZ only against root associated bacteria with which it had evolved. Future work will include similar studies of other North American Anthoxanthum species.

Justen Despain, Brigham Young University Life Sciences Mycobacterium ulcerans is an acid fast intracellular bacteria that is the causative agent of Buruli Ulcer, a disease endemic in tropical regions of Africa. The disease starts as a nodule that progresses into a necrotizing skin lesion. Treatment ranges from antibiotic therapy to surgical removal of surrounding skin to stop necrotization of the skin. Owing to the intracellular nature of M. ulcerans, antibiotic therapy can be an ineffective treatment option. It has been previously shown that mycobacteriophage can reduce extracellular bacterial load in mice infected with M. ulcerans. In this study, we propose a method of killing intracellular M. ulcerans by mycobacteriophage using avirulent Mycobacterium smegmaits as a vector.

Lee Leavitt, University of Utah Life Sciences The ventral respiratory column (VRC) is a region in the brainstem shown to control breathing patterns in mammals. Using activation and inhibition of neurons in this region, classes have been assigned based on response-combinations. Using a mouse model, cells from this region are dissociated, plated and incubated with a dye that indicates changes in cytoplasmic calcium levels. Hundreds of cells are measured while varieties of pharmacological agents are applied. Response-combinations provide a profile of the receptors found on these neurons. Previously, varieties of cell classes were shown to contain NMDA receptors (receptors linked to learning and memory). However, specific compositions of subunits within these receptors are not known. These receptors are ligand gated ion channels composed of four non-covalently bound proteins. Each subunit has a different activation profile determined by interactions of agonist and antagonists. Conantokins (peptides isolated from snail venom) and other compounds further afford understanding of the architecture the assigned cell-classes. This project has continued to classify the subunit compositions of NMDA receptors with the ultimate goal of understanding which NMDA receptor subunits are present in each class. This will provide valuable information on the VRC’s function, and will allow for pharmacological innervations to change behavior in this region.

Madison Landreth, Weber State University Life Sciences Enterococcus, a bacterial genus that normally inhabits the gastrointestinal tract of animals, can be pathogenic to humans, causing urinary tract infections, sepsis and other serious diseases. It is also one of the major causes of hospital acquired infections. One important complication of those infected with Enterococcus is the fact that these bacteria often have a high level of antibiotic resistance, making effective treatment of patients more difficult. While Enterococcus is a normal inhabitant of the gastrointestinal tract, it can survive outside its host in the environment, even in adverse conditions, such as the Great Salt Lake (GSL). In this experiment, hundreds of isolates of Enterococcus were collected from the Great Salt Lake, from various sites along the Weber River which flows into the GSL and from clinical sources. Isolates were tested for different phenotypic characteristics and for their resistant patterns against certain antibiotics. Preliminary results of the Kirby Bauer disk-diffusion assay demonstrated that 47% of enterococcal isolates from the Great Salt Lake were resistant to one or more of the five antibiotics compared to 98% of the clinical isolates. In contrast, in a previous study, as few as 15% of Enterococcus isolated from the fresh water sources were resistant to one or more of the five antibiotics. These data may have implications concerning the importance of anthropological impact on rates of antibiotic resistance in this genus.

Jeremy Rehm, Brigham Young University Life Science The recent surge of interest in personality differences between individuals of a single population or members of differing populations has generated numerous new hypotheses that may aid in elucidating patterns of ecology and evolution that were previously considered improbable. Two hypotheses relevant to fish biology relate the size of an organism from a certain predation environment to the level of boldness it exhibits. The first of these (predation hypothesis) predicts small individuals living with predators should not express boldness comparable to their larger counterparts, whereas the other (metabolic hypothesis) predicts the exact opposite. Our study investigated these hypotheses using two sister-taxa fish species in Panama (Brachyrhaphis roseni and B. terrabensis) that exhibit two size classes (large and small) and live in differing predation environments. Additionally, because males are smaller than females in both species, we could look at size-boldness relations within each species. The study, as in others, defined boldness as the amount of time for an individual to emerge from a shelter and into an unfamiliar territory. When the species are analyzed collectively, our results support previous findings that fish from high-predation environments tend to be bolder than those without predators; males tend to be bolder than females; and both mass and standard length positively correlate with boldness. However, within species analyses find that mass and standard length have no significant relation to boldness, and gender was only significant in the predation-exposed B.roseni, where males were bolder. These interesting findings contrast with previous studies, and lead us to question the value of these size-related hypotheses in the process of speciation and, ultimately, evolution.

Derrick Crawford, Brigham Young University Life Sciences Members of the FGF family of signaling factors are key components in distal outgrowth and patterning of the vertebrate limb. These factors are expressed and secreted by the apical ectodermal ridge (AER) on the distal margin of the limb. Blocking their function is known to truncate the limb skeleton. Conversely, replacing the AER with beads soaked in Fgf protein can rescue limb outgrowth and patterning. Our lab has demonstrated that one of the mechanisms whereby the Fgf/AER functions is to mediate directed outgrowth of the adjacent mesenchyme. As the AER regulates growth of mesenchyme toward itself, it would be predicted that the AER’s dimensions would be important for shaping the mesenchyme that it recruits. We have found that the shape of the AER changes over time in a manner that corresponds to the shape of limb elements as they form along the proximal distal axis. Further, mutants that exhibit defects in the dimensions of the AER show corresponding anomalies in the limb skeleton. Given these observations it would be predicted that an Fgf soaked bead being of fixed spherical dimensions would only be capable of forming a cylindrical, rod-shaped limb. A bead placed posteriorly fulfills this expectation whereas a bead placed apically does not. We provide a molecular explanation for this discrepancy. We have also manipulated the shape of the AER surgically and find that similar to beads the shape and the AP position of the AER dictates the shape of the forming limb skeleton.

Charlee Byers, Brigham Young University Life Sciences Agave utahensis (Utah agave) plays a critical role as a keystone species in its native habitat. A rise in frequent, intense fires across the range of these habitats threatens to eliminate Utah agave populations, and consequently limit its genetic diversity. Characterizing the genetic diversity of Utah agave and its subspecies will help in restoration efforts to protect the species. We constructed primers to amplify microsatellite markers of two subspecies of Utah agave, ssp. kaibabensis and ssp. utahensis. Using these markers, we determined the level of polymorphism within four populations of each of the two subspecies.

Sean Studstill, Weber State University Life Sciences Imidacloprid is a popular systemic insecticide that has been applied to our staple crops for two decades. According to the EPA, it is persistent in the environment and at risk of effecting non-targeted organisms. Imidacloprid is an insect neurotoxin; however it is also known to be toxic to various aquatic species in concentrations as low as 37 ppb. Ingestion of imidacloprid causes paralysis in organisms through the blockage of postsynaptic nicotinic cholinergic receptors. We sought to find out how toxic imidacloprid is to brine shrimp and what kinds of physiological reactions occur upon exposure.

Jake Gamboa, Brigham Young University Life Sciences It is estimated that approximately 350,000 people in the United Stated die annually from post-myocardial infarction arrhythmias. A majority of these people will undergo a surgery that results in partial or complete removal of the stellate ganglion and other nerve fibers of the pharyngeal space in an attempt to prevent over stimulation from the neurons to the area of dead heart tissue and, therefore, future arrhythmias. However, without a somatomototopy, it is unclear what physiological effects partial or full sympathectomies may have. We will create a three-dimensional map of the pharyngeal space nerve plexus which will, in turn, allow for a more accurate and precise surgery.

Josh Hall, Weber State University Life Sciences Reproduction in birds is extremely conservative with the vast majority of the birds adopting bird-egg contact incubation to maintain an appropriate microclimate for embryonic development (Deeming, 2004). The Great Salt Lake is a vital nesting site for American Avocets (Recurvirostra Americana) that shows extreme temperatures and hostile environments where nest success can be as low as 1 -14% (Cavitt, 2008). Constancy of incubation, i.e. the time that the eggs are in contact with an adult, is a major indicator of nest success and environmental conditions. Our goals were to examine some of the costs natural selection places on embryos and parents to maintain a constant embryo temperature. We hypothesized that incubation attentiveness would increase across the nesting cycle. Over 200 AMAV nests were surveyed. Thermal probes were used to record various nest microclimates at every minute. A pseudonest with painted chicken eggs was also created and a thermal probe was placed to measure the ambient temperature without any adult incubation. A motion sensitive camera was placed over nests to examine differences in parental care. Nests will be divided into three phases: early, mid, and late incubation. Thermal data will be analyzed using descriptive statistics and mean variance values to calculate how incubation constancy varied throughout these phases. We expect this data to tell us more on how natural selection is working on these populations and some possible theories of how this developed.

Goyeun Tun, University of Utah Life Sciences Chronic Fatigue Syndrome (CFS) and Fibromyalgia Syndrome (FMS) are disorders which their symptoms and treatments are not clearly known. CFS and FMS are not life threatening diseases; however, they can affect patients’ quality of life because they experience symptoms including exercise intolerance, need for bedrest, and debilitating chronic pain and fatigue with these disorders. The research from Dr. Light’s lab has shown that moderate exercise for 25 minutes causes changes in mRNA levels in CFS and FMS patients but not healthy controls. The objective of our study was to examine changes in white blood cell gene expression of CFS and FMS patients both on Lyrica and on placebo in a double-blinded, cross-over design (where each study subject was his or her own control) by using quantitative PCR gene expression analysis. The lab routinely analyzes blood samples for 48 different genes from study subjects and healthy controls collected before (baseline) and then 8, 24, 48 hours after exercise moderate exercise. My focus was on changes in expression of two ATP-responsive purinergic receptors, P2X7 and P2Y1, which have not been studied after exercise in CFS and FMS but have been associated with chronic inflammation and pain in animal models. White blood cell layers (buffy coat) were collected from samples, RNA was extracted and converted to cDNA. 384 well PCR plates were robotically loaded from 96 well source plates, then the PCR reaction was run in an ABI 7900 thermal cycler that tracks fluorescence in “real time” (real time qPCR). Analysis of results is in progress and will be reported on the poster.

Xin Wan, University of Utah Life Sciences Endothelial dysfunction exists in individuals with diet-induced obesity (DIO) and type 2 diabetes (T2DM). Markers of endothelial dysfunction include reduced phosphorylation (p) of endothelial nitric oxide (NO) synthase (eNOS) to total eNOS (p-eNOS:eNOS), and attenuated endothelium-dependent vasorelaxation. Free fatty acids (FFAs) are elevated in individuals with DIO and T2DM. Our laboratory has shown that when: (i) endothelial cells are incubated with saturated FFA palmitate; (ii) mice are infused with lard-oil; and/or (iii) when mice are fed with high-fat diet, protein phosphatase 2A (PP2A) binds directly with eNOS. When this occurs, the association among Akt-Hsp90-eNOS is disrupted, p-eNOS:eNOS is impaired, and endothelium-dependent dysfunction occurs. This is prevented using pharmacological and genetic approaches that limit production of FFA metabolite ceramide. It is unknown whether PP2A inhibition per se is protective. We hypothesized that arterial dysfunction in obese vs. lean mice is prevented by PP2A inhibition. Seven-week-old, male, C57B16 mice consumed standard (CON, n=20) or high-fat (HF, n=20) chow for 12-weeks. Subgroups (n=10) of CON and HF mice received IP injections of saline (vehicle; V) or Lixte Biotechnology 100 (LB1, 1 mg/kg/day) for the last 14-days. Preliminary experiments verified that LB1-treatment for 3 and 21 days decreases (p<0.05) arterial PP2A activity. HF mice gained weight and developed peripheral glucose intolerance vs. CON mice regardless of LB1 treatment. Endothelium-dependent vasorelaxation was impaired (p<0.05) in HF-V vs. CON-V mice, but dysfunction was less severe (p<0.05) in HF-LB1 mice. p-eNOS:eNOS was reduced (p<0.05) in arteries from HF-V vs. CON-V mice, but p-eNOS:eNOS was similar in arteries from HF-LB1 and CON-LB1 mice. Akt and Hsp90 co-immunoprecipitation with eNOS was impaired (p<0.05) in HF-V vs. HF-CON mice, but this was not observed in arteries from HF-LB1 and CON-LB1 mice. These findings suggest that PP2A activity suppression in vivo is sufficient to preserve endothelial function in obese mice.

J David Symons, University of Utah Life Sciences Autophagy plays a central role in cellular quality control by destroying damaged or excess proteins, lipids, membranes, and organelles that accumulate in response to deviations from homeostasis. The existence and role of autophagy in endothelial cells (ECs) and blood vessels has not been established. Autophagy can be quantified by assessing the ratio of the membrane bound conjugate of microtubule-associated protein light chain 3 (LC3-II) to the cytosolic non-lipidated conjugate LC3-1 (LC3-II:LC3-I) or GAPDH (LC3-II:GAPDH) via immunoblotting. We sought to determine the extent to which a variety of cellular stressors induces autophagy in ECs and intact blood vessels. LC3-II:LC3-I or LC3-II:GAPDH was elevated (p<0.05) (i) 450±6% (n=4) in ECs incubated for 2 h in amino acid (AA)-deplete vs. AA-replete media; (ii) 47±3% (n=3) in arteries from fasted (14 h) vs. fasted / refed (1 h) mice; (iii) 40±2% (n=3) in arteries from mice that completed acute exercise vs. sedentary controls; (iv) 38±1% in arteries from exercise-trained vs. sedentary mice under basal conditions (n=2 per group); and was decreased (p<0.05) (v) 57±8% (n=4) in arteries from ~30 month-old (i.e., old) vs. ~6 month-old (i.e., young) mice. Further, indices of autophagy were elevated (p<0.05) 101±6% in ECs exposed to 3 h x 500 uM palmitate vs. vehicle (n=4), and 50±6% in arteries from obese vs. lean mice (n=4 per group). Thus, autophagy is altered in ECs and blood vessels in response to physiological (e.g., fasting, acute exercise, exercise training, aging) and pathophysiological (acute lipotoxicity, diet-induced obesity) stimuli. Ongoing research will determine the functional role of vascular autophagy in health and disease.

Janweb Lagazo, Brigham Young University Life Sciences The scientific community and the general public have long been interested in the effects of water pollution. Most studies on water pollution have focused solely on industrial pollution, but have failed to consider the potential impact of pharmaceuticals that unintentionally accumulate in aquatic ecosystems via wastewater treatment effluents. The purpose of this study is to advance our understanding on how these wastewater effluents affect aquatic ecosystems in Utah. We quantified the concentration of select pharmaceuticals in Hobble Creek using mass spectrometry. Then we sampled above the treatment plant, at the effluent outlet, and downstream of the effluent to determine pre-effluent and post-effluent drug concentrations. We are currently using this preliminary data to investigate how common endocrine disrupting, anti-inflammatory, analgesic, and anti-anxiety drugs may potentially affect the aquatic ecosystem of the endangered Chasmistes liorus, commonly known as June sucker.

Dani Peterson, Brigham Young University Life Sciences Due to its long and complicated trajectory through the cranium, facial nerve VII (CN VII) can be damaged in surgeries, sometimes resulting in facial muscle paralysis. Surgical removal of acoustic neuromas and parotid tumors, in addition to surgical repair of the temporomandibular joint disorder are associated with a risk of damage to CN VII. In addition, insertion of auditory implants can damage the nerve, as can improper stimulation to the nerve after the implantation has occurred. We will create a three-dimensional (3D) model based off of data from dissection of the nerve in a human cadaver in order to give physicians a greater in vivo knowledge of the pathway of CN VII. We have dissected the lateral side of the right half of the head to the level of the parotid gland, identified the parotid plexus of CN VII, and followed its five branches. In addition, we are currently following the nerve through the internal auditory meatus on its pathway through the temporal bone. In preparation for the modeling MicroScribe technique described below, we have imaged the head using Magnetic Resonance Imaging (MRI) at BYU. These images will be used as a template for the nerve reconstruction model. After completing the dissection, we will track the nerve trajectory using a MicroScribe 3D Digitizer. The MicroScribe technique is used to create 3D computer models of any physical object. The user sets reference points and uses the stylus to trace data points of the object’s contours. Our final product will be a 3D spatial computer mapping of CNVII, as well as a mapping of the skull, parotid gland, and other landmarks to put the nerve model into context. We hypothesize that with our approach and MicroScribe technique, we will be successful in creating an accurate model of CN VII in the head.

Lauren Archibald, Brigham Young University Life Sciences Increased intake of selenium (Se) and soy have both been shown to reduce risk for prostate cancer, especially if these dietary treatments are combined. The purpose of this project is to determine how the timing of Se supplementation of either a low- or high-soy diet affects prostate cancer risk. [C57BL/6 X FVB] F1 TRAMP (TRansgenic Adenocarcinoma of Mouse Prostate) male mice were fed stock diets low or high in soy. Half of the mice received Se supplementation (4.0 mg Se/kg BW as Se-methylselenocysteine) by gavage 5 d/wk in a 2 X 2 factorial design. Se supplementation began at conception, 6 weeks, 12 weeks, or 18 weeks of age. The mice were then sacrificed at different stages of maturation (4, 12, 18, and 24 weeks). Our results showed that, at 12 weeks of age, urogenital tract weights, a measure of prostate proliferation and tumor volume, were significantly reduced by Se supplementation (p<0.001) and by soy (p=0.044), independent of time of dietary intervention. Histological scores of prostate cancer progression also showed a protective effect of Se supplementation (p=0.030). At this writing, statistical analysis of data from mice sacrificed at 18 weeks is in process. Data derived from 18-week mice, combined with our previous findings from 12-week animals, will allow us to chart the progress of prostate cancer in this model. In addition, results will show how dietary Se and soy may alter disease progression and how the timing of dietary intervention may determine its effects.