2014 Abstracts

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.

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.

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.

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.

Few animals are capable of using the creosote plant, Larrea tridentate, as food because of a high level of toxic secondary compounds. Some exceptions to this rule are Neotoma lepida and Dipsosaurus dorsalis which are both capable of sustaining themselves on this desert bush. In 2013, Magnanou et al helped identify heightened transcription of genes correlated with digestion of creosote in N. lepida. Building upon their findings, we explore whether the genes for digesting creosote are under an elevated evolutionary rate for D. dorsalis. We have obtained transcriptomes from whole blood of four Iguaninae species: Ctenosaura pectinata, D. dorsalis, Sauromalus ater, Cyclura lewisi yielding an average of 4 GB of DNA sequence data (~51,000,000 fragments) each. Using Velvet in Sequencher we assembled these data, recovering greater than 6000 unique RNA transcripts per transcriptome. We search through the contigs to identify genes in Iguaninae transcriptomes that are homologous to those showing differential expression in Lepida. Using BLAST, we retrieve homologous genes from the public NCBI database of Anolis carolinensis and other reptiles. Lastly we construct phylogenetic trees of each gene and investigate the rate of change along each reptile lineage.

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.

Millions of people are infected yearly with resistant pathogens, including MRSA (methicillin-resistant Staphylococcus aureus), a biofilm-forming pathogen that is often transferred to patients from contaminated surfaces. Therefore, improved methods to destroy biofilm-encapsulated pathogens or to prevent their initial formation are required. This research is focused on the development of a safe treatment against biofilms by integrating organic salts, or ionic liquids (ILs), into different surfaces. Textiles were integrated with ILs to prevent formation of biofilms/bacterial growth, and were also treated post-exposure to determine if the biofilms could be destroyed post-contamination. Effectiveness of newly designed ILs were tested via inhibition zone studies on LB agar plates, and post-treatment samples were analyzed via scanning electron microscopy for presence of bacteria. The bacteria tested included Pseudomonas aeruginosa, Staphylococcus epidermidis, and Escherichia coli. These microbes are similar to MRSA in that they form biofilms comprised of extracellular proteins, DNA and polysaccharides. Bacterial colonies encapsulate themselves with biofilms to provide protection from threats, including antibacterial drugs. By integrating ionic liquids into textiles, formation can be prevented by IL solvation and sequestering of the extracellular biofilm components, including the proteins and DNA. This research could have tremendous implications regarding defeating bacteria that are resistant to existing treatments due to biofilm encapsulation. Additionally, the results could lead to new antimicrobial textiles and new approaches to prevent adherence and growth resistant biofilm-encapsulated pathogens.

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.

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.

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.

Although the idea of life on other planets is mused over by many, the scientific study of the potential for extraterrestrial life did not begin until the mid-1950s (SETI, 2013). Since then, many technological advancements have been made that make the study of life on other planets simpler, however it is inherently difficult to study the potential for life in an environment that one cannot access. To address this issue, scientists look on Earth for extreme environments that mimic those found elsewhere in the universe.

Dopamine (DA) D2 receptor expression parallels DA levels in the brain and these autoreceptors on DA neurons been shown to be modulated by long-term ethanol exposure. We have previously demonstrated that VTA GABA neurons also express D2 receptors, and that DA and D2 receptor agonists markedly enhance the excitability of VTA GABA neurons, opposite to their well-known inhibition of DA neurons. Most importantly, D2 receptor antagonists block ethanol inhibition of VTA GABA neurons and D2 receptor expression in VTA GABA neurons down-regulates with chronic ethanol. This study evaluates short-term D2 receptor adaptation in VTA GABA neurons and in DA release in the nucleus accumbens (NAc) by acute ethanol. In electrophysiology studies in anesthetized rats, periodic iontophoretic application of DA, or the D2 agonist quinpirole, markedly enhanced VTA GABA neuron firing rate, which was initially inhibited by ethanol, but resulted in latent and marked rebound excitation 30-60 min following injection. Using fast scan cyclic voltammetry (FSCV), we evoked DA signals in the core of the NAc by electrical stimulation of the medial forebrain bundle at the level of the lateral hypothalamus (60 Hz, 24 pulses). Intraperitoneal (IP) administration of ethanol (1.0-3.0 g/kg) dose-dependently decreased the amplitude of the MFB-evoked NAc DA signal. IP administration of the D2 antagonist eticlopride (1 mg/kg) markedly increased (250%) the amplitude of the evoked DA signal. When ethanol was administered after eticlopride it increased the amplitude of the DA signal an additional 42%. These findings suggest that ethanol induced decreases in evoked DA release may be due to autoreceptor feedback. Work is in progress to evaluate the short-term expression of D2 receptors in VTA GABA neurons following acute ethanol and to evaluate the effects of ethanol-induced short and long-term adaptations in VTA GABA neuron D2 expression in mediating ethanol effects on DA release in the NAc.

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.

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.

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.

Peak density, which is the number of peaks and valleys in a specified spectral range of high-frequency (HF) ultrasound, correlates to breast pathology in lumpectomy specimens. It has been a question in both previous and current studies, however, whether the thickness of a sample has an independent effect on the peak density. The objective of this study was to discover any correlation, if any, between specimen thickness and peak density in HF ultrasound measurements (10-100 MHz). Phantoms were fabricated from a mixture of water, gelatin, and soluble fiber. Polyethylene microspheres (180-212 micrometer diameter) were embedded into half of the phantom specimens at 0.0003% concentration to simulate tissue heterogeneity. The other phantoms were devoid of microspheres to provide control measurements. Seventy two pitch-catch measurements were acquired in triplicate using 50-MHz transducers, a HF pulser-receiver, and a 1-GHz digital oscilloscope. The waveforms were analyzed to provide spectra and the resulting peak densities were determined. The results indicate that no significant correlation exists between specimen thickness and peak density. The coefficients of correlation for the microsphere and control specimens were 0.366 and 0.652, respectively. The peak density values were most consistent within the control specimens, ranging from 1 to 4. The peak densities for the microsphere phantoms had a greater range of values, varying from 1 to 8. It is believed that the wide variation in peak density for the microsphere phantoms was due to clustering of the microspheres. Future studies will include looking at previous phantom and tissue studies to further investigate the apparent lack of thickness-peak density correlation.

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.

Learning and memory occur due to adaptive brain changes in response to our environment. These changes are mediated by synaptic plasticity, particularly within the hippocampus, where spatial and declarative memories occur. Plasticity can either strengthen or weaken synapses, known as long-term potentiation (LTP) or long-term depression respectively. While many forms of synaptic plasticity are N-methyl-D-Aspartate receptor-dependent, recently endocannabinoids were identified to mediate several new forms of hippocampal synaptic plasticity. Endocannabinoids bind to receptors such as cannabinoid receptor 1 (CB1) and transient receptor potential vanilloid 1 (TRPV1), and mediate several forms of plasticity, including in the hippocampus. However, new research has demonstrated a non-CB1/TRPV1-dependent endocannabinoid synaptic plasticity in the hippocampus. While the receptor(s) involved is currently unknown, several potential candidate receptors that bind the endocannabinoid anandamide have been identified. These are orphan G-protein coupled receptors (GPRs) whose distribution in the brain and/or function is unknown. GPR55 is of particular interest as it activates second message systems, including increasing intracellular calcium. Using quantitative RT-PCR, electrophysiological and memory behavioral tasks we examined hippocampal GPR55 expression and function. GPR55 is indeed expressed in hippocampus of both rats and mice. Cellular expression is currently being examined and appears to be rare in interneurons and more likely expressed by pyramidal cells. Interestingly, application of the GPR55 agonist LPI (2 µM) to wild-type mice demonstrates a decrease of LTD in brain slices. This LPI effect was not noted in GPR55 knock-out mice in the presence of LPI. This data suggest GPR55 is physiologically relevant in the hippocampus. This is the first direct evidence we are aware of that a novel endocannabinoid receptor directly effects hippocampal LTD. Because neurodegeneration that affects memory is typically associated with an increase in LTD, this provides a potential target to slow the advance of diseases such as Alzheimer’s.

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.

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.

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.

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.

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.

Dermaptera is a comparatively small order of insects with approximately 1800 species placed in three suborders. While the majority of earwig species are placed within the suborder Forficulina and are free-living with forceps-like appendages, two dermapteran lineages have a very unusual morphologies and life histories. The viviparous Hemimerina live epizoically on giant rats in tropical Africa where they feed on fungi growing on the rats’ skin. Hemimerina lack eyes and wings and the cerci are filiform. The viviparous Arixenina are associated with bats in Malaysia and the Phillippines, and they feed on bat skin gland secretions. They have reduced eyes, are wingless, and possess straight cerci. The phylogenetic position of the suborders Arixenina and Hemimerina relative to Forficulina have previously been unclear; however preliminary analysis suggest the phylogenetic position of the suborders Arixenina and Hemimerina are nested within Forficulina, with ectoparasitism evolving multiple times within this order. We generated DNA sequence data from three nuclear (18S, 28S and H3) and two mitochondrial (COI and TUBA) genes for representatives of all three suborders and outgroups. A phylogeny was reconstructed to address the following questions: (1) Does Hemimerina + Arixenina form a monophyletic group and support a single origin of parasitism or are there multiple origins of parasitism? (2) Is Forficulina monophyletic with respect to these parasitic lineages? (3) Are morphological similarities shared by the ectoparasitic forms synapomorphic or homoplasious characters?

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.

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.

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.

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.

Interest in animal personalities, and particularly the effect that different environments have on personality, has increased dramatically over the past decade. Understanding how individuals vary in their behavior, and if there are consistent differences among populations from divergent selective environments, lays the foundation for studies focusing on the contribution of divergent behavior in species formation. To date, studies that have focused on how personalities differ across ontogenetic stages have failed to compare populations that occur in dramatically different environments. Our study attempts to fill this void by studying the ontogeny of personality in populations that have evolved in environments with different levels of risk (i.e., predation vs. no-predation). We tested the expression of different personality traits evolution across ontogeny (i.e., from juveniles to full grown adults) in two sister species of live-bearing tropical fish, Brachyraphis roseni and B. terrabensis. These species have evolved in different selective environments, with B. roseni having evolved in an environment where predators were present, while B. terrabensis evolved in an environment lacking predators. We assessed the boldness expression of individuals from several groups in populations, namely juveniles, small adults, and large adults. To measure boldness, we used an emergence test, and also an exploration and activity test (i.e., ratio of movement to idleness during an allotted time period). Our study provides evidence for an important relationship between predation environment and the evolution of personality traits across ontogeny.

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.

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.

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.

Agave utahensis acts as a keystone species across its native range in the Mojave Desert and Colorado Plateau (Gentry, 1982). As a keystone species, Utah agave contributes to soil formation along barren mountain ridges, and has provided starch-rich sustenance to Native American tribes. Furthermore, taxonomists consider each of the two subspecies, kaibabensis and utahensis, to have considerable morphological variation (Gentry, 1982) within their own unique ecological niches. Given the importance of Utah agave, the high degree of variation, and its unique ecological niches, there is surprisingly little information published regarding its physiological ecology. In fact, no effort has been made to determine the population densities of Utah agave due to the remoteness of the region and its difficult terrain (e.g., the Grand Canyon). Therefore, geospatial analysis tools specific to environmental niche modeling provide a powerful means through which these issues and knowledge gaps can be effectively addressed. My goal is to develop a species distribution model by joining known locations of Utah agave with climatic and environmental data in MaxEnt and ArcGIS software. Such a model can be used by others for further ecological field studies of Utah agave and its subspecies. Additionally, the approach I employ can be used as a pattern for mapping distributions of other important plant species in remote and difficult-to-access regions of the world.

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.

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.

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.

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.

The main purpose of the project is to compare the phylogeography of a species of stonefly (Klapopteryx kuschelli) and a species of dragonfly (Rhionaeshna variegata) in Patagonia. Specifically, the project will study how geography and behavior (i.e., their dispersal abilities) have affected their evolutionary histories. Of all the varied climates and geography in South America, comparatively few phylogeographic studies have been conducted with insects, especially in Patagonia. This study will provide an important foundation for a comparative phlyogeographic study of two insect groups inhabiting the same regions of Patagonia. Also, if funded, this project will give me an exclusive opportunity to interact with international research institutions in South America as well as their scientists.

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.

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.

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.

Breast cancer is still the most common cancer among women regardless of race or ethnicity. The focus of our research is to uncover the mechanism breast cancer cells use to escape the inherent limitations of the telomere and obtain immortality. The protective end of a chromosome, the telomere, degrades with each cellular division. The cellular response to telomere dysfunction is to activate programmed cell death. Therefore, this type of damage normally limits the proliferative potential of the cell and subsequently carcinogenesis.

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.

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.

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.

Leafy spurge (LS) is an aggressive Eurasian forb that has been successfully reduced in many areas in western North America through the biological control releases of flea beetles. Long term studies of this phenomenon are sparse. Three flea beetle species were released in the mid-1990s at a site dominated by LS in Richmond, Utah. This study assessed the long term effects of LS biocontrol on an ecological community at this site by addressing five questions: (1) Is LS abundance significantly lower now than in the 1990s? (2) What plant species are replacing LS and are they native or non-native? (3) Have the flea beetle populations persisted since their initial release? (4) What part does soil type play in which flea beetle species now dominate at the site? (5) In response to their unexpected presence, what role may long-horned beetles contribute to the long-term reduction of LS? It was found that LS abundance has significantly decreased from the 1990s; the dominant plant species are those of non-native grasses; flea beetles have persisted in significantly smaller numbers, with Aphthona lacertosa being the most abundant; and long-horned beetles appear to play a significant role in the reduction of sexual success of LS. The results of this project have implications for land managers when considering the vegetative response to LS biological control and the importance of long-horned beetles for long-term in managed, LS-reduced habitats.

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.

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.

The advancement of biologically derived alternatives to petroleum diesel fuel requires a multifaceted approach. At Utah State University we use an interdisciplinary team including the Colleges of Engineering, Agriculture & Applied Sciences, and Science in conjunction with industry partners to drive innovation in improving the science behind petroleum diesel alternatives. With increasing petroleum use, depleting reserves, increasing emissions standards, and other factors, there is need for petroleum diesel alternatives that are cost effective, offer improvement, and perform similarly to petroleum diesel. Our team has focused on the use of oleaginous microbes utilizing low value effluent and waste sources including sugars and CO2 to create biofuels. We have focused on a yeast, Cryptococcus curvatus, and a microalgae, Nannochloropsis salina which have shown high yields of fuel per cell mass. Using these microbes we have utilized USU’s own direct trans-esterification reaction to create sufficient quantities of biodiesel for engine performance and emissions testing, including a subset of ASTM tests characterizing the fuels from each organism. Our initial engine testing used petroleum diesel as a baseline in conjunction with commercial soybean biodiesel to establish the quality of our microbially derived biodiesel. Testing in stationary diesel engines and on the Bonneville Salt Flats has proven our microbial fuels perform similarly to soybean biodiesel and comparably to petroleum diesel. To further improve biological diesel replacements we have begun working to create green diesel, hydrocarbons from a biological source, using a novel method of hydrothermal liquefaction. Preliminary results of those tests are presented here. Through a multifaceted and interdisciplinary approach USU is successfully improving petroleum diesel alternatives from microbial sources including characterization of the properties of these fuels and is working to create the fuels at the scale necessary for exhaustive engine performance and emissions testing including ASTM testing of all important fuel properties.

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.