2013 Abstracts
Determination of Microbial Populations in a Synthetic Turf System
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.
Isolation of Hydrocarbon Metabolizing Microorganisms in the Great Salt Lake
Natalie Batty, Westminster College Biology Hydrocarbon metabolizing microbes play a critical role in bioremediation and bio-augmentation projects. In 1978, Brock et al. identified microbes in the Great Salt Lake (GSL) capable of metabolizing hydrocarbons, but since then these microbes have gone unstudied. This research has isolated and identified bacterial species capable of hydrocarbon metabolism from GSL. Though capable of survival in the high salt concentration of the Great Salt North Arm, we have demonstrated that these species are also capable of thriving in low salt concentrations. This research will continue with the characterization process to identify what hydrocarbon sources each species is capable of degrading.
The Role of Nup153 in Nuclear Lamina Assembly
Merima Beganovic, Westminster College Molecular Biology Nuclear Pore Complexes (NPC) create aqueous channels embedded in the nuclear envelope and are made from a network of proteins called nucleoporins (nups). The nucleoporin Nup153 has previously been found to be required for proper assembly of the nuclear lamina. In this study, a fragment of Nup153 was overexpressed in order to impede Nup153 function in T-Rex HeLa cells. After inducing the expression of the dominant negative fragment, I examined the localization of lamins A, B1 and B2, as well as SUN1, Emerin, and BiP, three proteins that mark cell membrane compartments. Lamins B1, B2, SUN1, and Emerin were found to mislocalize to the cytoplasm of the cells, and colocalization among the proteins was observed. Lamin A also had an abnormal phenotype unlike that seen with the B lamins, but indicative of a problem with integration of Lamin A into the nuclear lamina. Colocalization of the various lamin isoforms with membrane proteins such as SUN1 and Emerin indicates a problem with membrane assembly. The distinct localization of BiP, however, suggests that there is a “nuclear-like” membrane in the cytoplasm that either does not incorporate normally into the nuclear envelope as it forms or is newly-recruited to mislocalize the lamin proteins.
Survey for Helenalin in Utah Asteraceae Species
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
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.
Interaction of Chytrid Fungus and Normal Skin Biota in Amphibians
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
Gessica Stovall, Utah Valley University Exercise Science Introduction:
Identification of Novel Invertebrate Neurokinin Receptor Gene Sequence
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
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
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
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
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
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
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
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.
Exploring the Population Genetics of Artemia Franciscana in the Great Salt Lake
Graham Doherty, Westminster College Biology The Great Salt Lake is home to Artemia Franciscana, a species otherwise known as brine shrimp. These halophiles are able to live in extreme environments that have higher than average salt concentrations. Brine shrimp live their entire life cycles in the Great Salt Lake and can be found in both the north and south arm of the lake. Currently, the genetic information regarding the brine shrimp populations at different locations in the lake is underdeveloped. The Artemia populations live in different microenvironments throughout the lake with different physical characteristics and barriers between one another. The physical environment also affects the salt concentration in each microenvironment. Other studies have shown that the increased salinity of an environment accelerates the rate of change in the mitochondrial genome. We feel that different salinity levels in each microenvironment will create different amounts of genetic variation. Our goal is to determine if the distribution of genetic variation is different at different sites in the Great Salt Lake Artemia population.
Frequency Characteristics of Urban House Finch Songs
Dakota Hawkins, Westminster College Arts and Sciences Abstract. Previous studies have documented effects of urbanization on the behavior, reproduction and survival of wildlife. Specifically, noise pollution in urban areas has been known to mask communication among several avian species. In a previous study in Mexico City, House Finches increased the frequency (pitch) of their songs to help mitigate the effects of low frequency urban noise. To document the average minimum frequency of House Finch song in Utah, we recorded House Finches singing from May 2012 to August 2012. Three sample sites with 1 km radii were established in Salt Lake City, Utah while a fourth site was sampled in Logan, Utah. Ambient sound was recorded at locations where songs were recorded to measure urban noise. Average minimum song frequencies and ambient noise were calculated for three sites. Frequency measurements were not significantly different among the three urban populations. Future studies will compare the minimum frequency of these urban populations to nonurban populations and investigate syllable structure and use.
Basal Pterygote Relationships and Its Implications for the Evolution of Flight in Insects
Mark Tingey, Utah Valley University Biology The phylogenetic relationship of Ephemeroptera (mayflies), Odonata (dragonflies) and Neoptera (remaining winged insects) has been a problematic isse in insect evolution and systematics. Resolving their relationships is a critical step toward understanding insect diversification and the evolution of flight. Three hypotheses are evaluated so as to determine the phylogenetic placement of these three orders of insects: 1) Ephemeroptera sister to Odonata + Neoptera; 2) Monophyletic Paleoptera (Ephemeroptera + Odonata); and 3) Odonata sister to Ephemeroptera + Neoptera. Data from more than 260 taxa were collected from Genbank and assembled into a supermatrix representing 6 different molecular markers. Each of the genes were aligned and phylogenetic analyses were carried out utilizing a number of different methods including Bayesian, maximum likelihood, and parsimony frameworks in order to elucidate the relationships of these insect groups. The large number of taxa proved to be a daunting task, but provided new insights into the support for the different hypotheses. The implications for the evolution of flight were examined in light of the generated phylogenies.
Total Mercury and Methylmercury in Water and Brine Flies from Two Distinct Sites at the Great Salt Lake
Jeff Collins, Westminster College Biology The Great Salt Lake (GSL) has some of the highest mercury concentrations ever measured in surface waters. The accumulation of high levels of mercury, particularly its more toxic and readily biomagnified form, methylmercury, has been recorded in many species that inhabit or feed directly in the GSL, including waterfowl. However, no studies at the GSL, and very few studies elsewhere, have investigated the transfer of mercury from aquatic ecosystems to terrestrial ecosystems. We hypothesized that brine flies are able to accumulate mercury during their larval stages in the lake, then transfer this mercury to adjacent terrestrial ecosystems when they become flying adults. Concentrations of total mercury (HgT) and methylmercury (MeHg) were measured in samples of surface water and brine flies (larvae, pupae, and adults) collected once each month between February and July at Antelope Island, in the GSL. Samples were collected from two distinct sites: one along Gilbert Bay on the west side of the Island, and the other along Farmington Bay on the east. On average, HgT concentrations are greater in water samples from Farmington Bay (19.3 ± 9.0 ng/L unfiltered; 13.5 ± 10.1 ng/L filtered) compared to Gilbert Bay (8.5 ± 7.5 ng/L unfiltered; 9.2 ± 9.6 ng/L filtered). MeHg and HgT levels in brine flies displayed the opposite trend, with higher levels at Gilbert Bay (567 ± 123 ppb HgT; 425 ± 12 ppb MeHg) than at Farmington Bay (270 ± 60 ppb HgT; 208 ± 35 ppb MeHg). Potential explanations for the trends observed will be discussed.
Establishing Two-point discrimination of Cranial Nerve V
Kristie Williamson, Weber State University Athletic Training Context: Cranial Nerve V (CN V; Trigeminal nerve) is responsible for facial sensation. CN V has three separate branches which include ophthalmic, maxillary and mandibular. The ophthalmic nerve (V1) carries sensory information from the forehead; the maxillary nerve (V2) carries information from the cheek; and the mandibular nerve (V3) carries information from the chin. It is important to test all three areas of cranial nerve V to accurately assess full nerve function. Previous research has indicated that pressure threshold is not affected by age, however, normative values for two-point discrimination for CN V have not yet been established and may prove useful in concussion evaluation. Objective: Initial investigation aimed at determining gender-specific normative values for CN V two-point discrimination. Design: Prospective repeated-measures design. Setting: This study was performed in the athletic training facilities on the campus of a large Division I institution. Patients or Other Participants: Convenience sample of 106 healthy students enrolled at our institution (43 females, average age: 23.2; 63 males, average age: 22.1). Participants had no history of concussion within the last year, no facial scaring or plastic surgery and not a current smoker. Interventions: Measurement was taken using a two-point discriminator (Disk-Criminator) on the mid-forehead, mid-cheek, and mid-chin. Participants closed their eyes while each measurement was taken. Light pressure was applied (force equal to approximately 10-15 grams to produce blanching of the skin). Measurements were taken three times on each of three facial locations, sequentially. The smallest number in millimeters (mm) to correctly discriminate one/two points was recorded. The two pressure points were applied at exactly the same time. Main Outcome Measure: Discrimination score for each test location (mid-forehead, mid-cheek, mid-chin). Nested average was calculated within subjects for each test location; average was calculated between subjects for each test location. One-way between-subjects ANOVAs were conducted to evaluate potential differences in gender for each test location. Results: Between-subjects average for mid-forehead was 7.8mm (+/2.9), for mid-cheek was 10.4mm (+/2.6), and for mid-chin was 5.9mm (+/1.9). There was a significant effect for gender for cheek normative value (p<0.01); average discrimination score for males was 11mm, average for female was 9.7mm.
Sirt1-Mediated Suprression of Cell Death in Breast Cancer
Matthew Whited, Brigham Young University Biochemistry Several lines of evidence suggest that protein lysine acetylation pathways are deregulated in cancer (1). Moreover, deacetylase inhibitors are emerging as important anti-tumor therapeutics, suggesting that the forced reprogramming of protein-lysine acetylation is toxic to tumor cells. In this study we show that Sirt1, an NAD+-dependent Sirtuin deacetylase that promotes cancer cell survival, is aberrantly mislocalized to the cytoplasm of breast tumor cells. Moreover, the depletion of cytosolic Sirt1 by siRNA sensitizes breast tumor cells to paclitaxel-induced death. Previously, we developed a biotin-switch proteomics approach to identify cytosolic Sirt1 substrates (2). This approach yielded a variety of substrates with roles in metabolism, survival, and oxidative stress signaling. Our current work focuses on three of the proteins identified as Sirt1 substrates: SOD1, DJ-1, and 14-3-3z. SOD1 and DJ-1 both suppress oxidative stress-induced death, and high levels of 14-3-3z expression suppress chemotherapy-induced apoptosis and correlate with negative patient outcomes in breast cancer. Our preliminary results suggest that acetylation of DJ-1 and SOD1 suppress their anti-oxidant functions, while acetylation of 14-3-3z disrupts its binding to pro-survival proteins. Taken together, our data support a model in which cytosolic Sirt1 activates multiple pathways that work together to promote tumor cell survival.
Tissue Specific Isolation of Nucleosomes in Caenorhabditis elegans
Hyrum Shumway, Brigham Young University Microbiology and Molecular Biology Epigenetics is the study of factors of gene regulation that do not stem from the primary DNA sequence. One such example of epigenetic gene regulation is where DNA wraps around histone proteins to form nucleosomes. The positioning of nucleosomes is the first order of control for genic transcription. Wrapped DNA is less accessible for transcription compared to DNA that is nucleosome free. Model organisms for human epigenetics such as the nematode Caenorhabditis elegans are invaluable because of their ease in manipulation and because the components of nucleosomes (histone proteins and DNA) are highly conserved across phylogeny. The purpose of my particular research is to develop and validate a new protocol for tissue-specific isolation of nucleosomes through immunoprecipitation in young adult C. elegans. This protocol leverages green fluorescent protein fused to histones to research nucleosome positioning in the germ-line cells of C. elegans nematodes. When successful in our animal model, this procedure is the first step in allowing analysis of chromatin architecture of any tissue at any developmental or disease state including human cells. The widespread prevalence and implications of human disease is staggering in magnitude. Research is ongoing to decrease morbidity, increase prevention, and fight infection. With epigenetic tools, this vital research is benefitted and supplemented.
Antimicrobial Properties of Essential Oils Isolated from Anthoxanthum hirtum and A. odoratum Against Soil Bacteria
Harsh Kansagra, Southern Utah University Biology Anthoxanthum hirtum is a native grass with many traditional and modern uses, including human medicinal benefits. Populations are found locally in Utah, but at higher elevations, usually above 2500 m. Indigenous people used native sweetgrass in a variety of ways, including medicinally, as ceremonial incense, and in basketry. The active compound that elicits the sweet fragrance of the grass is produced by coumarin, a secondary metabolite used today both medicinally and commercially. Plants most often produce secondary metabolites, or essential oils, as a defense against pathogens, but these antimicrobial properties have not been investigated in A. hirtum. Our research used the Kirby-Bauer disk diffusion test to determine if closely related commercial diploid and polyploid sweetgrass strains (Anthoxanthum odoratum), as well as plants from native A. hirtum populations, produce zones of inhibition when tested against associated soil bacteria and fungi. Results of our research showed all species tested produced inhibition zones, but zone size varied in response to the secondary metabolites produced by each plant type. Despite this variation, these data suggest components of the essential oils may have antimicrobial properties. Results of this study increase our understanding of the antimicrobial properties of secondary metabolites produced by A. hirtum as well as the essential oils produced by commercial diploid and polyploid strains. Future studies will focus on identifying the chemical composition of each extract as well as the specific bacterial and fungal species associated with each plant.
The Role of Timing of Dietary Selenium and Isoflavone Supplementation in the Reduction of Prostate Cancer Risk Factors in TRAMP Mice
Ji Su Park, Brigham Young University Nutrition, Dietetics, and Food Science Selenium (Se) and soy have each been shown to reduce risk for prostate cancer when consumed at high levels. The purpose of this project was to define the molecular mechanisms of prostate cancer chemoprevention by Se and soy, and to describe how timing of dietary treatment modifies those effects. [C57BL/6 X FVB] F1 TRAMP (TRansgenic Adenocarcinoma of Mouse Prostate) male mice were fed stock diets high or low in soy, with or without a supplement of Se (4.0 mg Se/kg BW as Se-meth- ylselenocysteine) by gavage 5 d/wk in a 2 X 2 factorial design. Mice were exposed to different diets starting from conception, 6 weeks, or 12 weeks of age and were sacrificed at 18 weeks. Three-way ANOVA showed that supplemental Se increased serum and liver Se, with significant interactions with both time and soy intake. Selenium dosing decreased BW independent of soy intake and time of dietary intervention. Both Se and soy decreased epididymal fat pad weights, with Se’s effects being more pronounced in mice exposed to diets from conception than from 6 wk. Urogenital tract weights, a measure of prostate proliferation and tumor volume, were significantly reduced by Se supplementation (P<0.001) and soy (p=0.044), independent of time of dietary intervention. Histological examination of mouse prostates is in progress to determine dietary effects on disease progression. These data suggest that, in this model, chemopreventive efficacy of Se and soy does not differ between prenatal and early post-natal introduction.
The Effects of DNA Methylation on Nucleosome Positioning
Marcus Vranes, Brigham Young University Molecular Biology Recent studies have attempted to discover the correlation that exists between DNA methylation and nucleosome positioning, but none have explored the direct effect of DNA methylation on nucleosome formation and positioning. This proposed research will directly test the effects DNA methylation has on nucleosome positioning and whether the histone octamer has preferred sequences to which it binds, which will in turn add our understanding of gene expression and regulation. A better understanding of these concepts will help to aid efforts in gene therapy to better the quality of life of many who suffer from various genetic conditions.
Developing Recipes That Reduce The Fat Content In Baked Foods Where The Fat Substitutes Are Common Foods
Sarah Miller, Southern Utah University Agriculture and Nutrition Science The high intake of fat in the average American diet is a growing concern to the nutritional community. High levels of total fat intake increase cardiovascular disease (CVD) which is the number one killer in America. The successful development of low fat recipes that can be accessed and produced by the general population is valuable to the fight against obesity and CVD. If acceptable products can be made with lower fat content then it is possible to lower the fat intake in a person’s diet. This study is designed to test the acceptability of low-fat desserts using every day food products available for home use as fat substitutes. Four popular “name-brand” recipes were altered by removing 55-73% of the fat and replacing it with a common food item, either pureed white or pinto beans, applesauce or yogurt. Southern Utah University students were recruited via flyers, in class announcements, and word of mouth. Participants (n=56) were asked to taste and evaluate the four products. Afterwards they completed a short survey about their knowledge of low-fat food substitutes and acceptability of the products. Results and conclusions forthcoming.
Ability of Wolf Spider’s (Lycosidae) to Assess Their Nutritional Needs and Deficiencies
Sarah Miller, Southern Utah University Biology As humans, we have the ability to change our diets if we are deficient in any specific nutrient. Many animals have the ability to change their diet if they are deficient in nutrients. However, for many carnivores, it is generally thought that a prey item will have the same nutritional composition as the predator body composition. Carnivores are limited nutritionally in their diet. This means that they only get nutrients from the prey that they are able to locate to consume. This study is investigating prey choice of invertebrate predators (wolf spiders) when their prey item has been altered to be deficient in one specific nutrient (lipid or protein). I predict that the spiders will be able to choose the correct prey item for the nutrient in which they are deficient. The total weight of flies eaten in each treatment will be measured to determine if wolf spiders select prey items based on the nutrient content of the prey. Results will be analyzed using a nested ANOVA.
Urbanization and its Effects on Prey Preference in Wandering Garter Snakes (Thamnophis elegans vagrant)
Dillon Monroe, Southern Utah University Biology When an area is developed no part of that habitat is left unaffected. Urbanization creates many problems for animals by creating physical barriers, fragmenting habitat, introducing completely new habitat, and introducing species. Exotic species are often better competitors for resources and often competitively exclude native species, resulting in population declines and ecosystem disruption. Despite the negative effects of urbanization some species are able to adapt and survive in urban ecosystems. One such species is the wandering garter snake (Thamnophis elegans vagrans). This species is described as a generalist in habitat selection and is often seen in urban communities basking on sidewalks and pavement, living in gardens, and feeding on rodents, insects, and other species. One unique characteristic of this genus is that prey preference has been observed as being a genetic trait and possesses variation, and as such can be modified via natural selection. For my research project I looked at whether prey preference of T. elegans vagrans differs between snakes from three different locations with varying levels of human development. From these three locations in Utah, Tropic reservoir, Monroe (Sevier River), and Utah Lake, we obtained a total of 14 snakes and found that in this group of snakes there is no correlation of prey preference in snakes to level of development of the habitat.
Analysis of the Degradation of Flavor Volatiles in Single Origin High Cacao
Adrian Scottorn, Salt Lake Community College Natural Sciences Chocolate has long been the favorite snack of many, and recent studies have shown multiple positive health benefits of eating dark chocolate in particular, encouraging production and consumption of very high cacao content bars. We have quantified some of the changes that occur to the amount of known flavor compounds contained in a locally made artisan chocolate. The bars we chose were 70% minimum single origin cacao made with as few other ingredients as possible. By first measuring the amount of known flavor compounds, then stressing the bars in various ways, we looked in to what really happens when a quality chocolate bar is mistreated.
Early Parental Death, Genetic Variants and Risk for Alzheimer’s Disease: Building a Risk Profile from the Cache County Study on Memory, Health, and Aging
Michael Peterson, Brigham Young University Biology A person’s predisposition to Alzheimer’s Disease is known to be influenced by both genetic factors as well as environmental factors. One know environmental factor is that known to affect risk for disease is early parental death. The purpose of this research is to better understand the complex factors that influence the disease by analyzing the relationship between the environmental factor of early parental death with genetic variants known to influence the disease. We used extant data from the CCSMHA, an ongoing aging study including 89.7% (5092 of 5677) of all of the eligible residents of Cache County, Utah. This data includes information about environmental and psychosocial stressor of the subjects as well as information about physical examinations, metal screenings, and individuals’ genotypes at many loci that are known to be related to Alzheimers Disease. We used multivariate logistic regression to determine the effect of early parental death by SNP interactions on risk for AD. For the analysis we cleaned the data by removing SNPs less than a minor allele frequency of 0.01, a Hardy-Weinberg equilibrium value of 110-6, and a maximum missing snp call of 0.2. Individuals were also removed if genotyping rate was less than 0.2. After filtering we had 262 cases, 239 controls and 0 missing Final Results will be presented at the Conference.
Genome-Wide Association Study of Visinin-Like Protein Levels, an Endophenotype for Alzheimer’s Disease
Rachel Perry, Brigham Young University Life Sciences Previous studies have indicated that Visinin-like protein (VILIP) may be a powerful tool in predicting disease progression and guiding prognosis of Alzheimer’s disease (AD). Cerebral spinal fluid (CSF) was collected from hundreds of individuals with varying levels of AD. The CSF was then analyzed for levels of VILIP protein using Luminex technology. SNPs were genotyped using the Illumina OmniExpress chip. SNPs found to have a Hardy-Weinberg frequency less than 1×10-4 were not included, assuming that this variance was due to a genotyping error. SNPs and samples missing more than five percent of the data were also not included. Following the cleanup of the data, an association test using linear regression was performed. Covariates used in the analysis included age, gender, and covariates that accounted for population stratification (PC1 and PC2). Over one hundred SNPs were found with a p-value less than 1×10-5. The genomic inflation factor for the generated data was 1. One marker showed significance at the genome-wide level. We have identified a genetic marker that shows significant association with CSF VILIP levels. This finding may provide insight into genetic control of VILIP levels, which may be a useful in understanding the pathological processes involved in AD.
Anodic Stripping Voltammetric Analysis of Lead, Cadmium, and Copper in the Jordan River, Utah
Chris Thurman, Salt Lake Community College Natural Sciences This project is an ongoing multifaceted investigation of the Jordan River. The study’s intent is to monitor and asses the overall condition of the river from Utah Lake to the Great Salt Lake. Samples of river and pond water, typical vegetation such as cattails, and some soils have been collected with reference to sites along the river that may serve as sites for the introduction of waste and other contaminants. These samples have so far been examined with respect to temperature and dissolved oxygen at the sampling point. Two river samples, one pond sample, and plant sample have been analyzed by ICP-MS for 20 different metals. The levels of Lead, Cadmium, and Copper have been determined by anodic stripping voltammetry at the ppb level in a variety of other water samples from the river. The results of these measurements are disused and presented Geospatially.
The Association Between Pattern Separation and Depression
Matthew Schneider, Brigham Young University Physiology and Developmental Biology Alzheimer’s disease, the most common form of dementia, affects millions of people per year. Research has shown that Alzheimer’s affects the hippocampus brain region, which is involved in learning and memory. Understanding learning and memory functions is imperative to comprehending both healthy brain functions and Alzheimer’s disease. Many researchers seek to understand both the causes and treatments of the disease, but tangible information remains elusive. Studies thus far have shown that to encode memories, the brain must change neural synapses to either strengthen or weaken those pathways, a process known as synaptic plasticity. Using electrophysiology techniques on mouse hippocampal slices, this project will provide further insight on memory formation and regulation by imitating synaptic plasticity mechanisms. I will look at a specific cellular pathway involving the protein receptor GPR55, which has recently been shown to induce synaptic plasticity. By understanding how the GPR55 pathway functions, this research will contribute to the understanding and treatments of Alzheimer’s and other neurodegenerative diseases.
Analysis of Dental Parameters, Fluoride and pH, in Utah’s Waters, Drinks, and Foods
Lacie Cates, Salt Lake Community College Natural Sciences According to the Salt Lake Valley Health Department 1 the optimal level of fluoride is 0 .7 to 1.2 ppm in drinking water. Also acidity of many foods and drinks leads to tooth decay. In 2003 the State of Utah started adding fluoride to tap water. Then in 2008 Utah counties voted on addition of fluoride resulting in a variation of water treatment from county to county. At present the state has about 50% of the population receiving fluoride treated water with the aim of providing the 1 ppm fluoride level. It has also been reported that some counties are considering cutting back to about 0.7 ppm 2 . This study examined the pH and fluoride content of water samples from the major population counties in Utah. Fluoride levels ranged from 0.08 – 0.92 ppm. The pH values for these samples ranged from 6.26 -8.08. A further study of the fluoride levels and pH in bottled water and other drinks and foods such as fruit and cheese was conducted because many people in Utah do not drink tap water, particularly in regions of high water hardness or areas where taste and / or odor can be off putting. The pH values ranged from 2 to 8. The most acidic being colas and citrus based drinks and foods. The fluoride values varied from 0.03 to 0.47 ppm.
Adaptation of Staphylococcus Aureus to UV-C Light
Kristian Johnson, Dixie State University Biology Antimicrobial methods, such antibiotics and Ultraviolet (UV) irradiation, have been a means of suppressing prokaryote proliferation for nearly a century. Over the last several years, scientists have found that numerous strains of prokaryotes have developed resistance to antibiotics. Concurrently, the process of bacterial irradiation using UV-C is common practice in a variety of sterilization applications. As revealed in the seminal work by Chang et al. inactivation curves for Microorganisms such as Staphylococcus aureus (Staph) were established in 1985. Their values indicate survival rates based on Intensity, which is defined as the time of UV irradiance per unit area. Similar to the evolutionary evidence of antibiotic resistance, we are interested in the selective pressure UV-C has on Staph. By recapitulating Chang’s experiment nearly 30 years later, our preliminary results indicate an increased resistance to UV-C in Staph. In this experiment, we determine a current UV-C dose-dependent kill rate function for Staph.
Chytridiomycosis-resistant Frog Populations in Southern Utah
Chancen Hall and Nichkolas Hadley, Dixie State University Biological Sciences Batrachochytrium dendrobatidis (chytrid fungus) is prevalent worldwide, and the resulting chytridiomycosis has contributed to at least 168 amphibian species extinctions. In 2010, B. dendrobatidis was discovered in the greater Zion National Park area of southwestern Utah. Because few populations have shown resistance to chytridiomycosis, we decided to explore the effects of this disease on populations of Hyla arenicolor (canyon tree frog). We tracked the spread of B. dendrobatidis by testing skin samples taken annually from several different canyons and monitored population sizes. During the three years of our study, infected populations did not show subsequent population declines. This suggests that H. arenicolor population size in this region is unaffected by B. dendrobatidis. In the future, testing hypothesized explanations for surviving infection could help us identify populations not at risk and thus allocate conservation resources more efficiently.
GPR55: A Potential Enhancer of Learning and Memory in the Hippocampus
Rachel Schneider, Brigham Young University Neuroscience The ability to create distinct memories for very similar stimuli and events is called pattern separation. Pattern separation is thought to be dependent on neurogenesis (the birth of new neurons) in the dentate gyrus, a subregion of the hippocampus. Neurogenesis is reduced in depression, as is overall memory performance. It has been proposed that depression negatively impacts pattern separation abilities, however a link between depression and performance in pattern separation memory tasks has yet to be identified. Accordingly, we designed a study to investigate the relationship between pattern separation performance and level of depression. Eighty-two participants completed a pattern separation memory test and a set of questionnaires to gauge their level of depression. During the task, participants were presented with 600 images one at a time on a computer screen in a continuous recognition paradigm. Participants were asked to determine whether each image was new, old, or similar. Images seen for the first time during the task qualified as “new”, images that were repeated following a variable delay qualified as “old”, and images that were similar to previously presented stimuli, but not exactly the same, qualified as “similar”. A pattern separation score was calculated based on the proportion of correctly identified similar stimuli. We found a negative correlation between depression scores and pattern separation scores (r(82) = – 0.301, p < 0.01). This relationship held constant even when we controlled for other factors known to affect neurogenesis, such as exercise and anxiety levels. These results provide support for the theory that depression is negatively related to pattern separation performance, possibly due to a decrease in neurogenesis in the hippocampus.
Fire Retardant as an Environmental Risk Factor Contributing to Parkinson’s Disease
Michael Barney, Southern Utah University Biology Parkinson’s disease is caused by a decrease in dopaminergic neurons in the substantia nigra, which results in a loss of motor control. Although the exact causes of Parkinson’s disease are unclear, studies have shown that exposure to environmental contaminants causes death of cells in the substantia nigra (McCormack et al., 2002). The purpose of our study is to investigate the potential role of fire retardant as a risk factor for Parkinson’s disease. We subcutaneously injected 8 experimental mice with retardant solution (1 ml/60 g body weight) and 7 control mice with saline solution (1 ml/60 g body weight). Each mouse was given 8 injections over four weeks, after which the mice were sacrificed and brains were harvested. Frozen sections (40 mm thick) were mounted on slides and are being processed using anti-tyrosine hydroxylase, which will be visualized using a peroxidase reaction. The number of cells in the substantia nigra will be counted and compared between control and experimental groups. Our hypothesis is that exposure to fire retardant is a risk factor for Parkinson’s disease. If this hypothesis is supported, this would be the first study to show a link between fire retardants and Parkinson’s disease. This would have important implications for current forest fire fighting techniques.
Endocannabinoid Biosynthesizing Enzyme Expression in Hippocampal Stratum Oriens Neurons
Ryan Williamson, Brigham Young University Physiology and Developmental Biology The hippocampus is thought to mediate learning and memory by altering the strength of synapses within its circuitry. In many cases, this synaptic plasticity can be induced by intracellular signaling molecules. Lipid-based intracellular signaling molecules called endocannabinoids have been shown to modulate or mediate synaptic plasticity among hippocampal pyramidal cells and stratum radiatum interneurons; however, the role of endocannabinoids in mediating synaptic plasticity among interneurons in the stratum oriens is still unclear. Our goal was to determine whether stratum oriens interneurons have the machinery necessary for endocannabinoid production and, if so, whether this machinery is expressed in a sub-type specific manner. To do this, we used patch clamp electrodes to extract single cells from rat hippocampal slices and analyzed the expression of endocannabinoid biosynthetic enzyme mRNA using quantitative real-time PCR. In this analysis, we examined cellular expression of two interneuron markers, GAD65 and GAD67, as well as several calcium-binding proteins and neuropeptides to determine interneuron subtype. We also analyzed cellular expression of several endocannabinoid biosynthetic enzymes, including N-acyl phosphatidylethanolamine phospholipase D, diacylglycerol lipase alpha, and 12-lipoxygenase, as well as type I metabotropic glutamate receptors. Preliminary data suggests that stratum oriens interneurons express mRNA necessary for endocannabinoid biosynthetic enzymes. Additionally, we identified interneurons that coexpress mRNA for somatostatin and diacylglycerol lipase, suggesting that O-LM cells or another somatostatin-positive interneuron subtype may possess the enzymes necessary to produce the endocannabinoid 2-arachidonoylglycerol. Further work will allow us to examine how endocannabinoid biosynthetic enzyme expression correlates with other interneuron subtypes in the stratum oriens.
Validation of Cache County Genotype Data
Aaron Sharp, Brigham Young University Biology The Cache County study on memory, health, and aging has played a significant role in several studies. However, there is some potential skepticism in the scientific community about its sample. The population in Cache County is derived from a diverse group of founders, but it is perceived by some to be an isolated population. If so, conclusions discovered there might not apply to other populations. Our objective is to compare the Cache County data to a panel of genetic data—provided by the International HapMap Project and the Alzheimer’s Disease Neuroimaging initiative—that is known to be representative of typical European-American populations. Doing so will indicate whether the genetic diversity in the Cache County sample is characteristic of an isolate or not. Analysis will be done using the open source “Plink” analysis toolset, including the –cluster and –mds-plot computational algorithms. Using –cluster groups individuals according to identity by state distances. The –mds-plot algorithm creates a scatter-plot of the individuals in 2-dimensional space, identifying any systematic difference between the Cache County data and the general population. We expect that the Cache County data will be representative of general European-American populations, because of its diverse group of founders.