Life Sciences

Tielle Gallion, Brigham Young University Life Sciences Quality of life is highly dependent on how well the liver functions. Increases in liver-enzyme levels, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) are associated with liver damage. A common treatment for Relapse-Remitting Multiple Sclerosis (RRMS) is Copaxone. This treatment has shown to be effective in reducing the number of relapses (periods of disability), but despite its effectiveness there is a prevalence of side effects, including increased liver enzyme activity. The aim of this pharmacogenetic project is to look at the effects of Copaxone, a drug therapy used in treating RRMS, on a patient’s liver-enzyme levels. I will determine if a correlation exists between increased liver levels and a specific genotype present in RRMS patients. I also plan to utilize medical record extraction. We have identified clinic visits in which 1,050 patients are recorded as currently using Copaxone by creating an algorithm to extract this data from electronic medical records (EMRs). This records are part of the Vanderbilt University Medical Center BioVU database. With this information, I have determined the period of time when patients are taking the drug. I accomplished this by manually calculating start and stop dates of Copaxone and created a table with information for each individual. Laboratory values are stored in a database, and we are currently extracting liver levels for ALT, AST, ALP, and GGT during the identified time frames for respective patients. Once extraction of lab values is complete, I will perform a linear regression analysis in R, a statistical computing program, to determine if any correlation exists between RRMS patient genotypes and liver-enzyme levels. Patients have previously been genotyped on the ImmunoChip, which contains 196,524 SNPs and has undergone stringent quality control.

Matz Indergard, Paul Spruell, and Fredrich Govedich, Southern Utah University Life Sciences Dispersal of organisms allows them to colonize new habitats and may buffer against extirpation due to localized catastrophic events. For organisms inhabiting ephemeral environments, dispersal can be challenging, as suitable habitat is often small, isolated spatially, and unpredictable temporally. Fairy shrimp are small invertebrates that occupy ephemeral freshwater pools filled by rainfall. Fairy shrimp produce resting (resistant) eggs when conditions are not favorable. Upon the return of supportive conditions, these eggs will then hatch. Little is known about the methods of dispersal used by fairy shrimp. However, it has been suggested that the resting eggs are the most likely life stage to provide dispersal opportunities. We examined two proposed methods of egg dispersal for fairy shrimp (Brachinecta sp) from a series of ephemeral pools just north of Three Peaks near Cedar City, Utah. We hypothesized that dispersal could be attributed to prevailing winds or water runoff, which would disperse eggs in a direction corresponding to prevailing wind patterns or to outflow following the surface gradient. To conduct our test we established three reference points equally spaced in the linearly arrayed series of pools. We then collected soil samples at randomly determined locations around these center points to form an overlapping radial grid. We then added purified water to each of our samples and hatched any shrimp eggs that were present in the sediment. We then mapped the overall concentrations of shrimp in our sampling area. Results to date do not reveal an obvious pattern with prevailing wind direction and dispersal.

Benjamin Bitner, Jason Ray, Kyle Kener, and Brent Wright, Brigham Young University Life Sciences β-cell mass is lost in both major forms of diabetes. Mature β-cell have restricted proliferative capacity. Studies aimed at increasing β-cell mass frequently have the unwanted side effects of decreased glucose stimulated insulin secretion (GSIS) or increased apoptotic rates. Without functional β-cells, the body is unable to reduce elevated blood glucose, which results in hyperglycemia induced maladies. Enhanaced GSIS could be used as a treatment for diabetes. Overexpression of the β-cell transcription factor Nkx6.1 induces β-cell proliferation, enhances GSIS and protects against apoptosis. Nkx6.1 induces expression of VGF, which is necessary for enhanced GSIS. Microarray analysis of Nkx6.1 expressing primary rat islets demonstrates upregulation of c-Fos at 24 and 48 hours after adenoviral transduction. We have shown that c-Fos upregulates expression of VGF. Finally, preliminary data suggest that c-Fos modulates GSIS as demonstrated in INS-1 β-cell line, stable INS-1 knockdown cells transduced with lenti-sh-c-Fos and in primary islets. We present a model by which c-Fos is necessary for Nkx6.1 mediated enhanced GSIS by inducing expression of VGF.

Amanda Johnson, Rian Farr, and Autumn Woodfall, Dixie State University Life Sciences Autism affects approximately 1 in 68 children, and is one of the fastest growing developmental disorders in the United States. Studies suggest that autism may be a result of processes that occur during labor and delivery. Although Pitocin is widely used during labor and delivery in the United States, there is a paucity of research on the developmental outcomes of Pitocin use during labor and delivery in young children. Additionally, the components and metabolic fate that make up the compounded Pitocin are widely varied between suppliers. Samples of Pitocin were extracted to isolate the chemicals that comprised commercial available products. These were then analyzed by chemical and biological methods to determine the composition and chemical breakdown products. Understanding the components and breakdown of commercial Pitocin will help to determine the compounds potentially transferred to fetuses with Pitocin use during labor and delivery and the potential developmental outcome.

Coral Gardner and Don Long, Southern Utah University Life Sciences Irrigation of lawns and gardens in Cedar City, Utah is accomplished in two fundamentally different manners. In older neighborhoods, water is diverted from a natural stream (Coal Creek) into a series of canals and ditches for residential flood irrigation, compared to newer neighborhoods, which use sprinklers or similar devices from well water. The overall objective of this project is to better understand the chemical and biological changes that occur in irrigation and runoff waters in Cedar City. We are addressing the following three hypotheses. 1) Changes will be observed in water chemistry as surface water moves from Coal Creek through Cedar City. 2) Irrigation strategies influence water chemistry during periods of high precipitation. 3) Microbial community changes will be associated with differences in water chemistry. Water chemistry data including dissolved oxygen, conductivity, pH, alkalinity, dissolved organic matter and nitrate/nitrite were collected weekly at eight sites and during high precipitation events. Over the course of four months, there was a reduction in alkalinity levels among all sites. During high precipitation periods, nitrate was detected in newer neighborhoods. Dissolved oxygen and pH were at higher levels, while salinity and conductivity were lower in a reservoir site relative to irrigation canals. Future work will investigate bacterial community composition in Cedar City waters. We will isolate bacterial DNA from water samples and amplify the 16sRNA segment of DNA using the polymerase chain reaction. We will then correlate bacterial community composition to the water chemistry results described above.

Brianne Palmer, Utah State University Life Sciences Quaking aspen (Populus tremuloides) are declining in the interior west. Aspen are critical for the maintenance of wildlife habitat and are one of the few broadleaf trees in the western forest ecosystem. In western landscapes, it has recently been determined that a large proportion of aspen trees are triploid (three copies of each chromosome) and the remaining trees are diploid (two copies of each chromosomes). In this study we attempted to find differences in the physiology between the two cytotypes to determine future management strategies The size and density of stomata trees is likely to influence the survival of the species in water- and heat-stressed environments, since stomata control both photosynthesis rates and rates of water loss. Individuals with larger stomata or greater stomatal density may be efficient photosynthesizers but may be at risk for water loss during transpiration in environments with low precipitation and hot temperatures, such as those often seen during summers in the intermountain west. To determine if there is physiological differences between the cytotypes we measured the variation between stomatal sizes and densities between the cytotypes using cellulose acetate leaf impressions and microscope imagery. We collected leaves from twelve aspen stands (eight diploid and four triploid) representing the two cytotypes in Swan Flats and Fish Lake, Utah. From these analyses, we deduced that the variation in stomatal size and density is primarily among clones rather than among cytotypes. Further data collection and analyses will occur in the spring of 2015.

Austin Thompson and David Walton, Brigham Young University Life Sciences When we sustain a traumatic injury to the peripheral nervous system (PNS), our bodies elicit a series of responses to try to heal the acquired damage, including inflammation and repair processes. One of these responses is the increased expression of nerve growth factor receptor (NGFR), which helps to stimulate regeneration of the nerve. In a normal, healthy PNS, NGFR is rarely found. Following damage to the nerve, NGFR can be found in high levels around the damaged area. In our study, we are simulating traumatic injury to the sciatic nerve of rats in order to study the effects of regeneration after a local application of nerve growth factor (NGF). We are using both a physical crush model and a focal demyelination model to simulate the nerve injury. In the crush model, we are examining the effect of a crushed extracellular matrix (ECM) on degeneration and subsequent regeneration. In the focal demyelination model, we are investigating the effects of local demyelination with an intact ECM on degeneration and regeneration. In two additional experimental groups, we will perform an intraneural injection of NGF into the damaged sciatic nerve one week after the crush or a lysolecithin injection at the damaged site. We are examining the nerve both qualitatively using SEM and immunohistochemistry and quantitatively using electrophysiology. This allows us to understand the role of the ECM in regeneration, and its effect on the rate of regeneration. We hypothesize that the addition of NGF in combination with the increase of NGFR after injury will increase the rate of nerve regeneration. We expect regeneration to be faster in the focal demyelination model due to the presence of intact ECM than in the crush model where the ECM is damaged.

Simranvir Kaur, University of Utah Life Sciences The placenta is the major source of fetal serotonin during pregnancy, which is essential for fetal brain development. In Utah, approximately 13% of pregnant women take Selective Serotonin Reuptake Inhibitors (SSRIs) to treat depression, the use of which has been correlated to significantly lower serotonin levels in cord blood for newborns. Studies suggest association between maternal SSRI use and adverse outcomes such as preterm birth, cognitive deficit, and disruption of serotonergic systems. However the effect of SSRIs on placental gene expression, serotonin synthesis and transport in the placenta, is not known. This study evaluates the impact of maternal SSRI use on placental gene expression and levels of serotonin in the cord blood using a nested case-control observational study model. Biological samples will be collected until 20 cases (women taking SSRIs during pregnancy) and 20 appropriately matched controls have been enrolled into the study. Participants also complete an online questionnaire to measure depression and anxiety levels as well as document any medication they have taken during pregnancy. Data and sample collection for this study is still in progress. Once enough samples have been collected for batch analysis, we will complete RT-PCR and ELISA, expected in Spring 2015. Upon complete data analysis, we expect this study will help in targeting mothers who are at risk for adverse pregnancy outcomes and further provide suggestions for intervention.

Matthew Durrant and Shuqing Xu, Brigham Young University Life Sciences Researchers at the Max Planck Institute for Chemical Ecology have recently sequenced the Nicotiana attenuata (wild tobacco) genome. This genome affords researchers new opportunities to understand the evolution of this organism. One method for analyzing the evolution of specific genes in a given genome is referred to as phylostratigraphy, which makes use of large-scale BLAST sequence similarity searches. I designed a pipeline using the python programming language that implements a phylostratigraphic analysis to estimate the evolutionary age of all ~35,000 genes belonging to N. attenuata. By analyzing the large amounts of data produced by this BLAST search, each gene was assigned an estimated age through comparing the taxonomies of all other organisms that share similar protein sequences. This effectively answers the question “How old is this gene?” for each gene in the entire N. attenuata genome. Previous studies have produced microarray data that tracks the transcriptomic response of N. attenuata to an herbivore attack. By comparing the newly gathered gene evolutionary age information with this previously gathered microarray data, several new insights into the molecular signaling pathways of N. attenuata were made. It was found that at 1 hour following an herbivore attack, the transcriptome of N. attenuata is evolutionarily young, suggesting that the initial response to an herbivore attack recruits genes that have evolved more recently in the organism’s evolutionary history. At 5 hours after attack, however, there is a distinct decrease in the overall age of the N. attenuata transcriptome, suggesting that the organism is recruiting more ancient genes that are used to reconfigure the transcriptome of the organism. Beyond 5 hours, the transcriptome is once again relatively young, and it is clear that it has indeed been reconfigured to provide a more herbivore-specific defense response. This demonstrates a novel, evolutionary approach to analyzing signaling pathways in plants.

Laine Anderson, Utah State University Life Sciences Plague is maintained in complex epizootic and enzootic transmission cycles involving rodents and their fleas. Ground squirrels, prairie dogs, woodrats and their associated fleas have been identified as essential for bacterial maintenance. Oropsylla montana is of major interest due to the fact that it is distributed throughout the western U.S. where most human plague cases occur. Evidence suggests this species is the primary vector of plague to humans. Data on the genetic variation within and among populations of potential vectors of Yersinia pestis, including O. montana, is very limited. O. montana fleas were previously collected from 35 geographically distinct field sites from Colorado and New Mexico on the east, to California and Oregon on the west. Genomic DNA was extracted and mitochondrial and nuclear genetic data was sequenced to estimate phylogenetic relationships. The data collected from these studies will expand our knowledge of natural O. montana populations. We anticipate the genetic data collected from these flea populations will provide information that will assist with understanding human plague risk. Such information will, in turn, provide potentially significant insights into the ecology and epidemiology of plague in this region and is likely to suggest new strategies for monitoring and preventing this disease.

Kyle Kener, Brigham Young University Life Sciences Diabetes is characterized by the inability to maintain a normal blood glucose level caused by decreased insulin due to β-cell loss, or decreased insulin sensitivity in the liver, muscle, and adipose tissue. While β-cell death is a hallmark of T1D, β-cells are also destroyed as T2D progresses. Death of β-cells is eventually a hallmark of both forms of diabetes. This results in decreased functional β-cell mass, which is defined by the ability to secrete insulin while maintaining β-cell number through proliferation or decreased apoptosis. To resolve the decreased β-cell level, much research is being done regarding β-cell proliferation to increase pancreatic β-cell mass. However, another important step in this process is protecting β-cells from apoptotic mediated β-cell death. The β-cell transcription factor Nkx6.1 is sufficient to induce β-cell proliferation and increase protection against apoptotic insults. The Nkx6.1 target gene VGF is critical for protection against apoptosis. Our data demonstrates that Nkx6.1 upregulates expression of c-Fos. Furthermore, we show that c- Fos is sufficient to induce expression of VGF. In addition, our data demonstrates that expression of c-Fos is sufficient to protect β-cells from apoptotic insults. Our data demonstrates that c-Fos is the link between the Nkx6.1 and VGF, and that it’s expression is sufficient to protect rat pancreatic β-cells from apoptosis.

Madalynne Fedoruk and Darian Carey, Dixie State University Life Sciences For at least three decades, infection with chytrid fungus (Batrachochytrium dendrobatidis) has been a major cause of population decline in amphibians worldwide. This pathogen has been found in canyon tree frogs (Hyla arenicolor) in Zion National Park; previous studies suggested that population sizes decreased in frogs infected with the fungus; but our studies indicated that these frogs were resistant. We hypothesized that chytrid infection in the Zion Canyon tree frogs were not lethal, but other environmental factors caused populations to fluctuate. Canyon tree frogs were captured and swabbed in each of nine canyons in and around Zion National Park during the summer of 2014, and total frog populations were counted in each study canyon. The swabs were analyzed for the presence of chytrid DNA, and infection rates and population sizes were compared with data from 2011-2013. We found that the presence or absence of chytrid had no influence on population size in a particular canyon; population sizes were more dependent on precipitation. The Zion canyon tree frogs resist this normally lethal disease; how they evade it will be the goal of future studies.

Aimee Newsham, Dixie State University Life Sciences 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.

Jeff Mecham, Brigham Young University Life Sciences Breast cancer is diagnosed in one of every eight American women. But, a safe, effective treatment for cancer has yet to be developed. Resveratrol, a naturally occurring phenol found in the skin of grapes, shows promise to be a powerful but safe chemotherapeutic in a sea of otherwise damaging and toxic treatments. However, the mechanisms by which resveratrol operates are yet to be fully understood. Our project focuses on the mechanisms by which resveratrol induces cell death in breast cancer cells. We will focus on the mechanisms of the p53 pathway. The protein p53 operates as an internal housekeeper of the cell. When DNA is damaged, p53 assesses the damage and can cause cell death when needed to prevent the spread and replication of the damaged cells. Cancer cells often decrease the amount of p53, allowing them to replicate without hindrance. Cancer cells treated with resveratrol show an increase of the amount of intracellular p53 restoring the cells’ ability to induce cell death. Our project focuses on two apparent mechanisms by which this increase occurs: calcium signaling and the decrease of ubiquitination of P53. Ubiquitin is a protein used to mark other proteins for degradation. Other papers and studies have shown that resveratrol inhibits key players in the process of ubiquitinizing p53. We will focus on resveratrol’s effect on both the ubiquitinizing and deubiquitinizing machinery, including G3BP1, USP10, and MDM2. A better understanding of the mechanisms by which resveratrol leads to the targeted death of cancer cells is an important step towards better cancer treatments.

Jordan Tingey, Brigham Young University Life Sciences Diabetes is a serious condition that is increasing worldwide. Diabetes is characterized by lost β-cell mass and uncontrolled blood glucose levels. Pancreatic islet transplantation could be used to cure people with diabetes, however the lack of islets is a major obstacle to its use. If we could understand how to increase β-cell proliferation and glucose stimulated insulin secretion (GSIS) then we could increase success in pancreatic islet transplants. Nkx6.1 induces β-cell proliferation. Nkx6.1 mediated proliferation is dependent on expression of Nr4a1 and Nr4a3. Nr4a1 and Nr4a3 are orphan nuclear receptors. It is currently unknown what ligand induces their activation. Previous reports have shown that free fatty acids induces expression of Nr4a1 and Nr4a3 in muscle, liver and adipose tissue. We show that culture of our INS-1 832/3 β -cell line in the presence of 0.2 mM palmitate induces expression of Nr4a nuclear receptors. Furthermore, culture with 0.2 mM palmitate results in increased β -cell proliferation. Finally, using INS-1 cells cultured with palmitate, and INS-1 cells deficient for either Nr4a1 or Nr4a3 we demonstrate the effect on mitochondrial respiration. Our data demonstrate that free fatty acids that are present during diabetes may induces expression and enhance activation of the Nr4a nuclear receptors, thus resulting in enhanced β -cell proliferation.

Alex Bischoff, Connor Dodge, Sarah Livingston, Sterling Rosqvist, Tomonori Baba, Kyle Larsen,

Alysa Edwards, University of Utah Life Sciences Percutaneous devices (PDs) constitute foreign materials that penetrate through the protective skin barrier to provide connection between internal and external environments. It has been previously shown that the periprosthetic tissue at the PD-skin interface is under a continuous state of wound healing, which often results in epidermal downgrowth. This continuous downgrowth is detrimental to the long-term survival of these devices. To date, there are no effective methodologies available to either prevent or quantify the degree of epidermal downgrowth indicating a need to find effective markers to document the healing response around these devices. In this study, periprosthetic tissues from a previous pig- back study were subjected to two different evaluations: (1) standard histology (HandE) and (2) immunohistochemical staining (IHC). Healing responses around PDs made with different material types were examined using cytokeratin 6 and collagen 4 to determine the degree of wound healing and granulation tissue maturity. Varying exposure time and concentration of stains, staining procedures were optimized. The interfacial tissues were then analyzed using either a photo or a confocal microscope. Preliminary data (Figure 1) indicated that there were noticeable differences in the periprosthetic regions between the material types used. The IHC data confirmed that the periprosthetic tissue is a hyper cellular region with a high density of blood vessels (collagen 4) and migrating keratinocytes. This data further confirmed the morphological differences observed between implant types using standard histology. Continued analysis will quantify the amount of collagen 4 within the periprosthetic tissue using imageJ software. Semi-quantitative data from each implant type will then be compared to predict biocompatibility. This research has demonstrated that IHC staining could be a potential tool for understanding the healing cascades around the percutaneous device.

Derek Munday, University of Utah Life Sciences Cardiovascular diseases (CVD) are more prevalent in individuals with diet-induced obesity (DIO) and type two diabetes (T2DM). Examples of CVD include blood vessel dysfunction and systemic hypertension. Both of these pathologies are associated with a reduced ability of the inner lining of the blood vessel (the endothelium) to release a substance (nitric oxide) that causes the blood vessel to dilate. At present the mechanism whereby T2DM and DIO decrease the function of the enzyme (nitric oxide synthase; NOS) responsible for nitric oxide synthesis and release is unknown. Determining this mechanism is the current focus of our laboratory. Earlier we reported that the sphingolipid ceramide is elevated in cell models of lipotoxicity and in mice with DIO. Most recently in endothelial cells we showed that ceramide causes protein phosphatase 2A (PP2A) to bind directly with NOS which disrupts the interactions among proteins that are necessary for optimal NOS function. My overall project was concerned with determining whether this mechanism is operational in mice with DIO. Specifically, we tested the hypothesis that PP2A inhibition would preserve vascular protein- protein interactions required for optimal NOS enzyme function to an extent that arterial dysfunction and hypertension would not occur. Mice consumed a control (CON) or high fat (HF) diet for 12 weeks. During the last 2 weeks, cohorts of mice from each group were injected (IP) with saline (vehicle control) or the PP2A inhibitor LB1 (1.0 mg/kg/day). We observed that interactions among proteins required for optimal NOS enzyme function were disrupted in arteries from mice with DIO treated with saline but not with LB1. Furthermore, arterial dysfunction and hypertension existed in mice with DIO that received saline but not LB1. These results strongly suggest that PP2A activation contributes importantly to arterial dysfunction that exists in a pre-clinical model of DIO.

Scott Frodsham, Brigham Young University Life Sciences The goal of this study is to better understand if the genetic variants that strongly correlate with an increased risk of developing multiple sclerosis (MS) also increase the risk of developing diseases that commonly co-occur with MS. This relationship can be determined by comparing genetic data of patients diagnosed exclusively with MS to the genetic data of patients diagnosed with both MS and one of its comorbid diseases. Many electronic medical records (EMR) collected at medical institutions are made available for research purposes. The EMRs of individuals contained in the database that will be used for this study are linked to corresponding genetic information. Data extraction via computer algorithm will be executed to identify patients who, because of their respective diagnoses, will provide meaningful data for analysis. The case group for individuals diagnosed with just MS and have available genetic information consists of 1003 individuals. Applying a basic algorithm (ICD-9 billing codes) to this group has shown preliminary data on patients with MS and one other comorbidity as follows: Hypertension, 192 patients; anxiety, 17 patients; hypothyroidism, 84 patients; Type 1 diabetes, 24 patients; inflammatory bowel disease, 12 patients; migraine, 116 patients; restless leg syndrome, 14 patients; rheumatoid arthritis, 28 patients. The algorithms will be modified to find and include more patients for analysis. We will enhance patient identification by including medications and text keyword searches of clinical notes in the search. Genetic analysis will be performed on the final dataset.

Brooke Linney, Brigham Young University Life Sciences Recombinant DNA technology has become one of the most critical fields of research relating to biotechnology. Recombinant DNA can be used to obtain certain proteins or examine the effects of genes that we engineer, with many applications in medical research. As part of our lab’s use for recombinant DNA, we create a gene sequence to code for a certain protein, and then use heat-shocking transformation to stimulate Escherichia coli bacterial cells to incorporate the mutated DNA from the surrounding solution. As the bacterial cells then grow, they replicate the mutated plasmid that we introduced. This DNA can later be extracted from the bacterial cells and used for further synthesis, usually protein synthesis in our lab. The process of transforming bacterial cells with mutated DNA is directly affected by plasmid size. Transformation efficiency is maximized with smaller plasmids. One of the DNA plasmids we use to introduce mutations is the pET9a vector. This plasmid is a sequence of 4,341 base pairs, but by reducing the length of the plasmid, we can increase transformation efficiency. By restricting the size of the pET9a vector, we will also be able to introduce larger foreign DNA sequences than we would with the original pET9a vector. This presentation will explore the different methods of reducing sequence length to optimize the pET9a vector, mainly focusing on site-directed mutagenesis coupled with the use of restriction enzymes.

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

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

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.

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

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.

Sharisa Nay, Brigham Young University Life Sciences Gene therapy is a growing field of science with the potential to improve thousands of lives. With an eye toward improving the effectiveness and longevity of gene therapies, my project examines the preferential binding tendencies of the histone protein variants Htz1 and Htas1. Htz1 is the Ceanorhabditis elegans homolog of H2AZ, an important variant of the H2A histone. This protein has been shown as necessary for survival and as playing a role in the prevention of ectopic heterochromatin spread. Htas1 is another variant of H2A that plays a role in the increased transcription of sperm-producing genes. The preferred positioning of these variants on naked DNA is not yet known. Through a DNA extraction, nucleosome reconstitution, and DNA digest and sequencing, we will take these variants and examine where they are prone to localize within the N2 Bristol strain of c. elegans. This will allow us to include DNA sequences on our gene insertions possessing a high binding-affinity for these transcription-promoting histones. Thus, if we can identify the locations at which these variants will localize within DNA, we will be able to insert these preferred constructs into the genes used for gene therapy and thereby increase the effectiveness of gene therapies.

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

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

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.

Julia Hull, Weber State University Life Sciences Endophytes are fungi that live within aerial portions of plants for most or all of their life cycle without causing visible signs of disease. Gall forming aphids, Pemphigus betae, are highly competitive over gall site selection (Moran 1993), forming galls on the leaves of narrowleaf cottonwood and their hybrids. The favored gall location overlaps with areas of highest endophyte probability. I hypothesized that a negative correlation would exist between endophyte infection and aphid galling on leaves of backcross hybrid cottonwood trees.

Brianne Palmer, Utah State University Life Sciences Inversions have plagued the valleys in Utah resulting in built up pollution carpeting the cities and spreading into surrounding ecosystems. The ecological impact of these inversions is unknown. Inversion-based pollution events deposit nutrients and pollutants in the ecosystems. The purpose of this study is to determine if lichens, due to their unique physiology, preserve a record of inversion-based nitrogen deposition in order to assess the ecological impact of the inversions. In June and July of 2013 we collected 111 samples of lichens from sites exposed to the inversions, along Red Butte Creek and on an elevation gradient on Grandeur Peak in Salt Lake City. The collection focused on two species of lichens (Xanthomendoza montana and Xanthomendoza fallax) because they are easily identifiable and live in all the sampling sites. The samples were ground, run through a stable-isotope mass spectrometer and analyzed for the %N and δ15N. If lichens hold a record of inversion-based depositions there should be more nitrogen in lichens from the sampling sites closer to the pollution source (Salt Lake City). Once the data was complied, the resulting graphs showed no correlation between %N and distance from Salt Lake City between δ15N and the pollutant source. However, the isotopic variability within the samples was greater than expected and requires further research.

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

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

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

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

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

Kevin Nay, Southern Utah University Life Sciences Leech taxonomy has traditionally been based on morphological characters, but with new developments in DNA technology many taxonomists are starting to use genetic information in descriptions of new species. Leeches in southern Utah are poorly inventoried with respect to many other aquatic animals. There have been few morphological inventories of leeches and even fewer descriptions of the genetic diversity within leeches. Landscape genetics is a powerful tool used to understand geographic patterns of genetic diversity. Southern Utah has many naturally isolated bodies of water due to the climate and the dramatic changes in elevation in this part of the country. The landscape genetic study of leeches in southern Utah will provide us with a better understanding of genetic differentiation within southern Utah leeches. The mitochondrial DNA (CO I region) will be used to estimate genetic diversity and examine the relationships among individuals in two populations of leeches. I hypothesize that leeches in southern Utah will have greater genetic diversity then historically recognized from morphological studies suggesting a new species of leech. The study will lead to better understanding of the taxonomy and identification of southern Utah leeches.

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

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

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

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.

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

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

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

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.

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

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

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

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