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Antimicrobial Properties of Essential Oils Isolated from Anthoxanthum hirtum and A. odoratum Against Soil Bacteria

January 01, 2013 12:00 AM
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.

Analysis of Dental Parameters, Fluoride and pH, in Utah’s Waters, Drinks, and Foods

January 01, 2013 12:00 AM
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.

Chytridiomycosis-resistant Frog Populations in Southern Utah

January 01, 2013 12:00 AM
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.

Sirt1-Mediated Suprression of Cell Death in Breast Cancer

January 01, 2013 12:00 AM
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.

GPR55: A Potential Enhancer of Learning and Memory in the Hippocampus

January 01, 2013 12:00 AM
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.

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

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

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

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

Engineering Pathogen Specific High Affinity T-Cell Receptors

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

Endocannabinoid Biosynthesizing Enzyme Expression in Hippocampal Stratum Oriens Neurons

January 01, 2013 12:00 AM
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.

The Effects of DNA Methylation on Nucleosome Positioning

January 01, 2013 12:00 AM
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.

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

January 01, 2013 12:00 AM
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

January 01, 2013 12:00 AM
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 Effects of Temperature and Water Availability on the Germination of Bromus Rubens

January 01, 2013 12:00 AM
Rachel Nettles, Brigham Young University Plant and Wildlife Sciences Background/Questions/Methods

Subduing the Flu: New Alternatives to Amantadine

January 01, 2013 12:00 AM
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.

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

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

Baicalein and Light Stimulation as Clinical Therapies for Addiction

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

Fire Retardant as an Environmental Risk Factor Contributing to Parkinson’s Disease

January 01, 2013 12:00 AM
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.

Anodic Stripping Voltammetric Analysis of Lead, Cadmium, and Copper in the Jordan River, Utah

January 01, 2013 12:00 AM
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.

Analysis of the Degradation of Flavor Volatiles in Single Origin High Cacao

January 01, 2013 12:00 AM
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.

Adaptation of Staphylococcus Aureus to UV-C Light

January 01, 2013 12:00 AM
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.

Validation of Cache County Genotype Data

January 01, 2013 12:00 AM
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.

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

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

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

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

Facilitative and Competitive Interactions in Subalpine Aspen-Fir Forests

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

Implicit Bias in Early Healthcare Education

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Author(s): Claire Parker, David Jensen

Analyzing the estrus cycle in POMC-deficient mice

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Author(s): Mariah McDonald, Kaden Smith

L-DOPA, naloxone, and DOI decrease nicotine preference in C. elegans

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Author(s): Emily Mellon, Violet Czech, Kirsten Sumampong, Hailey Kim

Antimicrobial Activity of Cetaphil

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Authors: Lillian Gee, Makaylie Moore. Mentors: Lauren Brooks. Insitution: Utah Valley University. Cetaphil is a daily facial cleanser that claims to remove dirt, excess oils, and makeup. The human skin is home to a diverse community of bacteria, including beneficial bacteria that play a role in the skin's natural barrier function and immune defense, and pathogenic bacteria that may cause disease and infection. There is little research on the effectiveness of Cetaphil removing harmful bacteria such as Staphylococcus aureus, and commensal bacteria, like Staphylococcus epidermis. To compare the antimicrobial activity of Cetaphil on these two species, serial dilutions of both bacteria strains were made and then exposed to Cetaphil. Positive controls and the dilutions exposed to Cetaphil were plated on Tryptic Soy Agar plates and after incubation, bacterial growth was observed by counting the number of colony-forming units. Testing is beginning to show that Cetaphil is not only effective against S. aureus but is also effective against S. epidermis. This research is important for understanding how skincare affects harmful bacteria strains and the bacteria strains that are natural to the skin.