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2013 Abstracts

Characterizing the Role of HspB2 in Cardiac Metabolism and Muscle Structure Using Yeast and Mammalian Cells

January 01, 2013 12:00 AM
Whitney Hoopes, Brigham Young University Microbiology and Molecular Biology HspB2 is one of eleven known small Heat Shock Proteins (sHSP) that is expressed in human heart and skeletal muscle. In response to cellular stress, heat shock proteins play a vital role to help misfolded proteins and proteins susceptible to denaturation maintain their structure. Two members of the sHSP family, CryAB and HspB2, are both required for normal heart function and cardiac muscle integrity. CryAB-deficient mice have defects in cardiac muscle structure whereas HspB2-deficient mice display energy deficits (Rajasekaran et al. 2007). The contrasting phenotypes of CryAB and HspB2 suggest differential roles for these molecular chaperones in the heart. HspB2 has been found to localize with the mitochondria in several different cell lines and overexpression of this sHSP has been shown to support survival of cells against heat stress (Nakagawa, 2001). To understand the role and mechanism of HspB2 in cardiac muscle energy regulation, we have used a yeast two-hybrid (Y2H) system to uncover the novel protein binding partners specific to HspB2. From screening a human heart cDNA library, HspB2 interacted with approximately 10,000 out of 20 million plasmids. We have sequenced more than 1000 of these putative interactors and have identified over 100 unique proteins. Over 40% of these protein partners are involved in mitochondrial energy production and another 25% in cardiac muscle structure maintenance. In addition, we have identified an interaction between HspB2 and the related sHSP CryAB. We then compared this data with mitochondrial HspB2 binding partners identified by mass spectroscopy (MS) through a large-scale bioinformatics analysis and constructed a protein-protein network. Y2H dependency tests were conducted to verify interactions identified by both Y2H and MS. Following yeast verification, a subset of the interactions were confirmed in C9H2 cardiac cells through coimmunopurification. Our research describes the first protein-protein interaction network for any sHSP, supports a role for HspB2 in mitochondrial energy production and suggests a link between mitochondrial energy production/redox stasis and stressed cardiac muscle maintenance.

Impact of Ungulate Browsing on the Development and Resilience of Aspen Forests

January 01, 2013 12:00 AM
Christian Boekweg, Brigham Young University Plant and Wildlife Sciences It has been shown that the population densities of deer and elk in the Rocky Mountains are at an unprecedented high. The heavy browsing of said species (ungulates) on regenerating aspen suckers can be devastating, leading to homogenously aged aspen stands that now have reduced resilience to drought, fire or logging disturbance, and an increased susceptibility to pathogens. Lastly, the extensive, highly selective herbivory of the aspen suckers may cause a shift in forest composition away from the aspen tree to an increase in other, less palatable species. We selected 186 sites across the 3 national forests of Central and Southern Utah, and characterized stand composition using the point quarter method. Stands were defined by successional stages from early to late; aspen, mixed or conifer. We then used pellet counts to estimate animal density and evaluated the effect of animal density and stand type on the regeneration success of aspen suckers. The key result of our study is that high ungulate density is highly correlated with reduced sucker heights (p<0.001). This indicates that high ungulate density impedes aspen regeneration by preventing aspen from recruiting into the overstory. Our study suggests that closer monitoring of the long term effects of herbivory on aspen development and regeneration is necessary to ensure vigorous aspen forests.

Identification of Novel Serum Biomarkers for Alzheimer’s Disease Using an Integrated Serum Proteomics Method

January 01, 2013 12:00 AM
Jesse Cobell, Brigham Young University Biology Alzheimer’s disease (AD) is the sixth major cause of death in the U.S. However, at present, no diagnostically useful serum markers for AD have been identified. Hence, we used a novel serum proteomic approach to interrogate the low molecular weight proteome for serum biomarkers. This allowed for survey of around 5000 low MW species. To reduce ion suppression, an acetonitrile precipitation step was used to remove high abundance serum proteins. Protein-depleted sera from 58 cases and 55 controls were analyzed by cLC-ESI-QTOFMS/MS using reverse phase chromatography. Data were reviewed using Applied Biosystem’s Analyst-QS software to compile spectra. Differentially expressed peptides (cases vs. controls) were analyzed statistically using the Student’s t-test. This led to discovery of 36 candidate biomarkers. Additionally, we compared AD subjects with more severe disease (Clinical Dementia Rating (CDR) =3) with non-demented individuals (CDR=0) and found 23 biomarkers. Furthermore, on comparison of mild and moderate stage AD individuals (CDR = 0.5, 1, 2) with those with severe disease (CDR = 3), we found 24 biomarkers. Some of these biomarkers appeared more prominent in one gender. We then fragmented several of these biomarkers on an LTQ-Orbitrap XL hybrid mass spectrometer and cLC-ESI-QTOF-MS/ MS system using collision-induced dissociation to determine amino acid sequence analysis. We have identified 5 biomarkers and are in the process of identifying the remaining biomarker species. This serum proteomics approach found statistically different peptide abundances in subjects with AD. Additional biostatistical evaluations are underway to determine sensitivity and specificity of individual biomarkers and their combinations. Future studies will assess biomarkers according to disease stage and validate current biomarkers in blinded comparisons of other AD sera. This serum proteomics approach appears promising in locating and identifying clinically useful serum biomarkers of AD.

The Effects of Exercise on Synaptic Plasticity in the CA1 Region of the Hippocampus in Mice Who Experience Acute Stress

January 01, 2013 12:00 AM
David Marriott, Brigham Young University Physiology and Developmental Biology Acute stress has been shown to decrease Long-Term Potentiation (LTP) in the CA1 region of the mouse hippocampus. Additionally, stressed animals show signs of anxiety and suffer decreases in spatial memory tasks such as object recognition and maze navigation. Conversely, exercise has been shown to increase spatial memory task performance in mice, attenuate anxiety-like behaviors and enhance neurogenesis and LTP in the dentate gyrus. While the effects of stress and exercise have been examined independently, there is currently a lack of experimental evidence that connects how stress and exercise, when experienced by the same animal, might modulate LTP in the CA1 region of the hippocampus. In our ongoing study, mice have been separated into a control group, a stress group (restraint and tail-shock), and an exercise + stress group where mice have voluntary access to a running wheel (for 30 days) before undergoing the stress protocol. We hypothesize that exercised animals will experience a protective effect against the reductions in CA1 LTP. In the stress only group, preliminary data shows a modest stress effect on LTP, yet we are learning that factors such as controllability of the stressor or the ability to develop coping mechanisms might potentially attenuate

Modified Nucleosomes and the Effect on Positioning

January 01, 2013 12:00 AM
Tara Hammond, Brigham Young University Microbiology and Molecular Biology Genetic diseases, including Alzheimer’s, cystic fibrosis, and many cancers, can be detrimental to individuals and their families. Gene therapy can possibly cure these diseases by inserting a correct copy of the gene into the chromosome, upregulating good genes, or downregulating the harmful gene. When DNA is packaged into a cell, it wraps around histones-an octamer made up of two tetramers, each containing four different subunits to create nucleosomes. Where the nucleosome sits on the DNA sequence determines whether or not a gene can be transcribed. In heterochromatin, nucleosomes are denser and DNA is tightly packed, thus causing genes to not be transcribed. Euchromatin contains looser packed nucleosomes and therefore has higher transcription levels. This project seeks to determine if modified nucleosomes have DNA sequence preferences. We are working with histone H3 to tri-methylate lysine 4, which has been shown to correlate with euchromatin. The modified histone will be used to create octamers. C. elegans DNA will be added to modified histones and to unmodified histones and allowed to create nucleosomes. The wrapped DNA will be sequenced, allowing us to compare the modified and unmodified nucleosome DNA preference. The difference in preference will enhance our ability to know how to move nucleosomes, thus aiding in gene therapy.

APOE e4 Independent Associations in the APOE Gene Region with Cerebrospinal Fluid Levels of Amyloid Beta 42 in Alzheimer’s Disease

January 01, 2013 12:00 AM
Spencer Foutz, Brigham Young University Biology CSF AB42 levels are a biomarker for Alzheimer’s Disease. The APOE e4 allele associates with CSF AB42. Little is known about SNPs in the region independent of apoe e2/e3/e4 isoforms. By adjusting for the effect of these isoforms, statistical analysis uncovered new SNPS associated with CSF AB42. Information was used from 1338 individuals from four datasets, specifically: The WU-ADRC, ADNI, University of Washington, and UPENN. Samples included individuals with and without AD. The 169 SNPs used were extracted from the APOE region and surrounding 50 kb using 1000 Genome Software. Linear regression analysis was performed, adjusting for specific covariates. Adjustments were made for the APOE e2 and e4 alleles before repeating the analysis. Significant SNPs were tested in e3 homozygous individuals. Each series was separately analyzed and combined in a meta-analysis for confirmation. P-values, sample sizes, and effect sizes were used in the meta-analysis. Results from these analyses allowed us to conclude rs769449 is associated with lower levels of CSF AB42 and acts independent of the APOE e4 allele.

Facilitative and Competitive Interactions in Subalpine Aspen-Fir Forests

January 01, 2013 12:00 AM
Jason Bartholomew, Brigham Young University Plant and Wildlife Sciences After disturbances in plant communities (i.e. wildfire), there is a natural succession of plants in which plants colonize the empty area and are gradually replaced by more competitive species. In subalpine forests, the principle colonizers after wildfire are quaking aspen (Populous tremuloides) which are later replaced by subalpine fir (Abies lasiocarpa). It has been shown that aspen facilitate, or enable, the establishment of subalpine fir at their base. This study examines the aspen-subalpine fir interaction in order to better understand the dynamics of the shift from aspen to fir dominance. It is hypothesized that the fir in a facilitated pair eventually exerts a competitive influence on the aspen resulting in a decrease in aspen fitness. The growth rates of the two species were examined in different stand types (aspen, mixed and subalpine fir), as independent trees or in facilitated pairs, and in three separate size classes. Samples were collected by taking a core sample or cross-section from trees within the categories listed above. The age and annual growth rings were measured with a measuring stage. The annual growth rings were used to calculate basal area increase (BAI) which was used to determine growth rates. The results suggest the growth rate of aspen in facilitated pairs decreases as firs mature thereby decreasing fitness within the aspen population due to competitive influences from facilitated firs. This may explain the mechanism for the successional shift that can significantly impact indigenous animal populations and local fire cycles.

Linkage Analysis of Late Onset Alzheimer’s Disease Population in Search of Chromosomal Region Harboring Rare Causal Variants

January 01, 2013 12:00 AM
Kevin Boehme, Brigham Young University Biology Late Onset Alzheimer disease (LOAD) is caused by a complex combination of genetic and environmental factors. While multiple loci have been found associated with an increased risk of LOAD much of the heritability of the disease has yet to be accounted for. The prevailing thought now is that of rare variants playing an important role in LOAD. In this study we will use linkage analysis to identify novel regions of the genome that may harbor rare disease causing variants. Data for these analysis comes from 748 people (503 with LOAD) from the Cache County study on Memory and Aging. This unique population based sample provides great power for linkage as relatedness differs from siblings to distant relatives and complete pedigree information is available for all of the individuals. We will use LD-pruned SNP data from the Illumina Omniexpress BeadChip and pedigree data from the Cache County samples to perform linkage analysis. Quality control and LD-pruning will be con- ducted in PLINK while the Linkage analyses will be conducted using the MERLIN software. Our findings will be reported in the final poster presentation.

Engineering Pathogen Specific High Affinity T-Cell Receptors

January 01, 2013 12:00 AM
Bryce Anderson, Brigham Young University Microbiology and Molecular Biology Antigen presenting cells digest and display peptides from foreign and infected cells on the major histocompatibility complex (MHC) that are recognized by T-cells through their T cell receptor (TCR). The affinity of TCR:peptide-MHC interactions has been shown to be low however, and in order to effectively use a soluble TCR for therapeutics we need to engineer TCRs with increased affinity. To do this, we have designed a single chain TCR (ValphaVbeta) called LLO118 that is specific for a naturally occurring Listeria monocytogenes epitope. Using yeast display, stable mutants that expressed the LLO118 scTCR at higher levels than the wild type on the surface of yeast were isolated and sequenced. In order to improve affinity of LLO118 we are mutating amino acid residues in the complementarity determining regions, sites important for the TCR to bind with the peptide-MHC. We are generating unique libraries of yeast cells with TCRs that have potential affinity mutations and using fluorescently labeled peptide-MHC tetramers to select cells that have TCRs with higher affinity. By repeating this process with the cells that have higher affinity we are working to get a TCR that binds with much higher affinity than the wild type TCR. These high affinity TCRs are promising for further research in connecting them to a cytokine, greatly reducing systemic damage and other complications caused by administration of this cytokine throughout the body. Thus, our goal is to design a high affinity TCR fused to a cytokine that can be tested for therapeutic use in targeting specific cells in the immune response and improving T cell memory.

GPR55 Modulates the Cellular Mechanism Behind Memory Formation, Suggested Alternative Pathways Require Further Investigation

January 01, 2013 12:00 AM
Spencer Bell, Brigham Young University Physiology and Developmental Biology The hippocampus functions as the memory formation center of the brain. As memories are formed, brain cells in this area undergo changes by which connections between them are either strengthened or weakened, processes known as long-term potentiation (LTP) and long-term depression (LTD), respectively. Receptors located on these cells modulate these processes as they are activated by chemical signals known as neurotransmitters. We sought to examine the effects of a receptor known as G protein-coupled receptor 55 (GPR55) on LTP and LTD in the rodent hippocampus by applying agonists of the receptor, or chemicals that artificially activate it, to brain slices preserved in artificial cerebrospinal fluid. O-1602 is a purported synthetic agonist of GPR55. When we applied O-1602 to rat hippocampal slices during electrical induction of LTP, the magnitude of LTP was decreased when compared to controls. When LTD was induced in the rat hippocampus in the presence of O-1602, no significant difference was observed when compared to controls. Further experimentation involved the application of lysophosphatidylinositol (LPI), a naturally occurring GPR55 agonist, to genetically-engineered knock-out mice which lacked expression of GPR55. LPI is generally considered a more reliable agonist of GPR55, but caused enhanced LTP in wild-type mice which expressed GPR55 when compared with knock-out mice. This inconsistency and other inconsistencies in our data while using O-1602, while perhaps due to other physiological differences between rats and mice, may suggest the possibility that O-1602 activates a receptor other than GPR55. Our further research will seek to investigate this possibility.

Baicalein and Light Stimulation as Clinical Therapies for Addiction

January 01, 2013 12:00 AM
Brad Ackerson, Brigham Young University Neuroscience The highjacking by alcohol and drugs of abuse of the mesocortico-limbic system in the brain is responsible for addiction, specifically the ventral tegmental area (VTA) and its projecting dopaminergic neurons to the nucleus accumbens (NAc). Over the course of addiction, a hedonic response is developed from lower than normal levels of dopamine (DA) in which the individual pursues drug-seeking behavior. The current accepted treatment methods for addiction are replacement drug therapies, group therapy, or individual counseling – the prior being associated with additional side-effects and an inability to overcome the hedonic response of the addiction. The aim of this study was to evaluate alternative and natural therapeutics that produce long-term potentiation (LTP) of the neuronal systems involved in order to overcome addiction with minimal to no side-effects. Using fast-scan cyclic voltammetry (FSCV), the effects of baicalein, a flavonoid isolated from the root of Sculletaria Baicalensis, and low-level laser therapy (LLLT) on DA release in the NAc core were evaluated in vitro and in vivo in Wistar rats. Local stimulation evoked in vitro demonstrated that baicalein administration (10, 50, 100 uM) 30 minutes prior to 80 mM ethanol attenuated the DA inhibition of ethanol. DA signals were evoked in vivo in the core of the NAc by electrical stimulation of the medial forebrain bundle (MFB) at the level of the lateral hypothalamus (60 Hz, 60 pulses) in isoflurane anesthetized rats. Both the intraperitoneal (IP) administration of baicalein (1.0 mg/kg) and the administration of LLLT (25 Hz, 630 nm) 30 minutes prior to ethanol (2.0 g/kg) administration IP attenuated the DA inhibition of ethanol. These findings suggest that baicalein and LLLT may prove as effective clinical therapies for addiction.

Characterization of Yeast PAS Kinase Through Random Mutagenesis

January 01, 2013 12:00 AM
Kayla Bevard, Brigham Young University Microbiology and Molecular Biology Metabolic diseases, such as obesity and diabetes, are endemic in American society. Mutations in PAS kinase, a recently discovered sensory protein 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, when placed on a high fat diet, display phenotypes related to diabetes including resistance to weight gain, insulin insensitivity and triglyceride accumulation (Hao et al., 2007). PAS kinase consists of a sensory PAS domain that binds to and inhibits the kinase domain (Amezcua et al., 2002). Our model for PAS kinase activation involves the generation of a small molecule ligand that binds to the PAS domain and relieves this inhibition. We are currently engaged in several yeast genetic screens which will identify regions in the full length PAS kinase protein that are essential for either PAS kinase activation or for binding of its substrates. The first screen is based on the ability of PAS kinase, when overexpressed, to rescue 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). These mutations solidify our model for PAS domain inhibition and will 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 affinity of PAS kinase for its substrate, Pbp1. These mutations will help identify key regions of PAS kinase utilized in substrate recognition. Finally, we will compare the regions affected by our mutations with the regions of PAS kinase that we have found to be well-conserved throughout evolution. 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.

Copy Number Analysis to Determine Genetic Alterations in Paired Primary

January 01, 2013 12:00 AM
Sam Hawkins, Utah State University Biological Engineering Colorectal cancer (CRC) is one of the most frequently diagnosed cancers in women and men. It is often treatable if caught early. However, tumors may metastasize which can result in a poor prognosis. A better understanding of the tumorigenesis and evolution of metastatic tumors in CRC patients could lead to earlier diagnosis, pre-emptive screening, and a better outcome. Copy number analysis of primary tumor tissue has revealed genes associated with colon cancers, but a comparison between primary and metastatic tumors has never been done. Normal tissue, primary tumor tissue, and metastatic tumor tissue was collected from twentyfive individuals. Copy number alterations were determined by microarray data generated from Molecular Inversion Probe (MIP) technology (Affymetrix, Inc.) for copy number analysis using Nexus software (BioDiscovery, Inc.). Metastatic tumor samples show a greater rate of copy number alterations (CNAs) from the primary tumors and even more alterations from normal tissue samples. Certain regions of the metastatic genome show high rates of CNAs whereas the primary tumor genome does not. These areas are key regions for potential understanding into the molecular origins of metastatic tumors. Understanding specific regions and genes with CNAs in metastatic tumor samples may lead to further research in cancer genetics and possible target areas for pharmaceutical testing.

Assessing Pro-Inflammator Biomarkers as Earl Indicars of Influenza Disease

January 01, 2013 12:00 AM
Makda Gebre, Utah State University Biology The first pandemic of the 21st century was the influenza A (H1N1pdm09) virus that originated in Mexico and killed 12,000 people within the United States. It now circulates as a seasonal influenza virus causing mild symptoms in most but still killing some susceptible individuals. Influenza A (H1N1pdm09) virus leads to acute respiratory distress syndrome (ARDS) in high risk patients and has a mortality rate of 40-50% in those patients. ARDS patients usually die before any intervention since there is no clinical evidence that can be used to detect it. To detect ARDS early in disease progression, we need to search and develop biomarkers that can be tested in the patient’s blood or fluids. The three biomarkers tested as potential early disease indicators in this project included: C-reactive protein (CRP), serum amyloid A (SAA) and Transferrin. To test these biomarkers, mice were infected with the influenza A H1N1pdm09 virus and serum was collected from the mice at different time points. Serum collected was tested for amounts of CRP, SAA and Transferrin using an enzyme-linked immunosorbent assay (ELISA) and spectrophotometer. Control mice were mock infected and also tested for the biomarkers for comparison.

A New Approach to Creating Pradimicin-Type Antifungal/Antiviral Compounds

January 01, 2013 12:00 AM
Thomas Anderson, Utah State University Biology Pradimicin, a small molecule produced by the soil bacterium Actinomadura hibisica, is a promising candidate as a combined antifungal/antiviral therapeutic. It is active against a broad-spectrum of opportunistic, pathogenic fungi, interferes with the replication of influenza virus, and inhibits the reproduction of HIV-1. Toxicity and solubility problems have hindered past efforts to develop pradimicin as a therapeutic. Our research focuses on elucidating the bio-synthetic pathway of pradimicin in order to design and chemoenzymatically create pradimicin structural analogs with improved solubility and activity, and less toxicity. Several enzymes in pradimicin biosynthesis have been identified. We intend to characterize one of the key enzymes, PdmS, a putative glycosyltransferase, and to manipulate its gene to create novel, more efficacious pradimicin analogs. This project is funded for three years by the NIH NIAID (3 years). Methods: A gene knockout experiment was used to determine the role of PdmS in pradimicin production. Bio-synthetic precursors of pradimicin were subjected to bio-transformations in E. coli with recombinant genes for PdmS and another glycosyltransferase, OleD, to generate analogs with new sugar attachments. Analogs of pradimicin will be screened for bio-activity using standard microbroth dilution assay techniques. Confirmed results: The enzyme PdmS was identified and characterized as a glycosyltransferase. Expected results: newly created analogs of pradimicin exhibited minimal inhibitory concentrations of 10μg/mL against Candida albicans. Conclusion: Knockout of pdmS yielded the pradimicin aglycon, which confirmed the function of this glycosyltransferase and provides a start molecule for further structural modification to yield new analogs for bioactivity studies.

Real-Time Pathology with High-Frequency Ultrasound: A Feasibility Study using Bovine Tissues

January 01, 2013 12:00 AM
Monica Cervantes, Utah Valley University The central research question of this project was to determine if high-frequency ultrasound is sensitive to tissue pathology at the microscopic level. Previous studies on surgical specimens have shown that high-frequency ultrasound may be sensitive to a range of breast pathologies including fibroadenomas, atypical ductal hyperplasia, fibrocystic changes, and carcinomas. The ultrasonic parameters that were sensitive to pathology were the number of peaks (the peak density) of the first-order spectra of the waveforms (one forward Fourier transform), and the slope of the second-order spectra of the waveforms (two consecutive forward Fourier transforms). The ability to determine pathology rapidly and with minimal specimen preparation would make high-frequency ultrasound particularly well-suited for real-time use during cancer surgery to ensure all of the malignant tissue has been removed. The purpose of this research was to determine the sensitivity of the peak density and spectral slope to tissue microstructures other than those found in breast cancer. The results of this study would not only support the results from the breast cancer studies, but also extend those results to the detection of cancer and other diseases in a range of organs and tissues. The research methodology included the following steps. (1) Freshly excised bovine organs were obtained from a meat packaging facility, including the heart, liver, and kidney. (2) Specimens approximately 3x3x1 cm in size were dissected from the organs and tested immediately with ultrasound. (3) Both pitch-catch and pulse-echo waveforms were acquired from the samples. (4) The data were analyzed by determining the peak densities and spectral slopes. The results showed that the more heterogeneous tissues of the heart, the vascular structures (aorta, vena cava, etc.), displayed significantly higher peak densities than the muscle tissues. Similarly, the ureter, which has greater heterogeneities in its structure (larger and more varied), displayed significantly higher peak densities than the cortex and medulla tissues. No significant trends were observed for the liver tissue, or for the spectral slopes except for kidney medulla tissue. Heterogeneity and peak density in high-frequency ultrasonic spectra that may be useful for performing real-time pathology during cancer surgery.

Production of New Pyridine N-oxide by Bioconversion with Cunninghamella echinulata var. elegans

January 01, 2013 12:00 AM
David Gage, Utah State University Biological Engineering A new N-oxide was produced from 3-(N-Boc-aminomethyl)-5-bromopyridine by bioconversion with Cunninghamella echinulata var. elegans ATCC 9245, and its structure was established based on spectral data. The microbial N-oxidation is efficient and highly selective. The substrate was transformed into the product in 7 days.

Distinguishing Kentucky Bluegrass Varieties Using EST and Genomic Primers

January 01, 2013 12:00 AM
Kat Combs, Utah State University Plants, Soils, and Climate Kentucky bluegrass (Poa pratensis L.) is a commonly used turfgrass species with many varieties being sold around the world. However, those varieties are very difficult to tell apart morphologically. Our objective was to use genetic markers (primers) to identify varieties, even if they are visually similar. This is valuable to the turfgrass industry for plant variety protection. We also wanted to use this data to explore the apomictic tendency (clonal reproduction) of the varieties. We collected leaf tissue from 24 Kentucky bluegrass varieties, extracted DNA, and sequenced portions using 29 EST and 21 genomic primers. This data was used to determine genetic relationships using a neighbor-joining dendrogram. Similarities of the genetic sequences from the varieties were estimated using the DICE coefficient. We found more polymorphisms in genomic primers than in EST primers with high variability between the varieties. Both types of primers were robust enough to distinguish varieties and that each variety was unique and genetically distinguishable. In addition, we discovered some varieties had large amounts of variation within a variety. This was unexpected due to the usual apomictic nature of the species. The markers resulting from our research will be available to the turfgrass industry.

American Lichens: Do They Follow Latitudinal Gradients

January 01, 2013 12:00 AM
Robert Bradford, Utah Valley University Biology Most organisms exhibit latitudinal gradients in diversity (i.e., taxonomic richness decreases as latitude increases). Few studies have sought latitudinal gradients in lichens, especially in the midlatitudes. Our primary questions were: 1) do lichens along the west coast of the United States show latitudinal gradients? 2) If so, what is the rate of change and does the level of taxonomic richness affect this rate? We hypothesized that lichens would show a reverse latitudinal gradient in the region, as has been documented for lichens elsewhere in the northern hemisphere, but at a considerably smaller scale. This study fills in the gap in our understanding of lichen latitudinal gradients over large areas of North America. It also functions as a baseline for future climate change and conservation efforts. Our study area is bound at the south by the California-Mexico border (32.331° N) and at the north by the Washington-Canada border (47.178° N), and extends inland from the coastline to the crest of the Sierra Nevada Mountains (116.083° W, at its eastern -most point). We divided the region into 218 roughly equal-area (cite) grid cells using GIS, each bordered with latitudinal and longitudinal lines. We derived a list of all vouchered lichen specimens in each grid cell using Consortium of North American Lichen Herbaria, an online database. The data were synonymized, and species, generic, and familial richness were calculated for each grid cell. We found no correlation (R2 = 0.2306) between latitude and species richness, using the raw vouchered data. What we did find was a strong correlation (R2=0.9069) between sample density and species richness. These results are biased by sample density and do not reflect what is naturally occurring. We hypothesize that we can get an unbiased estimate of richness with MaxEnt models. Using the georeferenced lichen distributions and related climate data, we constructed species distribution models of all species with five or more occurrences (990 species). In GIS, we projected all 990 distribution models and our 218 grid cells together to calculate species richness for each cell at various thresholds (i.e. likelihood of occurrence at 10%, 20%, 30%, etc.).

Impact of Cations on the Aggregation,Dissolution,and Phytotoxicity of Metal Oxide Nanoparticles in a Sand-Wheat Growth System

January 01, 2013 12:00 AM
Trevor Hansen, Utah State University Biology The use of metallic nanoparticles (NPs) in various applications presents the possibility for environmental contamination by these NPs. CuO and ZnO NPs have dose-dependent phytotoxicity. However, NPs are modified by different environmental factors such as salts: chlorides of Na, K, and Ca are used on roads for de-icing, dust control, irrigation, water treatments, and fertilizer production. At low concentrations these salts promote plant growth, but elevated concentrations become phytotoxic. This study examines the impact of Na, K and Ca chlorides on the phytotoxicity of CuO and ZnO NPs for wheat grown in sand. Atomic force microscopy and dynamic light scattering showed dose-dependent aggregation of the NPs in the wheat rhizosphere. The salts also caused NP-aggregation, but the degree of dissolution of the NPs was salt specific. Release of Cu from CuO NPs in the planted sand was reduced by all salt treatments whereas Zn release from ZnO NPs was reduced by Na and K but not Ca. Overall, a phytotoxic synergistic effect was seen at lower salt concentrations, but at higher salt levels, toxicity was mediated mainly by the salts. Therefore, effects upon plant growth of NPs and salts will vary according to concentrations.

NANOGP8: Evolution of a Human-Specific Retro-Oncogene

January 01, 2013 12:00 AM
Aaron Fairbanks, Utah Valley University Biology NANOGP8 is a human (Homo sapiens) retrogene, expressed predominantly in cancer cells where its protein product is tumorigenic. It arose through retrotransposition from its parent gene, NANOG, which is expressed predominantly in embryonic stem cells. Based on identification of fixed and polymorphic variants in a genetically diverse set of human NANOG and NANOGP8 sequences, we estimated the evolutionary origin of NANOGP8 at approximately 0.9 to 2.5 million years ago, more recent than previously estimated. We also discovered that NANOGP8 arose from a derived variant allele of NANOG containing a 22-nucleotide pair deletion in the 3? UTR, which has remained polymorphic in modern humans. Evidence from our experiments indicates that NANOGP8 is fixed in modern humans even though its parent allele is polymorphic. The presence of NANOGP8-specific sequences in Neanderthal reads provided definitive evidence that NANOGP8 is also present in the Neanderthal genome. Some variants between the reference sequences of NANOG and NANOGP8 utilized in cancer research to distinguish RT-PCR products are polymorphic within NANOG or NANOGP8 and thus are not universally reliable as distinguishing features. NANOGP8 was inserted in reverse orientation into the LTR region of an SVA retroelement that arose in a human-chimpanzee-gorilla common ancestor after divergence of the orangutan ancestral lineage. Transcription factor binding sites within and beyond this LTR may promote expression of NANOGP8 in cancer cells, although current evidence is inferential. The fact that NANOGP8 is a human-specific retro-oncogene may partially explain the higher genetic predisposition for cancer in humans compared with other primates.

Insects of Capitol Reef National Park

January 01, 2013 12:00 AM
Matthew Ethington, Utah Valley University Biology Although the insect fauna of the Colorado Plateau region are somewhat well known, our specific understanding of the diversity in Capitol Reef National Park is sparse. Here we present an overview of the insect diversity of the park based on a number of undergraduate student-led excursions to collect specimens. The collections will contribute to an increased understanding of the parks insect diversity and will result in an appropriately curated natural history museum collection. Furthermore, an insect field guide for the park will be created in order to serve as an educational tool for visitors to the park. Field collecting efforts, consisting of traditional insect collecting methods (aerial nets, aquatic nets, light traps, etc.) will be used. The collected material will be curated, photographed, and stored in a natural history museum (Brigham Young University ML Bean Museum). This presentation will be a summary of material gathered in the last three years.

Rationally Modified Tumor Suppressor Protein p53: A Possible New Cancer Treatment

January 01, 2013 12:00 AM
Thomas Wallace, University of Utah Pharmaceutics and Pharmaceutical Chemistry The tumor suppressor p53 is mutated in more than 50% of all cancers, while the majority of remaining cancers contain mislocalized p53(1). p53 is part of a network of cellular pathways that regulate growth, cell cycle arrest, and clearly delineated apoptotic pathways(2). Restoring the function of p53 can be seen as an ultimate cancer treatment. Restoring p53 would activate an already existing cell pathway that prevents cells from becoming cancerous and cause cancer cells to go through apoptosis. Playing a role in tumor suppression has made p53 an attractive target for gene therapy. However, despite the potential of p53 as a powerful treatment, it is limited by the dominant-negative effect of dysfunctional mutant p53. This effect imposes one of the greatest limits on the effectiveness of a p53 based treatment. This project is focused on bypassing the dominant-negative effect of dysfunctional p53 over exogenous functional p53. The attempted solution was substitution of the p53 binding domain with a different but structurally analogous coiled-coil, based on a modified Breakpoint cluster region (Bcr) protein. By doing this, the dominant-negative effect of mutant p53 may be bypassed. The purpose of this project has been to synthesis and test rationally modified forms of p53 with modified Bcr coil that are introduced into cells via a plasmid to restore cellular p53 activity. In vivo cell tests have already shown the effectiveness of these constructs at causing higher rates of cell death in cancer cells and constructs are currently being refined to carry forward to xenograft model animal trails. The ultimate goal is to develop a treatment for human cancer patients where modified p53 will selectively cause apoptosis in cancer cells.

Characterization of Self-assembled Soft Material Composed of Waxes and Oils

January 01, 2013 12:00 AM
Chin Yiap Tan, Utah State University Nutrition and Food Sciences The long term objective of this research is to explore oil-based materials that can be used to replace trans-fats in the food industry. Vegetable oils such as olive (OO), corn (CO), soybean (SBO), sunflower (SFO), safflower (SAFO) and canola (CAO) were mixed with sunflower wax (SFW), paraffin wax (PW), and beeswax (BW) at different concentration levels (0.1%-100%). Crystal Morphology, oil stability, thermal behavior and viscoelastic properties were studied in this research. Results in our laboratory showed that the crystallization occurred sooner when the concentration of waxes increased. For example, crystallization in 0.1% of SFW in SFO occurred after 24 hours of incubation at 25°C, but when concentration increased to 0.25% the crystallization was observed at the first 15 minutes of measurement. In addition, a change in the thermal behavior of wax/oil soft materials was observed when the concentration of waxes increased. For example, the enthalpy change (H) in SFW/ SFO materials increased from 0.87 J/g at 1% of SFW to 21J/g at 10% of SFW. This increase in enthalpy indicated that the amount of crystal increased when the concentration increased. Overall, the crystallization behavior in wax/oil materials was affected by wax concentration and chemical compositions of oils and waxes.

Comparing Trophic Level Position of Invertebrates in Fish and Fishless Lakes in Arctic Alaska

January 01, 2013 12:00 AM
Katie Fisher, Utah State University Watershed Science Arctic lakes are very sensitive to the effects of climate change. It is important to understand the current food web dynamic and energy flow within these lakes to better understand how they will change in the future due to the effects of a rapidly changing climate. In order to understand the current conditions in arctic lakes, this project consists of an analysis of stable isotopes of carbon (13C) and nitrogen (15N) from invertebrates among fish and fishless lakes in arctic Alaska to compare their trophic level positions and primary energetic sources. In order to carry out this analysis, I collected pelagic invertebrates were collected from 6 different lakes, 3 of which have resident fish populations and 3 of which are fishless. Samples collected in 2011 were analyzed for stable isotope composition by a mass spectrometer at University of California Davis. Results from samples collected in 2012 are pending at Washington State University. I will analyze and correlate the stable isotope results with isotopic data collected from other related projects. With this analysis, I will create food webs to 1) assign trophic positions to each species in each lake and compare those positions across lakes, 2) assess the potential effect fish predation has on pelagic invertebrate community structure. I hypothesize that fish predation will determine zooplankton community structure (e.g., dominant taxa) and alter trophic linkages (e.g., secondary trophic level predation rates).

Fluorene as a Model Compound to Investigate Fire Induced Soil Water Repellency: A qualitative approach

January 01, 2013 12:00 AM
Vance Almquist, Utah State University Plants, Soils, and Climate Fire induced soil water repellency has been characterized across a variety of soils and landscapes as being a cause of watershed degradation and surface water pollution. The repellency is due to the condensation of volatile polyaromatic hydrocarbons onto soil particles. Although repellency is known to reverse, in some locations the reversal takes months; whereas, in other locations it may take years. Little is known about the reversal mechanisms and how they lead to such a large range of reversal times. Access to untreated fired-affected sites, can be unpredictable and samples vary greatly from site to site. Therefore, a model compound that mimics fire-induced soil hydrophobicity is needed to be able to systematically investigate soil hydrophobilicty reversal mechanisms. Fluorene is a relatively non-toxic, hydrophobic polyaromatic hydrocarbon. The behavior of fluorene coated sand grains was investigated under laboratory conditions using quartz sand. Moreover, its fluorescent properties could be used to non-destructively monitor its degradation over time. In this context fluorene was studied as a possible model compound for the study of mechanisms involved in the reversal of fire-induced soil hydrophobilicy. The compound was subjected to conditions known to degrade or reverse water repellency including temperature, UV-light, and moisture content. Changes in hydrophobicity were monitored using the ethanol drop test and fluorescent imaging. Digital image processing techniques with the public domain software, ImageJ, produced by the National Institute of Health, were used to analyze the images and generate spatial maps of treatment effects on fluorene degradation and hyrdophobicity reversal. Our results indicate that the hydrophobic fluorene-coated sand layers were degraded by treatments such UV light known to reverse hydrophobicity in fire-affected soils, thus suggesting that fluorene may serve as a suitable model compound for producing hydrophobic layers on course grained material.

Creating Claudin-16 Reporter Assays for Studying Epithelial Ovarian Cancer

January 01, 2013 12:00 AM
Miguel Cuevas and Joseph Wilkerson, Utah Valley University Biology Ovarian cancer is the leading cause of death from gynecologic malignancies in the United States and is the fifth leading cause of cancer death among American women. It is estimated that over 22,000 women in 2012 will be diagnosed with ovarian cancer in the United States and approximately 15,500 women will succumb to the disease. This is due to the fact that only 20 percent of cases are diagnosed before the cancer has spread to the peritoneal cavity. Currently, there are no reliable, standard screening tests; the only diagnostic test currently available is the CA125 tumor antigen blood test. This test is inadequate and not available as a general screening tool; additional diagnostics are required to effectively diagnose this disease. It has been previously shown that the tight junction protein Claudin-16, found only in normal kidney, is aberrantly expressed in epithelial ovarian tumors. Therefore, this protein is a good candidate for ovarian cancer diagnostics and targeted therapy. By identifying the promoter region that controls cldn-16 gene expression in ovarian tumors, we can create a luciferase reporter assay to identify cells that express Claudin-16 in culture. To do this, PCR-amplify of various upstream regulatory elements previously identified in kidney cell lines were subcloned into the pGL3 luciferase reporter vector. A higher amount of luminescence is present if the promoter sequence successfully up-regulates the luciferase gene in the vector. This is measured using a Dual Luciferase Assay to determine which promoter region is responsible for the over-expression of Claudin 16. Promoter activity was verified in kidney cell lines that normally express Claudin-16. Next, the assays will be repeated in ovarian cancer cell lines known to express Claudin-16 compared to cell lines that do not express the protein. The promoter assay will then be tested on a collection of ovarian cancer cell lines to determine if luciferase activity correlates with Claudin-16 expression. Once validated, we can test our construct as a cell based assay for identifying therapeutics that can lower Claudin-16 expression in ovarian cancer cells.

Molecular Phylogeny of Mayfly Family Baetidae (Ephemeroptera)

January 01, 2013 12:00 AM
Chase Barker, Utah Valley University Biology The Baetidae are an important group of Mayflies because of their position on the mayfly tree of life in that they are a key to understanding key evolutionary trends, such as the origin of wings and flight. Phylogenetic relationships of mayflies are still not very well known, however molecular and morphological data have begun to shed light on the relationships of these insects (Ogden et al. 2009). Recent molecular and morphological analyses have examined relationships of some of the lineages of Baetidae distributed in different regions of the world (Gattolliat et al., 2008; Kluge, 1997; Monaghan et al., 2005; Nieto, 2010;). Most of these studies suggest two subfamilies, Baetinae and Cloeoninae. This study represents one of the largest (in terms of data) and most diverse (in terms of taxa) phylogenetic analysis performed on the family Baetidae to date. The dataset consists of more than 50 taxa sequenced for six genes (Nuclear 18S rDNA; Nuclear 28S rDNA; Histone 3, Mitochondrial COI, Mitochondrial 16S rDNA; and Mitochondrial 12S rDNA). DNA sequences were aligned in Muscle (Edgar 2004) and tree reconstruction and nodal support was performed under Maximum Parsimony, Maximum Likelihood, and Baysian frameworks. The results indicate that the subfamilies were not strongly supported as monophyletic, contradicting conclusions from morphological data. Missing data in the data set might be contributing to low support across the tree.

Nanospecific Inhibition of Secondary Metabolism and Resistance Induction of the Soil Bacterium Pseudomonas chlororaphis 06 to ZnO Nanoparticles

January 01, 2013 12:00 AM
Jordan Goodman, Utah State University Biological Engineering Nanotechnology is revolutionizing imaging techniques, antibiotic therapy and cancer treatments. Nanoparticles (NPs) are also utilized in many commercial products such as sunscreens, paints, ceramics and semiconductors. Consequently, it is inevitable that NPs find their way into the environment. The effects that NPs have on agriculture and soil ecosystems are the focus of this project. Metal-oxide NPs such as ZnO are toxic to many bacterial pathogens but the beneficial root-colonizing isolate, Pseudomonas chlororaphis O6 (PcO6), has high tolerance. This bacterium represents an important group of micro-organisms that colonize plant roots improving their resilience to both abiotic and biotic stress. At sublethal dose, the ZnO NPs remodel the secondary metabolism of PcO6 in ways that could have an impact on agricultural ecosystems. Formation of antibiotic phenazines produced by PcO6 is strongly inhibited by ZnO NPs. Phenazines are important for PcO6 growth in biofilms, induction of tolerance mechanisms in the colonized plant, and antagonism of other rhizosphere pathogens. A role of NPs as a point source for soluble metal release is involved in these processes but does not explain the total effect of the NPs. The sublethal effects of the ZnO NPs on bacteria are similar to changes reported by sublethal doses of traditional antibiotics on human pathogens. These changes have a strong impact on fitness of bacterial pathogens, the most significant of which is induction of antibiotic resistance. Although many metal-containing NPs are being used in antimicrobial formulations, these alternative antibiotics too may be inducers of antibiotic resistance.

Photobiology: Optimizing Light Quality to Maximize Plant Growth and Development

January 01, 2013 12:00 AM
Kevin Cope, Utah State University Plants, Soils, and Climate Photosynthesis is driven primarily by radiation between 400 and 700 nm; however, not all wavelengths are equally efficient. Red light (600 to 700 nm) is 25 to 35% more efficient than blue light (400 to 500 nm) and 5 to 30% more efficient than green light (500 to 600 nm). Although blue light is less efficient than red light, it has been shown to be necessary for normal plant development in all tested crop species. The mixture of pigments in plant leaves allows them to absorb all colors of light. Both red and blue light are absorbed primarily in the upper leaf. Green light penetrates deep into the lower leaf and transmits to leaf layers below the upper leaf canopy. Accordingly, once the upper canopy is saturated with red and blue light, supplemental green light is beneficial in increasing whole plant photosynthesis. Although the effect of spectral quality on single leaves is well characterized, the effect on whole plant growth and development is poorly understood. The narrow spectral output of LEDs makes them particularly effective for photobiological studies. From our initial studies with radish, soybean, and wheat, we determined that blue light levels can be used to better predict plant development than red:far-red light ratios. We also found that plants require at least 80 μmol of blue photons m-2 s-1 in order to develop normally. Further studies are currently being conducted to determine the optimal ratio of red and green light for maximizing whole plant photosynthesis in lettuce and radish.

The Effectiveness of Meloidogyne incognita and M. arenaria Specific Primers for North American Populations

January 01, 2013 12:00 AM
Brianna Rhoads, Utah State University Biology Nematodes are microscopic roundworms and are known to be some of the most difficult agricultural pests to control. Most of the 700 different plant parasitic nematodes species infect the root system of their host plant, causing galling, stunting and allow secondary pathogens to infect the host. For diagnostic purposes, in order to identify species of root-knot nematodes, the use of species specific primers can speed up the identification; many farmers need fast and reliable identification to determine which crops can still be planted in root-knot nematode infested fields. The purpose of my project was to test species-specific primers developed in Europe and Asia for Meloidogyne incognita and Meloidogyne arenaria on North American populations of both species using PCR. The results showed that primers developed in Asia and Europe for M. incognita amplified North American populations but primers developed in Europe for M. arenaria did not amplify North American M. arenaria populations. Based on results, M. incognita specific primers are useful for diagnostic purposes in the United States.

Increasing Yield and Physical Properties of Dragline Spider Silk from Argiope Aurantia Through the Use of Synthetic Biology

January 01, 2013 12:00 AM
Ryan Putman, Utah State University Biological Engineering Spider silk is a biomaterial with extraordinary physical properties. It has a unique combination of tensile strength, elasticity, and even biocompatibility that has sparked interest in a wide range of disciplines. Potential implementations of spider silk include: medical advances (skin grafts, biomedical sutures, and artificial tendons/ligaments), automotive safety (seat belts, airbags), and military applications (parachutes, body armor). Although spider silk has a great variety of possible uses, collecting this product is not as easy as merely farming spiders and harvesting the silk. Spiders are naturally territorial and cannibalistic, thus alternative means of production are necessary to generate enough spider silk for realistic use. A non-pathogenic laboratory strain of Escherichia coli commonly used for research purposes will be modified to produce synthetic spider silk. Through the use of synthetic biology and molecular cloning techniques, recombinant DNA is inserted with the genetic code for dragline silk of the Argiope aurantia spider; once constructed, this DNA is transformed into E. coli. The goal is to take advantage of E. coli’s ability to be used as a “factory” for creating silk in a controllable and cost efficient system. Supplementation of additional tRNAs will be employed as a strategy to extend cell life and boost the overall spider silk protein yield. Preliminary results have been obtained that show the production of synthetic spider silk by the engineered E. coli. More research is being conducted to increase yields so that we can one day take advantage of this amazing biomaterial.

An Interactive, Taxonomy-Driven Tool for Genetic Sequence Database Mining

January 01, 2013 12:00 AM
Jarom Schow, Utah Valley University Biology DNA and protein sequence data from GenBank and other publicly available databases can be used to perform phlyogenetic analysis. However, the process of assembling data sets for taxa of interest using GenBank is a time consuming and labor-intensive manual process. To improve this process, we have developed a new set of software tools that identifies, organizes, and presents existing sequence data in a way to facilitate data set creation for organisms of interest. The software provides an interactive, taxonomy-driven user interface for viewing and selecting available gene sequence data and exporting it to common genetic analysis file formats. To identify available genetic data, the user selects one or more taxa (species, genus, family, etc.) of interest. The software then identifies all available sequence data for every member of the given taxa. The sequences are sorted by gene and taxon to determine availability and data coverage. Results are then displayed using a hierarchical taxonomy and list of sequence data organized by gene and availability. This enables the user to quickly identify which genes and taxa currently have the best coverage and select the desired data for export. A local database implemented with BioSQL and populated with sequence data from GenBank and taxa from the NCBI taxonomy database was used to access and organize the data. The software is written in C++ using the Qt framework for speed, robustness and cross-platform interoperability.

Determination of Microbial Populations in a Synthetic Turf System

January 01, 2013 12:00 AM
Jason Bass, Weber State University Microbiology There is growing concern regarding the contribution of infilled turf fields on increased athlete infections. Abrasions that occur on these fields create a portal of entry for pathogens such as Staphylococcus aureus. This study compares microbial populations on two infilled turf fields (year old vs. 6 year old turf). Infill material from both fields was sampled at three sites for 5 months during the football season (sidelines, middle of the field, and end of the field). Tryptic Soy Agar was used to determine total microbial load, MSA for S. aureus, and EMB Agar for coliforms such as Escherichia coli. Much higher microbial populations were found on the older turf field, a 1E+04 increase over similar locations on the new turf; suggesting microbial populations can accumulate in synthetic turf infill over time. On the older field the sideline has the highest counts with an average of 1.60E+08 CFUs/g of rubber infill. On the new synthetic turf, the area with the highest number of microorganisms was the end of the field rather than the sideline. This is probably due to where practices are held on the new turf field rather than actual game play. A high number of salt-tolerant, mannitol-fermenting bacteria (indicators of S. aureus) were also found (3.25E+02 CFUs/g on the new turf vs. 2.73E+03 CFUs/g on the old turf). Escherichia coli was isolated from the new turf, in an area of high usage. These results indicate that infill material can serve as a source for pathogens among athletes and that organisms accumulate over time posing a greater risk if proper cleaning is not routinely performed.

Survey for Helenalin in Utah Asteraceae Species

January 01, 2013 12:00 AM
Taylor Nelson, Weber State University Botany Arnica is a common form of complementary medicine used to treat bruises and sore muscles. Most arnica preparations are made as an oil or gel by soaking the flowers of Arnica montana (a member of the Asteraceae) in an oil, such as olive oil, to extract helenalin, the presumed active component. Helenalin, a sesquiterpene lactone, has been shown to inhibit transcription factor NF-?B, a factor which controls the expression of dozens of genes involved in inflammation responses (Rungeler et al. 1999). Due to arnica’s popularity, A. montana is becoming scarce in Europe (its native range) and is sometimes substituted commercially with A. chamissonis (Cassells et al. 1999). Therefore, this study focused on finding other sources of helenalin among the Asteraceae. A. chamissonis, A. cordifolia, A. latifolia, A. longifolia, A. mollis and Helianthella uniflora were collected from Alta, UT, and Helenium autumnale from Salt Lake City, UT, at elevations of approximately 10,500 feet and 4,200 feet, respectively. Sesquiterpenes were extracted from the flowers by dipping them in methylene chloride and separated by thin-layer chromatography. Helenalin and possible helenalin derivatives were identified by their reaction with vanillin and comparison to a pure helenalin standard. Flowers from all seven of the species contained helenalin, with Helenium autumnale containing the highest concentration and A. latifolia the lowest. Therefore, all of the species tested in this study have the potential to be used commercially to prepare arnica salves and thus allow the A. montana population to re-establish itself. An additional finding was that A. chamissonis grew the best under greenhouse conditions, which may explain its substitution in some commercial salves.

Research of an Upstream DNA Sequence Gene of Fremyella Diplosiphon Regarding Regulation of Gfp Gene Expression in Response to Light Intensity

January 01, 2013 12:00 AM
David Tregaskis, Utah Valley University Biology Little is known about the details regarding gene expression which accounts for the light harvesting pigments in bacteria; specifically in the cyanobacteria Fremyella diplosiphon. The purpose of our experiment is to identify the upstream sequence that controls the expression of the scytonemin light harvesting pigment in Fremyella diplosiphon. This experiment will help us better understand the regulation and expression of genes that control light absorption. To test our hypothesis, that there is a regulating upstream promoter sequence for light sensitivity in Fremyella diplosiphon, we will culture the bacteria and extract its DNA. PCR will be performed to isolate the upstream sequence of the gene from Fremyella. Each plasmid was designed by adding restriction sites that will allow the proper cloning of the PCR fragment. This sequence will be introduced into the pSUN 199 and pSUN202 plasmids. These plasmids contain the GFP gene that will be activated by the promoter. The plasmids will be transformed onto Fremyella and analysis of gene expression will be done under different light conditions. This experiment will be able to identify the upstream regulatory sequence of the Scytonemin gene in Fremyella.

Molecular Phylogeny of the Fused-back Mayflies (Pannota: Mayflies)

January 01, 2013 12:00 AM
Robert Putnam, Utah Valley University Biology Pannota, the fused back mayflies, is divided into two main lineages: Ephemerelloidea and Caenoidea. Most of the pannote families are relatively small, in terms of number of genera, with the exception of the larger family Ephemerellidae with 32 genera. Our objective was to 1) test the monophyly of the superfamilies; 2) investigate the relationships within the families of the Pannota as a whole. We combined DNA sequence data with newly generated data (total of ~6 genes) for around 50 taxa. We used standard PCR and DNA sequencing protocols to generate the data. The molecular data were aligned in Muscle and subsequently phylogenetic trees were reconstructed under Parsimony, Maximum Likelihood, and Bayesian frameworks. The results strongly supported the monophyly of the superfamilies, while the relationships of the families within Ephemerelloidea were not resolved.

Interaction of Chytrid Fungus and Normal Skin Biota in Amphibians

January 01, 2013 12:00 AM
Scott Nagao, Weber State University Microbiology Batrachochytrium dendrobatidis belongs to the fungal phylum Chytridiomycota (chytrids), and in 1999, was both identified as a new species and associated with causing chytridiomycosis in amphibians. Since its identification, this fungus has been implicated in rapid population declines in multiple amphibian species in North and Central America, Europe, and Australia. In adult amphibians, the fungus colonizes only the keratinized skin cells on the ventral and dorsal surfaces as well as on the webbing between the toes. Infection is associated with hyperkeratosis resulting in disruption of the skin’s osmoregulatory function, dehydration, electrolyte imbalance and death. There is data that indicates some bacteria that live on the epidermis of amphibians produce chemicals that inhibit the growth of B. dendrobatidis. However, this data is associated only with salamanders. There is no similar inhibitor data published for frog and toad species, and no data published on the identification of normal skin biota of frogs and toads. We have isolated over 175 bacterial isolates from 67 toads and frogs, extracted DNA, and used PCR to amplify the 16S ribosomal DNA gene. Amplicons have been sequenced to determine speciation based on genetics. This is the first report of normal bacterial skin biota in frogs. Further, a skin swab was also taken from these frogs and used to analyze which are infected with B. dendrobatidis via quantitative PCR screening. While the prevalence of fungal infection appears to be low, we have been able to correlate fungal-positive hosts with number and/or species of normal bacterial skin biota.

A Qualitative Study: The Role of Reflection and Service: Learning in an Exercise Science Class

January 01, 2013 12:00 AM
Gessica Stovall, Utah Valley University Exercise Science Introduction:

Correlation between Heart Rate, Estimated Heart Rate, and Rating of Perceived Exertion (RPE) During Running

January 01, 2013 12:00 AM
Emily Matis, Utah Valley University Exercise Science Introduction:

The Evaluation of the Balance and Stability Gains with Crutches using the Tru-Motion go Tip

January 01, 2013 12:00 AM
Jeffrey Pence, Utah Valley University Exercise Science Introduction:

Identification of Novel Invertebrate Neurokinin Receptor Gene Sequence

January 01, 2013 12:00 AM
Chelsie Thomas, Weber State University Zoology The invertebrate ribbon worm, Paranemertes peregrina, serves as a unique model in that it thrives in an environment exposed to drastic fluctuations in salinity during tidal interchange. Its ability to sustain homeostatic integrity is not well understood. With an aim to clarify this phenomenon, total RNA was isolated, and reverse transcription with polymerase chain reaction allowed us to serendipitously clone and elucidate a 488 base pair region of a gene coding for a Tachykinin Receptor (TKR), a subtype of G-protein coupled receptor (GPCR). This region shows 79% homology to the mouse TKR-2 mRNA sequence, and 44% homology to human Neuromedin-K receptor’s amino acid sequence. Specifically, we have isolated a portion containing a cytosolic carboxy-terminus that has classically been associated with palmitoylation or otherwise hydrophibicity-enhancing interactions. This process facilitates the docking of cytosolic subunits to the membrane in the assembly of GPCRs, acting as a regulatory component. In vertebrate models, much of these proteins are conserved. Given that only a few of these genes have been reported for invertebrates, this suggests a critical need for investigation of the evolution of TKRs as they relate to stress response. Additionally, this receptor poses questions about its potential role in pain, with the prospect of revealing insight about the long-elusive pain perception in invertebrates.

Assessing the Impact of Urbanization on Stream Health

January 01, 2013 12:00 AM
Morgan Anderson, Westminster College Environmental Studies Emigration Creek, part of the Salt Lake City watershed, runs through Westminster campus at two points: by Garfield school and through main campus. We assessed several measures of water quality, to understand the impacts of anthropogenic activities and urbanization on this stream ecosystem. Additionally, we identified areas for restoration with the ultimate goal of reintroducing the endangered native Bonneville Cutthroat trout. Beginning in June 2012, we monitored the creek monthly, and will continue for a full year’s time. We assessed the structure and stability of the stream along with dissolved oxygen, pH, temperature, and turbidity. The macroinvertebrate community composition was analyzed also as a biological indicator of stream health. Along both reaches lack of vegetative cover, lack of large rocks to support the lower banks, and scouring and deposition along the steam bottom were common. We found that for all water quality measures monthly average levels were in acceptable limits for trout populations except for dissolved oxygen. Dissolved oxygen dropped below optimal levels in July and August, most likely due to an increase in water temperature. We found that both reaches have moderate species diversity of macroinvertebrates, though the community is dominated by species that thrive in water highly polluted with organic waste. Preliminary results suggest that the current condition of Emigration Creek is not optimal for cutthroat trout, largely due to seasonally low dissolved oxygen levels and lack of preferred food. Continued monitoring, along with a comparative study of nearby creeks, will further our understanding of the impacts of urbanization and the restoration efforts needed to support native fish populations in the Salt Lake Valley.

Antimicobial Properties of Phytochemicals Against Multi-Drug Resistant Bacterial Pathogens

January 01, 2013 12:00 AM
Alisha Ryan, Weber State University Microbiology One of the major challenges facing U.S. military caregivers is the presence of multidrug resistant organisms in extremity wounds. The most frequently identified drug resistant strains of bacteria found in these wounds are Acinetobacter baumanni, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. Due to these organisms rapid increase in resistance to the commonly used drugs, it is crucial to discover and establish alternative methods for treating these microbial infections. Antibiotics are currently the most common treatment for infections by these pathogens, and there is little data on the evaluation of phytochemicals as potential chemotherapeutic agents that could take their place. We have screened 24 individual compounds from 9 major compound families to determine if plant-based phytochemicals could be explored further for use in treating bacterial infections in patients with military wounds. All compounds were tested to determine the minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC). Of the 24 compounds tested, 21 (87.5%) inhibited at least one strain used in this study, with only 1 of the 24 (4.2%) inhibiting all strains. There were 10 (41.7%) of the compounds that displayed MIC values less than 100 _g/ml. For compounds displaying MLCs, they ranged from 2.5 mg/ml to 78.13 g/ml. While there is much more research that needs to be done with each of these compounds, this work is a crucial first step in the drug discovery process. We believe that several of these may serve as potential novel inhibitors of these drug-resistant bacteria.

Serenity, A Drug Recovery Center for Women

January 01, 2013 12:00 AM
Brooke Nelson, Weber State University Interior Design According to the National Substance Abuse Index (2006), Utah women account for 31.7 % of the entire drug abusing population. Mind-altering substance abuse among women creates many unique problems in their lives according to Wesa and Culliton (2004). Additional research by Wesa and Cullliton shows this can affect their futures, finances, families, health, and relationships. Many of the women have other underlying problems that lead them to use mind altering substances to cope with these problems as researched by James (2011). In the Ogden, Utah area, mind-altering substances have created an epidemic that needs to be helped. Serenity, A Drug Abuse Recovery Center, will be an 18,000 square foot building located in Ogden, Utah. Serenity helps facilitate mental and physical healing to create an overall wellbeing for the women. Research has said that women are more likely to receive help for mental conditions than the substance according to Greenfield (2006). Mental disorders and substance abuse will be addressed at the recovery center. Individual and group therapy methods will be used because of their effectiveness shown by Greenfield. Research by Greenfield has shown that women in treatment centers have a greater chance of retention if dependent children are present. Serenity will provide a private room for a woman and two dependent children to stay at the center. The center will include nutrition education and dining area, and a children’s area. The children’s area will have reading, playing, learning, and therapy treating areas.

Dietary Protein Complexity Can Influence Re-feeding Syndrome

January 01, 2013 12:00 AM
Stephanie Hansen, Weber State University Zoology Re-feeding syndrome is the result of the rapid re-introduction of nutrients following a period of severe malnourishment or starvation. During starvation, critical nutrient absorption and anabolic pathways are shifted to conserve energy and maintain cellular function in intestinal enterocytes. Rapid reintroduction of nutrients can often overwhelm the remnant absorptive ability of the intestinal tract, leading to complications such as, failure to thrive and intestinal failure. We hypothesized that a polymeric diet would improve refeeding response compared to an elemental diet. We utilized the nematode Caenorhabditis elegans (C.elegans) as a minimalist model of refeeding. We induced a developmental state of starvation known as, dauer, and then transferred animals into either a polymeric diet or elemental diet. We measured body length daily for 5 days until the worms reached adulthood. Data were statistically analyzed using a general linear model of analysis of variance and expressed as mean+/-SEM. Our results indicate that polymeric diet (695.6+/-25.2 ?m) demonstrated at least 10% (p<0.05) improved growth compared to elemental diet (646.8+/-21.7), with polymeric worms reaching adulthood by day 2 (960.2+/-26.9), whereas elemental animals required 3 days to reach adulthood (894.2+/-26.9 day 2, 972.6+/-27 day 3). Additionally, we assayed intestinal proteolysis by feeding the worms green fluorescent protein (GFP) and measuring fluorescence along the length of the intestinal tract. Our findings demonstrate a decreased rate (p<0.05) of intestinal proteolysis among worms fed exclusively a polymeric diet, despite no significant change in the rate of fluorescent protein intake. Our data confirm our hypothesis that a polymeric diet improves growth and development over an elemental diet. In addition, by requiring some hydrolytic digestion, growth is improved. These results might help to improve clinical management of refeeding syndrome as well as rehabilitate patients suffering intestinal failure.

On Creating a Forensic Herbarium for Weber County,Utah

January 01, 2013 12:00 AM
Jackie Parker, Weber State University Botany Forensic researchers as well as law enforcement officials rely heavily on herbarium type specimens for field investigation. An electronic catalog could greatly assist forensic professionals by providing easy access to area specific specimens. The goal of this project was to compile a database and dichotomous key of native plants found in Weber County, Utah. Pollen and woody material resist degradation, making them perfect candidates for forensic analysis. Using the USDA plant database, all native flowering plants were identified (729 in total). Plant pressings, microscope slides, digital micrographs and line drawings were made of two-hundred native species to date. Pollen and woody tissue were gathered from representative plants and prepared for permanent herbarium collection. Woody tissue and pollen preparation had previously been optimized for best imaging results. Rudimentary dichotomous keys for both woody tissue and pollen were made using micrographs and line drawings to illustrate distinguishing features between plant species. As new species are prepared, they are included in the dichotomous key. The primary purpose of this collection is for it to be used by forensic investigators with limited botanical training. In the spring of 2013, field tests will be conducted to determine how effective the dichotomous key is when identifying unknown specimens. After the layout is finalized, the key will be presented to local law officials as an additional tool in the forensic repertoire.

Antibiotic Resistance of Enterococci Isolated from the Great Salt Lake and Fresh Water Sources

January 01, 2013 12:00 AM
Jennifer Jorgenson, Weber State University Microbiology 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 they often have a high level of antibiotic resistance, making effective treatment of patients more difficult. While it 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. In this experiment, isolates of Enterococcus were collected from the Great Salt Lake and from fresh water sources. These isolates were tested for different phenotypic characteristics and for their antibiotic resistant patterns against five antibiotics. The 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. In contrast only 15% of Enterococcus isolated from the fresh water source were resistant to one or more of the five antibiotics. This has implications for those who have recreational and occupational contact with the Great Salt Lake.

Isolation and Characterization of a Novel Bacteriophage from the Great Salt Lake that Infects Halomonas

January 01, 2013 12:00 AM
Lauren Johnson, Weber State University Microbiology Bacteriophages in aquatic environments play a significant role in bacteria population control, as well as recycling nutrients. The bacterial genus Halomonas is commonly found in the Great Salt Lake (GSL), but very little is known concerning its population dynamics. This euryhalophilic genus is highly versatile concerning its ability to grow in a wide range of substrates and environmental conditions including salt concentration. To better understand GSL microbial ecology, seven strains of Halomonas were isolated from the GSL and identified using 16S rRNA. Samples of South Arm GSL water were filtered twice through a 0.2 m filter, and tested against these Halomonas strains using soft agar overlays to detect Halomonas phage. Three strains exhibited plaque formation indicating the presence of phage. Halomonas phage isolates produced very small plaques, sometimes barely visible. Individual phages were isolated by vortexing agar plugs taken from single plaques in sterile saline then filtering through a 0.2 m filter. From host range streak plates, a single phage isolate (LJ17) appears to infect 4 closely related Halomonas strains. Electron micrographs of LJ17 phage indicate it has a small icosahedral head and perhaps a very short tail. There also appears to be a satellite phage that may be associated with LJ17. There are no reports of Halomonas phage isolated from the Great Salt Lake (GSL), although there are phages found for marine Halomonas strains. Successful isolation and characterization of novel GSL Halomonas phage, besides being critical for development of host/phage models, will also allow studies of GSL microbial ecology.

Student Perceptions and Knowledge of Evolution

January 01, 2013 12:00 AM
Tanner Mortimer, Utah Valley University Biology Nearly half of Americans reject evolution as an explanation of the origin of the diversity of life on Earth and rejection of evolutionary science has been shown to be correlated with a belief in a Christian God-creator (Gallup 2007; NSB 2010). The goal of this study is to explore the student perceptions and knowledge of Evolution at UVU in the non-majors and Biology majors introductory courses. Our study is designed to examine three hypotheses: Hypothesis 1 (Modern secularist): high levels of religiosity and fatalism should correlate with low levels of interest in science and scientific understandings of the world; Hypothesis 2 (Postmodern, postsecularist): no correlation between high levels of religiosity and fatalism, and interest in science and scientific understandings of the world; Hypothesis 3 (Social Psychology): that identifiable structural, contextual features will correlate with identifiable behaviors, irrespective of preexisting beliefs or previous behaviors. Due to the high % of LDS students that attend UVU, we are also interested in looking at the acceptance of Evolution among LDS before learning about the theory of Evolution (Pre-test) and after taking the introductory courses (Post-test). Preliminary results suggest that those students who had a strong sense of God-mediated control also had, in general, a negative attitude towards evolution.