2015 Abstracts
Influence of antibiotics on Gut Microbiota and Resultant Psychological Behavior
Shireen Partovi, Utah Valley University Physical Sciences The normal flora of microbiota that resides in our gastrointestinal tract acts as a community and provides a number of functions such as assisting with the breakdown of waste, protecting our tissues and organs from invading species, and playing a role in the immune response. These microorganisms may also play a role in altering our brain chemistry and changing our psychology and behavior. This mechanism is considered to be due to their ability to produce neurochemicals that mimic those produced by our own bodies. Therefore, it is possible that antibiotics may have a detrimental effect on our gut flora, thereby inducing a host of undesired side effects. These described side effects may include changes in normal psychological behavior, such as the presentation of anxiety or depression. This hypothesis is increasingly relevant as antibiotic resistance is rising due to poor education regarding bacterial infections and as the appeasement of insistent patients continues. This research aims to illustrate the harm that over-medicating may have on our behavior as a result of afflicted gut microbiota. This research will use peer reviewed journal articles that include studies and experiments to determine the influence of gut microbiota on brain chemistry and therefore psychology and behavior. Specifically, the exact implications of consistent antibiotic use and the impact on gut microbiota will be examined and a correlation will be presented. Therefore, the purpose of this research is to illustrate this problem in regards to the healthcare industry and the over-medication of patient.
Chemistry Laboratory Safety: Misconceptions among First-and Second-Semester General Chemistry Students.
Jennifer Melvin and Wendy Schatzberg, Dixie State University Physical Sciences The general chemistry laboratory can be a dangerous place, and the possible danger is amplified by the fact that general chemistry is frequently a student’s first chemistry laboratory experience. College students come from a variety of backgrounds with only some having had chemistry in high school, and therefore many students will not know beforehand what kind of precautions to take or what kind of action to take in case of an accident in the lab. Safety information is given briefly at the beginning of the semester, but the efficacy of this is uncertain. An online survey was conducted to gauge first- and second-semester general chemistry students’ knowledge about chemistry laboratory safety and the data gathered was analyzed. This survey was conducted at two secondary schools in two different countries. Statistical analysis will be presented. Results will allow discernment of misconceptions held by students over two semesters and between countries, and conclusions can be made about the effectiveness of current laboratory safety instruction methods.
A Comparative Study on the Uptake of Nutrients and Trace Metals of Two Plant Subspecies (P.australis and P.americanus) in Utah Lake
Ashley Ostraff, Utah Valley University Physical Sciences Utah Lake has a long history of being impacted by anthropogenic activities like, mining, agriculture, and surrounding industry. All of these activities have contributed to the runoff that feeds the lake, increasing the likelihood that this area contains high levels of trace metals, nitrogen, andphosphorus. Utah Lake contains two subspecies of phragmites, a wetland reed, one native (P. americanus) and one non-native (P. australis). P. australis is replacing the native species at an alarming rate. P. australis is known to have a deeper root system than the native subspecies, because of this we suspect that this allow access to a less competitive soil level giving this subspecies greater opportunity for nutrient and trace metal uptake. By comparing the root zone soils of both subspecies we hope to gather results that support this hypothesis. Examination of the roots will also showthe potential influence the soil conditions have on their growth and development. This study will compare nutrient and trace metal uptake of each subspecies to determine impact. Other factors that will be assessed include plant physiology, carbon to nitrogen ratio (C:N), bioconcentration factor (BCF) and total trace metal content in tissues of both species. Samples of P. americanus and P. australis will be collected at 9 locations in Utah Lake. Soil samples at the root zone of each plant will also be evaluated. Each sampl e will be digested in the Microwave Accelerated Reaction System and analyzed in the Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES) for C, N, P, K, Ag, Al, As, Ca, Cd, Cr, Cu, Fe, Hg, K, Mn, Na, Ni, P, Pb, Ti, and Zn. Results from this study will contribute valuable data to future efforts being used to preserve the biodiversity of the plants and animals that live in and around Utah Lake. The end goal of this student project is to be submitted to peer-reviewed scientific journals for publication and to be presented at academic and scientific conferences.
Quantifying Heavy Metal Pollution in Utah Lake via Root System Accumulation in Two Subspecies of Phragmites Australis and Subsequent Determination of Anthropogenic Relevance
Kevin Jackman, Utah Valley University Physical Sciences Phragmites australis is a non-native subspecies of wetland reed that was introduced to Utah Lake from Europe during early exploratory settlements and is now outcompeting native flora in the lake’s wetland ecosystem. Utah Lake is a repository for toxic heavy metals from diverse mining operations and industrial operations proximal to the water. International studies have shown Phragmites to have strong potential as a phytoremediator and a reliable biomonitoring species of polluted water and soil, yet no work in this regard has ever been performed in the state of Utah or on Utah Lake. It is by measuring the concentrations of arsenic, lead, and 12 other trace metals within the root and rhizome system of these plants that a measurement of the contamination of the lake can be made, and to determine a quantitative concentration and severity of contamination with regards to public health and safety. If these trace metals are present in excess in the lake and its soils, toxic, and harmful conditions are present and are an issue of health to the natural ecosystem of the lake, as well as the citizens recreating and working throughout the lake on a regular basis. Determination of atomic content evaluation will be performed by the Induced Coupled Plasma Optical Emission Spectrometer. Future work can then be proposed to remediate the lake, in an effort to improve the human and environmental condition of the area. This project has the interest of the Utah Department of Environmental Quality and relationships have been established for current and future cooperation. The aim of this project is to be published and presented on a peer-reviewed level in scientific journals and at conferences.
Expression of the C-terminal Domains of the Tight Junction Proteins Claudin-16, -3, and -4 to Identify Interacting Proteins in Epithelial Ovarian Carcinoma
Brandon Davies, Utah Valley University Physical Sciences Epithelial ovarian carcinoma (EOC) is the sixth most common cancer in US women. The long-term cure rates are low due to the lack of reliable biomarkers for early disease detection, resulting in advanced stage diagnosis. Approximately 75%-80% of ovarian cancers are diagnosed at stages IIIV with a 10% 5-year survival rate despite aggressive treatments. Claudin proteins are being studied as possible biomarkers as they are aberrantly overexpressed in EOC tumors. The Claudin family of proteins are a main component of tight junctions in the upper region of epithelial cells that act as gateways for the exchange of water and solutes while also helping determine the cell’s polarity and function. Changes in these proteins cause changes in phenotype and function of normal epithelial cells, such as proliferation control, trans-epithelial resistance, polarity, and solute transport. Claudin-16 is often aberrantly expressed in breast and ovarian cancer, while Claudins 3- and 4 are highly overexpressed in EOC. The location of these proteins is also correlated with oncogenic transformations and cell proliferation. Determining the specific characteristics of these Claudin proteins can prove to be of incredible benefit in cancer treatments. As these proteins are targeted during these therapies, these tight junctions may then send normal signals, which in turn can regulate the cell normally. The C-termini of the Claudins, which are cytoplasmically located, contain a known PDZ-binding motif and may interact with other junction proteins or with proteins involved in interesting signaling pathways. To identify these interacting proteins, we will use the Expresso T7 Cloning System (Lucigen Corp., Middleton, WI) to purify the Claudin-16, -3, and -4 C-terminal tails to use in pull-down assays. This process includes using affinity tags to capture the Claudin tails by FPLC, which can then be analyzed by SDS-PAGE and, ultimately, the corresponding genes cloned and sequenced. This study can potentially provide crucial information in relation to how members of the Claudin family interact with other proteins that are commonly found in tissues that are misregulated in cancer. With this data treatments can be improved to increase the responsiveness of ovarian cancer patients.
Spatially Understanding Utah’s Water Resources: A Story Told through Maps
Tim Beach, Utah State University Physical Sciences Successful management of the arid West requires a basic knowledge of available water resources, withdrawals, and existing management efforts. Utah, the second driest state in the country, is continuing its efforts to restore impaired water systems and teach the next generation to conserve water. Because of a fast-growing population, the diversity of Utah’s landscape, and strict regulations that govern water throughout the state, understanding the region’s current water condition can be difficult. Historically, the majority of this information has been stored in databases, represented by only numbers and statistics. In an effort to better conceptualize this data and increase the spatial understanding of Utah’s water, a set of maps have been produced. These maps were created using ESRI’s ArcMap software and Adobe Illustrator. Included are major lakes and rivers, average annual precipitation, average annual streamflow, total urban withdrawals per county, total agricultural withdrawals per county, and water management. Viewing visual representations of this information in multiple maps can aid in creating spatial inferences, and raise additional questions regarding aquatic restoration and management throughout the state. This set of maps can also be used to supplement physical science education at all school levels. Future work includes distributing these maps as posters to schools and water managers, as well making it a resource for teachers via websites and fact sheets.
Peak Density Histograms and Pathology Interpretations for High-frequence Ultrasonic Testing of Breast Cancer Surgical Specimens
Robyn Omer, Utah Valley University Physical Sciences Removal of all malignant tissue during lumpectomy is critical for preventing local recurrence of the breast cancer. Failure to remove all cancer results in 20-40% of lumpectomy patients returning for additional surgery. At Utah Valley University, a method is being developed to detect cancer during the initial surgery to ensure all of the cancer has been removed. Peak density, which is the number of peaks and valleys in a specified spectral range of a high-frequency (HF) ultrasound signal, correlates to breast pathology in lumpectomy specimens. The objective of this study was to determine if the histograms of peak density versus the number of measurements provide information on corresponding breast tissue pathology. High-frequency ultrasonic data were obtained from a blind study of surgical specimens obtained from 73 lumpectomy patients at the Huntsman Cancer Institute in Salt Lake City, Utah, and South Jordan, Utah. The data were normalized to remove bias between patients. The ultrasonic signals were converted to spectra using a Fourier transform. Peak densities were calculated from the spectra by counting the number of peaks and valleys in the 20-80 MHz range. This was achieved by counting where the slopes of the spectra (their derivatives) crossed zero. A histogram was created by assigning each peak density value to a bin, and then counting the number of measurements that fell within that bin. The histogram of the peak densities produced an asymmetric Gaussian-type distribution with a range of peak density values from 0 to 27 and a mode of 5. Using threshold values determined from a pilot study for differentiating pathology with peak density, it was determined that the peak of the distribution (5-6) corresponded to normal tissue pathology, the shoulders of the distribution (0-4 and 7-10) corresponded to abnormal pathologies, and the tail of the distribution (11-27) corresponded to malignant tissue types. These correlations matched the types of specimens tested, specifically tumors, margins, and lymph nodes. The correlations also provide a measure of the success of removing malignant tissue and achieving negative margins during lumpectomy procedures. Using histograms to analyze the data not only provides a new approach for differentiating tissue pathology, but also provides a statistical measure of the success of lumpectomy procedures performed by a specific surgeon or at a specific institution.
Measuring the Effect of Breast Density on High Frequency Ultra sound using Tissue Phantoms
Zachary Coffman, Utah Valley University Physical Sciences Breast density describes the proportion of connective tissue versus the fat tissue in the breast. Studies have shown that women with higher breast density are four to five times more likely to develop breast cancer than women with lower breast densities, (www.women.org/BreastCancer). Higher breast densities have proven to make current breast cancer imaging and detection more difficult. A pilot study done at the Huntsman Cancer institute showed that the ultrasonic parameter peak density, generated by high-frequency (HF) ultrasound (20-80 MHz), was sensitive to breast tissue pathology. The objective of this study was to determine the effect of breast density on ultrasound wave propagation from high frequency ultrasound using phantoms that mimic the histology of breast tissues. Phantoms were created from a mixture of distilled water, agarose powder, and 10X TBE stock solution. In order to simulate breast tissue histology and breast density, polyethylene microspheres were embedded into the phantoms in layers, totaling 4 layers per phantom. The polyethylene microsphere size (90-106 μm diameter) was kept constant within each phantom while the weight percent concentration of the microspheres varied (0.00g to 0.06g). Pitch-catch and pulse-echo measurements were acquired using 50-MHz transducers (Olympus NDT, V358-SU, 50 MHz, 0.635-cm diameter active element), a HF pulser-receiver (UTEX, UT340), and a 1-GHz digital oscilloscope (Agilent DSOX3104A). Glycerol (Genesis Scientific) was used as a coupling agent between the transducers and the phantoms. Spectra were derived from the data, giving peak density (the number of peaks and valleys in a specified spectral range), velocity, and attenuation values. The results showed that peak density did not start to show a trend until phantoms of 0.03g concentrations, where it increased from a value of 14.0 peaks (0.03g) to 18.7 peaks (0.06g). Velocity showed a statistically significant increase with greater polyethylene microsphere concentration, from 1508 m/s for 0.00g to 1536 m/s for 0.06g. No trends were observed for attenuation. These results indicate that higher levels of scattering centers in dense breast tissues will be detectable with high frequency ultrasound. This additionally shows that high frequency ultrasound may also be sensitive to greater amounts of connective tissue present in dense breast pathologies. High frequency ultrasound is sensitive to the weight percent of polyethylene microspheres. Future research is planned to further understand this relationship, including repeat studies and studies of phantoms containing chopped polyethylene fibers and triple the polyethylene microsphere concentrations to more closely simulate dense breast tissues.
Plant Fossil Locality in the Shinarump Member of the Chinle Formation (Upper Triassic) of Southwestern Utah (Washington County)
Melinda Hurlbut, Dixie State University Physical Sciences A newly discovered plant fossil locality in the Chinle Formation (Upper Triassic) in Washington County broadens the distribution of known Late Triassic plant fossil localities to include southwestern Utah. Previously reported plant localities in the Chinle Formation are known from southeastern Utah, Arizona, New Mexico, and equivalent strata from the Dockum Group in Texas. The majority of the known plant fossil localities are from higher stratigraphic units of the Chinle Formation. However, the new Washington County locality is from the older, basal unit, the Shinarump Member. The plant fossils identified from the new locality include fronds of the fern Phlebopteris sp., fertile leaves of the fern Cynepteris sp., and leaves of the bennettitalean Zamites sp. Undetermined species of conifers are represented by a partial branch and a three- dimensionally preserved cone containing well preserved seeds. The diversity of species found at the Washington County plant fossil locality correlates with other known floral assemblages from the Chinle Formation, suggesting general homogeneity of Chinle Formation floras. These plant fossil assemblages support interpretations of basal Chinle Formation paleoclimates as humid and warm.
High-Frequency Ultrasound (20-80 MHz) for Analyzing Breast Cancer Surgical Margins: A 73-Patient Clinical Study
Amy Fair Brother, Utah Valley University Physical Sciences Results from a 2010 pilot study indicated that multiple parameters in high-frequency (HF) ultrasound spectra (20-80 MHz) correlate to a range of tissue pathologies in surgical margins from breast conservation surgery (BCS). One of these parameters, peak density, was particularly effective at discriminating between normal, atypical, and malignant patholUtah Conference on Undergraduate Research 2015 100 ogies. Subsequently, Utah Valley University and the Huntsman Cancer Institute initiated a follow-up study to further investigate this approach. Objectives: The purpose of this study was to determine the sensitivity and specificity of HF ultrasound for differentiating malignant tissue from normal tissue in BCS surgical margins. Methods: A 73-patient blind study was conducted with conventional pathology used as the gold standard for assessing the HF ultrasound method. Specimens were delivered by the surgeon’s team immediately following resection and ultrasonically tested outside the surgical suite. The margins were approximately 3x20x20 mm, and were oriented using a small staple inserted by the surgeon in one corner and a stitch on one side. The margin was tested at 2-5 locations on the specimens using our methodology and then sent to pathology for their analysis. 498 specimens were tested from 73 patients. That corresponded to 1112 positions collected from the margins. The data was then analyzed for malignancy using peak density, and then correlated with the traditional pathology. Results: Results from the current study indicate that peak density can differentiate between malignant and nonmalignant pathologies with an accuracy of 73.8%. The correlation between pathology and peak density has a high level of statistical significance compared to random chance, with p = 0.000078 (Fisher’s Exact test). The results also provide data for improving the technique. For example, approximately 3 times more false positives were observed than false negatives, indicating the peak density threshold used for identifying malignant pathology is most likely too low and should be adjusted to a higher value. Conclusions: Results from this study showed that HF ultrasound has the potential to provide rapid, intraoperative evaluation of surgical margins, thereby decreasing the number of additional surgeries for patients and thus increasing the quality and efficacy of surgical treatment for breast cancer.
Towards The Synthesis Symmetrical and Asymetrical Bimetallic Complexes for Use in Studies of the Oxygen Reduction Reaction
Eric Johnson, University of Utah Physical Sciences The ability to reduce O2 in mild conditions holds many important implications such as: use as an economical fuel cell, pharmaceutical synthesis, biomass degradation and conversion of small molecules to fuels. We are building the [M(μ-OH) (oxapyme)M(H2O)]+ molecules and symmetrical counterparts for use in O2 reduction reactions (M = Cobalt, Nickel, Iron). The precursors to the [M(μ-OH)(oxapyme)M(H2O)]+ have been synthesized as follows. 2-[5-(2-Nitro-phnyl)-[1,3,4]oxadiazol- 2-yl]-phenylamine serves as the backbone of the complex, allowing for two distinct ligands to be attached to each side. Initial yields for this synthesis averaged at 6%. To be able to complete the synthesis this needed to be significantly raised. The literature procedure was modified in various ways until new reaction conditions were found that allowed for 40% yield. Other precursors include 2,2’-(1,3,4)Oxadiazole-2,5-diyl-bis-aniline which also serves as a ligand backbone but differs in that it allows for preparation of a symmetrical ligand have been synthesized with a 51% yield. The first ligand Bis-pyridine-2-ylmethyl-amino has been produced with a 60% yield. The second ligand Methyl-pyridine-2-ylmethyl-amino has been synthesized with an approximate yield of 75%. These yields are high enough to finish the synthesis of the ligand and subsequently coordinate the metals. Upon completion, the electrochemical properties of the compounds that differ in the metal composition and the ligand (symmetrical versus asymmetrical) will be determined using studies such as cyclic voltammetry. Once the metal and ligand that are most apt at oxygen reduction is determined, more advanced studies will be undertaken to identify the reaction mechanism and intermediates.
Invasive Aquatic Species Attachment Inhibited by Surfactant Paint
Kyle Marcus, Cami McKellar, Riley Pearce, Shay Beck, and Zenja Draca, Dixie State University Physical Sciences Invasive marine and freshwater species have a detrimental impact on aquatic ecosystems and are easily transferred between bodies of water as a result of unregulated settlement and attachment to commercial and recreational watercraft. This results in infestation of waterways and disruption of native organisms throughout an ecosystem. The quagga mussel (Dreissena rostriformis bugensis) is one example of such invasive species that has called for a focus on cross-contamination prevention. The goal is to use aquatic paint suitable for watercraft that prevents the settlement and/or attachment of various aquatic species such as algae, snails (Physa sp.), anemones (Aiptasia sp.), and quagga mussels. The paint is integrated with newly developed surfactants that act as an inhibitor to the adherence ability of invasive aquatic species. The surfactants are added to provide a barrier between the paint surface and the organisms creating an undesirable surface area for the organisms to attach. This barrier disrupts the specific naturally occurring chemical and physical processes that allow organisms to adhere to surfaces. The most innovative characteristic of the surfactant-based paint is its effectiveness on adherence inhibition without degradation, dissociation, or toxicity to the environment. Preventing adherence of invasive organisms to surfaces could have significant positive effects on the efficiency and cost of operations in marine environments and waterways.
A Plan for Complete Recycling of Stormwater on the Utah Valley University Main Campus, Orem, Utah
Paul Robertson, Utah Valley University Physical Sciences Evaluation of Utah Valley University’s stormwater plan reveals a simple system meant to collect stormwater into the city storm drains as quickly as possible. It is, however, vastly underdeveloped and many unspectacular summer and springtime storms have resulted in property damage, including those of nearby residents. The stormwater runoff has also collected concentrated amounts of hydrocarbons, nitrogen and heavy metals which are being fed directly into Utah Lake, acting as a significant source of pollution for the lake environment. The intentions of this project are to design a stormwater management plan that can withstand a 100 year, 24 hour event and prevent pollutants from entering the Utah Lake system. Mapping and modeling of the University’s storm drains will be accomplished using GIS as well as modeling for efficient retention sites on campus. Captured stormwater will then be used for a variety of functions here on campus and runoff into the adjacent lake will be reduced to insignificant values. Decisions regarding the ultimate implementation of this project will work in concordance with the University’s master plan of future development in order to realistically secure a reliable, low-maintenance system.
Patterning Supported Lipid Bilayers with Magnetic Tweezers
Madeline Parson, Travis Bulloch, and Tyler Argyle, Southern Utah University Physical Sciences Lipid bilayers have many important purposes in living cells. A lipid bilayer forms a barrier which separates the fluid inside the cell from the fluid surrounding the cell. The arrangement of components within cell membranes can be extremely important, particularly in cell communications. For example, when our immune system attacks certain pathogens, it recognizes them by specific proteins in the inner and outer regions. This can be thought of as forming a “bull’s-eye” shape. The ability to generate such patterns in bilayers might see applications in many areas of biology. Our goal is to take an initially un-patterned supported lipid bilayer (model cell membrane) and use magnetic tweezers as a delivery system to generate patterns. We hope to show pattern formation using fluorescent-labeled lipids within our bilayer. In the past, we have used streptavidin and biotinylated lipids with Oregon Green. Currently, we are pursuing a fluorescein/anti-fluorescein system that has been yielding much more promising results. We are currently using a fluorescent microscope to confirm that patterns are forming within the bilayer.
Mobility and Distribution of Trace Element Pollution in Sediments of the Utah Lake Outlet
Henintsoa Rakotoarisaona, Utah Valley University Physical Sciences The Jordan River is the only outlet of Utah Lake. Historically, this area has been impacted by urbanization, long term mining operations, industrial and agricultural activities resulting in potentially high levels of trace metal pollutants at the headwaters of the Jordan River. Since trace metals are known to be toxic at elevated levels, it is important to evaluate their concentration, distribution and mobility in this sensitive area in order to determine risk to wildlife, humans and downstream users of the Jordan River. Three core samples from 0 cm to 95 cm in depth were collected at the east (industrialized area), west (newly developed area) and north (an island barrier) sides of the outlet of Utah Lake. Each sample was digested in triplicates in the Microwave Accelerated Reaction System (MARS) using US Environmental Protection Agency Method 3052 and analyzed in the Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) for As, Cd, Cr, Mn, Pb, Cu, Zn, Co, and Ni. A multivariate analysis of variance (MANOVA) was used to analyze the data, with a boneferroni adjustment made for multiple comparisons. The results indicated that the East and North sediments of the Jordan River were significantly (P < 0.5) more heavily impacted by trace metal pollutants than the West side, with the North area accumulating higher levels of the majority of the trace metals or metalloid evaluated. Enrichment of Co, Cr, Pb and Zn was observed on the East sediment at concentrations reaching 10,821, 4.07, 13.7, 12.7, 119.4 mg kg-1, respectively. The most substantial elevation in the concentration of trace metals occurred with Cu in the North section of the lake, increasing from 349 mg kg-1 at 0-15 cm to 1383 mg kg-1 cm depth, showing high mobility, followed by Zn which increased from 46.7 mg kg-1 at 0-15 cm to 592 mg kg-1 30-45 cm depths. The enrichment of these trace metals in the East and North sediments at the outlet of Utah Lake pose a health risk to animals and humans who use these areas for recreational or agricultural purposes.
Correlation of Force and Peak Density during High-Frequency Ultrasound T esting: A Reliability Study
Benjamin Finch, Utah Valley University Physical Sciences The Breast Cancer Research Laboratory at Utah Valley University has been using high-frequency ultrasound to test the pathology of surgical margins from breast cancer conservation surgery. Their studies have shown that high-frequency ultrasound may be sensitive to a range of breast pathologies. The ultrasonic parameter that has been shown to be the most sensitive to pathology is the number of peaks (peak density) in the frequency spectra from the captured waveforms. During testing, the ultrasonic transducers apply an amount of force to the tissues that can vary depending on the researcher. The central question of this study was to determine the effect that the applied force has on the final peak density reading, and therefore the reliability of the results. In order to determine the correlation of force and peak density, an experiment was designed to measure the force applied to tissue by the ultrasonic transducer during testing and to simultaneously measure the applied force while collecting ultrasonic waveforms. An Arduino Uno R3 instrument was obtained as well as an Interlink Electronics FSR 406 force sensor pad. The Arduino was programmed to read the voltage from the FSR sensor and use that information to provide force (N) and pressure (N/cm2) data from the stage. Peak density readings from bovine mammary tissue (very similar to human breast tissue) were acquired using varying amounts of force. The results obtained exhibited significant changes in peak density with applied force. The results showed that if the applied force is under 1.18N, the peak density will fluctuate significantly and will therefore give inconsistent results. However, if the force applied is greater than 1.18N, the peak densities will maintain a relatively consistent form. In conclusion, if the force applied during testing is above 1.18N, the waveforms captured from high-frequency ultrasound testing on breast cancer tissues will provide consistent and reliable results, thus improving the quality of the data and accuracy of diagnosis.
Hip Joint Center Differences between Dual Fluoroscopy, Functional Hip and Coda—Establishing a Reference Standard
Michael Kutschke and Niccolo Fiorentino University of Utah Measurements of joint angles, moments, and forces can be sensitive to positional changes in the center of a joint. Due to its deep location, the hip joint center (HJC) can be difficult to determine. Regression equations and functional methods use skin markers and are commonly used to measure the HJC, yet introduce sources of error such as bony landmark identification, soft tissue artifact, and system resolution. Additionally, the accuracy of said methods cannot be established in-vivo without a reference standard. The purpose of this study was to implement a dual fluoroscopy (DF) and model-based tracking technique to quantify the subject-specific HJC position in-vivo. This reference standard (DF-HJC) was then compared to the HJC defined by regression equations (CODA) and a functional method (FHJC) using skin markers. Five subjects with normal hip anatomy provided informed consent to participate in this IRB approved study. A gait marker set was applied to the subject’s skin. The subject performed various activities involving the inferior limb while a 10-camera Vicon system measured the position of the skin markers. From these data, the CODA and FHJCs were identified. Simultaneously, the hip was visualized using a DF system, and the DF-HJC was identified using model-based tracking. The DF-HJC was then transformed to the Vicon coordinate system for comparison with the CODA and FHJCs. The average Euclidean distance (and standard deviation) for the five subjects from the DF-HJC was 2.0 (0.6) cm for CODA and 1.3 (0.4) cm for the FHJC. To our knowledge this is the first study to use DF and model-based tracking to measure the in-vivo position of the HJC. This technique affords greater accuracy in determining subject-specific bone geometry and visualization of deep hip anatomy, thus may be considered a reference standard for identifying the HJC.
Monitoring Angiogenesis in Early Chick Dev elopment Using High-frequency Ultrasound: Method Development and Preliminary Results
Cameran Mecham, Utah Valley University Physical Sciences Introduction:
High-Frequency Ultrasonic Measurement of Angiogenesis in Mice with Breast Tumors and Ligated Femoral Arteries
Michaelle Cadet, Utah Valley University Physical Sciences Breast cancer is the most common cancer among women in the United States. Tumor angiogenesis and its inhibition is an important aspect of oncology and the treatment of cancer. High-frequency ultrasound (10-100 MHz) is particularly sensitive to small vascular structures that are close in size to the ultrasound wavelength (15-150 _m). The ability to rapidly determine the degree of vascularization in small animals in vivo would provide a useful characterization tool for breast cancer studies. The objective of this study was to determine if direct ultrasonic measurements in the 10-100 MHz range could be used as a vascularization assay for breast tumors and other tissues. To accomplish this, six mice from the Huntsman Cancer Institute (Salt Lake City, Utah) with grafted breast cancer tumors (three control and three treated with an angiogenesis inhibitor called Avastin) were tested in vivo using through-transmission ultrasonic measurements. A second study was also performed at the Ludwig Boltzmann Institute for Experimental and Clinical Traumatology (Vienna, Austria), where the femoral artery in one hind leg of each of sixteen mice was ligated and tested over the time period of eight days. Eight of the ligated limbs were treated with vascular endothelial growth factor (VEGF) while the remaining eight ligated limbs were allowed to grow ischemic. The unligated limbs were controls. Results from the Huntsman Cancer Institute study indicated that breast tumors in Avastin-treated mice showed higher ultrasound velocities than control tumors. This can be ascribed to the vasculature in the nontreated tumors creating greater wave scattering in the tissue, thus decreasing the velocity. Results from the Boltzmann Institute study indicated that in mice with ligated femoral arteries, ultrasonic signals from ischemic limbs displayed a decrease in wave velocity over the test period as compared to the VEGF-treated limbs. However, both the ischemic and VEGF-treated limbs showed decreases in ultrasonic attenuation during the entire test period. Results from Avastin-treated mouse tumors and mouse limbs with ligated femoral arteries revealed that high-frequency ultrasound holds potential for measuring angiogenesis in vivo.
Peak Density and Attenuation as Complementary Parameters for Differentiating Breast Tissue Pathology
Nicole Cowan, Utah Valley University Physical Sciences Breast cancer is the second most prevalent cancer among women, affecting one out of eight women in their lifetime. The ability to differentiate between malignant and normal tissues during breast cancer surgery would enable the surgeon to remove all of the cancer from the affected region in the breast, thereby reducing the risk of recurrence and the need for subsequent surgeries. A pilot study conducted at the Huntsman Cancer Institute showed that high-frequency ultrasound (20-80 MHz), and in particular the ultrasonic parameter peak density, was sensitive to breast tissue pathology. The objective of this study was to determine the effect of tissue microstructure on peak density using phantoms that mimic the histology of breast tissue. Phantoms were created from a mixture of distilled water, Knox gelatin, and Metamucil fiber. In order to simulate breast tissue histology and terminal ductal lobular units, polyethylene microspheres were embedded into the phantoms in layers, totaling 4 layers per phantom. The volume percent of polyethylene microspheres was kept constant in each phantom while varying microsphere sizes (58-925 μm diameter). Pitch-catch and pulse-echo measurements were acquired using 50-MHz transducers (Olympus NDT, V358-SU, 50 MHz, 0.635-cm diameter active element), a HF pulsar-receiver (UTEX, UT340), and a 1-GHz digital oscilloscope (Agilent DSOX3104A). Glycerol (Genesis Scientific) was used as a coupling agent between the transducers and the phantoms. Spectra were derived from the data, giving peak density (the number of peaks and valleys in a specified spectral range) and attenuation values. In a previous study, histology- mimicking phantoms were fabricated where the weight percent of polyethylene microspheres was kept constant, but the microsphere diameter was varied. The former study showed a clear trend of higher peak density values for smaller diameters, but no trend for attenuation. In contrast, the phantoms from this study showed no trend in peak density, but a clear trend of higher attenuation values for larger microspheres. The results show that specific changes in tissue microstructure affect the parameters of peak density and attenuation differently. Changes in the number of scatterers and in their size, as in the previous study, affected peak density most significantly. In contrast, changes solely in the size of the scatterers, but not in their number, affected attenuation most significantly. These results are consistent with attenuation results for lobular carcinoma in the pilot study. These results show that peak density and attenuation are complementary parameters, and could be used together to characterize a variety of tissue pathologies
Effective Dissolution of Keratin Using Ionic Liquids to Extract Radioactiv e Materials
Brett Barton, Dixie State University Physical Sciences Dissolution using ionic liquids has been shown to be an efficient analytical method of dissolving keratin fibers. Using wool and hoof material from livestock, chloride-based ionic liquids were used to break down the hydrogen bonds important for keratin structure, making the extraction of constituent material much easier. Efficient methods of extraction allow for small samples to be analyzed while still providing high yields; efficient dissolution of keratin in a small sample size will be utilized to prevent any harmful effects on the animals. Ionic liquids are nondestructive solvents which allow for the safe extraction of organic substances. Denaturing keratin would help in the process of identifying any constituent radioactive materials. Efficient identification of radioactive material in livestock is crucial for maintaining health and quality of life.
Analytical Chemistry Applications for Ionic Liquids as Novel Solvents
Mattie Jones, Dixie State University Physical Sciences Current techniques for isolating components of samples found at crime scenes by their unique chemical properties are lengthy and often destroy important forensic evidence. New methods aimed at forensic analysis of sensitive, minute samples are critical to the intelligence community. In particular, successful extraction of dyes from materials found at crime scenes will provide innumerable benefits for matching, identifying, and finding origins of these materials and dyes. Ionic liquids possess the necessary chemical properties to ensure efficient extractions while maintaining the forensic signatures of the original materials. Ionic liquids, which are organic salts that are room temperature, provide a versatile solvent to achieve single-component extraction-separation-identification of forensic analytes. Following extraction, successful identification by infrared, absorption, and fluorescence spectroscopy has provided evidence of preserved quality and complete separation of material and dye. This novel approach to forensic analysis is advantageous particularly when sample sizes are extremely limited, but it can be readily scaled to larger applications. Developing a simple and affordable method of achieving specific molecular interactions provides a solution for often unidentifiable evidence in crimes. Harnessing the versatility of ionic liquids in a high-yielding recovery and efficient single-pot methods will enhance forensic abilities for the intelligence community and forensic investigators.
Species List of Insects in Capitol Reef National P ark
Robert Erickson, Utah Valley University Life Sciences Although the insect fauna of the Colorado Plateau region are somewhat well known, our specific understanding of the arthropod biodiversity in Capitol Reef National Park has been sparse. Objective: From the multiple insect surveys conducted in Capitol Reef National Park we intend to catalog the arthropod biodiversity into a species list. Methods: In addition to the previous collecting trips, we carried out collection efforts this past summer (2014). We used the Utah Valley University Capitol Reef Field Station as our home base. General insect collecting efforts were conducted around the field station and the nearby Pleasant Creek, near the public campgrounds, along trails, and in several other locations in the southern portion of the park. Additionally, we performed night collecting with a mercury vapor lamp trap on the nights we were in the park and utilized stationary malaise and aquatic larvae traps. The collected specimens were curated using methods of pinning, spreading, labeling, identifying, photographing, and organizing the insects. Results: The collections contributed to an increased understanding of the parks insect diversity and resulted in a curated natural history museum collection. A species list will be made available for the records of Capitol Reef National Park. More than 3000 specimens have been collected within the park. Furthermore, a booklet of the common insects for the park is in the process of being created in order to serve as an educational tool for visitors to the park and field station.
Evolution and Molecular Phylogenetics of Baetidae (Ephemeroptera)
Chase Barker, Utah Valley University Life Sciences Central Research Question:
Phylogeography of Desert Iguanas
Michael Packer, Utah Valley University Life Sciences Dipsosaurus dorsalis is a desert dwelling iguana species geographically distributed throughout the south-western United States, northern Mexico, and the Baja Peninsula. Throughout the Baja Peninsula, multiple species of herpetofauna exhibit a distinct north-south division, both morphologically and genetically, with no presence of any physical barriers. The goal of this study was to examine how the genetic structure of D. dorsalis has been influenced by past geological events, and whether this species exhibits a similar north-south division on the Baja peninsula. Additionally, this study aimed to see if the current taxonomy of Dipsosaurus is reflective of the evolutionary relationships across its entire geographic range. 100 tissue samples of D. dorsalis were collected across the species geographic range. DNA extraction from collected tissue samples and sequencing of nuclear loci MLH3 (~900bp), NT3 (489bp), as well as mitochondrial loci ND4 (~900bp) were performed to examine the distribution of genetic variation in D. dorsalis. The collected data was then used to construct phylogenetic trees for each locus, comparing individuals of Dipsosaurus to the geologic history of its geographic distribution. Although shallow, a maximum likelihood tree of the ND4 mitochondrial gene shows the phylogeographic separation of three distinct clades. The results do not support the designation of Dipsosaurus catalinensis as currently defined.
Gone Again: A story of Evolution, Mandibular Tusks, and Burrowing Mayflie
Stephanie Bartlett, Utah Valley University Life Sciences Our project investigates the phylogenetic relationships of the superfamily Ephemeroidea + Behningiidae. Found in waters worldwide, burrowing families are unique in that they have mandibular tusks that allow them to tunnel in the silt or gravel of riverbeds. Surprisingly, even without the mandibular tusks necessary for tunnel construction, the Behningiidae family is still found within these burrows as nymphs. Because the Behningiidae don’t have tusks, morphological research has lead to a phylogenetic classification of this family that our genomic investigation ultimately disputes. To begin this investigation, mayfly specimens were collected worldwide and prepared for DNA extraction. For each specimen genes were amplified via polymerase chain reaction and visualized on an agarose gel, before being sequenced and analyzed. The specific genes targeted for this analysis include; 12s mitochondrial rDNA, 16s mitochondrial rDNA, 18s nuclear rDNA, 28s nuclear rDNA, H3 nuclear protein coding, and CO1 mitochondrial protein coding. Datasets were supplemented with sequences acquired from Genbank. The ingroup consisted of approximately 30 samples. Phylogenetic relationships were estimated using Maximum Parsimony, Maximum Likelihood, and Baysian methods. We constructed phylogenetic relationships of burrowing mayflies using molecular DNA data analysis, when compared to morphological analysis we identified some important classification differences. As a result of our findings, we propose an alternative explanation for the evolution of mandibular tusks in burrowing mayflies (Ephemeroptera). The data support that burrowing mayflies first evolved tusks. Within the family Behningiidae, tusks were lost, while the burrowing lifestyle (i.e., living in a burrow to filter feed) was retained. This study represents the largest analysis to date for these insects and strongly supports the evolutionary trend of a gain and a loss of mandibular tusks during their evolution.
To Accept or Not Accept: A study of Evolution and Worldview Reconciliation in an Introductory Biology Course
Danny Ferguson, Utah Valley University Life Sciences This research examined the acceptance of evolution for introductory Biology students and the reasons why they don’t accept evolution, accept evolution, and why they change their minds over the semester. Previous studies examined student’s observations and knowledge of the evolutionary theory and found that the degree of conflict students perceived between religion and science was negatively correlated with their knowledge of evolution. Objective: The objective of this research was to better understand the student’s views of evolution and its integration into their worldly and religious views. Methods: We will conduct interviews with general Biology students in order to better understand the reasons why they don’t accept evolution, accept evolution, and why they change their minds over the semester. The interview questions are designed to investigate, in more detail than the previous surveys, the opinions of evolution and how they change over the course of the semester. The recordings will be transcribed and quantified by binning answers into categories. Given high % of students are LDS, we will ask a few additional questions to this portion of the population. Results: We found that as students knowledge of evolution increased, their conflict with religion decreased. The data demonstrate that there are three main reasons for this trend: 1) Evidence convinces the students to accept evolution; 2) Particularly for LDS students, knowledge of the official position of the Church, enables them to allow evolution to be a correct process in nature; and 3) the instructor as a role model (believer yet accepts evolution) can have a large influence.
Comparison of Macroinvertebrate Assemblages of Two Perennial Streams in Capitol Reef National Park
Jake Loveless, Utah Valley University Life Sciences Macroinvertebrate assemblage composition was assessed in two perennial streams, Pleasant Creek, and Sulphur Creek in Capitol Reef National Park, Utah, during the summer of 2014. Individuals collected were identified to genus. This information was used to compare assemblage composition, nutrient availability, and water quality between the two streams. Five collection trips were made (May 27, June 10,17, July 1,15). Sampling was conducted randomly using mesh kick nets with four samples being collected per site, per trip. Heavy rains caused flash flooding on July 15 prohibiting sampling, so a total of sixteen samples per site were taken. Samples were stored in 90% isopropyl alcohol until they could besorted and identified using a dissecting microscope. Water quality estimates were determined by taking the weighted average of the tolerance values, and the final classification was made using the family-level biotic index. Pleasant Creek showed the highest number of taxa present, fourteen, compared to six in Sulphur Creek. Both streams followed the predictions of the River Continuum Concept consisting of collector-dominated functional feeding groups, with Pleasant Creek showing a higher percentage of predators 35.7% to 16.6%. Tolerance values of collected taxa were used to estimate organic pollution indicating good water quality in Pleasant Creek, and fairly poor water quality in Sulphur Creek. The differences in the macroinvertebrate assemblage composition in this study were likely multi-causal. Stream size is a major factor influencing the structure of macroinvertebrate assemblages; in general, as stream size increases, more taxa are added. Pleasant Creek is a much larger stream with a well-established channel, while Sulphur Creek is shallow with depths never exceeding fifteen centimeters. The shallow channel of Sulphur Creek also made it prone to frequent flooding. Isolated floods have little impact on macroinvertebrate communities due to availability of aerial adults nearby to recolonize rapidly, however, frequent flooding may have long-term effects through extirpation of taxa with high mortality. The difference in water quality was the most surprising finding in this study. The fairly poor water quality of Sulphur Creek may also be a factor in the lack of genera found there. Organic pollution effects primary productivity in streams, and while primary productivity was not measured in this study, visual observations found very little algae or aquatic vegetation at Sulphur Creek.
Genetic Relationships among Populations of the Primary Flea Vector of Plague to Humans in the United States
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.
Bacterial Biofilm Disruption with the Utilization of Ionic Liquids
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.
Resveratrol as a Chemotherapeutic
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.
Understanding the Periprosthetic Microenvironment Surrounding Percutaneous Devices: Use Of Simple Immunohistochemical Techniques to Predict Healing Outcomes
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.
Protein Phosphatase 2A Inhibition Preserves Arterial Function in Obese Mice
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.
c-Fos and β-cell Proliferation
Jason Ray, Benjamin Bitner, Kyle Kener, and Brent Jackson, Brigham Young University Life Sciences Type 1 and Type 2 diabetes are increasing at an alarming rate. Both types of diabetes result in decreased functional β-cell mass, which is defined as the number of β-cells multiplied by their Glucose Stimulated Insulin Secretion rate. Decreased functional β-cell mass inhibits regulation of blood glucose levels. β-cells have an extremely low proliferation rate after adolescence, meaning the functional β-cell mass cannot naturally recover. Increasing functional β-cell mass could provide a cure for diabetes, either through pancreatic islet transplants or through enhancement of the endogenous β-cell population. Nkx6.1 has been shown to increase β-cell proliferation by inducing the nuclear receptors Nr4a.1 and Nr4a.3. We have shown that Nkx6.1 increases expression of the gene c-Fos, and that c-Fos induces expression of Nr4a1 and Nr4a3. Furthermore, we have shown that c-Fos is sufficient to induce proliferation of β-cells in the INS-1 832/3 cell line and in primary rat islets. Finally, using lenti-sh-c-Fos to create a stable c-Fos deficient stable cell line, we have demonstrated that Nkx6.1 mediated proliferation is modified by the lack of c-Fos. We propose a model by which c-Fos is a critical link between Nkx6.1 and Nr4a mediated β-cell proliferation.
The Role of PAS Kinase and Cbf1 in Glucose Metabolism
Jenny Pattison, Brigham Young University Life Sciences Sensory protein kinases are essential in the phosphorylation of many protein substrates, allowing them to control several metabolic functions and maintain cellular homeostasis. PAS kinase is a sensory protein kinase that is highly conserved and plays a crucial role in glucose homeostasis, however little is known about the molecular mechanisms behind its function. UGP1 is the only well-characterized substrate of PAS kinase, and its phosphorylation diverts glucose away from storage and towards cell wall biosynthesis. We have recently discovered another key substrate of PAS kinase that affects glucose metabolism in the cell, Centromere binding factor 1 (Cbf1). Cbf1 regulates genes involved in respiration, and we have shown that the phosphorylation of Cbf1 by PAS kinase inhibits Cbf1, decreasing respiration in yeast cells. We hypothesize that this is due to a decrease in mitochondrial mass in cbf1 deficient yeast. Further characterizing the effects of PAS kinase on Cbf1 will give further insight into how cells regulate their central metabolic functions, including respiration.
The Effects of Umbellularia Californica Essential Oil on the Cutaneous Vasculature of Frogs
Stephan Maman and Holden Wagstaff, Southern Utah University Life Sciences Most plant species produce chemical compounds called secondary metabolites that enhance fitness in a variety of ways. Many of these compounds are also physiologically active in vertebrates and have widespread medicinal uses. The most ubiquitous secondary metabolites are the terpenoids, many of which cause vasodilation of the aorta and mesenteric arteries. In this study, we examined the vasoactive effects of the essential oil of Umbellularia californica, which contains the terpenoid umbellulone. Oil obtained via steam distillation using aerial portions of U. californica was applied directly to cutaneous arterioles of frogs. Arteriole diameter was monitored both before and after oil application by video microscopy. Within seconds of application, the oil caused significant vasoconstriction that persisted until the oil was washed off. Our control, medical grade sesame oil, caused no observable effects when applied using the same protocols. These results are opposite to the vasodilatory effects of terpenoids on aortic rings and mesenteric arteries. This suggests that the vasoactive effects of umbellulone are different from other terpenoids, that the vasoactive effects of terpenoids differ depending on blood vessel type, or that application of the complete essential oil affects vasculature differently than application of the isolated terpenoid.
Changes in Urine pH: A Time Series Study
Michael Hinckley and Cayden Westwood, Dixie State University Life Sciences Clinical laboratory standards state that urine samples should be tested within 2 hours of collection. If testing is delayed beyond that time frame the sample needs to be refrigerated to inhibit urea conversion to ammonia and an increase in urine pH. Here medical lab science students investigated whether urine samples left at room temperature for 48 hours resulted in significant increases in urine pH with concomitant microbial growth. Five clean-catch urine specimens were obtained and promptly tested for pH levels using a sterile pH meter (EcoTestr, Oakton Instruments). Samples were then aliquoted into capped and uncapped containers that were left at room temperature. Samples were tested at 0, 8, 20, 24, 32 and 48 hours. The pH values from the uncapped and capped samples from these time periods were analyzed using a paired two tailed t-test. Results indicated one sample out of five was significant (P = .05). Initial samples were gram stained, then read to determine the presence of bacteria. Specimens were cultured on 5% Sheep Blood and MacConkey agar plates. Plates were read for bacterial growth at 24 hours incubation and growth was identified on two samples. At 48 hours, both uncapped and capped urine cultures were plated again and three samples demonstrated bacterial growth the following day. Microbial testing identified normal urogenital flora and pathogenic bacteria. Urine with pathogenic bacteria demonstrated significant increases in pH, while additional cultures with bacterial growth also increased but not with significance. Future studies could employ a larger sample size from both healthy and diseased individuals. Furthermore, identification of microbes that will thrive in acidic and alkaline pH would be of interest.
Trends in Birds Communities in Red Butte Canyon ov er the Past 22 Years
Kylynn Parker, University of Utah Life Sciences Red Butte Canyon (RBC) is a Research Natural Area administered by the US Forest Service in Salt Lake City, Utah. RBC is an undisturbed area and a haven for all types of birds. Most of the avian species found in this area are migratory, and either pass through or breed in the area. The overall aim of this project is to determine if there have been any notable changes in populations of species in the area over the past 22 years. The research question that is covered in this summary are the following: has the density and relative abundance of the top five most commonly detected avian species in Red Butte Canyon notably changed through time in Transect 1? Data was collected by Mark Leppert, PhD and Sherwood Casjens, PhD of the University of Utah. They recorded the number and species of birds that were both seen and heard in 10 different transects within RBC over the past 22 years (1991-2013) and 457 survey days. In 2013 and 2014, I compiled and entered all of the data into a database with the guidance of the researchers. For analysis, I focused on the five most commonly detected species in Transect 1. These species are Black-Capped Chickadee (Poecile atricapilla), Warbling Vireo (Vireo gilvus), Yellow Warbler (Dendroica petechia), American Robin (Turdus migratorius), and Lazuli Bunting (Passerina amoena). Detection trends were found by graphing the number of individual birds seen or heard in Transect 1 over the days since surveys began in 1991 and statistical evidence was found showing significant changes in species population size of these five most commonly detected species, especially in the case of the American Robin which exhibits a decline in detections in recent years.
Revision of Colossendeis colossea Wilson, 1881
Coral Gardner, Southern Utah University Life Sciences Sea spiders (pycnogonids) are a small group of exclusively marine arthropods which resemble terrestrial spiders. One family, the Colossendeidae can grow to a very large adult size, much larger than any other pycnogonid. Colossendeis colossea, the largest known species, has a leg span of up to 70 cm and is found in both very deep ocean waters and shallower water in the Antarctic. Since it has been very difficult to obtain and study live specimens of this species, very little is known of their biology and natural history. We have obtained a number of preserved specimens, all labeled C. colossea, from the United States National Museum and are asking the question, are all of these specimens actually C. colossea? To answer this question, the specimens are being compared to syntypes from the Museum of Comparative Zoology and also to the original type description (Wilson, 1881). Based on previous scientific papers on C. colossea and closely related species, the following are used to separate species within this genus: relative proportions of terminal segments of walking leg and of pedipalp; location and number of eyes and shape of eye tubercle; proboscis shape and orientation; number of spine rows on terminal segments of oviger and shape of oviger terminal claw; abdomen size and orientation. Based on these, our preliminary results demonstrate that some of the United States National Museum specimens are not C. colossea since they differ from the syntypes and type description in several of the above characteristics.
The Role of Pas Kinase in NAD Homeostasis
Daniel Barnett, Brigham Young University Life Sciences Approximately one-third of US adults have metabolic disease, increasing their risk for diabetes, cancers and neurodegenerative disease (www.ADA.org). At the heart of these diseases are imbalances in the cellular redox state. The cofactor NAD(P), commonly known as niacin, is required for over 300 essential reactions in the cell and is largely responsible for the cellular redox state. NAD kinase regulates the NAD to NADP ratio, an important ratio for controlling cellular redox state and central metabolism. Herein we provide evidence that PAS kinase, a nutrient sensing kinase required for glucose homeostasis, phosphorylates NAD kinase. We are currently investigating the effect of this phosphorylation on the function of NAD kinase both in vitro and in vivo by measuring NAD kinase activity and associated phenotypes. This research will increase our understanding of how cells regulate central metabolism. In addition, because PAS kinase is a nonessential protein, it may prove to be an invaluable treatment target for regulating NAD(P) levels and controlling cellular redox state. This may lead to therapeutic targets for cancer and metabolic diseases such as diabetes.
Aurora Kinase A is Critical for the Nkx6.1 Mediated Β-Cell Proliferation Pathway
Amanda Hobson, Carrie Draney, Andrew Stratford, Thomas Becker, Danhong Lu, Michelle Arlotto,
Is There a Correlation between Skin Temperature and Chytrid Survival on Hyla Arenicolor?
Brent Knoblauch and Marty Larsen, Dixie State University Life Sciences Chytrid fungus (Batrachochytrium dendrobatidis) is one of the major hypothesized theories behind global amphibian decline. Canyon Tree frogs (Hyla arenicolor) found in Zion National Park have been found to contain infected individuals, however population sizes have grown despite infection with the usually deadly fungus. The present study sought to demonstrate a correlation between skin temperatures and fungal proliferation, and investigated what enabled Hyla arenicolor to survive and reproduce with chytrid present. Ten frogs were sampled with sterile swabs from each of seven different study areas found in Zion National Park. Along with each swab a temperature was taken via infrared thermometer and recorded. DNA was also extracted from the swabs, and Batrachochytrium dendrobatidis- specific primers were used in a touchdown PCR protocol to determine infection rates. Swabs from specimens with higher temperatures were expected to display lower instances of infection. This would establish for the first time a clear demonstration that high skin temperatures were destroying the fungus in infected individuals in the wild.
Engineering High Affinity T-Cell Receptors Specific for Listeria Monocytogenes
Bryce Anderson, Kemais Ehlers, Deborah Johnson, and Stephen Persaud, Brigham Young University Life Sciences Antigen presenting cells digest and display foreign proteins from infected cells on the major histocompatibility complex (MHC) that is recognized by T cells via their T cell receptor (TCR). LLO56 and LLO118 are CD4+ helper T cells with TCRs specific for the same Listeria monocytogenes epitope. Despite differing by only 15 amino acids, these TCRs have dramatically different primary and secondary responses to infection. TCRs have very low affinity for peptide MHC. We determined to generate high affinity T cell receptors to test if T cell activation would be improved. We reasoned that the single chain LLO118 and LLO56 TCRs (Vβ2-linker-Vα2) could be subjected to directed evolution to generate mutants that are more stable and then used as a template for engineering high affinity T cell receptors. Single chain LLO118 and LLO56 were fused to the yeast surface protein Aga-2 and error prone PCR was used to generate mutagenic libraries. Stabilized single chain TCRs (scTCRs) were selected for using biotinylated Vβ2 and Vα2 antibodies and anti-biotin beads. First generation clones with increased stability compared to wild type were isolated for both LLO118 and LLO56. A second mutagenic library using the first generation mutants as templates was produced and the most stable clones were selected after temperature denaturation, permitting isolation of clones with increased stability. We are currently engineering high affinity T cell receptors by generating affinity libraries using site directed mutagenesis of the CDR3 regions. These libraries are sorted for their ability to bind to MHC tetramers and individual clones are tested using flow cytometry. Generation of pathogen specific high affinity TCRs will increase our understanding of how T cells are activated and could also provide infection specific diagnostics and therapeutics.
c-Fos Regulation of Glucose Stimulated Insulin Secretion in Be ta Cells
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.
The Putative Cannabinoid Receptor GPR55: Expression, Modulation of Hippocampal Plasticity and Behavior
Jacob Welch, Brigham Young University Life Sciences Learning and memory occur due to adaptive brain changes in response to our environment. These changes are mediated by synaptic plasticity, particularly within the hippocampus. Plasticity can either strengthen or weaken synapses, known as long-term potentiation (LTP) or long-term depression (LTD) respectively. While many forms of plasticity are NMDA-dependent, recently endocannabinoids were identified to mediate several new forms of hippocampal synaptic plasticity through the CB1 and TRPV1 receptors. However, research has demonstrated a non-CB1/TRPV1-dependent endocannabinoid synaptic plasticity in the hippocampus. Several potential candidate receptors that bind the endocannabinoid anandamide have been identified. These are among the orphan G-protein coupled receptors (GPRs) whose distribution in the brain and/or function is less well known. GPR55 is of particular interest as it activates second messenger systems. Using quantitative RT-PCR, electrophysiological and memory behavioral tasks we examined hippocampal GPR55 expression and function. GPR55 is expressed in hippocampus of both rats and mice. Cellular expression is currently being examined and appears to be rare in interneurons and more likely expressed by pyramidal cells. Interestingly, application of the GPR55 agonist LPI (2 μM) to wild-type mice demonstrates a significant enhancement of LTP in brain slices. This LPI effect was not noted in GPR55 knock-out (KO) mice, which exhibit significantly (p < 0.05) smaller LTP (146%) than wildtype (WT) (181%). GPR55 also appears to increase release probability (Sylantyev et al., PNAS, 2013), denoting a presynaptic role. Paired-pulse ratios are now being analyzed between GPR55 KO and WT mice to confirm this finding; however we did not note a change in EPSCs in CA1 in response to 2μM LPI. These data suggest GPR55 is expressed and physiologically relevant in the hippocampus. Because enhanced LTP is usually associated with better memory performance in rodents, this provides a potential target to enhance the cellular mechanism associated with memory formation.
Construction of a Weighing Lysimeter on the SUU Green Roof
Bryce Taylor and Ian Kesler, Southern Utah University Life Sciences Green infrastructure is the use of natural processes to manage stormwater runoff and has many positive effects on evaporative cooling, building energy demand, and wildlife habitat. Very little green infrastructure research has been conducted in arid environments such as Southern Utah. The scope of this research project is to establish a complete water budget of the green roof using a lysimeter (an acrylic box resting on a high-resolution weighing scale). The water budget will help in determining whether the cost of irrigating the green roof is off- set by the potential benefits to biodiversity and decreased energy demands.We have constructed a working lysimeter and will collect the required data by using: a high resolution scale, a tipping bucket to measure drainage, two soil moisture and temperature sensors and weather data from the campus weather station. A collection of mixed Sedum species vegetation will be planted in the lysimeter using a special green roof media to accurately simulate a greenroof environment. Both pitfall (used to capture grounded insects) and combination traps (used to capture airborne insects) will be used to measure the biodiversity. Data collection will begin during the winter of 2014 and continue indefinitely.
Patterns of Urban Forest Composition in Utah’s Growing Mountain Communities
Anne Thomas and Richard Gill, Brigham Young University Life Sciences Providing water for expanding urban communities in the western United States is a growing concern for city planners and governmental agencies. Landscaping can claim up to 50% of the urban water budget but also has the most potential for water conservation. Landscape water use is highly variable, however, because of species-specific differences in tree water use and because of decision-making by city planners and residents in maintaining trees. The objective of this study is to improve our ability to predict urban forest water use by identifying differences in tree basal area and diversity between neighborhoods that have arisen at different periods of development in Heber Valley, Utah. We classified neighborhoods as established, exurban (rural housing), commercial, or new tract based on age, location, and lot size. We performed a stratified random survey with twenty lots in each category and collected diameter and species data for each tree in the lot. Some of the patterns we observed were easily anticipated, such as higher basal area per hectare in the older, established neighborhoods relative to newer tract housing. Surprisingly, the number of individual trees per hectare in tract and established neighborhoods is very similar. Perhaps of more interest is the low species richness of tract housing compared to exurban communities. Because exurban communities are being replaced by tract housing there is evidence that tree diversity will be lost. Another important aspect of community structure in urban forests is the ratio of conifers to broadleaf trees because of fundamental differences in water use patterns. Conifers comprised twenty-five percent of the basal area in exurban and thirty-five percent in established neighborhoods, as opposed to five percent in tract. Our data suggest that tree diversity is likely to decrease while water demand is likely to increase with changes in urban forests in the coming decade.
Entomophilic Nematode Distribution and Diversity in Beaver Dam Wash
Westen Archibald, Trevor Smart, and Emily Forsyth, Brigham Young University Life Sciences The Beaver Dam Wash National Conservation Area is considered an important biodiversity hotspot for the Western United States. The wash is an ecotone on the Northeastern edge of the Mojave Desert and is directly adjacent to both the Colorado Plateau and the Great Basin. Thus, this area contains various species of flora and fauna from each of the surrounding biomes. Prior research of the Beaver Dam Wash has documented high aboveground biodiversity for a desert ecosystem however no research has been conducted on belowground diversity. For this reason we chose to characterize the diversity and distribution of entomophilic nematodes, as they are commonly used as a biological indicator to the surrounding ecosystems. We hypothesized that because plant and insect biodiversity is high here, we would also find a diversity of entomophilic nematodes that is higher than the surrounding areas. To test this hypothesis we collected soil samples from 15 sample sites representative of the diversity of the different ecological communities. We tested for patterns of codistribution between entomophilic nematodes and environmental variables, such as plant cover, proximity to water, presence of organic matter, elevation, ecosystem classification, and soil chemistry. We extracted nematodes from the soil samples and sequenced the 28s rDNA region of representative individuals from each sample. Nematode diversity was low; one species was broadly distributed, X others were more patchily distributed. No correlation was found between above ground factors or soil chemical properties (ppm phosphorous, percent organic matter, and percent soil moisture). We conclude that the distribution of entomophilic nematodes in this ecological confluence is to some degree stochastic and uncoupled from aboveground diversity or belowground soil conditions.
Yeast PAS Kinase as a Model for Cellular Metabolic Control
Katherine Harris, Brady Evans and Thomas Andros, Brigham Young University Life Sciences Metabolic diseases, such as obesity and diabetes, have become endemic and the need for better treatments is rising. Mutations in PAS kinase, a recently discovered sensory kinase, have been shown to cause Maturity Onset Diabetes of the Young (MODY) in humans (Semplici et al., 2011). In addition, PAS kinase deficient mice display many phenotypes related to diabetes including resistance to weight gain, insulin insensitivity and triglyceride accumulation in response to a high-fat diet (Hao et al., 2007). Despite its importance in metabolism, little is known about the regulation of PAS kinase. PAS kinase consists of a sensory PAS domain that binds to and inhibits a protein kinase domain (Amezcua et al, 2002). We are currently engaged in several yeast genetic screens which will allow identification of regions in the full length PAS kinase that are essential for activation or for binding its substrates. The first screen is based on the finding that PAS kinase overexpression rescues a temperature-sensitive mutation in Tor2, the tor2(ts). We have isolated both point mutations and truncations in PAS kinase which alleviate the tor2(ts), suggesting they are hyperactive alleles. These mutations identify novel regions involved in PAS kinase regulation. Our second screen uses the yeast 2-hybrid to select for both point mutations and truncations that increase the ability of PAS kinase to bind its substrates. These mutations will help identify key regions of PAS kinase utilized in substrate recognition. Finally, we have identified regions of PAS kinase that are well-conserved throughout evolution and will compare these regions with the regions affected by our mutations. This study will be the first reported mutagenic analysis of PAS kinase. Analysis of these specific genetic regions will help elucidate the molecular mechanisms involved in the regulation and function of PAS kinase, a key player in the development of metabolic disease.
Ventral Tegmental Area Dopamine and GABA Neurons: Physiological Properties and Expression of Mrna for Endocannabinoid Biosynthetic Enzymes and Type I Mglurs
Scott Newton, Brigham Young University Life Sciences The ventral tegmental area (VTA) is known to controls the processing of rewarding and addictive behaviors. The VTA contains dopamine (DA) cells, which release DA to downstream targets in response to rewarding stimuli, and GABA cells, which modulate DA cell activity. Therefore, both cell types are involved in associative reward learning. Synaptic plasticity plays an important role in adaptive reward signaling within the VTA. Endocannabinoids mediate or modulate synaptic plasticity at synapses within the reward circuit. However, the source of endocannabinoids within the VTA is not well understood. Therefore, our goal was to describe the distribution of endocannabinoid biosynthetic enzyme mRNA within VTA neurons. We extracted single VTA neurons via whole cell patch clamp and used single-cell real-time quantitative PCR to identify DA and GABA neurons based on mRNA expression of cell-type specific targets. Additionally, electrophysiological properties such as action potential frequency and sag potential amplitude were examined between the two cell types. Concurrent with established observations, slower firing frequencies were observed in DAergic neurons, however overlap was identified between these two cell types. VTA neurons were then probed for endocannabinoid/ biosynthetic enzyme mRNA, such as N-acyl-phosphatidylethanolamine-specific phospholipase D (NAPE- PLD), diacylglycerol lipase α (DAGLα), and 12-lipoxygenase. We also probed for type I metabotropic glutamate receptor (mGluR) mRNA, as endocannabinoid synthesis requires mGluR activation. Our data demonstrate that endocannabinoid biosynthetic enzyme mRNA is expressed in both DAergic and GABAergic cells with concurrent expression of type I mGluRs. Next, to ensure mRNA expression was representative of protein content, slices were stained using immunohistochemistry for GAD67, DAGLα, NAPE-PLD and type I mGluRs. Positive labeling for these targets was observed in VTA neurons, supporting our RT-PCR results. Collectively, these data suggest DAergic and GABAergic cells of the VTA have the capability to produce endocannabinoids and potentially alter synaptic plasticity involved in reward and addiction.