2015 Abstracts
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.
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.
Evolution and Molecular Phylogenetics of Baetidae (Ephemeroptera)
Chase Barker, Utah Valley University Life Sciences Central Research Question:
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.
ATP Production in Serpentinization-Driven Ecosystems
Alex Hyer, University of Utah Life Sciences Geochemical reactions associated with the process known as serpentinization can generate copious quantities of organic carbon and chemical energy that support life, but these reactions also greatly increase the pH of serpentinization sites. High-pH environments hinder ATP production because the low proton concentrations at high pH can result in a reversed proton gradient across cell membranes. Organisms present at serpentinization sites are not well-characterized, and adaptations to their alkaliphilic lifestyle are unknown. Bacillus pseudofirmus OF4 was not isolated from serpentinization sites, but it is a well- studied alkaliphile that has several distinct adaptations for ATP production in high pH environments. Its sequence for ATP synthase, the molecular motor responsible for ATP generation in the cell, contains several distinctive mutations including an AXAXAXA motif that increases the C-ring stoichiometry, a P51XXE54XXP57 motif that creates a distinct kink in the outer helix of the C-subunit, and a V21 mutation granting rotamer freedom to E54. In this study, we search for similar mutations in metagenomic databases containing DNA sequences collected from serpentinization sites. Analysis of alignments from the programs BLAST and Exonerate indicate that V21 is present in several alignments, but P51 is found in only a single alignment from serpentinization sites. The AXAXAXA motif is non- existent in our databases, but the more common GXGXGXG motif is present with alanine replacements occurring periodically. In general, these results indicate that our DNA sequences from high-pH sites of serpentinization are more reminiscent of typical neutrophiles than of the alkaliphile B. pseudofirmus OF4. Therefore, serpentinization- adapted organisms may harbor other adaptations to high pH such as local pmf regulation. Future work will test for quantitative correlations between environmental pH measurements and the incidence of mutations in ATP synthase in order to identify novel adaptations to high pH in serpentinization-driven ecosystems.
The Impact of Maternal Ssri Use on Serotonin Synthesis and Protein Transporters in the Placenta and Newborn Serotonin Levels
Simranvir Kaur, University of Utah Life Sciences The placenta is the major source of fetal serotonin during pregnancy, which is essential for fetal brain development. In Utah, approximately 13% of pregnant women take Selective Serotonin Reuptake Inhibitors (SSRIs) to treat depression, the use of which has been correlated to significantly lower serotonin levels in cord blood for newborns. Studies suggest association between maternal SSRI use and adverse outcomes such as preterm birth, cognitive deficit, and disruption of serotonergic systems. However the effect of SSRIs on placental gene expression, serotonin synthesis and transport in the placenta, is not known. This study evaluates the impact of maternal SSRI use on placental gene expression and levels of serotonin in the cord blood using a nested case-control observational study model. Biological samples will be collected until 20 cases (women taking SSRIs during pregnancy) and 20 appropriately matched controls have been enrolled into the study. Participants also complete an online questionnaire to measure depression and anxiety levels as well as document any medication they have taken during pregnancy. Data and sample collection for this study is still in progress. Once enough samples have been collected for batch analysis, we will complete RT-PCR and ELISA, expected in Spring 2015. Upon complete data analysis, we expect this study will help in targeting mothers who are at risk for adverse pregnancy outcomes and further provide suggestions for intervention.
The Musical Phiquence: Finding Phi in Musical Progressions
Sergio Arellano, Snow College Mathematical Sciences Why is music so pleasant to? In Western music, we have used what is called the “tempered scale” for centuries, and even though it has gone through changes, it is still largely based around the mathematical principles that Pythagoras created two thousand years ago. Is it possible that the explanation to this phenomenon has to do with the mathematical basis of music, which is unconsciously perceived by the brain? The human brain is known for detecting the underlying mathematical patterns present in many non-music related disciplines, such as visual arts. Independent of the music world, there is a proportion called the Golden Ratio or Phi. It is found in art and geometry, because human eyes tend to find that this proportion produces beauty. This is not a coincidence; this proportion is found everywhere around us, especially in nature. With this in mind, it makes sense to search for the Golden Ratio in the tempered scale to explain the mind’s intuitive appreciation of music. This research discovered the surprising fact that there is a Phi relationship between the first and third degree of the major scale, in terms of the frequency of sounds. The relationship is this: the sum of the frequencies of the notes of the minor III chord divided by the sum of the frequencies of the major I chord in any given major key, tends to be Phi. Another very surprising result was the importance of the number 24 in the frequencies of the major scale. These two particularities help to shed light on why human ears have an untaught comprehension of music; the underlying perception of mathematical relationships by the mind are related to the natural appreciation of it.
Study the Stability of Steady Solutions for a Model of Mutualism
Amy Gifford and Brennon Bauer, Southern Utah University Mathematical Sciences Mutualism is the way two organisms of different species exist in a relationship in which each individual benefits from the activity of the other. We study a mathematical model of mutualism. The stability of the steady state solutions of this system will be analyzed. Also, we give some numerical experiments that verify the theoretical results for those steady solutions.
Counting Integer Points in Scaled Polytopes
Christopher Vander Wilt and Daniel Gulbrandsen Utah Valley University Let nP denote the polytope obtained by expanding the convex integral polytope P⊂R^d by a factor of n in each dimension. Ehrhart [1] proved that the number of lattice (integer) points contained in nP is a rational polynomial of degree d in n. What happens if the polytope is expanded by not necessarily the same factor in each dimension? In this talk a partial answer to this question will be provided, using powers of n as different factors to expand the polytope. It will be shown that the number of lattice points contained in the polytope formed by expanding P by multiplying each vertex coordinate by such a factor is a quasi-polynomial in n. Quasi-polynomials are a generalization of polynomials, where the coefficients of the quasi-polynomials are periodic functions with integral period. Furthermore, particular cases where the number of such lattice points is a polynomial will be presented. In addition, the period of these quasi-polynomials as well as the Law of Reciprocity will be addressed. At the end, future work will be discussed. [1] E. Ehrhart, “Sur les Poly`edres Rationnels Homoth ´etiques `a n Dimensions,” C. R. Acad. Sci. Paris 254 (1962)
The Divisibility of p^(n)-1 for p>5, p a Prime Number
Jason Adams, Nathan Jewkes and Tyrell Vance, Southern Utah University Mathematical Sciences We will study the divisibility of p^(n)-1 where p is a prime number larger than 5 and n is a positive integer. We will generalize the result by considering the case where n is odd and two cases where n is even. We show that when n=2^(k), k an integer greater than 1, 2^(k+2)∙3∙5 is a factor of p^(n)-1. We also show that when n=2^(m)∙l for m a positive integer greater than 1 and l an odd positive integer greater than 1, 2^(m+2)∙3∙5 is a factor of p^(n)-1.