2014 Abstracts

Kyle Hansen, University of Utah Life Sciences Our brain processes physical stimuli from the world around us to shape how we perceive our environment. In the olfactory system, this processing begins with olfactory sensory neurons that relay information about distinct odorants to a region of the brain called the olfactory bulb. Olfactory sensory neurons express the same odorant receptors converge onto structures within the olfactory bulb called glomeruli. Here, we map lateral interactions between glomeruli to better understand how olfactory information from the environment is processed within the olfactory bulb. Using GCaMP6, a fluorescent calcium indicator, in awake mice we can observe optogenetically-evoked patterns of lateral connectivity between glomeruli in the olfactory bulb. The findings from this study will help clarify potential interglomerular connection schemes and provide understanding of how local circuitry in the olfactory bulb transforms the representations of sensory information before it is relayed to higher brain areas.

Brady Evans, Brigham Young University Life Sciences Nutrient-sensing kinases, such as AMPK and mTOR, play a key role in regulating cellular metabolism. They recognize nutrient levels within the cell and distribute nutrients accordingly. The failure to coordinate metabolic processes can lead to diseases such as diabetes, obesity and cancer. PAS kinase is another member of the nutrient-sensing kinase group that regulates glucose distribution in the cell. Despite its clear importance, little is known about the mechanisms regarding PAS kinase activation and its function. Studies have shown that mice without the PAS kinase gene are resistant to weight gain and maintain insulin sensitivity when placed on a high fat diet. Weight gain and insulin resistance are associated with increased rates of diabetes, cancer and other metabolic diseases. AMPK is currently the key target for the treatment of diabetes and has been found to be a cancer target as well. The yeast homologue, Snf1, is necessary for activation of yeast PAS kinase. Further studies have shown that PAS kinase is activated under the same respiratory conditions as Snf1. The aims of this project is to determine if Snf1 directly phosphorylates PAS kinase and to determine the effects of this phosphorylation. We have found that PAS kinase is activated quickly when cells are placed under Snf1-activating conditions. In addition, Snf1 is necessary and sufficient for this activation. Snf1 also copurifies with PAS kinase suggesting this activation is direct. PAS kinase purified from wild type verses Snf1-deficient cells shows phosphorylation on two key residues, S1020 and S1035. In addition, Snf1 directly phosphorylates PAS kinase in vitro. Together these findings suggest that Snf1 directly phosphorylates PAS kinase in order to regulate cellular glucose homeostasis

Joe Passman, University of Utah Life Sciences Neurodegenerative disorders and amyloidosis are thought to be a consequence of the misfolding of intrinsically disordered proteins (IDPs). In non-pathogenic states, IDPs are thought to manipulate their configurational ensembles (CEs) – through partially folding and/or forming secondary structure – to create binding sites for the multiple interaction partners necessary for cell-signaling, recognition, and regulation. An obstacle to accurate in vivo understanding of non-pathogenic mechanisms is that little is known about the impact of the crowded cellular environment on the global (i.e. partial folding) and local structural characteristics (i.e. secondary structure) of IDPs. It is difficult to understand why an IDP may misfold if little atomistic structural understanding exists regarding IDP functional mechanisms in non-adherent physiological states.

Shwan Javdan, University of Utah Life Sciences Each year in the United States, over 22,000 new cases of ovarian carcinoma are diagnosed and 14,000 women die from its progressive cancer stages. Anthracyclines, a class of common chemotherapy drugs, have long been the primary treatment for this and many other cancers, but they often leave patients with cardiotoxicity, hepatotoxicity, and other adverse effects. Polymer-drug conjugates using poly[N-(2-hydroxypropyl)methacrylamide] (poly-HPMA) are nanosized, water-soluble constructs that accumulate passively in solid tumors by the enhanced permeability and retention effect as well as actively by cell surface targeting methods. As such, they have exhibited reduced toxicity in the body. The goal of this study was to develop a novel, targeted HPMA copolymer-drug conjugate for the treatment of ovarian carcinoma. To accomplish this, an antibody fragment called Fab’, targeted to ovarian carcinoma cell surface antigen OA3, was bound to an HPMA copolymer-gemcitabine conjugate. Gemcitabine is a nucleoside analog used in chemotherapy that has demonstrated considerable effectiveness in recent years. The copolymer-gemcitabine conjugate was successfully developed following the synthesis of all the requisite components, including Fab’, diblock chain-transfer agent (di-CTA), N-(2-(2-pyridyldithio)ethyl)methacrylamide (PDTEMA), and polymerizable backbone-degradable gemcitabine derivative. Syntheses of di-CTA and PDTEMA were optimized and then characterized by high-performance liquid chromatography, H-NMR, and mass spectrometry. Monomers were combined by reversible addition-fragmentation chain-transfer polymerization, and then the Fab’ fragment was bound to the polymer backbone via a disulfide exchange reaction with PDTEMA. The targeted copolymer-drug conjugate was then verified by fast-protein liquid chromatography as well as ultraviolet-visible spectroscopy. A number of in vitro tests have demonstrated the properties of an efficient drug delivery system. Flow cytometry displayed active accumulation of the copolymer-gemcitabine conjugate to OVCAR3 cell surfaces. Furthermore, IC50 measurement found promising drug efficacy comparable to modern anthracyclines. Future work will involve in vivo evaluation of the conjugate’s therapeutic efficacy in nude mice bearing OVCAR3-xenografts.

Preston Manwill, Southern Utah University Life Sciences Plants produce compounds, secondary metabolites, which can be harnessed for medicinal uses. Caryophyllene, a secondary metabolite produced by a variety of higher plants, has shown promise as an analgesic and anti-inflammatory. Caryophyllene synthase is the final enzyme in the biosynthetic pathway that produces caryophyllene. The gene and enzyme responsible for caryophyllene production have been observed in crop plants and model organisms, but few studies have investigated caryophyllene production in indigenous plants. We investigated the production of caryophyllene in Ericameria linearifolia (Narrowleaf goldenbush) because it is a shrub native to southwest North America. The first portion of this research examined sequence similarities between caryophyllene synthase gene orthologs in E. linearifolia and other plant genera. Genomic DNA was isolated from E. linearifolia leaves collected from plants growing in southern Utah. Literature reviews and database inquiries have provided DNA sequences for similar synthases, from which primers will be designed to copy and sequence the E. linearifolia caryophyllene synthase gene, confirming the orthologs presence. Following confirmation of the gene, mRNA will be extracted from leaf material and oligo(dT) primers used to synthesize cDNA. Subsequent cDNA amplification and cloning of products into a vector will allow for transformation into a protein-producing bacterium. These final steps make the downstream procedures of protein production, structure determination, and functional characterization of the protein possible. Comparison with enzymes that produce similar organic compounds may result as well, and would provide insight that could improve the biosynthesis of medicinal compounds.

Preston Manwill, Sourthern Utah University Life Sciences Plants produce myriad secondary metabolites (essential oil) that play a role in ecological interactions. Species evolve unique mixtures of organic compounds due to strong selective pressures that act on metabolic pathways. Few studies have investigated the antimicrobial properties of secondary metabolites that evolved against soil microbes associated with native plants. The essential oil of Mojave goldenbush (Ericameria linearifolia), a shrub native to the Intermountain West, was characterized and its bacteriostatic abilities against root associated and non-root associated bacteria morphs were examined in this study. We hypothesized E. linearifolia oil would act as a greater growth inhibitor against native soil bacteria with which it has evolved than against bacteria from non-associated soils. Secondary metabolites were extracted using steam distillation and analyzed with gas chromatography mass spectroscopy. A Kirby-Bauer assay tested presence, size and retention of inhibition zones (IZ) produced against soil bacteria. The essential oil of E. linearifolia contained twenty secondary metabolites, two of which, limonene and sabinene, constituted greater than fifty percent of the oil. Presence and retention of IZ produced by E. linearifolia metabolites varied significantly by community association and bacteria morph. The hypothesis was supported by IZ that were largest against bacteria cultures isolated from soil that surrounded Mojave goldenbush roots and least effective against cultures isolated from non-root associated soil. Our findings suggest that secondary metabolites of E. linearifolia have evolved to specifically prevent negative interactions with bacteria most associated with the plant.

Cameron Sargent, Brigham Young University Life Sciences Although viruses present a potential solution for treating and studying many harmful diseases, they have yet to be utilized successfully, largely due to an overall lack of understanding of the interactions between viruses and their hosts. This study, however, demonstrates the efficacious usage of comparative genomics in further elucidating viral reproduction mechanisms, a step in overcoming this hurdle. Following the sequencing and annotation of genomes of the B4 subcluster of mycobacteriophage, a previously unstudied selection of viruses, comparative genomics analysis identified regions of proteins from these viruses that are highly conserved, or highly similar. One of these B4 phage proteins was then compared on the basis of its predicted folded structure and its amino acid composition to a protein with the same function from another phage, whose structure and DNA binding mechanism was determined via NMR spectrometry. Gene sequence alignments identified the region of highest conservation in the B4 phage. Protein folding then revealed that this region created a structure that was analogous to the functional region of the protein studied elsewhere, indicating that it too recognizes, binds to, and modifies DNA in the same manner. Furthermore, another DNA-binding motif with high conservation among B4 phage was observed in both the folded B4 proteins and the protein analyzed via NMR spectrometry. This comparison procedure not only further elucidated the function of an important gene in B4 phage but also displayed that comparative genomics, a resource-effective and computer-based approach, can successfully identify the critical functional regions of a protein. This study therefore provides a novel procedure for further understanding the mechanisms of virus-host interactions, which in turn facilitates the successful application of viruses in treating and studying diseases.

Eliza Warren, Brigham Young University Life Sciences We have previously demonstrated that ethanol inhibition of medium spiny neurons (MSNs) in the nucleus accumbens (NAc), and γ-aminobutyric acid (GABA) neurons in other ethanol-sensitive brain areas, is mediated by GABA(B) receptors. The aim of this study was to evaluate the involvement of GABA and glutamate (GLU), and in particular GABA(A), GABA(B) and NMDA receptors, in mediating ethanol inhibition of dopamine (DA) release in the NAc. Using fast scan cyclic voltammetry (FSCV), we evaluated the effects of ethanol on DA release in the NAc core of C57/BL6 and CD-1 mice. In the slice preparation, local stimulation evoked robust, frequency-dependent DA release in the NAc, with maximal release at 20 Hz. Ethanol decreased DA release with an IC50 of 60 mM in C57Bl6 mice. In anesthetized C57BL6 mice, ethanol decreased DA release with an IC50 of 2.0 g/kg. Superfusion of the GABA(B) receptor antagonist CGP55845 enhanced DA release 85%, while neither the GABA(A) receptor antagonist bicuculline, the GABA(A) receptor agonist muscimol, the GABA(B) receptor agonist baclofen, nor the NMDA GLU antagonist APV significantly affected DA release. CGP55845 blocked ethanol inhibition of DA release, while neither bicuculline nor APV altered ethanol’s effects. GABA inhibited DA release at 0.5-10 uM but not at 0.1-1 mM. Compared to ethanol effects on DA release in C57BL6 mice, CD-1 GAD GFP knock-in mice, which only express GAD at 50% levels, were significantly less sensitive to ethanol. As DA release was sensitive to low concentrations of GABA, CGP55845 may act as a GABA(A) rho receptor antagonist, and neither bicuculline nor baclofen had any effect on DA release, ethanol may be acting on extrasynaptic GABA(A) rho receptors on DA terminals to inhibit DA release in the NAc.

Ryan Watkins, Univeristy of Utah Life Sciences The lymphatic system is responsible for controlling systemic fluid buildup. Lymphangiogenesis is a dynamic process involving sprouting and maintaining new lymphatic vasculature. Vascular endothelial growth factor C (VEGF-C) is known as a key growth factor through VEGF receptor 2 and 3 (VEGFR2 and VEGFR3). Interestingly, the cornea expresses VEGF-C but is alymphatic. We found that the soluble isoform of VEGFR3, which lacks tyrosine kinase domains, is responsible for the alymphatic nature of the cornea by sequestering endogenous VEGF-C. Although soluble VEGFR3 or soluble VEGFR2 can be useful for inhibition of VEGF-C derived lymphangiogenesis, they also bind VEGF-A. Inhibition of VEGF-A suppresses blood vessel formation, damaging tissue and creating additional side effects. The development of a new anti-lymphangiogenic drug, that only blocks VEGF-C, has many implications: preventing tumor metastasis and reducing rejection rates of tissue and organ transplants. VEGF-C mainly binds to VEGFR3 domain 2 and VEGFR2 domain 3. To develop an anti-lymphangiogenic drug that specifically binds VEGF-C the binding domains were inserted into a vector that produces a recombinant protein (VEGF-C trap) that sequesters VEGF-C, suppressing lymphangiogenesis. Mice in a cornea transplantation model were treated with VEGF-C trap. After 8 weeks, 60% of the treated cornea survived (no rejection) compared to 10% in the empty vector control group. A 60% transplant survival rate is one of the highest rates compared to other single treatment methods. Blood and lymph vessel area was calculated and showed a decrease in lymph vasculature but not blood vessel. Suggesting only VEGF-C activity was affected. In a tumor metastasis model, nude mice will be injected subcutaneously with cultured MCF-7 cells which have been transfected with VEGF-C trap or an empty vector. We expect to see a decrease of lymphatic vasculature in the mice injected with MCF-7 cells containing the VEGF-C trap and ultimately less metastasis.

Preston Manwill, Southern Utah University Life Sciences Members of Anthoxanthum (sweetgrass) have a history of ceremonial and medicinal use. Coumarin, a secondary metabolite produced by the grass, is an anticoagulant and antimicrobial agent. Antimicrobial properties of commercially available European A. odoratum metabolites have been nominally investigated, but no North American species have been studied. Additionally, European sweetgrass is purported to be available in both diploid and polyploid strains. Our research objectives were to: 1) determine if ploidy level strains do exist, and 2) investigate secondary metabolite production and evolution in commercial strains and A. hirtum, a Great Basin native. Using epidermal casts and fuschin staining, differences in guard cells and nuclei were surveyed. Secondary metabolites from the A. odoratum strains and A. hirtum were obtained through steam distillation and a vacufugation protocol that concentrated hydrosols. GC/MS analyses characterized and quantified secondary metabolites. The secondary metabolites coumarin, dihydrobenzofuron, and dihydroactinidiolide were identified, with the first and latter greatest in A. hirtum. Significant guard cell differences between strains, as well as species, were observed. Two ploidy strains of A. odoratum were suggested. A Kirby-Bauer assay tested presence, size and retention of inhibition zones (IZ) produced against soil bacteria. Gram-staining initially characterized bacterial morphs. Secondary metabolites from the diploid strain were most effective against all bacterial morphs, but polyploid metabolites also generated and retained IZ against diploid root associated and non-root associated morphs. The native sweetgrass produced IZ only against root associated bacteria with which it had evolved. Future work will include similar studies of other North American Anthoxanthum species.

Justen Despain, Brigham Young University Life Sciences Mycobacterium ulcerans is an acid fast intracellular bacteria that is the causative agent of Buruli Ulcer, a disease endemic in tropical regions of Africa. The disease starts as a nodule that progresses into a necrotizing skin lesion. Treatment ranges from antibiotic therapy to surgical removal of surrounding skin to stop necrotization of the skin. Owing to the intracellular nature of M. ulcerans, antibiotic therapy can be an ineffective treatment option. It has been previously shown that mycobacteriophage can reduce extracellular bacterial load in mice infected with M. ulcerans. In this study, we propose a method of killing intracellular M. ulcerans by mycobacteriophage using avirulent Mycobacterium smegmaits as a vector.

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.

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.

Ethel Tackie-Yarboi, Westminster College Life Sciences The brine shrimp, Artemia franciscana are an important source of nutrients for many of the over 15 million migratory birds that visit Great Salt Lake (GSL) in Utah each year. As well as being a source of food, the brine shrimp are an intermediate host to cestode parasites that infect many of the bird species. Although the effects of cestode infection in brine shrimp and the contribution of brine shrimp to cestode circulation in birds have been studied, little research has yet addressed the rate of cestode infection or molecular phylogeny of these cestodes. We collected brine shrimp from three sites in GSL and tested individuals for cestode infection using previously identified and newly designed cestode specific PCR primers that amplify the 18S ribosomal RNA gene. Our preliminary results showed that the Spiral Jetty site from the North arm of GSL had a 2% infection rate, the Antelope Island site from the South arm had a 45% infection rate, and the shrimp collected at the Black Rock site, also in the South arm, had a 32% infection rate using our cestode specific PCR assays. We also have preliminary DNA sequencing results that demonstrate that we are amplifying cestode DNA, which is most closely related to members of the genus Hymenolepis. Our data suggest that there may be a higher prevalence of cestode infection in the shrimp in the South arm than those from the North arm of the Great Salt Lake. Our goal is to extend our study in order to better identify the species of cestodes that infect brine shrimp using further DNA sequencing and to expand our infection rate samples to better estimate the percentage of the brine shrimp population that is infected by the cestode parasites, as well as test samples from several years.

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.

Audrey Butler, Utah Valley University Life Sciences High-frequency (HF) ultrasound has been shown to be sensitive to a range of breast pathologies, and is being explored for the intra-operative assessment of lumpectomy margins. This sensitivity is believed to arise from microstructure-dependent interactions of ultrasound in the tissue. The objectives of this study were to develop breast tissue phantoms with microstructures that accurately mimic the histology of normal and malignant tissue, and to determine the effects of these microstructures on HF ultrasonic spectra (10-100 MHz). Phantoms were created from a mixture of water, gelatin, and soluble fiber. To simulate various breast tissue histologies, polyethylene beads, polyethylene fibers, and nylon fibers with a range of diameters were embedded into phantoms. Microstructures ranging from randomly dispersed beads to bead-fiber constructs resembling terminal ductal lobular units (TDLUs) were modeled and tested. Pitch-catch and pulse-echo measurements were acquired using 50-MHz transducers, a HF pulser-receiver, and a 1-GHz digital oscilloscope. Spectra were derived from the data and peak densities were determined from the spectra. Peak density, which is the number of peaks and valleys in a specified spectral range, has been shown to correlate with tissue complexity. Preliminary results from dispersed beads (58-925 µm diameter) of constant volume concentration (0.8%) indicated that the smaller beads produced higher peak densities than the larger beads with a consistent and statistically significant trend. These results substantially improve upon previous phantom studies and upon results from original breast cancer studies, demonstrating the strength of the HF ultrasound response to tissue microstructure. The higher peak densities can be attributed to either the higher number of scatterers for small beads or the size of scatterer in relation to the ultrasonic wavelength. These and other results from more advanced histologically accurate microstructures modeling TDLUs will be discussed.

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.

Evan Campbell, Brigham Young University Life Sciences Asthma is a chronic allergic disorder manifest by airway restriction due to inflammation, bronchoconstriction, and increased respiratory mucous secretion. As many as 300 million people worldwide are affected by asthma and its prevalence is increasing primarily in countries experiencing urbanization and Westernization. Asthma is currently the most common chronic illness among children in the U.S., and the third leading cause of hospitalization for children aged 0 – 15 yrs. Reservoir dust collection and area air sampling are the two primary methods of measuring allergen levels in house dust. Allergen sensitization leading to asthma is thought to occur prior to age six while the immune system is still naïve. In the case of the dust mite allergen Der p1, the exposure window may be as early as age two. However, little evidence is available to establish a dose–response relationship between inhalation exposure and early immunological sensitization to allergens. Temperature and relative humidity play a major role in dust mite survival and proliferation and indoor humidity above 50-60% in arid environments has been shown to support dust mite populations. Evaporative “swamp” coolers cool air by adding humidity to it and can create favorable environments for dust mite survival. We are working to determine how much swamp coolers contribute to dust mite levels in Utah by quantitating dust mite allergen levels in homes with and without swamp coolers. In order to do this we are developing sensitive ELISA and quantitative PCR methods to allow us to determine levels of exposure even when low levels of dust are collected.

Logan Warner, Utah Valley University Life Sciences High-frequency ultrasound (20-80 MHz) has been found to be sensitive to a range of pathologies in excised breast tissue before fixation in formalin or other formaldehyde analogues. Formalin fixation, however, may alter the structure and rigidity of a sample so that data gathered using high-frequency ultrasound after fixation may no longer be viable for research purposes. This limits the amount of time researchers may conduct tests, so preservation via simple sugars is being considered. Numerous studies have been conducted using sucrose, trehalose, or glucose as cryoprotectants for cells and simple tissues. The objective of this study was to test the sensitivity of high-frequency ultrasound to changes in the microstructure, stiffness, and cellular integrity of tissue samples due to cryopreservation with these sugars. Domestic pig heart tissue was placed in aqueous solutions of sucrose, trehalose, and D-(+)-glucose. The specimens were refrigerated and observed over time using high-frequency ultrasound to detect tissue damage. The results of this study suggest that cryopreservation with sugars will not only allow more time for researchers to conduct ultrasonic tests on surgical specimens, but also that high-frequency ultrasound could potentially be used as an assay to measure tissue degradation in preserved living tissues such as transplant organs.

Hillary Hansen, University of Utah Life Sciences Type 1 diabetes is an autoimmune disease where pancreatic β-cells are destroyed, resulting in insulin deficiency. Generating new β-cells from stem cells for treating diabetes will benefit from understanding their development in vivo. Pancreatic β-cells, along with all other pancreatic lineages arise from multipotent pancreatic progenitor cells (MPCs). Previous studies demonstrate that the structural and signaling protein β-catenin is required for the development of the exocrine acinar lineage. β-cells still differentiate in the absence of β-catenin, however, β-cell mass is dependent upon β-catenin. We determined that this dependency reflects a role for β-catenin in the maintenance of MPC patterning as well as for expansion of the progenitor pool. Whether our observed effects are due to the signaling or structural function of β-catenin remains unknown, and is the focus of this research. Using mouse genetics we are able to separate the structural and signaling functions of β-catenin. Eliminating both functions in PBKO (full knockout) mice produces decreased β-cell mass and irregular patterning. Decreased β-cell mass is also observed in PBsKO (signaling deficient) mice, though patterning remains unaffected. This suggests that pancreas growth is dependent upon canonical Wnt/β-catenin signaling, and that maintaining progenitor identity requires the structural role of β-catenin. Elucidating distinct roles for β-catenin could be used to drive stem cell-derived MPCs to expand and differentiate to the desired pancreatic cell fate.

John Hancock, Brigham Young University Life Sciences LLO56 and LLO118 are CD4+ T cells specific for the same Listeria monocytogenes epitope. Despite their TCRs differing by only 15 amino acids, LLO118 and LLO56 have dramatically different primary and secondary responses to Listeria monocytogenes infection. We reasoned that LLO56, the single chain TCR (Vβ2-linker-Vα2) could be subjected to directed evolution to generate mutants that are more stable and bind to peptide-MHC with higher affinity. Single chain LLO56 was fused to the yeast surface protein Aga-2 and error prone PCR was used to generate mutagenic libraries. A first generation stabilized single chain TCR (scTCR) was selected using biotinylated Vβ2 and Vα2 antibodies and anti-biotin beads. The first generation LLO56 mutant expressed LLO56 on the surface of yeast at higher levels than wild type by flow cytometry. To produce mutants with additional stability, a second-generation mutant was generated by combining multiple stability mutations isolated in a number of first generation clones.

Madison Peterson, Utah Valley University Life Sciences High-frequency (HF) ultrasound in the 20-80 MHz range has recently been found to be sensitive to pathology in tissue margins from breast cancer surgery. In order to improve the resolution and sensitivity of this method, however, transducers need to be employed that have piezoelectric elements that are smaller than those currently in use. The purpose of this study was to determine if small element transducers (Blatek pachyometer, 50 MHz, element diameter < 2 mm) produce similar results as those obtained from large element immersion transducers (Olympus NDT, V358-SU, 50 MHz, 6.35-mm diameter active element). Ultrasonic tests were performed on 10 phantom breast samples made of Knox gelatin base and soluble fiber (Metamucil), five of which contained chopped nylon fibers and five which contained polyethylene microspheres. Pulse-echo and through transmission measurements using a HF square-wave pulser/receiver (UTEX, UT340) and a digital storage oscilloscope (Agilent, DSOX3104A, 1 GHz, 4 analog channels) were acquired from a total of 3 sites per phantom in triplicate, first testing all specimens with the large transducers then again with the small transducers. Specimens were marked with India ink for location and accuracy of testing. The density of peaks in the ultrasonic spectra of the small transducers paralleled those of large transducers. Results from HF ultrasonic measurements of phantom breast tissue obtained from small transducers compared to the large transducers indicate that they produce statistically comparable peak densities. In breast conservations surgery it is crucial to excise all cancerous tissue to prevent recurrence. This method could provide in vivo cancer detections in margins and allow for more precise excision of tumors and cancerous tissue preventing the need for subsequent surgeries and thus, less risk, reduced pain and suffering, lower costs and better outcomes for breast cancer patients.

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.

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.

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.

Colby Pearson, Utah Valley University Life Sciences Many lichens synthesize unique secondary metabolites, such as atranorin (AT), which may serve as photo-protection against harmful UV radiation. Our study investigates changes in metabolism and internal levels of atranorin in lichens under various light conditions and atranorin supplementation. We will expose 60 Physcia adscendens (Fr.) H. Olivier lichen thalli to one of three different light levels (UV +PAR; PAR only; and neither UV nor PAR). Half of our experimental thalli will be supplemented with AT. Lichen biomass, AT concentration via HPLC, and photosynthetic and respiration rates will be measured before and after the experiment to measure metabolic responses of both supplemented and non-AT-supplemented thalli under each light level.

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.

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.

Kurt Williams, Brigham Young University Life Sciences Macrophages play an important role in innate and adaptive immune responses, inflammation, and tissue repair and are characterized by two distinct phenotypes: classically-activated (M1) and alternatively-activated (M2) macrophages. M1 macrophages are characterized by a pro-inflammatory phenotype and are involved in production of pro-inflammatory cytokines and aggressive engulfment, whereas M2 macrophages are characterized by an anti-inflammatory phenotype and are involved in production of anti-inflammatory cytokines (e.g. IL-10) and tissue repair. Macrophage engulfment of apoptotic cells leads to polarization toward the M2 phenotype and is thus “immunologically silent”. Additionally, there is evidence that tumor-associated macrophages (TAMs) tend toward an M2 phenotype and as a result offer protection from an immune response in the tumor microenvironment. To further investigate the role of necrotic and apoptotic cells in regulating macrophage polarization, we cultured human macrophages with necrotic, apoptotic, or standard viable Raji cells and fluorescent beads and performed an engulfment assay. In a preliminary study we found that macrophages cultured with apoptotic cells showed a decrease in engulfment levels compared to macrophages cultured with necrotic cells. Macrophages cultured with standard viable Raji cells had the lowest levels of engulfment compared to macrophages cultured with apoptotic cells or necrotic cells. Thus, in our initial experiments macrophages cultured with necrotic cells appear to have a more “M1” phenotype, whereas macrophages cultured with apoptotic cells appear to have a more “M2” phenotype. Further experiments are necessary to validate this preliminary data and further characterize the capabilities of necrotic and apoptotic cells to differentially polarize macrophages. If these observations are replicated, it has potential applications in cancer biology and therapeutics, atherosclerosis, diabetes, autoimmunity, and other diseases with an inflammatory component.

Clair Bidez, Westminster College Life Sciences Changes to stream ecosystems due to urbanization are known to degrade riparian ecosystems through multiple stressors including increased erosion and sedimentation, expansion of impervious surface leading to altered flow regimes, degradation of riparian vegetation and habitat, and nutrient contamination. Ultimately, such degradation can inhibit ecosystem services such as contaminant filtration and nutrient cycling. This study examined the effects of urbanization on the function of riparian communities in three streams in the Salt Lake Valley watershed. We attempted to characterize these changes through monthly monitoring in urban and non-urban reaches of the same streams. Specifically, we measured periphyton biomass (as measured by chlorophyll-a) as a known indicator of nutrient pollution and a proxy of riparian health. In addition, we monitored water quality parameters including temperature, dissolved oxygen, and dissolved nitrate concentration. The urban reaches of the streams contained 3.6 to 9.3 times the dissolved nitrate concentration as their non-urban counterparts. Mean stream periphyton biomass was positively correlated with mean stream nitrate concentration for each reach. These findings point to urbanization as a potential source of ecosystem degradation in the Salt Lake Valley. They suggest that long-term monitoring is warranted, along with an in-depth investigation into the ultimate mechanisms responsible for the degradation.

Eric Middleton, University of Utah Life Sciences One of the most impressive examples of homing among birds is demonstrated by Columba livia, or the rock pigeon. In racing or carrier breeds, birds can routinely travel over 100 miles and reliably find their way back to their home loft (Pratt, 1954). Bred from feral rock pigeons, these racers and carriers have been selected for their increased ability to home. While much research has been conducted on the mechanisms of homing, the heritability of homing is not very well understood. Furthermore, it is unclear if homing is predominantly learned or innate (Melhorn, Haastert, Rehkamper, 2010). The existence of homing breeds demonstrates that homing is heritable, but the degree to which this is the case or what traits in particular are improved with selective breeding are more uncertain. To attempt to better understand the heritability of homing, we will compare the homing ability of pigeons who are siblings to each other, and to unrelated pigeons. We first captured 90 wild pigeons from areas around Salt Lake City and allowed them to breed freely. We tracked the ancestry of all of the pigeons born from the feral birds and banded each of the birds in order to reliably distinguish which birds were related. We are now in the process of teaching the captive bred birds to home. Once this is completed, we will release the birds at varying distances from the loft. To quantify homing ability, we will measure angle of displacement from the loft when the birds first orient themselves and being homing, the time it takes to home, and which birds successfully make it home to the loft. This data will then be used to compare the homing ability of siblings to unrelated pigeons.

Lee Leavitt, University of Utah Life Sciences The ventral respiratory column (VRC) is a region in the brainstem shown to control breathing patterns in mammals. Using activation and inhibition of neurons in this region, classes have been assigned based on response-combinations. Using a mouse model, cells from this region are dissociated, plated and incubated with a dye that indicates changes in cytoplasmic calcium levels. Hundreds of cells are measured while varieties of pharmacological agents are applied. Response-combinations provide a profile of the receptors found on these neurons. Previously, varieties of cell classes were shown to contain NMDA receptors (receptors linked to learning and memory). However, specific compositions of subunits within these receptors are not known. These receptors are ligand gated ion channels composed of four non-covalently bound proteins. Each subunit has a different activation profile determined by interactions of agonist and antagonists. Conantokins (peptides isolated from snail venom) and other compounds further afford understanding of the architecture the assigned cell-classes. This project has continued to classify the subunit compositions of NMDA receptors with the ultimate goal of understanding which NMDA receptor subunits are present in each class. This will provide valuable information on the VRC’s function, and will allow for pharmacological innervations to change behavior in this region.

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.

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.

Cynthiann Heckelsmiller, Weber State University Life Sciences Heavy foot traffic compacts soil, leading to decreased gas exchange, water permeability, and reduced nutrient cycling. Plant communities are defined by the availability of nutrients, water, and other resources.

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.

Sean Studstill, Weber State University Life Sciences Imidacloprid is a popular systemic insecticide that has been applied to our staple crops for two decades. According to the EPA, it is persistent in the environment and at risk of effecting non-targeted organisms. Imidacloprid is an insect neurotoxin; however it is also known to be toxic to various aquatic species in concentrations as low as 37 ppb. Ingestion of imidacloprid causes paralysis in organisms through the blockage of postsynaptic nicotinic cholinergic receptors. We sought to find out how toxic imidacloprid is to brine shrimp and what kinds of physiological reactions occur upon exposure.

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.

Josh Hall, Weber State University Life Sciences Reproduction in birds is extremely conservative with the vast majority of the birds adopting bird-egg contact incubation to maintain an appropriate microclimate for embryonic development (Deeming, 2004). The Great Salt Lake is a vital nesting site for American Avocets (Recurvirostra Americana) that shows extreme temperatures and hostile environments where nest success can be as low as 1 -14% (Cavitt, 2008). Constancy of incubation, i.e. the time that the eggs are in contact with an adult, is a major indicator of nest success and environmental conditions. Our goals were to examine some of the costs natural selection places on embryos and parents to maintain a constant embryo temperature. We hypothesized that incubation attentiveness would increase across the nesting cycle. Over 200 AMAV nests were surveyed. Thermal probes were used to record various nest microclimates at every minute. A pseudonest with painted chicken eggs was also created and a thermal probe was placed to measure the ambient temperature without any adult incubation. A motion sensitive camera was placed over nests to examine differences in parental care. Nests will be divided into three phases: early, mid, and late incubation. Thermal data will be analyzed using descriptive statistics and mean variance values to calculate how incubation constancy varied throughout these phases. We expect this data to tell us more on how natural selection is working on these populations and some possible theories of how this developed.

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.

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.

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.

Janweb Lagazo, Brigham Young University Life Sciences The scientific community and the general public have long been interested in the effects of water pollution. Most studies on water pollution have focused solely on industrial pollution, but have failed to consider the potential impact of pharmaceuticals that unintentionally accumulate in aquatic ecosystems via wastewater treatment effluents. The purpose of this study is to advance our understanding on how these wastewater effluents affect aquatic ecosystems in Utah. We quantified the concentration of select pharmaceuticals in Hobble Creek using mass spectrometry. Then we sampled above the treatment plant, at the effluent outlet, and downstream of the effluent to determine pre-effluent and post-effluent drug concentrations. We are currently using this preliminary data to investigate how common endocrine disrupting, anti-inflammatory, analgesic, and anti-anxiety drugs may potentially affect the aquatic ecosystem of the endangered Chasmistes liorus, commonly known as June sucker.

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.

Dani Peterson, Brigham Young University Life Sciences Due to its long and complicated trajectory through the cranium, facial nerve VII (CN VII) can be damaged in surgeries, sometimes resulting in facial muscle paralysis. Surgical removal of acoustic neuromas and parotid tumors, in addition to surgical repair of the temporomandibular joint disorder are associated with a risk of damage to CN VII. In addition, insertion of auditory implants can damage the nerve, as can improper stimulation to the nerve after the implantation has occurred. We will create a three-dimensional (3D) model based off of data from dissection of the nerve in a human cadaver in order to give physicians a greater in vivo knowledge of the pathway of CN VII. We have dissected the lateral side of the right half of the head to the level of the parotid gland, identified the parotid plexus of CN VII, and followed its five branches. In addition, we are currently following the nerve through the internal auditory meatus on its pathway through the temporal bone. In preparation for the modeling MicroScribe technique described below, we have imaged the head using Magnetic Resonance Imaging (MRI) at BYU. These images will be used as a template for the nerve reconstruction model. After completing the dissection, we will track the nerve trajectory using a MicroScribe 3D Digitizer. The MicroScribe technique is used to create 3D computer models of any physical object. The user sets reference points and uses the stylus to trace data points of the object’s contours. Our final product will be a 3D spatial computer mapping of CNVII, as well as a mapping of the skull, parotid gland, and other landmarks to put the nerve model into context. We hypothesize that with our approach and MicroScribe technique, we will be successful in creating an accurate model of CN VII in the head.

Lauren Archibald, Brigham Young University Life Sciences Increased intake of selenium (Se) and soy have both been shown to reduce risk for prostate cancer, especially if these dietary treatments are combined. The purpose of this project is to determine how the timing of Se supplementation of either a low- or high-soy diet affects prostate cancer risk. [C57BL/6 X FVB] F1 TRAMP (TRansgenic Adenocarcinoma of Mouse Prostate) male mice were fed stock diets low or high in soy. Half of the mice received Se supplementation (4.0 mg Se/kg BW as Se-methylselenocysteine) by gavage 5 d/wk in a 2 X 2 factorial design. Se supplementation began at conception, 6 weeks, 12 weeks, or 18 weeks of age. The mice were then sacrificed at different stages of maturation (4, 12, 18, and 24 weeks). Our results showed that, at 12 weeks of age, urogenital tract weights, a measure of prostate proliferation and tumor volume, were significantly reduced by Se supplementation (p<0.001) and by soy (p=0.044), independent of time of dietary intervention. Histological scores of prostate cancer progression also showed a protective effect of Se supplementation (p=0.030). At this writing, statistical analysis of data from mice sacrificed at 18 weeks is in process. Data derived from 18-week mice, combined with our previous findings from 12-week animals, will allow us to chart the progress of prostate cancer in this model. In addition, results will show how dietary Se and soy may alter disease progression and how the timing of dietary intervention may determine its effects.

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.

April McMurray, Brigham Young University Life Sciences Diabetes Mellitus Type 2 (DMT2) is a lifestyle-related disease where the body does not produce enough insulin or the cells are unreceptive to it, and it is now the most common form of diabetes. Individuals who do not control the disease can suffer serious complications such as limb amputation, damage to the eyes, kidneys, nerves, heart, and it can be very costly. This problem is particularly serious in Tonga; the prevalence is almost twice as high as that in the United States. The purpose of this research project was to determine to what extent the cultural, economic, and educational factors contribute to such high prevalence. In May I traveled to Tonga with the nursing students from Brigham Young University to conduct my research. I distributed surveys to patients and medical staff in the diabetes clinic in the Vaiola hospital in Nuku’alofa, Tonga. The surveys had questions related to their socioeconomic status, understanding and attitudes of diabetes, as well as patient management practices. While I was there, I also kept extensive field notes on observations related to my research, which provided supplemental information regarding the Tongan lifestyle that was difficult to gather from the surveys. Preliminary analysis indicates that there has been a very small, positive shift in understanding and attitudes towards DMT2, but economic- and culture-based habits still impede Tongans from managing the disease effectively. There were several limitations to this study: small sample numbers, lack of resources, some resistance from Tongan medical personnel, and particularly cultural barriers made it difficult to gather enough information to come to significant conclusions. However, the research does give insight concerning potential future studies and interventions to help the people of Tonga treat this disease.

Benjamin Gann, Utah Valley University Life Sciences (E)-2,4-bis(p-hydroxyphenyl)-2-butenal (2-Butenal) was shown to inhibit various inflammatory responses by inhibiting NF-kB pathway. A pull-down assay proved 2-butenal to bind to IKKb and was proposed as an active site kinase inhibitor through molecular docking experiment. However, 2-butanal has a highly conjugated aldehyde group that makes it very unstable. Therefore, we have designed more stable 2-butenal analogues and prepared them using Heck reaction. Molecular docking experiment shows that many of them have a greater affinity to IKKb.

Jeffrey Leavitt, Utah Valley University Life Sciences Central Research Question: Heptageniidae is a large family within the order Ephemeroptera (mayflies). This family consists of over 500 described species. Recently a study was done across 200 of the species to break them up into subfamilies, and genera. The studied concluded that there are 29 genera and three subfamilies Ecdyonurinae, Heptageniinae, and Rhithrogeninae (Wang, 2004). Furthermore, Ogden et al. (2009) proposed that the families Arthropleidae and Pseudironidae were derived heptageniid lineages. The phylogenetic relationships of Heptageniidae, Arthropleidae, Pseudironidae, to other closely related families are inconclusive. We propose to study these three families and the three subfamilies of Heptageniidae in detail via molecular systematics.