2013 Abstracts
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.
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.
Impairment of Withholding a “Pre-Potent” Response In Rats With METH-induced Neurotoxicity
Lee Leavitt, University of Utah Biology Chronic methamphetamine (METH) abuse leads to structural and functional damage in the brain, which likely contributes to cognitive and behavioral dysfunction. Recent data suggest an association between METH abuse and impaired inhibitory control over behavior; that is an impaired ability to inhibit inappropriate actions or thoughts. However, the extent to which METH-induced neurotoxicity is responsible for such impairment remains to be determined. Previously, we reported that rats with METH-induced partial dopamine (DA) and serotonin (5-HT) loss in striatum and prefrontal cortex (PFC) exhibited impaired response inhibition in the form of perseverative responding. Furthermore, levels of serotonin transporter (SERT) binding in PFC correlated with perseverative responding. Herein, we have examined another dimension of response inhibition impulsive action, which is an inability to withhold a “pre-potent” response in rats with METH-induced neurotoxicity. Rats were trained to perform a stop-signal task (SST). Once rats achieved stable responding (>80% correct response) on both “Go” and “Stop” trials, they were treated with saline (0.9%), a neurotoxic regimen of METH (4 x 10 mg/kg, 2-hr intervals, s.c.) under normal ambient temperature conditions (“neurotoxic” METH group) or the METH regimen under cooling conditions (“normothermic” METH group). One week after the treatment, rats were again tested on the SST. After behavioral tests were done, animals were sacrificed and brains removed for determination of monoamine loss. The results showed: 1) the “neurotoxic” METH-, but not the “normothermic” METH or saline-treated rats, showed a 40-60% loss of SERT and dopamine transporter (DAT) binding in PFC and striatum; 2) the “neurotoxic” METH group showed normal behavioral performance in “Go” trials of the SST relative to the “normothermic” METH group or saline controls; 3) the “neurotoxic” METH group exhibited impaired withholding of a “pre-potent” response, as reflected by increased numbers of errors on the “Stop” trials of the SST. Overall, these data suggest that impaired inhibitory control over behavior (i.e., increased impulsive action) can arise as a consequence of METH-induced neurotoxicity to central dopamine and serotonin systems. Supported by NIH grant DA 024036
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.
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.
A Preliminary Assessment of Mercury Concentrations in a Terrestrial Songbird on Antelope Island
Heather Reynolds, Westminster College Biology The presence of mercury in a food chain can have harmful effects, including altering behavior, on organisms. Mercury is typically found in aquatic ecosystems, however recently is also recognized as a potential problem in terrestrial ecosystems. High levels of mercury have been found in the Great Salt Lake in a form able to bioaccumulate up the food chain. If the aquatic ecosystem is linked to the terrestrial ecosystem, then some of the highest predators included in this food chain may be songbirds that eat spiders. We quantified mercury in the blood of the Loggerhead Shrike, Lanius ludoviciaus, on Antelope Island. Upon being caught, the birds were measured, color banded, and a blood and feather sample taken from each of the 15 shrikes to be tested for mercury. Mercury concentrations ranged from 0.96 to 4.00 ppm, with a mean 1.14+/0.31 ppm. Sub-lethal effects in songbirds from another study were seen with concentrations ranging from 2.0-3.5 ppm. Two shrikes we tested exceeded that range. There was high variability in concentrations of mercury, which might be related to distance from shore. These preliminary data demonstrate that some mercury from the aquatic ecosystem is bioaccumulating in the terrestrial ecosystem on Antelope Island, and there may be harmful levels in the birds which could lead to change in behavior, and eventually decline in population. Future studies will investigate the costs of mercury contamination.
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.
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.
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.
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.
“Killing Me Softly”: The Economic Toll of Forest Fire Repression
Adam Olson, Brigham Young University Plant and Wildlife Sciences Forests systems of the Central Rocky Mountains rely on an intricate balance of natural disturbance cycles in order to develop properly. Forest fires are one such disturbance, however, certain fire characteristics, particularly fire severity, can vary widely across forest landscapes. In our study, we examined the influence of fire severity on aspen regeneration as well as aspen defense against wildlife and livestock browsing. Our results indicate that high or moderate burn severity is more favorable to aspen regeneration and survival. These areas of severe burn result in a higher density of aspen suckers, more vertical and lateral growth, greater chemical defense concentrations, and less browse damage than the neighboring plots of low or no burn. This data suggests that fire severity should be taken into account when considering plant regeneration and susceptibility to browse damage in burned landscapes.
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.
The Effect of Male Dominance on Female Mate Choice in Poecilia gillii
Michael McEntire, Brigham Young University Biology Female mate choice (intersexual selection) and male dominance interactions (intrasexual selection) can each play important roles in sexual selection. These two mechanisms tend to be discussed in isolation. The goal of this study is to explore the interaction between these two forms of sexual selection. To test this idea, we focused on the livebearing tropical fish system Poecilia gillii. We grouped males into similarly sized pairs and observed them for a week to determine which male was dominant. These pairs were then presented to females in mate choice trials to ascertain female preference. We also photo- graphed the males to determine coloration. We found that females were unable to detect dominant males without viewing the physical contest and that carotenoid coloration bore no effect on female preference. Females tended to choose the male to their left, suggesting the preferential use of their right eyes in making decisions on mate choice.
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.
Characterizing the Role of HspB2 in Cardiac Metabolism and Muscle Structure Using Yeast and Mammalian Cells
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
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.
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.
Identification of Novel Serum Biomarkers for Alzheimer’s Disease Using an Integrated Serum Proteomics Method
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.
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.
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.
The Effects of Exercise on Synaptic Plasticity in the CA1 Region of the Hippocampus in Mice Who Experience Acute Stress
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
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
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.
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.
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.
The Effects of Temperature and Water Availability on the Germination of Bromus Rubens
Rachel Nettles, Brigham Young University Plant and Wildlife Sciences Background/Questions/Methods
Association of the CETP Gene with Cognitive Decline and Dementia in the Cache County Study
Caitlin Munger, Brigham Young University Biology Alzheimer’s is a fatal, non-treatable neurodegenerative disease and the most common cause of dementia. While no one gene has been found to determine the development of Alzheimer’s, past studies have established a strong hereditary influence on Alzheimer’s. So far, only 5 genes have been found which replicably contribute to the genetic risk of developing Alzheimer’s. However, the gene for Chlolesteryl Ester Transfer Protein (CETP) has been identified as a possible new contributor to the genetic risk factor. In order to test this association we obtained data on over 4000 subjects studied in the Cache County Study on Memory, Health and Aging over a 15-year period. This data included DNA samples, cognitive decline rates and incidence of dementia–particularly Alzheimer’s Disease. DNA samples were SNP genotyped using quantitative PCR. The SNP genotypes and corresponding phenotypes for each subject were then analyzed for association usingmixed linear models and for survival, or the amount of time until the disease appeared, using Cox proportional hazard models. We found a correlation between the V405I SNP and a decreased rate of cognitive decline. We found that for each additional G the rate of decline decreased by 0.6 points per year on the MMSE test. The identification of CETP as a player in the genetic risk for Alzheimer’s and dementia will provide much needed information on the genetic factors involved in dementia and allow for possible future therapeutic targets.
Subduing the Flu: New Alternatives to Amantadine
Joseph Moulton, Brigham Young University Physiology and Developmental Biology With the advent of recent mutations in the influenza A viral genome, drugs that previously blocked the proton flux responsible for disassembly of the viral envelope and exposure of viral RNA to the transcriptional machinery of the host cell have become ineffective. Our study of the M2 hydrogen ion channel responsible for this flux has led to a vastly-increased under- standing of the mechanisms behind the conductance activity and potential blockage of these transmembrane tetramers. By embedding M2 proton channel subunits of the S31N mutant strain into liposomal bilayers and suspending these bilayers in the buffers and ionic gradients characteristic of the intracellular environment, we have been able to simulate and observe nor- mal functioning of the influenza A virus. Using these liposomal bilayers, we have developed a series of experimental protocols to test a variety of amantadine- and rimantadine-related drugs for successful blockage of M2 S31N proton conductance. Our research presentation will be centered around the mechanisms of this channel and the favorable results that we have obtained from many of these drugs.
Hydrogeochemistry, Geothermometry, and Structural Setting of Thermal Springs in Northern Utah and Southwestern Idaho
Brennan Young, Utah State University Geology Thermal springs in northern Utah and southeastern Idaho mostly lie near active or inactive Basin-and-Range normal faults. They are dynamic systems, and the character of some has changed drastically since work as early as the 1980’s (Blackett and Wakefield, 2002; IDWR, 2001). We examined and sampled 60 thermal springs and most samples met criteria for cation geothermometers, or mathematical-geochemical tools used to estimate the maximum temperature of hydrothermal reservoirs. Of the 60 springs, 51 met criteria for the Na-K-Ca geothermometer and the remaining nine springs did not meet the criteria for the K-Mg, Na-K, Na-K-Ca, or Na-K-Ca-Mg cation geothermometers used in this project (Fournier and Truesdell, 1973; Fournier and Potter, 1979; Giggenbach, 1988). Of those 51 springs, only one is considered to be in partial equilibrium with the thermal reservoir, and estimates a reservoir temperature of 79°C (Giggenbach, 1988). Though the majority of springs exhibit a chemical signature of having mixed with shallow groundwater (Giggenbach, 1988), the Na-K-Ca geothermometer gives the most reliable results for springs in northern Utah and southeastern Idaho, but only for springs with surface temperatures exceeding 30°C and with greater than 1000 ppm total dissolved solids (TDS). Geothermometer results for these springs yield reservoir temperature estimates between 193 and 249°C.
Gravitational Atom Consisting of Electron or Proton Orbiting a Small Black Hole
Daniel Gray, Utah Valley University Physics Mini black holes (BH) of various mass could be left over in space from the early expansion Big Bang phase (so called primordial BHs). As a result of interaction of those BHs with interstellar hydrogen they could form a bound system with an electron or a proton (or both). What would such system look like? Would it be stable, metastable, or would BH quickly consume the orbiting particle? How much is life time of such “gravitational atom”? If such system is stable then what is the size of it; how much is the bonding energy of its ground state (=ionization potential energy) and how much are the energies of its exited states? Are those atoms “gravitational atoms” observable? What other properties do they have? Based on known physics we try to analyze the behavior of such exotic systems and answer the above questions for black holes of various masses.
Search For Shrodinger Type Structuring In Keplerian System
Samuel Leventhal, University of Utah Physics The uncertainty existing within the scientific community as to why quantum mechanics (QM) behaves as it does comes from the fact there exists no mathematically sound approach for deriving the postulates of QM. It is the purpose of our research to present a derivation for the postulates of QM through the theory of Scale Relativity (SR), followed by a search for physical signatures of SR in the mechanics of celestial bodies. The construction of SR is based on an extension of the relativity principle to scale transformations coupled with a loss of differentiability. Our first paper presents the derivation of QM through scale relativity. During the SR derivation we also show fundamental qualities of QM, such as the presence of complex numbers in state functions. Lastly, the seemingly unrelated behaviors between relativity and quantum phenomena are shown a single mathematical formulation, only to change form due to scale. The new resolution variable within the adapted Schrodinger equation allows it to become applicable to macroscopic scales allowing us to look at large scale mechanics for signs of SR. Gravitation being scale invariant leads it to be a perfect candidate for experimental purposes. Our second paper investigates whether or not celestial bodies, formed by chaotic gravitational structuring, obey the properties of a Schrodinger equation dependent on the Keplerian potential. If so SR implies solar systems would form along probability distributions predicted by the square magnitude of the Schrodinger-Keplerian wave equation. In theory a planets probability distribution would depend on discrete variables, denoted orbital rank, n=n. In search for SR it is sufficient to see if planets tend to have orbital ranks near integer values. We start by calculating the orbital ranks within various solar systems, followed by testing whether the accumulation of planets’ rank near integer values is a probable event. To test this we take the squared difference between the calculated rank and the nearest integer. As a result we are able to test how likely orbital structuring will be discrete. Our results show a strong certainty that orbital rank is likely to accumulate near integer values.
Peroxide Resistance in CMC4 Deletion in Saccharomyces Cerevisiae
Sally Hansen, Utah Valley University Chemistry Cytochrome c oxidase (CcO) or complex IV is the terminal component of the electron transport chain. In eukaryotic organisms, CcO is composed of 12-13 subunits. The core of eukaryotic CcO contains three mitochondrially encoded subunits that comprise the catalytic core of the complex and several gene products encoded for by the nucleus (1). Essential to the redox function of CcO are several critical cofactors: two hemes and two copper centers (2). The crystal structure of CcO has led us to several insights about its structural components and catalytic activity (3). However, a large set of nuclear gene products are essential for CcO activity that are not part of the structural machinery of the complex (4,5). These components have been implicated in various stages of CcO assembly including, heme processing and insertion (6), CuA and CuB site delivery and insertion (7), subunit processing and subunit assembly (8,9). Among them are well characterized CcO assembly factors that involve the biogenesis of the CuA and CuB sites in CcO. Sco I, Cox17, Cox 11 and Cox23 are all essential to CcO activity and have properties that appear to be critical to the maturation of the CuA and CuB sites(10-13). Cmc4 appears to be involved in cytochrome c oxidase biogenesis. Peroxide phenotypes have been linked to cytochrome c oxidase assembly (14). Saccharomyces cerevisiae strains lacking CMC4 were found to exhibit peroxide resistance when compared wild type parental stains. Resistance was seen in liquid culture and in media containing glucose and glycerol. These results may indicate that cytochrome c oxidase assembly is altered in CMC4 deletion stains.
Raman Spectroscopy Methods to Detect Monomolecular Layers on Oxide Surfaces
Natascha Knowlton, University of Utah Chemistry The chemical derivatization of oxide surfaces (silica, alumina, glass) is critical to the development of separation media, sensing surfaces, or biocompatible interfaces. Presently, there are few analytical methods that allow the detection and characterization of functionalized monolayers on these surfaces. Raman scattering spectroscopy can provide useful structural information in the form of vibrational spectra of molecules of interest, and it is compatible with oxide substrates. Raman scattering, however, is a very weak effect so that its application to detecting monolayers is challenging. In this work, two approaches to detecting and characterizing molecular layers on oxide surfaces with Raman spectroscopy are compared. First, gold colloidal nanoparticles are deposited onto the surface of interest, which enhance the Raman scattering near the gold surface by surface-plasmon resonance. This technique is suitable for ex situ analysis of monolayers on planar surfaces. Secondly, monomolecular layers can also be detected by Raman scattering on porous oxide supports such as alumina or silica without any optical enhancement due to the very high surface area of these materials. Detection in porous particles is compatible with in situ monitoring of surface derivatization reactions. These two methods are compared for monitoring of reactions of silane-coupling agents and their subsequent functional group transformations on glass and silica surfaces.
Examination and Characterization of Coal Creek Runoff as well as the Impact of Cedar City, Utah
Alyssa Brown, Southern Utah University Physical Science Coal Creek is a perennial stream that runs through Cedar City, Utah and drains into Rush Lake. The basin is a closed basin, meaning that most of the surface water will eventually become ground water, the main source of drinking water for Cedar City. Because of the increase of urbanization of this region in par- ticular, there is a higher probability of pollutants entering the water source and greatly affecting the quality of the drinking water. Most pollutants are characterized as nonpoint source pollution, which is defined by the Environmental Protection Agency (EPA) as, “land runoff, precipitation, atmospheric deposition, drainage, seepage or hydrologic modification” and is the leading cause of water quality problems (Environmental Protection Agency, 2012). Common sources of land runoff, as defined by the EPA, include septic waste, lawn and garden fertilizers, improperly disposed chemicals, automobile fluids, vehicle emissions, and road deicers (Environmental Protection Agency, 2012). In order to characterize the pollutants found in samples and determine the concentration of ions of interest, ion chromatography was used. By determining the identity and concentrations of particular ions, the presence of nonpoint pollution, and the source, may be determined. This study may lead to better indications of where major sources of drinking water pollutants are originating, illustrating the effect that urbanization and rapid population growth have on the quality of both surface and ground water. This will allow better characterization of pollutants and pollution sources in the future.
Hydrogen Peroxide Phenotypes of Saccharomyces Cerevisiae Knockouts
Joshua Jeffs, Utah Valley University Chemistry The electron transport chain (ETC) is a system within a cell that couples electron transfer between a donor and a receptor. The ETC is made up of several components, specifically: complexes one through four, a coenzyme Q, and a cytochrome complex. Cytochrome C oxidase (CcO), also known as complex IV, uses electrons and H_ ions to reduce molecular oxygen to water. CcO is made up of several subunits or proteins that are responsible for the function of CcO. The creation of CcO involves multiple steps that include many different gene products or proteins. (3) Of these proteins several are from a specific type of family called cysteine-X _-cysteine motifs (twin CX_C). The cysteine residues in the cysteine pairs are each spaced by nine residues. The best characterized protein of this group is Cox17, a copper-binding protein that plays a role in copper transfer to CcO. There are 14 potential proteins that are twin-Cx_C motifs (1). There were four knock out strains that showed resistance when plated on YPD with varying concentrations of hydrogen peroxide: 0.010%, 0.014%, 0.018%, and 0.022%. Strains lacking CMC4, MIC14, MIC17, and PET191 all showed resistance to hydrogen peroxide compared to the parental strain. Strains lacking MDM35 and COX23 showed sensitivity to hydrogen peroxide. These result may indicate a tie to impaired or incomplete cytochrome c oxidase assembly.
Effects of a Blood Fixing Agent on Bluestar Enhanced Blood Stains
Chelsie Greer, Weber State University Chemistry Luminol with its chemiluminescense properties can be used at crime scenes to develop and document bloodstains. By itself, luminol cannot be used because it destroys the blood stain pattern details. Using a fixing agent before applying luminol will chemically fix the detail of the bloodstain, preserving the pattern for analysis. 5-Sulfosalicylic acid (5-SSA) is used as a fixing agent and can be applied to bloodstains before the luminol to adhere the blood to the surface, but it interferes with the luminosity. We investigated the degradation effects of luminol and 5-SSA to develop a better method to use both chemicals together. Blood patterns were placed onto glass, systematically treated with a series of Bluestar (a commercial, luminol-based, blood detection reagent), 5-SSA and/or buffered rinse solutions then, photographed immediately. The luminol intensity was measured by processing digital photographs of the luminescent blood stain through Image J to examine the individual pixels of the photographs. The effect of the treatment on the pattern fastness was measured by making fingerprint impression in blood, and having fingerprint analyst score the treated prints on their second level detail quality. Results will be presented to show if an intermediate rinse between the application of the blood fixing reagent and the application of luminol can mitigate some of the negative effects of this procedure.
Characterizing Sky Variability for Multi-Messenger Astronomy
Rachel Nydegger, Utah State University Physics Multi-messenger astronomy employs both electromagnetic and gravitational-wave detectors to paint a richer picture of celestial objects, providing more depth and information. The interferometers utilized for gravitational-wave observations receive input from very broad fields of view on the sky, typically a few square degrees. To have simultaneous electromagnetic observations (typically less than one square degree) requires innovative techniques for the telescopes to find the origin of radiation. One idea is to “tile” the view of the interferometer, using multiple telescopes to simultaneously point at different areas of the field to observe the source. One difficulty of this observing paradigm is distinguishing random electromagnetic variable sources from a gravitational-wave counterpart. To better understand this problem, this project repeatedly observes a single field on the sky. Each observation is analyzed to count the number of sources that appear in the field as a function of brightness. Repeating this process over time will yield the frequency of random optical transients, as well as characterize the population and brightness distribution of variables in the field. Future work will extend this observation campaign to cover different galactic latitudes.
Space Travel with Ultrarelativistic
Jeremy Redd, Utah Valley University Physics Future space propulsion systems will likely use annihilation of matter and antimatter in propulsion system. Annihilation of matter and antimatter is not only the energy source of ultimate density 9×10^16 J/kg but also allows to utilize ultimate exhaust speed the speed of light c thus potentially allows to accelerate a payload to ultrarelativistic velocities. Such velocities make interstellar and even intergalactic travel possible in the lifetime of one generation only (20-30 years). In our presentation we discuss advantages and disadvantages of interstellar travel with relativistic and ultra-relativistic velocities. Using relativistic Tsiolkovsky rocket equation we also discuss the feasibility of achieving relativistic velocities with annihilation powered photon engine and technical challenges to build such engine.
Simulation of UV Radiation Degradation of Polymers on MISSE-6 in the Low Earth Orbit Environment
Kelby Peterson, Utah State University Physics The State of Utah Space Environment & Contamination Study (SUSpECS) experiment flown on the Materials International Space Station Experiment 6 (MISSE-6) was an experiment designed to examine the consequences of the space environment on various materials used in space-component design. SUSpECS was comprised of approximately 180 samples that were suspended from the side of the International Space Station (ISS) for 18 months and returned to allow for pre- and post-flight comparisons. The sample with the most evident changes was a thin film of polyethylene terephthalate (PET) MylarTM coated with Vapor Deposited Aluminum (VDA). The post-flight analysis showed evidence of atomic oxygen erosion of the VDA layer, UV-induced discoloration of the polymer, and a crater created by a micrometeoroid impact. This presentation focuses on the UV-induced discoloration and laboratory tests to simulate these effects. The UV tests expose similar polymers to varying intensities of vacuum UV radiation from deuterium lamps over a condensed time span and quantify the discoloration of the polymers through comparison of the UV/Vis/NIR reflection spectra. The results from the UV simulation are used to determine the approximate time period of the UV exposure for the SUSpECS sample and in turn the erosion rate of the VDA layer.
Less Cookbook and More Research: The Total Syntheses of JBIR-94 and JBIR-125, Student-Designed Research Projects Conducted in a Sophomore Organic Chemistry Lab
Cathy Crawford, Utah State University Chemistry and Biochemistry In the spirit of many chemistry instructors’ longstanding interest in making teaching labs less “cookbook-like” and more research-driven, we recently restructured our second-semester sophomore organic chemistry lab to include a synthesis project that was chosen, designed, and carried out by students. Students were given the incentive of co-authorship on any publications resulting from their work. This led to the development of total syntheses of JBIR-94 and JBIR-125, new antioxidative/anticancer compounds with radical-scavenging potencies comparable to those of a-tocopherol, the active constituent in Vitamin E. Our presentation will summarize our progress and findings, and includes our progress on bioactivity studies conducted on the JBIR’s and their synthetic precursors.
Dissonant Modes of Bottle-shaped Thermoacoustic Prime Movers Part 2: Hysteresis of Mode Transitions
David Pease, Utah Valley University Physics Transition regions to higher resonant modes of a bottle-shaped thermoacoustic prime mover (neck: 5.39 cm long, 1.91 cm ID; variable cavity with a sliding piston: up to 38 cm long, 4.76 ID) were studied. As the piston is extended, lengthening the cavity, starting from the neck, a transition of the dominant frequency from the fundamental to the first overtone occurs. However, when the length is then shortened, transition back to the first mode does occur at the same piston position, revealing hysteresis. Within the window of hysteresis for the cavity length, either state of the fundamental or first overtone is possible. Transition regions to higher modes continue as the length of the cavity is increased. The position and width of the hysteresis was studied for the first two transition regions as a function of input power and stack volume filling factor. Input powers studied were between 12.0 and 16.5 W and volume filling factors for the stack were about 3.0, 3.7 and 4.9%. The transition regions occurred with cavity lengths between 12.6 and 14.0 cm for the first transition and between 25.0 and 27.8 cm for the second transition. Preliminary results indicate that the transition region occurs shallower in the cavity and the hysteresis widens as the input power is increased. The hysteresis is wider for the second transition region. Decreasing the stack mass causes an increase of the hysteresis width, but has no strong effect on the hysteresis depth.