Life Sciences

Ashtyn Smith, Westminster College Life Sciences Methylmercury (CH3Hg) is a neurotoxin that accumulates in lakes and streams due to the action of microorganisms, which can produce this biologically relevant organic form from elemental mercury (Hg). Therefore, the activities of microorganisms become key to understanding the balance of Hg and CH3Hg in the movement through the food chain in any ecosystem. Many species of microorganisms are resistant to Hg and can thrive in polluted waters. Recent studies have shown that Hg resistance in microbes can stem from one of two gene pairs, merAB or hgcAB. The merAB system allows the organism to covert CH3Hg into elemental Hg. Conversely, the hgcAB system coverts Hg into CH3Hg. Thus, it is important to determine how the microbial community of Great Salt Lake, Utah is affecting the CH3Hg concentrations in the lake. In order to determine the genotype of the lake’s halophiles, “salt-loving” organisms, microorganisms were collected from the deep brine layer in eight areas of the lake. The microorganisms were then isolated and cultivated on increasing concentrations of HgCl2. Halophiles from these samples have been isolated on 25 ppm HgCl2 at various salinities, demonstrating a robust resistance to Hg. PCR amplification and genetic sequencing will be used to determine the gene mechanism of mercury resistance (merAB or hgcAB) as well as the 16S rRNA gene, which will aid in identification of the species. Should this study identify GSL microorganisms that exhibit the merAB genotype, these organisms could potentially be utilized as bioremediators of the CH3Hg pollution in the lake.

April Moorer, Weber State University Life Sciences Corexit is a dispersant used in the gulf of Mexico as a reactive measure to counteract the oil spill of April of 2010. Studies reveal that toxicity is produced and has impacted marine life. Research shows that reproductivity is diminished as a consequence. Paralysis, tumor development, and fatalities are also proven to occur. The purpose of this experiment is to study effects on the physiological structure of brine shrimp at various life stages resulting from the exposure to toxicity induced by corexit.

J Andrew Chappell, Utah Valley University Life Sciences In a joint effort with Huntsman Cancer Institute at the University of Utah, students from Utah Valley University are using high-frequency (HF) ultrasound to test the pathology of surgical margins from breast cancer conservation surgery. The method, developed by Dr. Timothy E. Doyle, provided significant results in a NIH-funded 2010 feasibility study. The results of the study indicated that peak density, the number of peaks and valleys in the HF ultrasonic spectrum, correlates to breast tissue pathology. This technology would allow surgeons to test – in the operating room – whether a surgical margin was clean or if cancer still remained in the margin. This advancement would decrease the amount of return surgical visits a patient must undergo, reduce costs for patients and hospitals, reduce breast cancer recurrence rates, and ultimately increase the survival rate of patients with breast cancer. During the ultrasonic testing, the students work in a team of four in a room outside of the surgical suite. Specimens are brought in by the surgeons’ team and tested immediately following resection. The margins are approximately 3x20x20 mm in size, and are oriented using a small staple inserted by the surgeon in one corner and a stitch on one side. The margin is tested at specific locations depending on the size of the margin and then sent to pathology for analyses. Pathological results and HF ultrasound results will be compared for correlation at the end of the study, which is expected to last about one year. The study will include approximately 80 patients, 360 tissue samples, 1400 tested locations, and 4,300 data points. The goal of the study is to evaluate the accuracy of the method in determining margin pathology. If successful, the method will be moved into clinical trial.

Bryan D Merrill, Brigham Young University Life Sciences The spore-forming bacteria Paenibacillus larvae causes American Foulbrood (AFB), a highly contagious disease that is lethal in honey bee larvae. P. larvae is the most serious pathogen affecting honey bees. Its increasing antibiotic resistance has led to more research in characterizing bacteriophages which infect and destroy P. larvae. Of the 13 P. larvae phages that have been described in publications, six have been fully sequenced and are currently available for genomic analysis. To understand how P. larvae phages are related to each other and to other phages, the computer program Phamerator was used to analyze more than 100 phage genomes and group phage genes into “phams” based on similarity. Through analysis of grouped phage genes (structural proteins, terminases, recombinases, etc.) we can better understand in vivo replication strategies and evolutionary history of these P. larvae phages.

Hailey Shepherd, Dixie State University Life Sciences Previous studies of canyon tree frogs (Hyla arenicolor) in Zion National Park have shown that some populations test positive for a dangerous fungus in the Chytridiomycosis family. This fungus has been linked to large population losses worldwide in many keystone amphibian species, but appears to have no effect on populations of H. arenicolor. Since Chytrid fungal growth is inhibited at high temperatures, we hypothesized the frogs are able to rid themselves of the fungus because they bask in the sun. During the summer of 2013 we swabbed frogs in multiple slot canyons to test for the presence of the fungus. We also recorded skin temperatures of the frogs we swabbed. Skin temperatures were as high as 38 C°, which is above the previously established lethal threshold of 28 C° for Chytridiomycosis. Our data support the idea that these frogs may be able to rid themselves of infection by allowing skin temperatures to raise enough to become intolerable for this fungus. This is the first known evidence of wild amphibian populations showing a behavior that may clear the infection. If further research supports these findings, it could lead to more effective allocation of limited conservation resources.

Danielle Holliday, University of Utah Life Sciences Intrauterine growth restriction (IUGR) induces visceral obesity in adulthood, specifically among males. In male rat offspring, IUGR increases visceral adipose tissue (VAT) over subcutaneous adipose tissue (SAT). VAT and SAT functions are regulated by estrogen signaling, and suppressed estrogen signaling contributes to obesity development. Estrogen signaling is composed of estradiol and estrogen receptor alpha (ERα) and beta (ERβ). Estrogen receptors regulate the expression of several obesity related genes, such as lipoprotein lipase (LPL). However, the effects of IUGR on estrogen serum levels and signaling in the adipose tissue are unknown.

Camrin Rivera, Westminster College Life Sciences The Great Salt Lake (GSL) of Utah is home to organisms that are able to survive and reproduce in waters that can have a salt concentration of over 30%. Oil Jetty is located on the saltier north arm of the GSL and is the location of a natural hydrocarbon seep that flows into the lake. Previous work showed that organisms capable of degrading hydrocarbons can be cultured from this area (Ward & Brcok, 1978). It has also been shown that the salt concentration has an inverse relationship with these organisms’ ability to degrade hydrocarbons (Ward & Brock, 1978). This previous study gives insight of the hydrocarbon degrading capabilities of microorganisms in the GSL; however, the identification and characterization of individual hydrocarbon-degrading microorganisms has yet to be completed. The purpose of this study is to identify GSL microorganisms capable of degrading hydrocarbons under high salinities by sequencing of the 16s rRNA gene. Future studies will include the characterization of these microorganisms in order to find isolates whose hydrocarbon degrading abilities could be utilized for bioremediation and biotechnologies.

Cameron Haas, Brigham Young University Life Sciences The influenza A virus contains a proton-selective ion channel, A/M2, through which acidification of the cell is induced. A/M2 is a homotetramer (consists of four identical helices) consisting of 97 residues and activated by low pH levels. Mutations in the amino acid sequences may induce resistance to channel inhibiting drugs. It is believed that residues 26, 27, 30, 31, and 34 are the major contributors of drug resistance, but other nearby residues may prove important as well. The A/California/04/2009 version of the influenza virus is sensitive to the drug AK-11, while its M2 channel is not. The A/Udorn/307/1972 with the S31N mutation M2 channel has been shown to have reduced sensitivity to amantadine compared to its wild type. While both contain a D (aspartic acid) at residue 21, A/Puerto Rico/8/1934 has a mutation from D to G (glycine). The A/Puerto Rico/8/1934 virus A/M2 contains mutations S31N and V27T and has shown sensitivity to the AK-11 drug, but the mechanism of inhibition of the A/M2 channel has not been verified. In these experiments we will be identifying sensitivity to AK11 of A/Udorn/307/1972 with the S31N mutation as well as inducing double mutations with S31N at residues 27 and 21 in the A/M2 from the virus and measure sensitivity by electrophysiological recordings in oocytes of Xenopus laevis. By doing so we may identify the role of these residues in drug resistance and the effects of these amino acid mutations, while verifying the A/M2 channel as the mechanism of acidification inhibition and drug sensitivity. We hypothesize that either D21G, V27T or both mutations causes drug sensitivity in M2 S31N, explaining the sensitivity of A/Puerto Rico/8/1934 to AK-11.

Ashley Nelson, Brigham Young University Life Sciences The motivational effects of opiates and ethanol switch from a dopamine (DA)-independent to a DA-dependent pathway when the animal is in a drug-dependent state. A corresponding change occurs in ventral tegmental area (VTA) GABA(A) receptors in opiate-dependent animals, which switch from a GABA-induced hyperpolarization of VTA GABA neurons to a GABA-induced depolarization. The aim of this study was to evaluate VTA GABA neuron excitability, GABA synaptic transmission to VTA GABA neurons and GABA-mediated DA release in the nucleus accumbens (NAc) under ethanol-naïve and dependent conditions. To accomplish these studies, we used standard whole-cell and attached-cell mode electrophysiological techniques to evaluate acute and chronic ethanol effects on VTA GABA neurons in GAD GFP mice, which enabled the visual identification of GABA neurons in slice preparation. In naïve animals, superfusion of ethanol (IC50 = 30 mM) and GABA(A) receptor agonist muscimol (IC50 = 100 nM) decreased VTA GABA neuron firing rate in a dose-dependent manner. Compared to saline-injected controls, in animals made dependent on ethanol by twice daily injections of 2.0 g/kg ethanol, neither ethanol nor muscimol significantly affected VTA GABA neuron firing rate on average. We and others have found that ethanol decreases DA release at terminals, as measured by fast scan cyclic voltammetry. We have recently reported that ethanol inhibition of DA release at terminals in the NAc of ethanol-naïve animals is mediated by GABA, possibly from VTA GABA neurons that project to the NAc. We evaluated the effects of ethanol on DA release in the same ethanol-dependent animals. Compared to controls, superfusion of ethanol did not significantly affect DA release. Together, these findings suggest that VTA GABA neurons undergo a switch in GABA(A) receptor function with chronic ethanol, which results in a corresponding switch in DA release, perhaps resulting from adaptations in VTA GABA neuron input to the NAc.

Taylor Woodward, Brigham Young University Life Sciences The prevailing view is that enhancement of dopamine (DA) transmission in the mesocorticolimbic system, consisting of DA neurons in the ventral tegmental area (VTA) that innervate the nucleus accumbens (NAc), underlies the rewarding properties of alcohol and nicotine (NIC). Dopamine neurotransmission is regulated by inhibitory VTA GABA neurons. We have shown previously that VTA GABA neurons are excited by low-dose ethanol, but inhibited by moderate to high-dose ethanol. The aim of this study was to evaluate the role of []6 nicotinic cholinergic receptors (nAChRs) in ethanol effects on VTA GABA neurons as well as DA release in the NAc. In electrophysiology studies, superfusion of ethanol enhanced the frequency, but not amplitude, of mIPSCs recorded in acutely dissociated VTA GABA neurons from GAD GFP mice. The []6 nAChR antagonist []-conotoxin P1A did not affect mIPSCs, but prevented the ethanol (30 mM)-induced increase in mIPSC frequency. While microdialysis studies show that ethanol enhances DA release in the NAc, we and others have found that ethanol decreases DA release at terminals using fast scan cyclic voltammetry (FSCV). We have reported that ethanol inhibition of DA release at terminals in the NAc of ethanol-naïve animals is mediated by GABA. Using FSCV in the slice preparation, ethanol inhibited DA release in the NAc. Superfusion of the []6 nAChR antagonist []-conotoxin MII did not affect DA release, but prevented ethanol inhibition of DA release. Taken together, these findings suggest that ethanol enhancement of GABA inhibition of VTA GABA neurons is mediated by []6 nAChRs located on GABA terminals to other VTA neurons, affecting DA release in the NAc. Results from this study could provide a pharmacologic rationale for considering drugs that act selectively on nAChRs as therapeutic agents for the treatment of alcohol dependence and alcohol and NIC co-dependence.

Chelsea Moody, Dixie State University Life Sciences A relatively new pathogen, chytrid fungus, has been a major cause of decline in amphibians worldwide. This pathogen has been found on Canyon Tree Frogs in Zion National Park. Earlier data suggested that chytrid did not affect populations of Canyon Tree Frogs in Zion National Park. We predicted that since the populations weren’t impacted by the presence of the fungus, there must exist a mechanism by which the frogs rid themselves of the infective agent. To test this prediction, we captured and swabbed ten frogs in each of nine canyons in Zion National Park during the summer of 2013. Swabs were sent to the San Diego Zoo to be analyzed for the presence of chytrid DNA. Chytrid infection rates and population sizes were compared with data from previous years. As expected, the data showed that two of the three infected canyons experienced declines in the percentage of frogs testing positive for the fungus. Thus, these frogs must possess a mechanism to survive this pathogen, whereas most amphibians do not. Our results may prove helpful to other biologists seeking to understand how amphibians can survive this pathogen.

Whitney Badal, Brigham Young University Life Sciences Light therapy has been utilized to treat alcoholism and opiate-dependent rats as well as ameliorating symptoms of Parkinson’s disease. As both addiction and Parkinson’s Disease (PD) are often associated with decreased dopamine transmission in the striatum, it is likely that light therapy is able to increase dopamine release. A similar technique called near-infrared light treatment has also been shown to be effective in mice in restoring the function of dopamine cells in the substantia nigra pars compacta (an area associated with PD). A possible explanation for this is that light catalyzes the formation of neuromelanin. It is likely that neuromelanin is a neuroprotective cellular agent that is able to reduce damage caused by reactive oxygen species. Using UV-IR spectrophotometry, we show that in the presence of photostimulation, dopamine (0.3-30 uM) oxidizes and polymerizes into neuromelanin. Since hydrogen peroxide catalyzes this formation of neuromelanin, it is likely that this is a radical-polymerization reaction, suggesting that neuromelanin may be a radical scavenger. Additionally, the presence of the selective iron chelator desferrioxamine, the calcium chelator EGTA, or lack of calcium in the artificial cerebral spinal fluid markedly reduces the formation of neuromelanin. Using fast scan cyclic voltammetry in mouse horizontal and/or coronal brain slices, dopamine release in the nucleus accumbens core was enhanced by light exposure, in particular UV and short-wavelength visible light. These findings indicate that both iron and calcium are necessary for melanization in neural tissues and that light-induced melanization enhances dopamine release, suggesting a physiological role for melanization in synaptic transmission.

Corey Wolf, Westminster College Life Sciences The yeast species Saccharomyces cerevisiae is used in numerous industries including brewing, baking, and winemaking. In recent years an increased demand for low ethanol beers has pushed breweries to develop a beer that is both rich in flavor and low in alcohol content. The current strategies employed to decrease the ethanol concentration in beer are costly, time intensive, and result in less flavorful beers. In this study, a method for decreasing alcohol content in the brewing process through the use of metabolic inhibitors of Alcohol Dehydrogenase is tested. Alcohol Dehydrogenase is the enzyme responsible for the conversion of acetaldehyde, an intermediate in the metabolic pathway of yeast, into ethanol. Two competitive inhibitors of Alcohol Dehydrogenase, Pyrazole and Fomepizole, were applied to small scale fermentations in varying concentrations in an attempt to decrease the overall ethanol yield of the beer. The application of either inhibitor resulted in an initial decreased rate of fermentation overall, measured as a change of sugar and ethanol concentration. In the later stages of fermentation, the rate of ethanol production returned to normal, suggesting the inhibitors were either catalyzed or removed by the cell. While this method for manipulating the products of fermentation did not yield the desired effects, this study did highlight the importance of the fermentation pathway for maintaing healthy Saccharomyces cerevisiae cultures. This research may be helpful in understanding the complete metabolic pathway of brewer’s yeast, with further application to organisms with conserved pathways.

Marilize Van der Walt, Utah State University Life Sciences Urbanization is accompanied by a lot of changes to the landscape that have the potential to affect the native species inhabiting the area. If animals are chronically exposed to these anthropogenic disturbances and are unable to acclimatize, changes in circulating glucocorticoid hormones may cause adverse effects to the animal’s health, such as an impairment in innate immune activity. One such disturbance is human-induced noise. Using the side-blotched lizard, Uta stansburiana, because of their localized habitat and inability to escape such disturbances within the urban environment, we looked at the field and lab components of noise as a stressor. We first measured decibel levels in urban and rural field sites in their natural habitats in St. George, Utah, and conducted a lab study exposing U. stansburiana to either a synthesized urban sound recording or no sound recording (control) for nine days. We collected blood samples and measured circulating corticosterone and testosterone concentrations and bactericidal ability to determine if there are endocrine and immune alterations in response to increased noise decibels. Our results show that lizards exposed to urban noise experienced increased corticosterone levels indicative of stress. Furthermore, bactericidal ability was indirectly affected by noise through significant correlations between body condition and corticosterone and testosterone concentrations. These results indicate that an increase in ambient decibel levels acts as a stressor to animals in urban areas. By dissecting out an individual component of a complex stressor we can better understand the effects of urbanization as a whole.

Austin Spence, Utah State University Life Sciences This study investigated the physiological effects of localized habitat disturbances around two well-studied hibernacula of the wandering gartersnake Thamnophis elegans. After three years of monitoring snake morphology and physiology at several hibernacula, several disturbance events occurred, including log clearing, stream bank disturbance, and vegetation removal. Individuals from three populations, two with disturbed hibernacula and one control population with no disturbance, were collected during the spring emergence immediately following the disturbance. Blood samples were collected upon capture and following a uniform stressor to measure baseline and post-stress physiological conditions. The samples were analyzed using a radioimmunoassay to measure corticosterone levels and a bacterial killing assay to measure innate immunocompetence. Baseline and post-stress corticosterone levels were higher in both populations with disturbance events compared to the control population. The bacterial killing ability of the site with the most anthropogenic activity was lower than the control site, indicating immunocompromise. Data are currently being analyzed to assess differences within the same populations between years with and without disturbance events. Pre-disturbance data are a rare and useful commodity and allow us to facilitate a better understanding of the various effects of anthropogenic change on natural populations. This study was funded through the Undergraduate Research and Creative Opportunity Grant from Utah State University.

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

Daniel Hall, Brigham Young University Life Sciences OBJECTIVE(S): Predictive analytics (PA) is increasingly being used in the delivery of healthcare. Whether PA can improve patient handoffs on a busy surgical service is unknown. This study aims to determine if predictive models for acute cellular rejection (ACR) episodes and biliary complications after orthotropic liver transplantation (OLT) can be built in order to improve patient care.

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

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

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

Claudia Gonzalez, Brigham Young University Life Sciences Cortisol has been shown to be a potential bio-marker as it discriminates between individuals with and without depression (Rush et al., 1996 and Ising et al., 2007). However, cortisol has not been used to predict variation in temperament extremes that lead to pathological behaviors in adulthood. In order to examine the relationship between cortisol and temperament extremes, data from the bio-behavioral assessment (BBA) was used. The BBA data base includes data collected from over 2,700 infant rhesus macaques located in California National Primate Research Center (CNPRC). During the BBA four blood samples per subject are obtained and later assayed for plasma cortisol levels. In this study, the plasma cortisol response levels were looked at in a holistic form encompassing all individual cortisol samples. The four points of plasma cortisol concentrations were used to extract patterns of response per subject which provided classifications for each of the monkeys. The pathological patterns of cortisol response were characterized by abnormal plasma cortisol levels in response to Dexamethasone suppression testing and adrenocorticotropin ACTH injections. The variability in plasma cortisol patterns was then compared to BBA temperament ratings of vigilance, gentle, nervousness and confidence. These results showed that 12 of the 26 possible patterns of response were significantly (p<.05) related to each of the temperament ratings of vigilance, gentleness and confidence. Thus cortisol response patterns can be used both as biomarkers for vigilance, gentleness and confidence, and as potential predictors for pathological behaviors in adulthood.

Mena Davidson, Westminster College Life Sciences Urban stream syndrome is the phenomenon of stream degradation as streams run from their sources through urban areas, which is exhibited by nutrient loading, decreased dissolved oxygen, changes in channel structure, and increased turbidity and rapid flow events. This can have a direct negative effect on our recreation, drinking water, and the ecosystems surrounding the streams. To discover if urban stream syndrome occurs in the Salt Lake Valley, we monitored pH, turbidity, dissolved oxygen, dissolved nitrates, and collected macroinvertebrates in three streams monthly. We analyzed total abundance, species diversity, and percent pollution sensitive species to assess the macroinvertebrate communities, as known indicators of water quality. Preliminary data shows a significant decrease in percent sensitive species in the urban areas as compared to the nonurban areas over 10 sampling periods. We found decreased biodiversity and fewer sensitive species in urban areas, although we found no difference in total abundance. These findings indicate that urbanization in the Salt Lake Valley leads to degradation in riparian ecosystems and suggest that further investigation is needed to identify the mechanisms leading to this degradation.

Michaelle Cadet, Utah Valley University Life Sciences Utah County, Utah has an estimated population of 540,000 residents and is considered to be a non-attainment area for criteria pollutants such as PM-10 and CO. High levels of these contaminants may increase the risk of respiratory diseases. Additional environmental issues exist including water contamination and eutrophication of Utah Lake. These environmental issues are frequently on the news and warnings are issued by the Department of Environmental Quality notifying citizens of the potential health concerns associated with environmental pollution. With these announcements, it is expected that Utahans are acutely aware of environmental issues, particularly, in the academic settings. The purpose of this study was to investigate the status of environmental awareness among students at Utah Valley University and to correlate the results to socio-demographic categories. Utah Valley University, located in Utah County, is a public institution of higher education with approximately 30,000 registered students. Data for this study was collected through a survey using a multistage sampling technique with population stratified based on colleges and schools within the university. Sample size included approximately 1,000 students. The survey contained 12 environmental questions relating to recent local, national and international media exposure. Standards and protocol of the International Research Board were employed. We hypothesize that married, educated, religious males will positively correlate with higher levels of environmental awareness, but will be less than atheists. Additionally, those identifying with the Republican Party will have lower levels of awareness. No significant difference will be found between majors. Furthermore the young, poor and ethnic populations will be less aware than their more affluent counterparts. Results of this study will be used to inform residents of environmental issues and the associated health concerns. Additionally, this study will be used to inform legislators about the importance of environmental education in the community.

Robert Erickson, Utah Valley University Life Sciences Although the insect fauna of the Colorado Plateau region are somewhat well known, our specific understanding of the arthropod biodiversity in Capitol Reef National Park is sparse.

Jordan Fenlon, University of Utah Life Sciences Tsr, the serine chemoreceptor for E. coli, is a transmembrane protein with a periplasmic sensing domain and cytoplasmic adaptation and kinase control domains. The focus of my research project is Tsr residue A413, located in the cytoplasmic tip of the receptor’s kinase control domain. The project involves characterization of mutant Tsr proteins containing amino acid replacements at residue 413. Based on previous work in the Parkinson lab, this residue is thought to play a key role in Tsr signaling by regulating the dynamic motion of the tip.

Michael Gillespie, Brigham Young University Life Sciences It is vital to understand the anatomical microstructure of tendons and ligaments in order to ascertain their specific qualities and functions. Recent developments in micro-scribe 3D digitization are highly effective in revealing these intricacies. A necessary component to creating 3D fiber maps from this technology is the ability to distinguish between individual tissue fibers with the naked eye. However, this is a very difficult task with most tendons and ligaments. We developed a paste made of blue dye and powdered sugar that when applied, fits in-between these fibers and contrasts the specimen color. This exposes the fine architecture, making individual fibers much more visible and thus able to be 3D digitized. With these 3D fiber maps now available, tendon and ligament microstructure can be viewed in greater detail than previously possible. This technique was applied to human cadaveric calcaneal tendon and quadriceps tendon. It was discovered that the fibers of human calcaneal tendon have higher overlap relative to the rigidly parallel fibers of the quadriceps tendon. This further understanding carries implications regarding the advancement of biomechanical models, artificial reconstruction, and surgical repair of these tissues. It also highlights the need for further investigation into the microstructure differences among tendons and ligaments.

David Vogelsang, Brigham Young University Life Sciences More than one-third of U.S. adults (35.7%) are obese (CDC, 2013a), and since 1980, obesity among adolescents has risen from 5% to 18% (CDC, 2013b). Unless we do something to combat the growing obesity epidemic, we are consigning ourselves and future generations to a lifetime of heart disease, diabetes, cancer, and psychological distress (CDC, 2013a). Cooking Anatomy Academy (CAA) promotes healthy eating and cooking among parents and students in the Polynesian community to raise awareness about the growing obesity epidemic. Our primary focus is introducing parents and students to healthy, great tasting foods and easy to prepare meals (Brown, 2011). Our secondary focus is to teach the simple anatomy and physiology important to understanding healthy nutrition. CAA is composed of seven, one hour lessons that are being offered as an afterschool program at Mana Academy Charter School. We’ve developed the CAA curriculum to incorporate many of the nutrition guidelines on MyPlate.gov, and focus on moderation, variety and raw/unrefined foods. To study the impact of Cooking Anatomy Academy, we will take a qualitative approach and use journal entries to collect data. Participant journal entries will answer prompts like, “based on what you learned today, what will you have for a snack tomorrow,” or, “how many fruit servings should you have each day?” CAA mentors will record any positive or negative feedback they receive during each lesson. From the data we collect, we hope to see that CAA is helping participants make healthy food choices and increasing their obesity awareness.

Lance Deeter, University of Utah Life Sciences Continuous-flow left ventricular assist devices (LVADs) are used in advanced heart failure patients either to bridge them to transplantation or as a permanent-destination therapy. We determined whether chronic exposure to non-pulsatile blood flow and acute increases in coronary perfusion pressure associated with LVAD implantation would influence arterial function. Arteries from a transmural biopsy of the left-ventricle were obtained from ten male patients (54±4 years old) at the time of LVAD implant (n=17, 184±25 µm i.d.) and 239±51 days later upon LVAD explant (n=21, 281±22 µm i.d.). Lmax tension was determined and dose-response curves to potassium chloride (KCl, 10-100 mM) were performed using isometric tension techniques. Next, bradykinin (BK, 10-6 to 10-10 M) and sodium nitroprusside (SNP, 10-4 to 10-9 M) concentration-response curves were completed on vessels precontracted to ~65% of maximal tension development. Maximal BK-induced vasorelaxation was greater (p<0.05) at explant (85±5%) vs. implant (59±9%), while SNP evoked responses (~90%) were similar between time-points. These findings suggest coronary endothelial function is improved by LVAD implantation. Heart failure was precipitated by a myocardial infarction in six of the ten patients. These are referred to as “ischemic” patients whereas the remaining four are “non-ischemic” patients. We hypothesized that coronary vascular responses would be improved by LVAD implantation to a greater extent in ischemic vs. non-ischemic patients. In ischemic patients maximal BK-induced vasorelaxation was greater (p<0.05) in coronary arteries obtained at explant (87±6%, n=14, 305±30 μm i.d.) vs. implant (53±11%, n=12, 204±33 μm i.d.). In non-ischemic patients maximal BK-induced vasorelaxation was similar in arteries obtained at explant (79±9%, n=7, 232±21 μm) and implant (72±17%, n=5, 135±13 μm). SNP responses were similar (~90%) between groups at implant and explant. Collectively, our data suggest that LVAD implantation improves endothelium-dependent vasorelaxation in ischemic but not in non-ischemic patients.

Josh Harvey, Brigham Young University Life Sciences Soil CO2 flux represents an important pathway of carbon transfer from ecosystems to the atmosphere. Soil CO2 flux can be altered by global warming-driven changes in seasonal temperature and water availability. Subalpine ecosystems have high levels of carbon in their soils that are stabilized by low temperatures and low microbial activity during long and snowy winter seasons. Subalpine ecosystems can be important sinks for carbon, storing carbon that otherwise would be in the atmosphere contributing to global warming. In our study we show how changes in temperature and water availability during springtime increase the levels of subalpine carbon output. So long as the carbon outputs outweigh carbon inputs, increases in soil flux would amplify global warming. The amplification of global warming would loop back to affect soil fluxes again (by raising temperatures, melting snow earlier, and changing precipitation patterns) thus creating a positive feedback system. Understanding what feedbacks are present in a climate system and their underlying mechanisms will improve our forecasts of changes in atmosphere chemistry and temperature.

Brittany Hammontree, Dixie State University Life Sciences Depending on metabolic conditions or dietary preferences, people can often become deficient in critical vitamins and minerals. For example, a number of people are deciding to become vegetarians, and vitamin B12 deficiencies could become a huge epidemic, as this essential vitamin is only obtained through meat products. This issue was the driving force to look deeper into new ionic liquid materials and how they could be used as a transport agent for vitamin B12, along with other vitamins. Ionic liquids are organic salts that are currently being explored in many scientific fields due to their unique properties. However, using ionic liquids as a transporter in transdermal applications has yet to be explored Developing new mechanisms of administering nutrients via transdermal processes can increase the bioavailability and effectiveness of vitamins and minerals that often cannot survive oral administering due to the acidity and molecular absorption via the stomach. This research focused on finding the right ionic liquid with high solubility of the individual vitamins. Several ionic liquids were developed, and the different vitamins were tested for solubility levels. This greater solubility allows for maximum exposure of the vitamin during transdermal delivery. In particular, two different vitamins were tested – vitamin K and Protoporphyrin, a chemical analog to vitamin B12. Additionally, the effect of these ionic liquids on the physiology of the blood and plasma as it enters the body past the skin layers is critical to understand. In addition to transdermal applications of vitamins, transport of these vitamins to other countries and remote locations could have tremendous implications. Ionic liquids tend to increase shelf life of solutes, and the availability to provide these materials during medical missions or service trips would be increased substantially, particularly in more remote settings.

Daniel Loveland, Brigham Young University Life Sciences The monoamine oxidase A (MAOa) gene has been shown to be associated with various social behaviors and disorders such as: aggression, depression, and anxiety (Meyer et al., 2006; Kinnally et al., 2010; Newman et al., 2005); and the MAOa gene interacts with environmental influences to produce its phenotypic effects (Newman et al., 2005; Kinnally et al., 2010). The MAOa gene encodes the enzyme monoamine oxidase A, which is the main enzyme to break down the monoamines into their respective metabolites. An orthologous repeat variant of the MAOa genotype seen in humans has been found in the rhesus macaque: a 5 repeat (R), a 6R and a 7R. This study investigates the influence MAOa genotypes have on central monoamine functioning as measured by cisternal cerebrospinal fluid (CSF) monoamine metabolites associated with behavioral dysfunction (dopamine metabolite: homovanillic acid-HVA, norepinephrine metabolite: 3-methoxy-4-hydroxyphenylgycol-MHPG, and serotonin metabolite: 5-hydroxyindoleacetic acid-5-HIAA). Cisternal CSF was obtained from 136 30-day old infant male rhesus macaques with varying genotypes and rearing backgrounds. We expected to find a rearing by genotype (GxE) effect on the monoamine systems with differences between mother-reared subjects when compared to subjects reared without mothers in peer-only groups. We found significant variability between genotypes; results also showed early rearing modulated this genotypic effect on brain chemistry. This supports our hypothesis that GxE interactions influence monoamine metabolite concentrations, suggesting a possible relationship of GxE interactions on social disorders such as aggression, depression and anxiety.

Derek Harris, Dixie State University Life Sciences The presence of extracellular DNA (eDNA) in various environmental and biological media has become the subject of growing interest in the field of research. In media such as bacterial biofilms, it has been shown to play a vital role in their structure and antimicrobial properties. Existing methods for extraction of pure eDNA from these media are complex and problematic; particularly from biological media where cells containing genomic DNA are also present. Novel surfactants have been developed, whose miscibility and polarity are easily tuned to suit a variety of conditions necessary for eDNA extractions. They can accomplish extraction of pure eDNA through concurrent hydrophilic and hydrophobic interactions in a single step, while remaining unreactive with the surrounding media or lysing cells and exposing genomic DNA. We have shown by spectrophotometric quantification that these surfactants extract measurable amounts of DNA into a water-immiscible solvent layer, which can then be removed from the media. The DNA can then be further amplified and purified for analysis. Further refinement of extraction methods utilizing these surfactants could prove a tremendous asset to research attempting to elucidate the possible genetic content of eDNA and the mechanisms behind its often crucial role in environmental and biological media.

Dusting Day, Brigham Young University Life Science Atherosclerotic vascular disease is the leading cause of morbidity and death in the United States. Approximately 1.4 million surgical procedures are required every year for treatment of vascular disease and its subsequent issues. While saphenous vein and internal mammary artery grafts are most commonly chosen by physicians, many patients who are in need of arterial grafts have vessels that are not ideal for grafting because of damage to the vessels or disease. This introduces the necessity for synthetic blood vessel grafts that function precisely as natural vessels in vivo. Our blood vessel research team has entered the tissue engineering field in its most exciting effort: the scalable rendering of cell-seeded vascular constructs with rapid prototyping machines or 3D printers. We have built and are modifying a 3D printer to deposit living endothelial and smooth muscle cells into vascular structures. Using agar, alginate, or collagen gels as placement media, cells can be arranged in shapes resembling multilayered artery tubules and proliferate to form functional arteries. The endothelial layer and smooth muscle layer of cells interact to secrete a natural extracellular matrix (ECM) between them. We have successfully cultured endothelial cells and are perfecting our technique of harvesting aortic smooth muscle cells for culture. These cells will be encapsulated in a gel we have optimized for cell adhesion and proliferation and will then be printed with our rapid prototyping machine into the shape of a blood vessel. After proper cell growth and secretion of the ECM we will subject our synthetic graft to tensile strength testing, thrombosis tests, and eventually implantation into an animal for observation of any immunogenic effects. Our project’s success would bring an array of new treatment options through biomedical engineering that would save many lives of those who suffer from cardiovascular disease.

Chance Christensen, Utah State University Life Sciences Affective disorders such as depression, phobias, schizophrenia, and post-traumatic stress disorder impair the ability to time in the seconds-to-minutes range, i.e., interval timing. According to the Relative Time-Sharing (RTS) model, presentation of task-irrelevant distracters during a timing task results in a delay in responding suggesting a failure to maintain subjective time in working memory, possibly due to attentional and working memory resources being diverted away from timing. Given that some anxiolytic medications have beneficial effects on attention and working memory, e.g., decreasing emotional response to negative events, we hypothesized that they would result in a decreased effect of distracters on the timekeeping abilities. We investigated the effect of acute administration of anxiolytic medication when anxiety-inducing task-irrelevant distracters were presented during an interval timing task, using methods similar to Matthews et. al. (2012) Frontiers in Integrative Neuroscience 6(111): 1-12. Results are discussed in relation to the brain circuits involved in RTS of resources, and the pharmacological management of affective disorders.

Brett Gardiner, Brigham Young University Life Sciences Normal human anatomy used in the classroom is not reflective of variations confronted in pathology subjects. Current commercialized models are not products of real data, rather representations of it. While learning complicated medical anatomy, students take an enormous stride from the anatomy lab to situational surgical settings. 3D models can bridge this gap in medical education without patient risk, particularly for the brain where surface regions have strong associations to specific physiological activity. Subject specific models are especially advantageous for comprehending real surface morphology of neurologic diseases. Using rapid prototype technology, we have developed an accessible process to produce physical 3D models from specific MRI data of stroke and Alzheimer’s subjects. The neuroanatomical abnormalities modeled from real data by our 3D printouts will educate students on the anatomical variations encountered in an authentic clinical scenario of neuropathology. Our project consists of three phases: (1) image acquisition, (2) post-processing imaging data with segmentation, and (3) 3D printing. By delineating cortical regions we are providing a unique multidimensional facet of clinically accurate data not before available to the classroom. This powerful and versatile technique can allow students and professionals to visualize the inherently complicated structures as seen in clinical neuropathology. From students in the classroom, lawyers in the courtroom or preoperative surgical explanations, these customizable models will resemble real anatomical information. Through rapid prototyping of specific subject data, unique variations in pathology can be reviewed outside of the clinical setting. Beyond its potential use by teachers, lawyers and doctors can benefit from a 3D production to enhance their explanations of anatomical variations from specific pathological subject data.

Ashley Calder, Utah Valley University Life Sciences An estimated 50-80% of dementia patients suffer from Alzheimer’s disease (AD). Currently there is no test to diagnose AD except post mortem. Recent papers indicate that AD affects the cytoskeleton and cellular structure through mutations that alter structural proteins, and that dysfunction of the cytoskeleton may play a pivotal role in AD and other neurodegenerative diseases. In particular, specific genetic components of AD affect microtubule and actin filaments that control endocytosis, exocytosis, the shape and size of the neuron, vesicular transport along neurites (dendrites and axons), and fibril formation. The goal of our research is to determine if breast cancer molecular subtypes can be used as a model for AD. Breast cancer is comprised of five molecular subtypes that contain different molecular structures depending on mutations specific to each subtype and the proteins being synthesized. These mutations and their expressed proteins change the characteristics of the cytoskeleton and resulting properties of the cell such as size, shape and stiffness. Both computer simulation and experiment have demonstrated that high-frequency ultrasound in the 10-100 MHz range is sensitive to these properties. For this study, ultrasonic tests were conducted on monolayer cell cultures of breast cancer cell lines of different subtypes. Ultrasonic waveforms were analyzed by transforming them into their corresponding spectra. The positions, widths, and shapes of the spectral peaks were compared and correlated to model results using a pattern recognition algorithm. Preliminary results indicate that cell stiffness and size can be determined from the measurements. Further analyses of these and additional data will determine if ultrasound is sufficiently sensitive to differentiate between the molecular subtypes of breast cancer. Results from these analyses, future studies with neuron cell cultures, and application of the results to the development of a minimally invasive, in vivo method for accurately diagnosing AD will be discussed.

Tamara Fox, Weber State University Life Sciences Clostridium botulinum has been implicated in cases of infant botulism across the United States. It is recommended that infants under the age of one year not be fed honey because of the presence of C.botulinum spores. The goal of this project is to determine whether honey produced in small and large apiaries in Utah contain varying amounts of toxin producing C. botulinum. Honey samples will be collected from hives maintained in Utah and tested for the presence of toxin producing strains of C. botulinum. Samples will be dissolved and centrifuged to isolate the spores and then superheated to release the DNA. Testing will then be done through a multiplex polymerase chain reactions (PCR) using primers specific for 16s rRNA, Clostridia species, and toxins A, B, E, and F. The presence and type of toxin producing Clostridia species will be compared with a Chi-Squared Test of Independence. Research will be completed by February of 2014 and we expect small apiaries will have a lower frequency of toxin producing C. botulinum strains than large apiaries and that toxin phenotype will vary between the two groups. The results will increase understanding on the variance of C. botulinum in Utah honey and will contribute to further research on this topic.

Eric Swenson, Dixie State University Life Sciences Underwater invasive species such as mussels and algae attach to submersible substrates that are advantageous for growth, and often include recreational boats that are frequently moved from one waterway to another. This can lead to rapid and uncontrolled spread of the invasive species. Through the use of newly developed ionic surfactants, growth and attachment of species on submersible surfaces can be inhibited. New surfactants have been incorporated into the final coating of typical product sealants to prevent the attachment of various marine organisms. By integrating the surfactant into the coating, an increase in the usefulness and longevity of effective inhibition is expected, as compared to commercially available alkali anti-fowling agents that are applied after the sealing process. It is hypothesized that the adherence properties possessed by marine organisms will be compromised by the introduction of ionic surfactants through changes in the chemical makeup of the exposed surfaces. The ultimate goal is to develop a substrate surface in which there is no affinity for attachment and/or causes fatality to the organism. If the desired outcome is reached, the potential benefits that ensue could create drastic improvements in areas of invasive species prevention and management in the desert southwest fresh-water lakes. Additionally, extension of the surfactant application to metal surfaces could have significant impacts on efficiency in water treatment systems, as well as cargo shipping in saltwater environments.

David Baird, Utah Sate University Life Sciences We obtained from the Bureau of Land Management (BLM) 30 years of monthly waterfowl population surveys completed at Pariette Wetlands in the Uintah Basin, Utah between 1980 and 2010. The Pariette Wetlands are the largest wetlands managed by the BLM within the lower-48 states and are comprised of 4,033 acres of land. Pariette Wetlands is surrounded by about 6,000 square miles of land where oil and gas production is the major activity. This waterfowl refuge is a significant location for migrating waterfowl species within the Pacific Flyway and provides important summer habitat for resident waterfowl. Our objectives were to determine what the trends were for waterfowl population abundance, occupancy, and species richness.

Chase Barker, Utah Valley University Life Sciences Central Research Question: Phylogenetic relationships of mayflies are still not very well known, however molecular and morphological data have begun to shed light on the relationships of these insects (Ogden et al. 2009). Our central question is to elucidate the phylogenetic relationships within the mayfly family Baetidae.

Molly Van Engelenhoven, Utah State University Life Sciences For thousands of years farmers have known that soil organisms can destroy crops but only in the past ten years have researchers begun to understand that complex interactions among soil organisms and plants can determine plant diversity and productivity. Much of the research in this young field of study has come from qualitative greenhouse experiments and separately from simple mathematical models. Here, we report findings from a six-year plant-soil feedback (PSF) field experiment that assessed the effects of soil organisms on the growth of twelve plant species common to the intermountain west. Initially, twelve native and exotic species were randomly assigned to 1550 experimental plots in a field. After four years of growth, the plants were killed. Next, the same plant species were randomly assigned to the 1550 plots again to test how each plant species responded to the soil microbial communities ‘cultivated’ by each other plant species. Two years later plant growth responses to different soil types were measured. Most plants showed large growth differences on different soil types. For example, Bromus tectorum grew twice as much on Pseudoroegneria spicata-cultivated soils than on self-cultivated soils. When the plant growth responses observed in this experiment were used in a mathematical model, PSFs were found to play a critical role in determining which plants were likely to be dominant in a community and how productive these different plant communities would be. In summary, this large-scale experiment provides strong evidence for the role of PSFs in plant productivity and diversity. Results have implications for natural methods of effective native plant restoration, biofuel production and weed control.

Laurel Thompson, Utah Valley University Life Sciences Breast cancer is divided into subtypes which are defined by their proteomics, histology, and genetic expression profile. Current methods, therefore, are aimed at testing these, and include DNA microarrays, immunohistochemical staining, and proteomic analysis. These methods are effective classifiers, but are not easily transferable to real-time clinical applications, such as the determination of cancerous status during operation or when taking a biopsy. The determination of molecular subtype by other means would be a significant advancement in cancer detection and treatment. We have made some preliminary studies that suggest high-frequency ultrasound may be sensitive to variations among the cancer subtypes as manifest in cell cultures through their cytoskeletal protein structure, which has a distinct spectral signature. The object of this study was to explore the basis for this variation through a combination of experimental and theoretical analysis. We used first-principal modeling methods and compared the model spectra generated from these to spectra obtained in the cell culture lab. Variations in bulk modulus, cell position and size were modeled and combined with experimental spectra in principal component analysis (PCA), and the Euclidean distances between each principal component of the experimental were found as they relate to the theoretical principal components. A graphical method similar to heat maps used for gene expression profiling was then developed to display the relative distances (similarities) between spectra. The program was tested by comparing experimental spectra of three breast cancer cell lines to model spectra. The results indicate the properties and thus molecular subtypes of breast cancer cells could potentially be determined by comparing their measured spectra to model spectra using a feature classification program such as PCA and that these classifying features can be displayed in a convenient graphical representation according to their spectral similarities.

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

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

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

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

Caitlin Carter, Utah Valley University Life Sciences High-frequency (HF) ultrasound (20-80 MHz) has been previously used to detect differences in microstructures and cell materials of different breast tissues types. These differences were used to distinguish between benign and malignant pathologies in different breast tissues. This same technology is predicted to be able to improve methods of detecting changes in cellular activity before changes in pathology take place. The purpose of this study was to use HF ultrasound to detect changes in the actin cytoskeleton, extracellular matrix (ECM), and integrin signaling, therefore differentiating the molecular subtypes associated with these changes in cell biomechanical properties. It is predicted that these cellular changes will also be associated with changes in the ultrasonic properties of breast cancer cells. The ability to rapidly and inexpensively detect the genetic changes or molecular subtypes of breast cancer would greatly impact and personalize patient treatment as well as provide more precise surgical removal of malignant and premalignant tissue. In order to test this hypothesis, four different breast cancer molecular subtypes including luminal A, luminal B, Her2+, and basal (triple negative) were grown as monolayer cell cultures. These subtypes were chosen because of their range of aggressiveness (luminal A as least aggressive and basal as most). After growth at different seeding levels, cell cultures were tested with a HF ultrasound system using a 50 MHz, 6.35-mm diameter immersion transducer and pulse-echo transmission. The data was compared to simulations using multipole expansions which predicted ultrasonic scattering based on possible variations in the biomechanical properties of malignant cells. The analyzed data showed differences in the spectra of waveform signals between each tested breast cancer molecular subtype. It is anticipated that this technique would provide an efficient and cost-effective method for differentiating between different molecular subtypes of breast cancer.

Teresa St. Pierre Nufer, Brigham Young University Life Sciences Acute stress has been shown to decrease Long-Term Potentiation (LTP) in the CA1 region of the mouse hippocampus. Stressed animals also 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 with stress group where mice have voluntary access to a running wheel (for 30 days) before undergoing the stress protocol.

J David Symons, University of Utah Life Sciences Cardiovascular complications (e.g., arterial dysfunction) are more prevalent in patients with type 2 diabetes (T2DM). Patients with T2DM have elevated levels of circulating free fatty acids (FFAs). We have shown that when bovine aortic endothelial cells (BAECs) are treated with the physiologically relevant FFA palmitate, protein phosphatase 2A (PP2A) activity increases, phosphorylated endothelial nitric oxide (NO) synthase (eNOS) to total eNOS (p-eNOS:eNOS) decreases, and metabolites of NO production decrease. NO is an important endothelial-derived relaxing factor that is vasculoprotective. As such, FFA-induced, PP2A-mediated reductions in p-eNOS:eNOS and NO production might explain why vascular complications are more common in pathologies associated with lipotoxicity e.g., T2DM and diet-induced obesity. Recently we showed in BAECs that if PP2A is inhibited using okadaic acid (OA), palmitate-induced increases in PP2A activity, and reductions in p-eNOS:eNOS and indices of NO production are negated. We sought to translate these finding from BAECs to the intact organism. However, OA cannot be used in vivo. Lixte Biotechnology 1 (LB1) is a PP2A inhibitor that has been used in vivo in the context of cancer research. The purpose of this study was to determine the efficacy of LB1 under our experimental conditions, with the long-range goal of using LB1 in mice. Further, we used this opportunity to optimize the measurement of NO directly using electron paramagnetic resonance spectroscopy (EPR). BAECs were treated for 3 h with vehicle (V), 500 µM palmitate (P), 4 µM LB1, or P + LB1 (n=10 per treatment). P increased (p<0.05) PP2A activity (50±12%), and decreased (p<0.05) p-tyr307:PP2A (29±9%; redundant indicator of increased PP2A activity), p-eNOS:eNOS (30±3%), and NO production (27±9%). All P-induced effects were prevented by concurrent treatment with LB1. Future experiments will determine whether chronic treatment of mice with LB1 is capable of suppressing PP2A activity in intact arteries.

Brian Ballard, Brigham Young University Life Sciences Antigen presenting cells digest and display proteins from foreign and infected cells on the major histocompatibility complex (MHC) which can then be recognized by T-cells through their T cell receptor (TCR). LLO56 and LLO118 are CD4+ T cells specific for the same Listeria monocytogenes epitope but show dramatically different primary and secondary responses to infection. Because TCRs have very low affinity for MHC we would like to create a high affinity T cell. We reasoned that the single chain LLO118 TCR (Vβ2-linker-Vα2) could be subjected to directed evolution to generate mutants that are more stable and then used as a template for engineering high affinity T cell receptors. Single chain LLO118 was fused to the yeast surface protein Aga-2 and error prone PCR was used to generate mutagenic libraries. The first generation stabilized LLO118 single chain TCR (scTCR) was selected using biotinylated Vβ2 and Vα2 antibodies and anti-biotin beads and it expressed LLO118 at higher levels than wild type by flow cytometry. To produce mutants with additional stability, a second mutagenic library using the first generation mutants as templates has been produced and the most stable clones will be selected after temperature denaturation, permitting isolation of clones with increased stability for generating high affinity pathogen specific scTCRs. After engineering a high affinity T cell our research will further understanding on TCRs and the MHC and could also serve as a resource for creating a therapeutic drug.