Life Sciences
MafA’s Ability to Enhance ë_-cell Function
Aaron Leifer, Jasmine Banner, Collin Christensen, Trevor Lloyd, Kenneth Call, Brigham Young University Approximately 9.4 percent of the United States is affected by type 1 or type 2 diabetes. Diabetes results from the body’s inability to maintain healthy blood glucose levels due to the loss of pancreatic ë_-cells (insulin secreting cells) or from the body’s insulin sensitive cells becoming insulin resistant. Both type 1 and type 2 diabetes results in a loss of functional ë_-cells. The current treatments for diabetes are insulin injections or transplants, many times requiring up to three donors per transplant. Neither option is an optimal cure: insulin injections do not cure the disease, and transplants are not available to the majority of people. We propose that being able to replicate ë_-cells in-vivo would allow us to provide a cure to diabetes. ë_-cells stop reproducing (proliferating) soon after birth except in a few occasions such as obesity and pregnancy, leading us to believe that there are key gene(s) that induce cell proliferation when activated. Finding these gene(s) would present a viable cure, being able to grow ë_-cells in-vivo for transplantation or even injection. The gene MafA is present in mature ë_-cells and previous research has revealed its vital role in the pancreas. MafA is turned on around embryonic day 15.5 and steadily increases expression up until the cell becomes a mature ë_-cell. The time period when MafA is turned on corresponds with when a ë_-cell is proliferating and developing leading us to believe that MafA is crucial to finding a cure. Here we show the effect of MafA overexpression on INS1 832/13 ë_-cell proliferation, survival, and insulin secretion.
Assessing the Role of Temperature in Dengue Fever Outbreak Dynamics with Wolbachia Transinfection Control Methods.
Colton Smith, Dixie State University
Modeling Decidual NK cell transfer of Granulysin to Trophoblast Cells using stimulated Peripheral Blood NK cells
Kayleigh Ingersoll, Brigham Young University
Deletion of Nr4a nuclear receptors on ë_-cells in hyperlipidemia
Weston Elison, Brigham Young University
Predisposed Success as an Athlete: Genes that Give an Edge
Tason Turek, Dixie State University
Expression, Purification, and Activity of Recombinant Human SKIP
Somer Doody, Utah Valley University
The Effects of Palmitate on Beta Cell Insulin Secretion
Trevor Lloyd; Mason Poffenbarger; Austin Ricks; Andrew Barlow; Zoey Fishburn, Brigham Young University
Virus-like Particle Enzyme Encapsulation
Joshua Wilkerson; Seung-Ook Yang; Parker J. Funk; Steven K. Stanley, Brigham Young University
Micropropagation of two edaphic endemic species, Lepidium ostleri and Eriogonum soredium
Alyson DeNittis, Utah Valley University
The Effects of Hyperlipidemia on Pancreatic Beta Cells
Diabetes affects over 30 million Americans and 185,000 Utahn’s. Type 1 and Type 2 diabetes are characterized by decreased functional β-cell mass and insulin production. Diabetes also results in increased circulating glucose and fatty acid levels, which damage and destroy β-cells over time. Our study will shed further light on the effects of palmitate, the most commonly made fatty acid in the liver, on hyperlipidemia. In this study we test the specific effects of chronic palmitate exposure on various cell lines acclimated to 0.15 mM, 0.3 mM, and 0.5 mM concentrations of palmitate. We demonstrate the effects of progressive long-term exposure to palmitate on β-cell proliferation and resistance to apoptosis. We demonstrate mechanistic changes that result in the observed phenotypes. Our goal in this study is to explore how β-cells adapt to exposure to hyperlipidemia, and to define interventions to protect β-cells from the harmful effects of hyperlipidemia.
Does Pdx1 Enhance Functional ë_-cell Mass?
Aaron Leifer, Jasmine Banner, Collin Christensen, Trevor Lloyd, Kenneth Call, Brigham Young University Diabetes Mellitus has become a worldwide epidemic affecting over 400 million people. Both type 1 and type 2 diabetes result from the body’s inability to produce or respond to insulin in order to regulate blood sugar. In both cases, the insulin secreting ë_-cells in the pancreatic Islets of Langerhans have become endangered and in many cases non-functional. The function of these ë_-cells is defined by their ability to multiply and maintain a steady number, to defend against induced cell death and ultimately to secrete insulin. Since ë_-cell production reaches its peak during fetal development, this would suggest that diabetics have an inactive pathway to produce functional ë_-cells. However, recent studies have identified key transcription factors that aid pancreatic progenitors in becoming functional ë_-cells. Pdx1 is a transcription factor that is active throughout the ë_-cell pathway and found in mature ë_-cells. Research has identified Pdx1 as a key component in helping both ë±-cells and ë_-cells proliferate and even in reprogramming ë±-cells to become functional ë_-cells. Additionally, Pdx1 has been identified to help ë_-cells effectively secrete insulin. We present data demonstrating the effect of Pdx1 adenoviral over-expression on three independent markers of functional ë_-cell mass: 1) cell proliferation, 2) cell survival, and 3) insulin content and secretion. Defining the effect of Pdx1 on each of these parameters will provide further data to explore therapeutic interventions for diabetic patients.
Barriers to the proliferation of aged ë_ cells through overexpression of Nkx6.1
Parker Booren, Nathanael Jensen, Talon Aitken, Samuel Grover, Jackie Crabree, Brigham Young University Diabetes continues to grow at a rapid rate, affecting the lives of both young and old. Both Type 1 and Type 2 diabetes lead to eventual ë_ cell depletion (and subsequent decrease in insulin secretion). This can be treated through ë_ cell transplantation from the pancreata of cadavers. Currently, collecting sufficient ë_ cells for one diabetic patient requires pancreata from multiple cadavers. If proliferation can be induced in a donor’s aged ë_ cells, transplantation would become more effective as one donor now becomes sufficient to serve one or two patients. Nkx6.1 is a transcription factor that increases insulin secretion and induces proliferation of young rat ë_ cells (5 weeks) through the upregulation of its target genes: VGF, Nr4a1 and Nr4a3. Aged rat ë_ cells (5+ months) fail to proliferate after overexpression of Nkx6.1. We have also shown that upregulation of Nkx6.1’s target genes is disrupted in these aged ë_ cells. This may be due to changes in expression of a binding partner necessary for Nkx6.1’s upregulation of these target genes or to changes in Nkx6.1 posttranslational modifications that impede binding partner interactions in aged ë_ cells. We present data from co-immunoprecipitation and mass spectrometry experiments that reveal the presence or absence of Nkx6.1’s binding partner in young and aged ë_ cells. Furthermore, we present mass spectrometry results of Nkx6.1 posttranslational modification from young and old ë_ cells. This data will increase understanding on the ability of Nkx6.1 to upregulate its target genes in an aged ë_ cell.
Effect of salinity on the hatching of Branchinecta lindahli Packard, 1883.
Nayla Rhein, Southern Utah University
Creating a Monster: Attachment Theory in Mary Shelley’s Frankenstein
Sam Passey, Lyndsey Graig, Christine Fiscer, RonJai Staton, Jeremy Scritchfield, Barbara Balbas, Amy Harmon, Craig Demke, Joey Jergins, Tim Bywater, and Dannelle Larsen Rife, Dixie State University Life Sciences Research in human development suggests relationships are vital for physiological and emotional well-being across the lifespan. Attachment theory is foundational for relationships and is intrinsic in human nature as it is represented through words of novelists. Attachments are developed within the first year of life based on caregivers’ appropriate, contingent, and prompt responses to the infant’s cues. Avoidant attachment develops when the infant receives minimal responses to his or her cues. John Bowlby proposed the attachment relationship between the infant and parent creates an internal working model (IWM). This IWM sets the foundation of all subsequent close relationships throughout the lifespan. Individuals who have avoidant attachment representations are dismissive of, and lack security in relationships. Living in a time where women were marginalized, segregated, and many lacked formal education, Mary Shelley effectively produced a popular work of fiction in the early 1800s. Shelley was a keen observer of relationships long before Attachment Theory was developed in the 1960s. Psychobiographical methods were used to examine Shelley’s Frankenstein as a case study of Attachment Theory. Results suggest Shelley’s Frankenstein depicts basic components of attachment theory, and “Frankenstein,” the monster character, exemplifies avoidant attachment. Through his dismissive and proximity seeking behaviors, the monster characterizes Bowlby’s description of avoidant attachment. Lacking relationships during critical periods for development of empathy, the monster loses the ability to feel remorse. This critical examination of early British literature as a case study for Attachment Theory lends retrospective support for the understanding of human relationships.
Improvement of Care in the Surgical Intensive Care Unit through Family Feedback
Avani Latchireddi, Wade Mather, and Joseph Tonna, University of Utah Life Sciences Purpose This research project is to assess patient satisfaction and feedback around care provided in the Surgical Intensive Care Unit (SICU) at the University of Utah Hospital with the goal of iterative improvement of care. Research Question/Hypothesis We hypothesize that by assessing patient satisfaction of ICU care, we can implement directed changes targeting patient-identified concerns. Methodology A survey of 37 questions, based on a validated national survey of family satisfaction with ICU care (FSICU-24) was put together addressing issues ranging from emotional care to technical aspects of the SICU experience on a whole. It is administered to the family member who was most involved in the patients care in the Surgical ICU after transfer out of the ICU. All the data is securely maintained and analyzed through a REDCap database for the purposes of quality improvement. Observations Over the initial weeks of administration, a few observations for improvement opportunities have been repetitive. Many patients and family members highly appreciate their attending doctors but cannot keep track of their names with the many teams of doctors. Having a time frame in which the doctor would arrive on rounds such that the family member can be present would be very helpful. The family members of patients sometimes feel uncared for in the SICU. Many would appreciate having someone show them the cafeteria or simply ask them if they need anything in particular. The plan of the day sheet (checklist as well as a list of the medical plan the team intends to follow) is often not given and/or explained to the patient and their family. Conclusion The following changes will be considered for feasibility of implementation. Surveys will be continuously administered in order to observe the effect the implemented changes have had. For example, changes might include the nurses explaining the plan of the day sheet to the patient and their family after the doctor has stated the plan of care; having picture cards of doctors with their name and photo would help patients and families better identify their caregivers; a volunteer could go around the ICU once a day and ask if the family has any needs. The expectation is to see improved patient and family satisfaction in those selected areas.
A Framework for Validating Modeled Air Quality Data in Health Research
Nicole Burnett, Peter Mo, Naresh Rajan, Randy Madsen, Ram Gouripeddi, and Julio Facelli, University of Utah Life Sciences Background In the Salt Lake Valley there are three permanent Environmental Protection Agency (EPA) certified air quality monitoring stations that intake air samples and produce results of the air quality in the proximity of the monitor station. Due to the fact that the monitors only represent a small area of the 500- square-mile Salt Lake Valley, there are spatial gaps when using the air quality monitoring data for epidemiological studies. Since for certain studies health researchers may require a higher resolution spatiotemporal air quality grid [1], we need to devise new approaches to provide air quality data that could meet the epidemiological studies requirements. Research Methodology Modeled air quality data available from the EPA, has higher spatial and temporal resolution than data from monitoring stations, but it needs experimental validation and uncertainty quantification (UQ) in the Salt Lake Valley. We can achieve these validation and UQ goals through statistical comparisons of measured air quality data at the same location and time as the modeled air quality data. The air quality model that we primarily used is the one that the EPA has developed for the Centers for Disease Control and Prevention’s (CDC) National Environmental Public Health Tracking Network [2]. This is a model that uses a Hierarchical Bayesian Space Time Modeling approach [3]. This model was validated on the east coast of the United States so it is unknown how effective is in taking into account the terrain of the Salt Lake Valley. Modeled PM2.5 data in a 12×12 kilometer continuous grid resolution for the years 2007 and 2008 were compared against measured data of the same timeframe and location. The measured data was obtained from EPA’s Air Quality System (AQS) Datamart [4]. The statistical comparisons performed using these the two data sets were done using daily and monthly PM2.5 averages for the years 2007 and 2008 using MySQL, MATLAB and R. Conclusion & Significance We have developed a prototype for comparing and validating modeled air quality data against measured air quality data for the Salt Lake Valley. We found the modeled data fits the measured data fairly well. We will expand our work by developing a validating framework that will include a library of data modeling algorithms such as, The Complex Terrain Dispersion Model Plus Algorithms for Unstable Situations (CTDMPLUS) [5] and Yanosky’s [6], which could be selected by the user. The framework will be developed using OpenFurther, and then integrated with biomedical data [7]. The framework will be integrated into the PRISMS project [8] as part of the informatics infrastructure for studying the effects of air quality on pediatric asthma. References M. Z. Al-Hamdan, W. L. Crosson, A. S. Limaye, D. L. Rickman, D. A. Quattrochi, M. G. Estes, J. R. Qualters, A. H. Sinclair, D. D. Tolsma, K. A. Adeniyi, and A. S. Niskar, “Methods for Characterizing Fine Particulate Matter Using Ground Observations and Remotely Sensed Data: Potential Use for Environmental Public Health Surveillance,” J. Air Waste Manag. Assoc., vol. 59, no. 7, pp. 865-881, Jul. 2009. “Air Quality Data for the CDC National EPHT Network | Human Exposure and Atmospheric Sciences | US EPA.” [Online]. Available: http://www.epa.gov/heasd/research/cdc.html. [Accessed: 18-Sep-2014]. N. J. McMillan, D. M. Holland, M. Morara, and J. Feng, “Combining numerical model output and particulate data using Bayesian space-time modeling,” Environmetrics, p. n/a-n/a, 2009. “AirData | US Environmental Protection Agency.” [Online]. Available: http://www3.epa.gov/airdata/index.html. [Accessed: 20-Oct-2015]. Environmental Protection Agency, “Revision to the Guideline on Air Quality Models: Adoption of a Preferred General Purpose.” Environmental Protection Agency, 09-Nov-2005. J. D. Yanosky, C. J. Paciorek, F. Laden, J. E. Hart, R. C. Puett, D. Liao, and H. H. Suh, “Spatio-temporal modeling of particulate air pollution in the conterminous United States using geographic and meteorological predictors,” Environ. Health, vol. 13, no. 1, p. 63, Aug. 2014. R. Gouripeddi, N. Sundar Rajan, R. Madsen, P. Warner, and J. C. Facelli, “Federating Air Quality Data with Clinical Data,” presented at the 2014 AMIA Annual Symposium Proceedings, 2014. “Pediatric Research Using Integrated Sensor Monitoring Systems | National Institute of Biomedical Imaging and Bioengineering.” [Online]. Available: http://www.nibib.nih.gov/research-funding/prisms. [Accessed: 20-Oct- 2015]. Acknowledgements Grants: UU Air Quality Program, U54EB021973, NCRR/NCATS UL1RR025764, 3UL1RR025764-02S2, AHRQ R01 HS019862, DHHS 1D1BRH20425, UU Research Foundation. CHPC at UU.
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.
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.
The Role of Palmitate in Upregulating Nr4a1 and Nr4a3
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.
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.
Biodiversity Mapping of North America: A Comparison of Datatypes
Kari Norman, Utah State University Life Sciences The identification of important areas for biodiversity is essential for affective allocation of limited conservation resources. Since Myers’ seminal biodiversity hotspot paper in 2000, great strides have been made in more accurate global mapping. While global mapping identifies important patterns in large-scale biodiversity, conservation management rarely if ever occurs on the same scale. Finer scale mapping is therefore essential to make research applicable for on the ground decision-making. This project focuses on North America, a continent that holds no priority areas when included in a global analysis. Using citizen scientist data of multiple vertebrate and plant taxa, we created maps of species richness and significantly rare species. In addition, we created similar maps based on range map data and compared to determine if the two data types produced different biodiversity priority areas. The results of this study provide informative maps about the locations of species, their vulnerability, and how human action may impact them, as well as underline the importance of understanding the data behind their construction.
Isolation and Characterization of Novel Lytic Phage to Treat Methicillin- Resistant Staphylococcus Aureus
Tyler White, Kyle Jensen, Jacob Hatch, Trevor Wienclaw, Brian Hair, and Aaron Trent, Brigham Young University Life Sciences The bacterium Staphylococcus Aureus (SA) is a common commensal organism of the human nose and skin that can lead to diseases such as pneumonia, endocarditis, and meningitis. These SA infections are usually remedied via antibiotic treatment with methicillin. However, over the course of frequent exposure to various antibiotics, the bacteria have evolved resistance to methicillin to create resistant strains (MRSA) that is completely resistant to this drug and many others (leaving vancomycin as the last viable option). As a result, the need for an antibiotic alternative treatment for this infection is becoming increasingly crucial especially in hospitals where nosocomial transmission of the bacteria is prevalent. Phage are bacterial-specific viruses that have shown promise as anti- bacterial agents for human bacterial pathogens. Thus far, we have isolated 18 different samples of phage that lyse MRSA and 51 strains of S. Aureus (over 20 of which are MRSA). We are currently testing all phage samples against these bacterial strains to determine which phage possess a broad tropism to kill many SA/MRSA isolates. We are also conducting experiments to determine the relative lytic ability of each phage. Lytic phage with broad tropism and/or strong lytic ability will have their genomes sequenced in order to verify their respective novelty. Novel phage will be aggregated to form a highly virulent cocktail that can be used to treat a broad spectrum of SA and MRSA infections.
C-Fos Protects Β-Cells from Apoptosis
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.
The Role of Cdk2, Cy lclin E1, and CIP/K IP Inhibitors in Aged Islets
Brent Wright, Brigham Young University Life Sciences Diabetes is one of the leading causes of death among Americans and is a major health concern worldwide. Nearly one in four Americans aged 65 or older are diabetic. Type 1 and Type 2 diabetes both result in reduced functional β-cell mass, which regulates the storage and secretion of insulin. Increased functional β-cell mass could essentially cure diabetes. We have shown that Nkx6.1 overexpression induces proliferation of 2-month-old primary rat β-cells but fails to induce replication of 8-month-old primary β-cells, as measured by 3H- thymidine incorporation is age-dependent. Cell cycle activator and inhibitor mRNA levels were measured in young and aged untreated islets and islets transduced with AdCMV-BGal or AdCMV-Nkx6.1. This data demonstrated a significant increase in mRNA expression of cell cycle inhibitors p21 and p57 of the CIP/KIP family in young islets transduced with Nkx6.1. However, p21 and p57 mRNA showed no significant increase in aged islets. Cdk2 and cyclin E1 mRNA expression showed a similar trend for young and aged islets. The increased expression of Cdk2, a necessary factor for transition from G1 to S phase, could provide possible explanation for increased proliferation in young islets. Fluctuating mRNA levels of key cell cycle components in aged islets, provides a possible explanation for the decreased effectiveness of Nkx6.1 in inducing proliferation in aged islets.
Are Canyon Tree Frog (Hyla arenicolor ) Populations in Zion National P ark Affected by Chytrid Fungus (Batrachochytrium Dendrobatidis)? Four Y ear Study Says No!
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.
Effects of Resveratrol on PMCA in Breast Cancer Cells
Joshua Peterson, Brigham Young University Life Sciences Cancer kills millions of people every year. Cancer occurs when cells proliferate at an excessive rate and do not die as regularly functioning cells do. In cancer cells, the mechanism that initiates apoptosis (cell death) is inhibited. These cells eventually multiply to the point where they interfere with physiological function and cause death. Therefore, one of the aims of cancer research is to find treatments that initiate apoptosis in cancer cells. Many current chemotherapeutic (anti- cancer) treatments are toxic to all mitotic cells, rather than to cancer cells alone. Studies have shown that resveratrol, which is found in grapes, peanuts, and berries, facilitates apoptosis in cancer cells without causing apoptosis in regular cells. The apoptotic activity of resveratrol in tumor cells is dependent on a large, sustained increase in cytoplasmic calcium ion concentration. In properly functioning cells, plasma membrane Ca2+- ATPase (PMCA) pumps excess calcium from the cytosol to the extracellular space. PMCA prevents toxically high levels of calcium and maintains cytoplasmic calcium homeostasis. Using live cell microscopy to monitor intracellular calcium ion concentration, we explore the direct and indirect effects of resveratrol on PMCA activity in MDA-MB-231 (a breast cancer cell line).
Comparison of Dust Mite Antigen Levels in Utah Homes with Swamp Coolers versus Homes with Air Conditioners
Morgan Christiansen, Taylor Hoybjerg, and Ryan Cook, Brigham Young University Life Sciences Asthma is a chronic allergic disorder manifest by airway restriction due to inflammation, bronchoconstriction, and increased respiratory mucous secretion. As many as 300 million people worldwide are affected by asthma. It is becoming increasingly prevalent, especially in countries experiencing urbanization and Westernization. Asthma is currently the most common chronic illness among children in the U.S. and the third leading cause of hospitalization for children aged 0 – 15 yrs. Reservoir dust collection and area air sampling are the two primary methods of measuring allergen levels in house dust. Allergen sensitization leading to asthma is thought to occur prior to age six while the immune system is still naïve. In the case of the dust mite allergen Der p1, the exposure window may be as early as age two. However, little evidence is available to establish a dose–response relationship between inhalation exposure and early immunological sensitization to allergens. Temperature and relative humidity play a major role in dust mite survival and proliferation—indoor humidity above 50-60% in arid environments has been shown to support dust mite populations. Evaporative “swamp” coolers add moisture to cool the air and this increased humidity can create favorable environments for dust mite survival. The purpose of this research is to quantify the levels of dust mite antigen in homes with air conditioners and homes with swamp coolers in Utah Valley to determine the role humidity plays in the abundance of dust mite antigens. We are testing the hypothesis that homes with swamp coolers exhibit higher levels of antigen due to heightened humidity levels.
Radon Testing
Breanna Torgersen, Hailey Seaver, Abigail Gunn, Siena Davis, and Jim Johnston, Brigham Young University Life Sciences Exposure to radon gas (222Rn) is the second leading cause of lung cancer in the U.S. Because 222Rn is a colorless, odorless, and tasteless gas, it must be tested for to recognize its existence in a home. Studies show that many homeowners know little about radon, and do not test their homes as recommended. This study was undertaken to measure Utah County residents’ knowledge about radon, and to understand factors associated with radon testing. Utah County residents (N = 200) are currently being surveyed as they exit the vital records office at the Utah County Health Department (UCHD). Subjects complete a 51-item survey measuring demographics, radon knowledge, and social cognitive theory-based constructs related to radon testing. Preliminary data (n = 65) shows subjects’ mean radon knowledge score was 1.68 (33%, SD = 26.24%) on a 5-item test. There was a significant relationship between radon testing and self-efficacy (OR 1.76, 95% CI: 1.2–2.6, p = 0.007). Subjects with higher self-efficacy for radon testing were more likely to report that they had tested their home at least once. Data collection is still underway on this study, and all results reported here are preliminary.
Effect of Aggregation at a Winter Feeding Station on Intestinal Parasite Load in Elk (Cervus canadensis)
Morgan Hughes, Utah State University Life Sciences Elk feeding stations are used throughout the Western US as a means to prevent depredation on private lands (Smith,2001). Many of the unintended effects of such artificial congregation remain unexamined. In many species, increased densities result in increased parasite loads (Dietz, 1988) adding physiological stress to individual animals and reducing the economic value of the animal to sportsmen (Choquette, 1956). Through laboratory analysis of fecal float samples, I will monitor changes in the number of parasite eggs for elk at Hardware Ranch feeding station over the winter season. This is to discover if there may be negative implications of feeding stations which should be further examined. Increased prevalence of parasites could also indicate an increased danger for transmission of other diseases which are a threat to domestic livestock (Williams, 2002).
Investigation of a Recent Rise of Dual Amantadine-Resistance Mutations in the Influenza A M2 Sequence
Matthew Durrant and Dennis Eggett, Brigham Young University Life Sciences The drugs amantandine and rimantadine were previously used to treat influenza A infections in humans. In 2005, a mutation in the Influenza A M2 channel conferred resistance to these drugs, rendering them obsolete against treating influenza. The S31N amantadine-resistance mutation in the influenza A M2 sequence currently occurs more frequently in nature than the S31 wild type. Overcoming this mutation with new drug compounds is the focus of Influenza A M2 channel researchers. However, there are several other identified mutations that have also been shown to confer resistance to these drugs. These other mutations are thought to occur only in small frequencies in nature. A statistical analysis of 1,007 unique M2 protein sequences shows an enhanced frequency for the S31N/V27A dual amantidine resistant mutation in recent years, especially in swine, compared to expected frequencies based on the occurrence rates of individual mutations in wild type (S31) M2. The development of the S31N/V27A variant in the Midwestern US swine may be a harbinger of novel human strain development. At the same time, the different propensities for the V27A as compared to the V27T dual mutant may reflect differences in viral fitness or protein energetics, and this information could be exploited to focus drug development so as to reduce further drug insensitivity. V27A/S31N is a possible path forward for the evolution of M2, which may convey a new level of drug resistance and should receive attention in drug design.
RRMS Patient Genotype Correlated with Copaxone-Induced Hepatotoxicity
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.
Dispersal of Fairy Shrimp Eggs through Wind and Runoff
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.
Potential Toxicity of Synthetic Administered Pitocin
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.
Three-Dimensional Rapid Prototyping of Vascular Substitutes for Medical Applications
Alex Bischoff, Connor Dodge, Sarah Livingston, Sterling Rosqvist, Tomonori Baba, Kyle Larsen,
Changes in Water Chemistry in Cedar City Irrigation System
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.
Variance in Stomatal Size and Density between Triploid and Diploid Quaking Aspen (Populus Tremuloides) in Utah
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.
Neuroregeneration: Potentiation of Nerve Growth Factor Receptor in Peripheral Nerves
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.
The Evolutionary Hourglass of an Herbivore Induced Transcripto me in N. Attenuate
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.
A Genetic Analysis of the Multiple Sclerosis Disease Course as Influenced by Comorbid Diseases
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.
Optimization of the pET9a Vector
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.