Life Sciences

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

Merrill Christensen, Brigham Young University Life Sciences Selenium (Se) has established chemopreventive efficacy against prostate cancer, the second leading cause of cancer death among men in the US. Currently, most studies only employ one single chemical form of Se, even though different forms of Se act through varied mechanisms to achieve their anticancer effects. In this study, we propose that systematically combining multiple forms of Se will produce an optimal combination of Se compounds to inhibit in vitro prostate cancer cell growth. As proof of principle, we (1) synthesized Se nanoparticles (nano Se), (2) determined the IC50s of methylseleninic acid (MSA), sodium selenite, and nano Se in PC-3 cells, (3) utilized mixture designs and response surface methodology to direct our combination experiments with these three compounds, (4) tested the fifteen combinations determined in the previous step, and (5) constructed a polynomial model to derive the optimum combination of MSA, sodium selenite, and nano Se to inhibit PC-3 cell growth. Our results showed that combining different forms of Se compounds enhanced its chemopreventive effect. Future studies will expand the use of three Se-containing compounds to four and demonstrate similar effects in xenograft mice.

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

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

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

Albert Pope, Utah Valley University Life Sciences 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.

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

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

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

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

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

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

Alysa Fratto, Westminster College Life Sciences 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.

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

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

Robyn Kira Omer, Utah Valley University Life Sciences 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.

Dasom Kim, Brigham Young University Life Sciences 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.

Mark Soffe, Utah State University Life Sciences SSBs are DNA binding proteins that are essential components of cells and play key roles in DNA replication, repair, and recombination. Here we utilize two biochemical properties associated with the E. coli SSB protein to develop a novel procedure to purify proteins using a resin-free strategy. 1. E. coli SSB binds to single stranded DNA (ssDNA) with extremely high affinity (K = 1013 M-1), indicating very tight binding. 2. It is also a unique protein with respect to its purification – it is possible to obtain greater than 95% pure SSB from the total cell lysate without using any sort of column or resin, utilizing polyethyleneimine (PEI) and ammonium sulfate precipitation. Our design uses SSB as an affinity/solubility tag to enhance the solubility and expression of difficult-to-purify proteins, and allows for the simple, resin-free purification using PEI and ammonium sulfate precipitation. There also may be a possibility to co-express protein dimers and possibly tetramers using this method. Constructs have been made that include the SSB gene, along with the ability to fuse any gene of interest, as well as a TEV Protease cleavage sequence allowing for proteolytic cleavage after gene expression. Two genes of interest have been cloned in thus far—TEV protease and Rad51. In this proposal I outline experiments to develop this strategy further and test our proof of principle concept and its application to a broader set of target proteins.

Teressa Paulsen, University of Utah Life Sciences 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.

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

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

Jacob Anderson, Utah Sate University Life Sciences 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.

Michael Morgan, Utah State University Life Sciences 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.

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

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

Jacob Bassett, Utah Valley University Life Sciences Many discoveries regarding the complex interplay between biological pathways within a cell begin with attempts to link new observations to scrupulously studied cellular mechanisms. Our lab is studying the soluble inter-mitochondrial space protein mdm35, which has been observed to facilitate the function of ups1 & 2 proteins, as they in turn regulate the mitochondria’s phosphatidic acid metabolism. In addition to this phenotype, our lab has observed a copper resistance at 0.17% on YPD plate and 0.14% in liquid cultures of S. cerevisiae lacking the mdm35 protein, when compared to the Wild Type strain. Our investigation measures the levels of expression in the cup1 and ctr1 promoters in an attempt to link this observation to a potential mechanism contributing to this resistance.

Sam Hawkins, Utah State University Biological Engineering Colorectal cancer (CRC) is one of the most frequently diagnosed cancers in women and men. It is often treatable if caught early. However, tumors may metastasize which can result in a poor prognosis. A better understanding of the tumorigenesis and evolution of metastatic tumors in CRC patients could lead to earlier diagnosis, pre-emptive screening, and a better outcome. Copy number analysis of primary tumor tissue has revealed genes associated with colon cancers, but a comparison between primary and metastatic tumors has never been done. Normal tissue, primary tumor tissue, and metastatic tumor tissue was collected from twentyfive individuals. Copy number alterations were determined by microarray data generated from Molecular Inversion Probe (MIP) technology (Affymetrix, Inc.) for copy number analysis using Nexus software (BioDiscovery, Inc.). Metastatic tumor samples show a greater rate of copy number alterations (CNAs) from the primary tumors and even more alterations from normal tissue samples. Certain regions of the metastatic genome show high rates of CNAs whereas the primary tumor genome does not. These areas are key regions for potential understanding into the molecular origins of metastatic tumors. Understanding specific regions and genes with CNAs in metastatic tumor samples may lead to further research in cancer genetics and possible target areas for pharmaceutical testing.

Makda Gebre, Utah State University Biology The first pandemic of the 21st century was the influenza A (H1N1pdm09) virus that originated in Mexico and killed 12,000 people within the United States. It now circulates as a seasonal influenza virus causing mild symptoms in most but still killing some susceptible individuals. Influenza A (H1N1pdm09) virus leads to acute respiratory distress syndrome (ARDS) in high risk patients and has a mortality rate of 40-50% in those patients. ARDS patients usually die before any intervention since there is no clinical evidence that can be used to detect it. To detect ARDS early in disease progression, we need to search and develop biomarkers that can be tested in the patient’s blood or fluids. The three biomarkers tested as potential early disease indicators in this project included: C-reactive protein (CRP), serum amyloid A (SAA) and Transferrin. To test these biomarkers, mice were infected with the influenza A H1N1pdm09 virus and serum was collected from the mice at different time points. Serum collected was tested for amounts of CRP, SAA and Transferrin using an enzyme-linked immunosorbent assay (ELISA) and spectrophotometer. Control mice were mock infected and also tested for the biomarkers for comparison.

Thomas Anderson, Utah State University Biology Pradimicin, a small molecule produced by the soil bacterium Actinomadura hibisica, is a promising candidate as a combined antifungal/antiviral therapeutic. It is active against a broad-spectrum of opportunistic, pathogenic fungi, interferes with the replication of influenza virus, and inhibits the reproduction of HIV-1. Toxicity and solubility problems have hindered past efforts to develop pradimicin as a therapeutic. Our research focuses on elucidating the bio-synthetic pathway of pradimicin in order to design and chemoenzymatically create pradimicin structural analogs with improved solubility and activity, and less toxicity. Several enzymes in pradimicin biosynthesis have been identified. We intend to characterize one of the key enzymes, PdmS, a putative glycosyltransferase, and to manipulate its gene to create novel, more efficacious pradimicin analogs. This project is funded for three years by the NIH NIAID (3 years). Methods: A gene knockout experiment was used to determine the role of PdmS in pradimicin production. Bio-synthetic precursors of pradimicin were subjected to bio-transformations in E. coli with recombinant genes for PdmS and another glycosyltransferase, OleD, to generate analogs with new sugar attachments. Analogs of pradimicin will be screened for bio-activity using standard microbroth dilution assay techniques. Confirmed results: The enzyme PdmS was identified and characterized as a glycosyltransferase. Expected results: newly created analogs of pradimicin exhibited minimal inhibitory concentrations of 10μg/mL against Candida albicans. Conclusion: Knockout of pdmS yielded the pradimicin aglycon, which confirmed the function of this glycosyltransferase and provides a start molecule for further structural modification to yield new analogs for bioactivity studies.

Monica Cervantes, Utah Valley University The central research question of this project was to determine if high-frequency ultrasound is sensitive to tissue pathology at the microscopic level. Previous studies on surgical specimens have shown that high-frequency ultrasound may be sensitive to a range of breast pathologies including fibroadenomas, atypical ductal hyperplasia, fibrocystic changes, and carcinomas. The ultrasonic parameters that were sensitive to pathology were the number of peaks (the peak density) of the first-order spectra of the waveforms (one forward Fourier transform), and the slope of the second-order spectra of the waveforms (two consecutive forward Fourier transforms). The ability to determine pathology rapidly and with minimal specimen preparation would make high-frequency ultrasound particularly well-suited for real-time use during cancer surgery to ensure all of the malignant tissue has been removed. The purpose of this research was to determine the sensitivity of the peak density and spectral slope to tissue microstructures other than those found in breast cancer. The results of this study would not only support the results from the breast cancer studies, but also extend those results to the detection of cancer and other diseases in a range of organs and tissues. The research methodology included the following steps. (1) Freshly excised bovine organs were obtained from a meat packaging facility, including the heart, liver, and kidney. (2) Specimens approximately 3x3x1 cm in size were dissected from the organs and tested immediately with ultrasound. (3) Both pitch-catch and pulse-echo waveforms were acquired from the samples. (4) The data were analyzed by determining the peak densities and spectral slopes. The results showed that the more heterogeneous tissues of the heart, the vascular structures (aorta, vena cava, etc.), displayed significantly higher peak densities than the muscle tissues. Similarly, the ureter, which has greater heterogeneities in its structure (larger and more varied), displayed significantly higher peak densities than the cortex and medulla tissues. No significant trends were observed for the liver tissue, or for the spectral slopes except for kidney medulla tissue. Heterogeneity and peak density in high-frequency ultrasonic spectra that may be useful for performing real-time pathology during cancer surgery.

David Gage, Utah State University Biological Engineering A new N-oxide was produced from 3-(N-Boc-aminomethyl)-5-bromopyridine by bioconversion with Cunninghamella echinulata var. elegans ATCC 9245, and its structure was established based on spectral data. The microbial N-oxidation is efficient and highly selective. The substrate was transformed into the product in 7 days.

Kat Combs, Utah State University Plants, Soils, and Climate Kentucky bluegrass (Poa pratensis L.) is a commonly used turfgrass species with many varieties being sold around the world. However, those varieties are very difficult to tell apart morphologically. Our objective was to use genetic markers (primers) to identify varieties, even if they are visually similar. This is valuable to the turfgrass industry for plant variety protection. We also wanted to use this data to explore the apomictic tendency (clonal reproduction) of the varieties. We collected leaf tissue from 24 Kentucky bluegrass varieties, extracted DNA, and sequenced portions using 29 EST and 21 genomic primers. This data was used to determine genetic relationships using a neighbor-joining dendrogram. Similarities of the genetic sequences from the varieties were estimated using the DICE coefficient. We found more polymorphisms in genomic primers than in EST primers with high variability between the varieties. Both types of primers were robust enough to distinguish varieties and that each variety was unique and genetically distinguishable. In addition, we discovered some varieties had large amounts of variation within a variety. This was unexpected due to the usual apomictic nature of the species. The markers resulting from our research will be available to the turfgrass industry.

Robert Bradford, Utah Valley University Biology Most organisms exhibit latitudinal gradients in diversity (i.e., taxonomic richness decreases as latitude increases). Few studies have sought latitudinal gradients in lichens, especially in the midlatitudes. Our primary questions were: 1) do lichens along the west coast of the United States show latitudinal gradients? 2) If so, what is the rate of change and does the level of taxonomic richness affect this rate? We hypothesized that lichens would show a reverse latitudinal gradient in the region, as has been documented for lichens elsewhere in the northern hemisphere, but at a considerably smaller scale. This study fills in the gap in our understanding of lichen latitudinal gradients over large areas of North America. It also functions as a baseline for future climate change and conservation efforts. Our study area is bound at the south by the California-Mexico border (32.331° N) and at the north by the Washington-Canada border (47.178° N), and extends inland from the coastline to the crest of the Sierra Nevada Mountains (116.083° W, at its eastern -most point). We divided the region into 218 roughly equal-area (cite) grid cells using GIS, each bordered with latitudinal and longitudinal lines. We derived a list of all vouchered lichen specimens in each grid cell using Consortium of North American Lichen Herbaria, an online database. The data were synonymized, and species, generic, and familial richness were calculated for each grid cell. We found no correlation (R2 = 0.2306) between latitude and species richness, using the raw vouchered data. What we did find was a strong correlation (R2=0.9069) between sample density and species richness. These results are biased by sample density and do not reflect what is naturally occurring. We hypothesize that we can get an unbiased estimate of richness with MaxEnt models. Using the georeferenced lichen distributions and related climate data, we constructed species distribution models of all species with five or more occurrences (990 species). In GIS, we projected all 990 distribution models and our 218 grid cells together to calculate species richness for each cell at various thresholds (i.e. likelihood of occurrence at 10%, 20%, 30%, etc.).

Trevor Hansen, Utah State University Biology The use of metallic nanoparticles (NPs) in various applications presents the possibility for environmental contamination by these NPs. CuO and ZnO NPs have dose-dependent phytotoxicity. However, NPs are modified by different environmental factors such as salts: chlorides of Na, K, and Ca are used on roads for de-icing, dust control, irrigation, water treatments, and fertilizer production. At low concentrations these salts promote plant growth, but elevated concentrations become phytotoxic. This study examines the impact of Na, K and Ca chlorides on the phytotoxicity of CuO and ZnO NPs for wheat grown in sand. Atomic force microscopy and dynamic light scattering showed dose-dependent aggregation of the NPs in the wheat rhizosphere. The salts also caused NP-aggregation, but the degree of dissolution of the NPs was salt specific. Release of Cu from CuO NPs in the planted sand was reduced by all salt treatments whereas Zn release from ZnO NPs was reduced by Na and K but not Ca. Overall, a phytotoxic synergistic effect was seen at lower salt concentrations, but at higher salt levels, toxicity was mediated mainly by the salts. Therefore, effects upon plant growth of NPs and salts will vary according to concentrations.

Thomas Wallace, University of Utah Pharmaceutics and Pharmaceutical Chemistry The tumor suppressor p53 is mutated in more than 50% of all cancers, while the majority of remaining cancers contain mislocalized p53(1). p53 is part of a network of cellular pathways that regulate growth, cell cycle arrest, and clearly delineated apoptotic pathways(2). Restoring the function of p53 can be seen as an ultimate cancer treatment. Restoring p53 would activate an already existing cell pathway that prevents cells from becoming cancerous and cause cancer cells to go through apoptosis. Playing a role in tumor suppression has made p53 an attractive target for gene therapy. However, despite the potential of p53 as a powerful treatment, it is limited by the dominant-negative effect of dysfunctional mutant p53. This effect imposes one of the greatest limits on the effectiveness of a p53 based treatment. This project is focused on bypassing the dominant-negative effect of dysfunctional p53 over exogenous functional p53. The attempted solution was substitution of the p53 binding domain with a different but structurally analogous coiled-coil, based on a modified Breakpoint cluster region (Bcr) protein. By doing this, the dominant-negative effect of mutant p53 may be bypassed. The purpose of this project has been to synthesis and test rationally modified forms of p53 with modified Bcr coil that are introduced into cells via a plasmid to restore cellular p53 activity. In vivo cell tests have already shown the effectiveness of these constructs at causing higher rates of cell death in cancer cells and constructs are currently being refined to carry forward to xenograft model animal trails. The ultimate goal is to develop a treatment for human cancer patients where modified p53 will selectively cause apoptosis in cancer cells.

Chin Yiap Tan, Utah State University Nutrition and Food Sciences The long term objective of this research is to explore oil-based materials that can be used to replace trans-fats in the food industry. Vegetable oils such as olive (OO), corn (CO), soybean (SBO), sunflower (SFO), safflower (SAFO) and canola (CAO) were mixed with sunflower wax (SFW), paraffin wax (PW), and beeswax (BW) at different concentration levels (0.1%-100%). Crystal Morphology, oil stability, thermal behavior and viscoelastic properties were studied in this research. Results in our laboratory showed that the crystallization occurred sooner when the concentration of waxes increased. For example, crystallization in 0.1% of SFW in SFO occurred after 24 hours of incubation at 25°C, but when concentration increased to 0.25% the crystallization was observed at the first 15 minutes of measurement. In addition, a change in the thermal behavior of wax/oil soft materials was observed when the concentration of waxes increased. For example, the enthalpy change (H) in SFW/ SFO materials increased from 0.87 J/g at 1% of SFW to 21J/g at 10% of SFW. This increase in enthalpy indicated that the amount of crystal increased when the concentration increased. Overall, the crystallization behavior in wax/oil materials was affected by wax concentration and chemical compositions of oils and waxes.

Katie Fisher, Utah State University Watershed Science Arctic lakes are very sensitive to the effects of climate change. It is important to understand the current food web dynamic and energy flow within these lakes to better understand how they will change in the future due to the effects of a rapidly changing climate. In order to understand the current conditions in arctic lakes, this project consists of an analysis of stable isotopes of carbon (13C) and nitrogen (15N) from invertebrates among fish and fishless lakes in arctic Alaska to compare their trophic level positions and primary energetic sources. In order to carry out this analysis, I collected pelagic invertebrates were collected from 6 different lakes, 3 of which have resident fish populations and 3 of which are fishless. Samples collected in 2011 were analyzed for stable isotope composition by a mass spectrometer at University of California Davis. Results from samples collected in 2012 are pending at Washington State University. I will analyze and correlate the stable isotope results with isotopic data collected from other related projects. With this analysis, I will create food webs to 1) assign trophic positions to each species in each lake and compare those positions across lakes, 2) assess the potential effect fish predation has on pelagic invertebrate community structure. I hypothesize that fish predation will determine zooplankton community structure (e.g., dominant taxa) and alter trophic linkages (e.g., secondary trophic level predation rates).

Vance Almquist, Utah State University Plants, Soils, and Climate Fire induced soil water repellency has been characterized across a variety of soils and landscapes as being a cause of watershed degradation and surface water pollution. The repellency is due to the condensation of volatile polyaromatic hydrocarbons onto soil particles. Although repellency is known to reverse, in some locations the reversal takes months; whereas, in other locations it may take years. Little is known about the reversal mechanisms and how they lead to such a large range of reversal times. Access to untreated fired-affected sites, can be unpredictable and samples vary greatly from site to site. Therefore, a model compound that mimics fire-induced soil hydrophobicity is needed to be able to systematically investigate soil hydrophobilicty reversal mechanisms. Fluorene is a relatively non-toxic, hydrophobic polyaromatic hydrocarbon. The behavior of fluorene coated sand grains was investigated under laboratory conditions using quartz sand. Moreover, its fluorescent properties could be used to non-destructively monitor its degradation over time. In this context fluorene was studied as a possible model compound for the study of mechanisms involved in the reversal of fire-induced soil hydrophobilicy. The compound was subjected to conditions known to degrade or reverse water repellency including temperature, UV-light, and moisture content. Changes in hydrophobicity were monitored using the ethanol drop test and fluorescent imaging. Digital image processing techniques with the public domain software, ImageJ, produced by the National Institute of Health, were used to analyze the images and generate spatial maps of treatment effects on fluorene degradation and hyrdophobicity reversal. Our results indicate that the hydrophobic fluorene-coated sand layers were degraded by treatments such UV light known to reverse hydrophobicity in fire-affected soils, thus suggesting that fluorene may serve as a suitable model compound for producing hydrophobic layers on course grained material.

Miguel Cuevas and Joseph Wilkerson, Utah Valley University Biology Ovarian cancer is the leading cause of death from gynecologic malignancies in the United States and is the fifth leading cause of cancer death among American women. It is estimated that over 22,000 women in 2012 will be diagnosed with ovarian cancer in the United States and approximately 15,500 women will succumb to the disease. This is due to the fact that only 20 percent of cases are diagnosed before the cancer has spread to the peritoneal cavity. Currently, there are no reliable, standard screening tests; the only diagnostic test currently available is the CA125 tumor antigen blood test. This test is inadequate and not available as a general screening tool; additional diagnostics are required to effectively diagnose this disease. It has been previously shown that the tight junction protein Claudin-16, found only in normal kidney, is aberrantly expressed in epithelial ovarian tumors. Therefore, this protein is a good candidate for ovarian cancer diagnostics and targeted therapy. By identifying the promoter region that controls cldn-16 gene expression in ovarian tumors, we can create a luciferase reporter assay to identify cells that express Claudin-16 in culture. To do this, PCR-amplify of various upstream regulatory elements previously identified in kidney cell lines were subcloned into the pGL3 luciferase reporter vector. A higher amount of luminescence is present if the promoter sequence successfully up-regulates the luciferase gene in the vector. This is measured using a Dual Luciferase Assay to determine which promoter region is responsible for the over-expression of Claudin 16. Promoter activity was verified in kidney cell lines that normally express Claudin-16. Next, the assays will be repeated in ovarian cancer cell lines known to express Claudin-16 compared to cell lines that do not express the protein. The promoter assay will then be tested on a collection of ovarian cancer cell lines to determine if luciferase activity correlates with Claudin-16 expression. Once validated, we can test our construct as a cell based assay for identifying therapeutics that can lower Claudin-16 expression in ovarian cancer cells.

Ashley Stengel, University of Utah Biology Agricultural herbivores, such as cattle, often encounter plants containing toxins. One class of toxins, tannins, bind to proteins and inhibit digestive enzymes from acting to liberate nutrients. In this way, tannins prevent optimal absorption of nutrients. To overcome this challenge, some herbivores host bacteria with the ability to degrade tannins. Currently, there is a push to find novel microbes capable of aiding animals in detoxifying these compounds. Therefore, we aimed to isolate tannin-degrading bacteria from the desert woodrat (Neotoma lepida), a wild, herbivorous rodent that feeds largely on a tannin-rich shrub, creosote bush (Larrea tridentata). Woodrat feces were cultured on tannin-treated media, and bacteria capable of degrading tannins were further characterized with DNA sequencing. Results revealed that from 9 isolates, three species of tannin-degrading bacteria were present: Enterococcus faecalis, Bacillus subtilis, and Escherichia coli. Further characterization was performed through measurement of tannase activity. The activity of each isolate varied significantly between bacterial species as well as within a species. Ultimately, I hypothesize that these tannin-degrading bacteria facilitate the ingestion of tannin-rich plants by woodrats. Additionally, this research exemplifies how wild herbivores, such as the desert woodrat, provide an essential source of tannin-degrading microbes that could be introduced into domestic herbivores in order to improve agricultural practices.

Chase Barker, Utah Valley University Biology The Baetidae are an important group of Mayflies because of their position on the mayfly tree of life in that they are a key to understanding key evolutionary trends, such as the origin of wings and flight. Phylogenetic relationships of mayflies are still not very well known, however molecular and morphological data have begun to shed light on the relationships of these insects (Ogden et al. 2009). Recent molecular and morphological analyses have examined relationships of some of the lineages of Baetidae distributed in different regions of the world (Gattolliat et al., 2008; Kluge, 1997; Monaghan et al., 2005; Nieto, 2010;). Most of these studies suggest two subfamilies, Baetinae and Cloeoninae. This study represents one of the largest (in terms of data) and most diverse (in terms of taxa) phylogenetic analysis performed on the family Baetidae to date. The dataset consists of more than 50 taxa sequenced for six genes (Nuclear 18S rDNA; Nuclear 28S rDNA; Histone 3, Mitochondrial COI, Mitochondrial 16S rDNA; and Mitochondrial 12S rDNA). DNA sequences were aligned in Muscle (Edgar 2004) and tree reconstruction and nodal support was performed under Maximum Parsimony, Maximum Likelihood, and Baysian frameworks. The results indicate that the subfamilies were not strongly supported as monophyletic, contradicting conclusions from morphological data. Missing data in the data set might be contributing to low support across the tree.

Jordan Goodman, Utah State University Biological Engineering Nanotechnology is revolutionizing imaging techniques, antibiotic therapy and cancer treatments. Nanoparticles (NPs) are also utilized in many commercial products such as sunscreens, paints, ceramics and semiconductors. Consequently, it is inevitable that NPs find their way into the environment. The effects that NPs have on agriculture and soil ecosystems are the focus of this project. Metal-oxide NPs such as ZnO are toxic to many bacterial pathogens but the beneficial root-colonizing isolate, Pseudomonas chlororaphis O6 (PcO6), has high tolerance. This bacterium represents an important group of micro-organisms that colonize plant roots improving their resilience to both abiotic and biotic stress. At sublethal dose, the ZnO NPs remodel the secondary metabolism of PcO6 in ways that could have an impact on agricultural ecosystems. Formation of antibiotic phenazines produced by PcO6 is strongly inhibited by ZnO NPs. Phenazines are important for PcO6 growth in biofilms, induction of tolerance mechanisms in the colonized plant, and antagonism of other rhizosphere pathogens. A role of NPs as a point source for soluble metal release is involved in these processes but does not explain the total effect of the NPs. The sublethal effects of the ZnO NPs on bacteria are similar to changes reported by sublethal doses of traditional antibiotics on human pathogens. These changes have a strong impact on fitness of bacterial pathogens, the most significant of which is induction of antibiotic resistance. Although many metal-containing NPs are being used in antimicrobial formulations, these alternative antibiotics too may be inducers of antibiotic resistance.

Kevin Cope, Utah State University Plants, Soils, and Climate Photosynthesis is driven primarily by radiation between 400 and 700 nm; however, not all wavelengths are equally efficient. Red light (600 to 700 nm) is 25 to 35% more efficient than blue light (400 to 500 nm) and 5 to 30% more efficient than green light (500 to 600 nm). Although blue light is less efficient than red light, it has been shown to be necessary for normal plant development in all tested crop species. The mixture of pigments in plant leaves allows them to absorb all colors of light. Both red and blue light are absorbed primarily in the upper leaf. Green light penetrates deep into the lower leaf and transmits to leaf layers below the upper leaf canopy. Accordingly, once the upper canopy is saturated with red and blue light, supplemental green light is beneficial in increasing whole plant photosynthesis. Although the effect of spectral quality on single leaves is well characterized, the effect on whole plant growth and development is poorly understood. The narrow spectral output of LEDs makes them particularly effective for photobiological studies. From our initial studies with radish, soybean, and wheat, we determined that blue light levels can be used to better predict plant development than red:far-red light ratios. We also found that plants require at least 80 μmol of blue photons m-2 s-1 in order to develop normally. Further studies are currently being conducted to determine the optimal ratio of red and green light for maximizing whole plant photosynthesis in lettuce and radish.

Tyler Shimko, University of Utah Biology Differences in the genomes of organisms control an organism’s ability to deal with and adapt to environmental stresses. In this project, two strain isolates of the nematode Caenorhabditis elegans were analyzed using high-throughput assays measuring growth and offspring production to determine the genes that confer a greater resistance to the herbicide paraquat. Paraquat acts by interfering with electron transport mechanisms within the cells of living organisms. This mechanism not only allows it to act as an effective herbicide, but also causes it to pose a considerable risk to the health of animals, including livestock and humans. Using statistical genetics, regions of the genome were identified that are likely responsible for differences in growth rate and fecundity observed in the two strain isolates when grown in paraquat. Near-isogenic lines and extra-chromosomal arrays were then created to isolate these portions of the genome in a control genetic background. This approach allowed us to be able to attribute any differences in the two traits to the genes contained within the intervals. After analyzing the body size data, representing the growth of the animals over 72 hours, we were able to draw a preliminary conclusion that an interval on chromosome V may have a small but significant effect on growth determination. As a result of this project, a specific interval was identified that may be responsible for a greater growth rate, three near-isogenic lines were created, and 34 extra-chromosomal arrays were generated. This work will be used in the future to identify the gene(s) responsible for the greater growth rate and fecundity observed in some animals exposed to the herbicide paraquat. These results will allow us to draw conclusions about the roles that these genes, and others like them, play in an organism’s ability to cope with environmental stresses.

Brianna Rhoads, Utah State University Biology Nematodes are microscopic roundworms and are known to be some of the most difficult agricultural pests to control. Most of the 700 different plant parasitic nematodes species infect the root system of their host plant, causing galling, stunting and allow secondary pathogens to infect the host. For diagnostic purposes, in order to identify species of root-knot nematodes, the use of species specific primers can speed up the identification; many farmers need fast and reliable identification to determine which crops can still be planted in root-knot nematode infested fields. The purpose of my project was to test species-specific primers developed in Europe and Asia for Meloidogyne incognita and Meloidogyne arenaria on North American populations of both species using PCR. The results showed that primers developed in Asia and Europe for M. incognita amplified North American populations but primers developed in Europe for M. arenaria did not amplify North American M. arenaria populations. Based on results, M. incognita specific primers are useful for diagnostic purposes in the United States.

Ryan Putman, Utah State University Biological Engineering Spider silk is a biomaterial with extraordinary physical properties. It has a unique combination of tensile strength, elasticity, and even biocompatibility that has sparked interest in a wide range of disciplines. Potential implementations of spider silk include: medical advances (skin grafts, biomedical sutures, and artificial tendons/ligaments), automotive safety (seat belts, airbags), and military applications (parachutes, body armor). Although spider silk has a great variety of possible uses, collecting this product is not as easy as merely farming spiders and harvesting the silk. Spiders are naturally territorial and cannibalistic, thus alternative means of production are necessary to generate enough spider silk for realistic use. A non-pathogenic laboratory strain of Escherichia coli commonly used for research purposes will be modified to produce synthetic spider silk. Through the use of synthetic biology and molecular cloning techniques, recombinant DNA is inserted with the genetic code for dragline silk of the Argiope aurantia spider; once constructed, this DNA is transformed into E. coli. The goal is to take advantage of E. coli’s ability to be used as a “factory” for creating silk in a controllable and cost efficient system. Supplementation of additional tRNAs will be employed as a strategy to extend cell life and boost the overall spider silk protein yield. Preliminary results have been obtained that show the production of synthetic spider silk by the engineered E. coli. More research is being conducted to increase yields so that we can one day take advantage of this amazing biomaterial.

Anindita Ravindran, University of Utah Exercise and Sport Science Obesity predisposes individuals with Type II Diabetes to cardiovascular complications such as impaired blood vessel function. Due to the elevation of free fatty acids (FFAs) in obese individuals, ceramide, a lipid metabolite, accumulates and might contribute to the inability of a blood vessel to constrict or relax appropriately. Vessel dysfunction is partly caused by the inability of the endothelium, the innermost protective lining of blood vessels, to synthesize and release nitric oxide (NO). Our data indicate that ceramide impairs endothelial NO synthase (eNOS), the enzyme that synthesizes NO. In order to study mechanisms by which ceramide might impair eNOS, it is important to measure cellular ceramide production in response to pharmacological and genetic manipulations. Previously we used P-32 radioactive assays to measure ceramide accumulation. However, the use of radioactivity is expensive, potentially hazardous, and waste disposal is an environmental concern. Therefore, I sought to import a less harmful, more cost effective, yet accurate technique of measuring ceramide production by immunofluorescence (IF). IF allows ceramide to be tagged with a primary antibody which can be detected by a secondary antibody conjugated with a fluorescent dye. I have observed that 250, 500, and 750 uM palmitate (pal) incubation for 3 h increases (p<0.05) endothelial cell ceramide accrual in a dose-dependent manner. Further, a FFA-independent method to alter ceramide accrual i.e., 3 h incubation of cells with N-oleoylethanolamine, also elevates (p<0.05) ceramide production. Importantly, I have shown that 500 uM palmitate-induced ceramide accrual can be prevented (p<0.05) by two structurally dissimilar inhibitors (10 uM myriocin, 1mM L-cycloserine) of the rate-limiting enzyme responsible for ceramide biosynthesis i.e., serine palmitoyl transferase (SPT). None of these inhibitors impairs cell viability. These data indicate that IF is an accurate and reproducible method whereby ceramide accrual can be quantified in endothelial cell systems.

Jason Chen, University of Utah Biology Both human language and birdsong are learned sensorimotor processes that require exposure to adult communication signals in order to be reproduced. The social aspect of the development of these vocal signals is distinctive because interaction is not required for the development of vocal signals in other organisms. Human infants isolated from interaction will not learn to speak normally. The need for interaction is also characteristic of the ontogeny of vocal signals in the zebra finch (Taeniopygia guttata), but only zebra finch males develop vocal signals, in the form of song. Juveniles raised in isolation will develop highly atypical song characterized by prolonged, monotonic syllables and irregular rhythms. When juveniles are tutored by these isolated birds in one-on-one conditions their song drifts towards normally raised birds. It is unclear how tutoring, as a social interaction, influences this observation. We hypothesized that different social experience influences birdsong. We predicted the song of juveniles raised in isolation with an experienced female to be closer to normal song than that of those raised in isolated sibling groups, or those raised in isolation. Current data suggest that the syntax and temporal features of song from juveniles raised in isolation with experienced females is indeed closer to normal song than that of isolated juveniles.

Harsh Kansagra, Southern Utah University Biology Anthoxanthum hirtum is a native grass with many traditional and modern uses, including human medicinal benefits. Populations are found locally in Utah, but at higher elevations, usually above 2500 m. Indigenous people used native sweetgrass in a variety of ways, including medicinally, as ceremonial incense, and in basketry. The active compound that elicits the sweet fragrance of the grass is produced by coumarin, a secondary metabolite used today both medicinally and commercially. Plants most often produce secondary metabolites, or essential oils, as a defense against pathogens, but these antimicrobial properties have not been investigated in A. hirtum. Our research used the Kirby-Bauer disk diffusion test to determine if closely related commercial diploid and polyploid sweetgrass strains (Anthoxanthum odoratum), as well as plants from native A. hirtum populations, produce zones of inhibition when tested against associated soil bacteria and fungi. Results of our research showed all species tested produced inhibition zones, but zone size varied in response to the secondary metabolites produced by each plant type. Despite this variation, these data suggest components of the essential oils may have antimicrobial properties. Results of this study increase our understanding of the antimicrobial properties of secondary metabolites produced by A. hirtum as well as the essential oils produced by commercial diploid and polyploid strains. Future studies will focus on identifying the chemical composition of each extract as well as the specific bacterial and fungal species associated with each plant.

Diego Cordero, University of Utah Biology Many zygotes and spores of brown algae are photosensitive and establish a developmental axis in accordance with directional light cues. Ectocarpus siliculosus is being advanced as a genetic and genomic model organism for investigating brown alga development, and this report investigates photopolarization of the growth axis of mitospores. When exposed to unidirectional light, mitospores photopolarized and established a growth axis such that germination was preferentially localized to the shaded hemisphere of the spore body. The roles of the microtubule cytoskeleton and endomembrane cycling in the photopolarization process were investigated using pharmacological agents. Disruption of microtubule dynamics progressively reduced the percentage of mitospores that photopolarized, while inhibition of vesicle secretion blocked photopolarization nearly completely. Chronic treatment with these pharmacological agents severely affected algal morphogenesis. Microtubules in mitospores and algal filaments were imaged by confocal microscopy. Mitospores contained a radial microtubule array emanating from a centrosome associated with the nuclear envelope. At germination, the radial array gradually transitioned into a longitudinal array with microtubules extending into the emerging apex. At mitosis, spindles were aligned with the growth axis of cylindrical cells in the filament, and the division plane bisected the spindle axis. These studies demonstrate that dynamic membrane cycling and microtubule assembly play fundamental roles in photopolarization, and provide a foundation for future genetic and genomic investigations of this important developmental process.