Life Sciences

Michael Gillespie, Brigham Young University Life Sciences It is vital to understand the anatomical microstructure of tendons and ligaments in order to ascertain their specific qualities and functions. Recent developments in micro-scribe 3D digitization are highly effective in revealing these intricacies. A necessary component to creating 3D fiber maps from this technology is the ability to distinguish between individual tissue fibers with the naked eye. However, this is a very difficult task with most tendons and ligaments. We developed a paste made of blue dye and powdered sugar that when applied, fits in-between these fibers and contrasts the specimen color. This exposes the fine architecture, making individual fibers much more visible and thus able to be 3D digitized. With these 3D fiber maps now available, tendon and ligament microstructure can be viewed in greater detail than previously possible. This technique was applied to human cadaveric calcaneal tendon and quadriceps tendon. It was discovered that the fibers of human calcaneal tendon have higher overlap relative to the rigidly parallel fibers of the quadriceps tendon. This further understanding carries implications regarding the advancement of biomechanical models, artificial reconstruction, and surgical repair of these tissues. It also highlights the need for further investigation into the microstructure differences among tendons and ligaments.

Bradley Prince, 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, where spatial and declarative memories occur. Plasticity can either strengthen or weaken synapses, known as long-term potentiation (LTP) or long-term depression respectively. While many forms of synaptic plasticity are N-methyl-D-Aspartate receptor-dependent, recently endocannabinoids were identified to mediate several new forms of hippocampal synaptic plasticity. Endocannabinoids bind to receptors such as cannabinoid receptor 1 (CB1) and transient receptor potential vanilloid 1 (TRPV1), and mediate several forms of plasticity, including in the hippocampus. However, new research has demonstrated a non-CB1/TRPV1-dependent endocannabinoid synaptic plasticity in the hippocampus. While the receptor(s) involved is currently unknown, several potential candidate receptors that bind the endocannabinoid anandamide have been identified. These are orphan G-protein coupled receptors (GPRs) whose distribution in the brain and/or function is unknown. GPR55 is of particular interest as it activates second message systems, including increasing intracellular calcium. Using quantitative RT-PCR, electrophysiological and memory behavioral tasks we examined hippocampal GPR55 expression and function. GPR55 is indeed 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 decrease of LTD in brain slices. This LPI effect was not noted in GPR55 knock-out mice in the presence of LPI. This data suggest GPR55 is physiologically relevant in the hippocampus. This is the first direct evidence we are aware of that a novel endocannabinoid receptor directly effects hippocampal LTD. Because neurodegeneration that affects memory is typically associated with an increase in LTD, this provides a potential target to slow the advance of diseases such as Alzheimer’s.

Audrey Butler, Utah Valley University Life Sciences High-frequency (HF) ultrasound has been shown to be sensitive to a range of breast pathologies, and is being explored for the intra-operative assessment of lumpectomy margins. This sensitivity is believed to arise from microstructure-dependent interactions of ultrasound in the tissue. The objectives of this study were to develop breast tissue phantoms with microstructures that accurately mimic the histology of normal and malignant tissue, and to determine the effects of these microstructures on HF ultrasonic spectra (10-100 MHz). Phantoms were created from a mixture of water, gelatin, and soluble fiber. To simulate various breast tissue histologies, polyethylene beads, polyethylene fibers, and nylon fibers with a range of diameters were embedded into phantoms. Microstructures ranging from randomly dispersed beads to bead-fiber constructs resembling terminal ductal lobular units (TDLUs) were modeled and tested. Pitch-catch and pulse-echo measurements were acquired using 50-MHz transducers, a HF pulser-receiver, and a 1-GHz digital oscilloscope. Spectra were derived from the data and peak densities were determined from the spectra. Peak density, which is the number of peaks and valleys in a specified spectral range, has been shown to correlate with tissue complexity. Preliminary results from dispersed beads (58-925 µm diameter) of constant volume concentration (0.8%) indicated that the smaller beads produced higher peak densities than the larger beads with a consistent and statistically significant trend. These results substantially improve upon previous phantom studies and upon results from original breast cancer studies, demonstrating the strength of the HF ultrasound response to tissue microstructure. The higher peak densities can be attributed to either the higher number of scatterers for small beads or the size of scatterer in relation to the ultrasonic wavelength. These and other results from more advanced histologically accurate microstructures modeling TDLUs will be discussed.

Josh Harvey, Brigham Young University Life Sciences Soil CO2 flux represents an important pathway of carbon transfer from ecosystems to the atmosphere. Soil CO2 flux can be altered by global warming-driven changes in seasonal temperature and water availability. Subalpine ecosystems have high levels of carbon in their soils that are stabilized by low temperatures and low microbial activity during long and snowy winter seasons. Subalpine ecosystems can be important sinks for carbon, storing carbon that otherwise would be in the atmosphere contributing to global warming. In our study we show how changes in temperature and water availability during springtime increase the levels of subalpine carbon output. So long as the carbon outputs outweigh carbon inputs, increases in soil flux would amplify global warming. The amplification of global warming would loop back to affect soil fluxes again (by raising temperatures, melting snow earlier, and changing precipitation patterns) thus creating a positive feedback system. Understanding what feedbacks are present in a climate system and their underlying mechanisms will improve our forecasts of changes in atmosphere chemistry and temperature.

Logan Warner, Utah Valley University Life Sciences High-frequency ultrasound (20-80 MHz) has been found to be sensitive to a range of pathologies in excised breast tissue before fixation in formalin or other formaldehyde analogues. Formalin fixation, however, may alter the structure and rigidity of a sample so that data gathered using high-frequency ultrasound after fixation may no longer be viable for research purposes. This limits the amount of time researchers may conduct tests, so preservation via simple sugars is being considered. Numerous studies have been conducted using sucrose, trehalose, or glucose as cryoprotectants for cells and simple tissues. The objective of this study was to test the sensitivity of high-frequency ultrasound to changes in the microstructure, stiffness, and cellular integrity of tissue samples due to cryopreservation with these sugars. Domestic pig heart tissue was placed in aqueous solutions of sucrose, trehalose, and D-(+)-glucose. The specimens were refrigerated and observed over time using high-frequency ultrasound to detect tissue damage. The results of this study suggest that cryopreservation with sugars will not only allow more time for researchers to conduct ultrasonic tests on surgical specimens, but also that high-frequency ultrasound could potentially be used as an assay to measure tissue degradation in preserved living tissues such as transplant organs.

Hillary Hansen, University of Utah Life Sciences Type 1 diabetes is an autoimmune disease where pancreatic β-cells are destroyed, resulting in insulin deficiency. Generating new β-cells from stem cells for treating diabetes will benefit from understanding their development in vivo. Pancreatic β-cells, along with all other pancreatic lineages arise from multipotent pancreatic progenitor cells (MPCs). Previous studies demonstrate that the structural and signaling protein β-catenin is required for the development of the exocrine acinar lineage. β-cells still differentiate in the absence of β-catenin, however, β-cell mass is dependent upon β-catenin. We determined that this dependency reflects a role for β-catenin in the maintenance of MPC patterning as well as for expansion of the progenitor pool. Whether our observed effects are due to the signaling or structural function of β-catenin remains unknown, and is the focus of this research. Using mouse genetics we are able to separate the structural and signaling functions of β-catenin. Eliminating both functions in PBKO (full knockout) mice produces decreased β-cell mass and irregular patterning. Decreased β-cell mass is also observed in PBsKO (signaling deficient) mice, though patterning remains unaffected. This suggests that pancreas growth is dependent upon canonical Wnt/β-catenin signaling, and that maintaining progenitor identity requires the structural role of β-catenin. Elucidating distinct roles for β-catenin could be used to drive stem cell-derived MPCs to expand and differentiate to the desired pancreatic cell fate.

Michael Naegle, Brigham Young University Life Sciences Dermaptera is a comparatively small order of insects with approximately 1800 species placed in three suborders. While the majority of earwig species are placed within the suborder Forficulina and are free-living with forceps-like appendages, two dermapteran lineages have a very unusual morphologies and life histories. The viviparous Hemimerina live epizoically on giant rats in tropical Africa where they feed on fungi growing on the rats’ skin. Hemimerina lack eyes and wings and the cerci are filiform. The viviparous Arixenina are associated with bats in Malaysia and the Phillippines, and they feed on bat skin gland secretions. They have reduced eyes, are wingless, and possess straight cerci. The phylogenetic position of the suborders Arixenina and Hemimerina relative to Forficulina have previously been unclear; however preliminary analysis suggest the phylogenetic position of the suborders Arixenina and Hemimerina are nested within Forficulina, with ectoparasitism evolving multiple times within this order. We generated DNA sequence data from three nuclear (18S, 28S and H3) and two mitochondrial (COI and TUBA) genes for representatives of all three suborders and outgroups. A phylogeny was reconstructed to address the following questions: (1) Does Hemimerina + Arixenina form a monophyletic group and support a single origin of parasitism or are there multiple origins of parasitism? (2) Is Forficulina monophyletic with respect to these parasitic lineages? (3) Are morphological similarities shared by the ectoparasitic forms synapomorphic or homoplasious characters?

John Hancock, Brigham Young University Life Sciences LLO56 and LLO118 are CD4+ T cells specific for the same Listeria monocytogenes epitope. Despite their TCRs differing by only 15 amino acids, LLO118 and LLO56 have dramatically different primary and secondary responses to Listeria monocytogenes infection. We reasoned that LLO56, the single chain TCR (Vβ2-linker-Vα2) could be subjected to directed evolution to generate mutants that are more stable and bind to peptide-MHC with higher affinity. Single chain LLO56 was fused to the yeast surface protein Aga-2 and error prone PCR was used to generate mutagenic libraries. A first generation stabilized single chain TCR (scTCR) was selected using biotinylated Vβ2 and Vα2 antibodies and anti-biotin beads. The first generation LLO56 mutant expressed LLO56 on the surface of yeast at higher levels than wild type by flow cytometry. To produce mutants with additional stability, a second-generation mutant was generated by combining multiple stability mutations isolated in a number of first generation clones.

Brian Ballard, Brigham Young University Life Sciences Antigen presenting cells digest and display proteins from foreign and infected cells on the major histocompatibility complex (MHC) which can then be recognized by T-cells through their T cell receptor (TCR). LLO56 and LLO118 are CD4+ T cells specific for the same Listeria monocytogenes epitope but show dramatically different primary and secondary responses to infection. Because TCRs have very low affinity for MHC we would like to create a high affinity T cell. We reasoned that the single chain LLO118 TCR (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 was fused to the yeast surface protein Aga-2 and error prone PCR was used to generate mutagenic libraries. The first generation stabilized LLO118 single chain TCR (scTCR) was selected using biotinylated Vβ2 and Vα2 antibodies and anti-biotin beads and it expressed LLO118 at higher levels than wild type by flow cytometry. To produce mutants with additional stability, a second mutagenic library using the first generation mutants as templates has been produced and the most stable clones will be selected after temperature denaturation, permitting isolation of clones with increased stability for generating high affinity pathogen specific scTCRs. After engineering a high affinity T cell our research will further understanding on TCRs and the MHC and could also serve as a resource for creating a therapeutic drug.

Lance Deeter, University of Utah Life Sciences Continuous-flow left ventricular assist devices (LVADs) are used in advanced heart failure patients either to bridge them to transplantation or as a permanent-destination therapy. We determined whether chronic exposure to non-pulsatile blood flow and acute increases in coronary perfusion pressure associated with LVAD implantation would influence arterial function. Arteries from a transmural biopsy of the left-ventricle were obtained from ten male patients (54±4 years old) at the time of LVAD implant (n=17, 184±25 µm i.d.) and 239±51 days later upon LVAD explant (n=21, 281±22 µm i.d.). Lmax tension was determined and dose-response curves to potassium chloride (KCl, 10-100 mM) were performed using isometric tension techniques. Next, bradykinin (BK, 10-6 to 10-10 M) and sodium nitroprusside (SNP, 10-4 to 10-9 M) concentration-response curves were completed on vessels precontracted to ~65% of maximal tension development. Maximal BK-induced vasorelaxation was greater (p<0.05) at explant (85±5%) vs. implant (59±9%), while SNP evoked responses (~90%) were similar between time-points. These findings suggest coronary endothelial function is improved by LVAD implantation. Heart failure was precipitated by a myocardial infarction in six of the ten patients. These are referred to as “ischemic” patients whereas the remaining four are “non-ischemic” patients. We hypothesized that coronary vascular responses would be improved by LVAD implantation to a greater extent in ischemic vs. non-ischemic patients. In ischemic patients maximal BK-induced vasorelaxation was greater (p<0.05) in coronary arteries obtained at explant (87±6%, n=14, 305±30 μm i.d.) vs. implant (53±11%, n=12, 204±33 μm i.d.). In non-ischemic patients maximal BK-induced vasorelaxation was similar in arteries obtained at explant (79±9%, n=7, 232±21 μm) and implant (72±17%, n=5, 135±13 μm). SNP responses were similar (~90%) between groups at implant and explant. Collectively, our data suggest that LVAD implantation improves endothelium-dependent vasorelaxation in ischemic but not in non-ischemic patients.

Ashtyn Smith, Westminster College Life Sciences Methylmercury (CH3Hg) is a neurotoxin that accumulates in lakes and streams due to the action of microorganisms, which can produce this biologically relevant organic form from elemental mercury (Hg). Therefore, the activities of microorganisms become key to understanding the balance of Hg and CH3Hg in the movement through the food chain in any ecosystem. Many species of microorganisms are resistant to Hg and can thrive in polluted waters. Recent studies have shown that Hg resistance in microbes can stem from one of two gene pairs, merAB or hgcAB. The merAB system allows the organism to covert CH3Hg into elemental Hg. Conversely, the hgcAB system coverts Hg into CH3Hg. Thus, it is important to determine how the microbial community of Great Salt Lake, Utah is affecting the CH3Hg concentrations in the lake. In order to determine the genotype of the lake’s halophiles, “salt-loving” organisms, microorganisms were collected from the deep brine layer in eight areas of the lake. The microorganisms were then isolated and cultivated on increasing concentrations of HgCl2. Halophiles from these samples have been isolated on 25 ppm HgCl2 at various salinities, demonstrating a robust resistance to Hg. PCR amplification and genetic sequencing will be used to determine the gene mechanism of mercury resistance (merAB or hgcAB) as well as the 16S rRNA gene, which will aid in identification of the species. Should this study identify GSL microorganisms that exhibit the merAB genotype, these organisms could potentially be utilized as bioremediators of the CH3Hg pollution in the lake.

Dusting Day, Brigham Young University Life Science Atherosclerotic vascular disease is the leading cause of morbidity and death in the United States. Approximately 1.4 million surgical procedures are required every year for treatment of vascular disease and its subsequent issues. While saphenous vein and internal mammary artery grafts are most commonly chosen by physicians, many patients who are in need of arterial grafts have vessels that are not ideal for grafting because of damage to the vessels or disease. This introduces the necessity for synthetic blood vessel grafts that function precisely as natural vessels in vivo. Our blood vessel research team has entered the tissue engineering field in its most exciting effort: the scalable rendering of cell-seeded vascular constructs with rapid prototyping machines or 3D printers. We have built and are modifying a 3D printer to deposit living endothelial and smooth muscle cells into vascular structures. Using agar, alginate, or collagen gels as placement media, cells can be arranged in shapes resembling multilayered artery tubules and proliferate to form functional arteries. The endothelial layer and smooth muscle layer of cells interact to secrete a natural extracellular matrix (ECM) between them. We have successfully cultured endothelial cells and are perfecting our technique of harvesting aortic smooth muscle cells for culture. These cells will be encapsulated in a gel we have optimized for cell adhesion and proliferation and will then be printed with our rapid prototyping machine into the shape of a blood vessel. After proper cell growth and secretion of the ECM we will subject our synthetic graft to tensile strength testing, thrombosis tests, and eventually implantation into an animal for observation of any immunogenic effects. Our project’s success would bring an array of new treatment options through biomedical engineering that would save many lives of those who suffer from cardiovascular disease.

David Money, Brigham Young University Life Sciences Interest in animal personalities, and particularly the effect that different environments have on personality, has increased dramatically over the past decade. Understanding how individuals vary in their behavior, and if there are consistent differences among populations from divergent selective environments, lays the foundation for studies focusing on the contribution of divergent behavior in species formation. To date, studies that have focused on how personalities differ across ontogenetic stages have failed to compare populations that occur in dramatically different environments. Our study attempts to fill this void by studying the ontogeny of personality in populations that have evolved in environments with different levels of risk (i.e., predation vs. no-predation). We tested the expression of different personality traits evolution across ontogeny (i.e., from juveniles to full grown adults) in two sister species of live-bearing tropical fish, Brachyraphis roseni and B. terrabensis. These species have evolved in different selective environments, with B. roseni having evolved in an environment where predators were present, while B. terrabensis evolved in an environment lacking predators. We assessed the boldness expression of individuals from several groups in populations, namely juveniles, small adults, and large adults. To measure boldness, we used an emergence test, and also an exploration and activity test (i.e., ratio of movement to idleness during an allotted time period). Our study provides evidence for an important relationship between predation environment and the evolution of personality traits across ontogeny.

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.

David Vogelsang, Brigham Young University Life Sciences More than one-third of U.S. adults (35.7%) are obese (CDC, 2013a), and since 1980, obesity among adolescents has risen from 5% to 18% (CDC, 2013b). Unless we do something to combat the growing obesity epidemic, we are consigning ourselves and future generations to a lifetime of heart disease, diabetes, cancer, and psychological distress (CDC, 2013a). Cooking Anatomy Academy (CAA) promotes healthy eating and cooking among parents and students in the Polynesian community to raise awareness about the growing obesity epidemic. Our primary focus is introducing parents and students to healthy, great tasting foods and easy to prepare meals (Brown, 2011). Our secondary focus is to teach the simple anatomy and physiology important to understanding healthy nutrition. CAA is composed of seven, one hour lessons that are being offered as an afterschool program at Mana Academy Charter School. We’ve developed the CAA curriculum to incorporate many of the nutrition guidelines on MyPlate.gov, and focus on moderation, variety and raw/unrefined foods. To study the impact of Cooking Anatomy Academy, we will take a qualitative approach and use journal entries to collect data. Participant journal entries will answer prompts like, “based on what you learned today, what will you have for a snack tomorrow,” or, “how many fruit servings should you have each day?” CAA mentors will record any positive or negative feedback they receive during each lesson. From the data we collect, we hope to see that CAA is helping participants make healthy food choices and increasing their obesity awareness.

Derek Harris, Dixie State University Life Sciences The presence of extracellular DNA (eDNA) in various environmental and biological media has become the subject of growing interest in the field of research. In media such as bacterial biofilms, it has been shown to play a vital role in their structure and antimicrobial properties. Existing methods for extraction of pure eDNA from these media are complex and problematic; particularly from biological media where cells containing genomic DNA are also present. Novel surfactants have been developed, whose miscibility and polarity are easily tuned to suit a variety of conditions necessary for eDNA extractions. They can accomplish extraction of pure eDNA through concurrent hydrophilic and hydrophobic interactions in a single step, while remaining unreactive with the surrounding media or lysing cells and exposing genomic DNA. We have shown by spectrophotometric quantification that these surfactants extract measurable amounts of DNA into a water-immiscible solvent layer, which can then be removed from the media. The DNA can then be further amplified and purified for analysis. Further refinement of extraction methods utilizing these surfactants could prove a tremendous asset to research attempting to elucidate the possible genetic content of eDNA and the mechanisms behind its often crucial role in environmental and biological media.

Austin Pearce, Brigham Young University Life Sciences Agave utahensis acts as a keystone species across its native range in the Mojave Desert and Colorado Plateau (Gentry, 1982). As a keystone species, Utah agave contributes to soil formation along barren mountain ridges, and has provided starch-rich sustenance to Native American tribes. Furthermore, taxonomists consider each of the two subspecies, kaibabensis and utahensis, to have considerable morphological variation (Gentry, 1982) within their own unique ecological niches. Given the importance of Utah agave, the high degree of variation, and its unique ecological niches, there is surprisingly little information published regarding its physiological ecology. In fact, no effort has been made to determine the population densities of Utah agave due to the remoteness of the region and its difficult terrain (e.g., the Grand Canyon). Therefore, geospatial analysis tools specific to environmental niche modeling provide a powerful means through which these issues and knowledge gaps can be effectively addressed. My goal is to develop a species distribution model by joining known locations of Utah agave with climatic and environmental data in MaxEnt and ArcGIS software. Such a model can be used by others for further ecological field studies of Utah agave and its subspecies. Additionally, the approach I employ can be used as a pattern for mapping distributions of other important plant species in remote and difficult-to-access regions of the world.

Daniel Loveland, Brigham Young University Life Sciences The monoamine oxidase A (MAOa) gene has been shown to be associated with various social behaviors and disorders such as: aggression, depression, and anxiety (Meyer et al., 2006; Kinnally et al., 2010; Newman et al., 2005); and the MAOa gene interacts with environmental influences to produce its phenotypic effects (Newman et al., 2005; Kinnally et al., 2010). The MAOa gene encodes the enzyme monoamine oxidase A, which is the main enzyme to break down the monoamines into their respective metabolites. An orthologous repeat variant of the MAOa genotype seen in humans has been found in the rhesus macaque: a 5 repeat (R), a 6R and a 7R. This study investigates the influence MAOa genotypes have on central monoamine functioning as measured by cisternal cerebrospinal fluid (CSF) monoamine metabolites associated with behavioral dysfunction (dopamine metabolite: homovanillic acid-HVA, norepinephrine metabolite: 3-methoxy-4-hydroxyphenylgycol-MHPG, and serotonin metabolite: 5-hydroxyindoleacetic acid-5-HIAA). Cisternal CSF was obtained from 136 30-day old infant male rhesus macaques with varying genotypes and rearing backgrounds. We expected to find a rearing by genotype (GxE) effect on the monoamine systems with differences between mother-reared subjects when compared to subjects reared without mothers in peer-only groups. We found significant variability between genotypes; results also showed early rearing modulated this genotypic effect on brain chemistry. This supports our hypothesis that GxE interactions influence monoamine metabolite concentrations, suggesting a possible relationship of GxE interactions on social disorders such as aggression, depression and anxiety.

Brittany Hammontree, Dixie State University Life Sciences Depending on metabolic conditions or dietary preferences, people can often become deficient in critical vitamins and minerals. For example, a number of people are deciding to become vegetarians, and vitamin B12 deficiencies could become a huge epidemic, as this essential vitamin is only obtained through meat products. This issue was the driving force to look deeper into new ionic liquid materials and how they could be used as a transport agent for vitamin B12, along with other vitamins. Ionic liquids are organic salts that are currently being explored in many scientific fields due to their unique properties. However, using ionic liquids as a transporter in transdermal applications has yet to be explored Developing new mechanisms of administering nutrients via transdermal processes can increase the bioavailability and effectiveness of vitamins and minerals that often cannot survive oral administering due to the acidity and molecular absorption via the stomach. This research focused on finding the right ionic liquid with high solubility of the individual vitamins. Several ionic liquids were developed, and the different vitamins were tested for solubility levels. This greater solubility allows for maximum exposure of the vitamin during transdermal delivery. In particular, two different vitamins were tested – vitamin K and Protoporphyrin, a chemical analog to vitamin B12. Additionally, the effect of these ionic liquids on the physiology of the blood and plasma as it enters the body past the skin layers is critical to understand. In addition to transdermal applications of vitamins, transport of these vitamins to other countries and remote locations could have tremendous implications. Ionic liquids tend to increase shelf life of solutes, and the availability to provide these materials during medical missions or service trips would be increased substantially, particularly in more remote settings.

Eric Middleton, University of Utah Life Sciences One of the most impressive examples of homing among birds is demonstrated by Columba livia, or the rock pigeon. In racing or carrier breeds, birds can routinely travel over 100 miles and reliably find their way back to their home loft (Pratt, 1954). Bred from feral rock pigeons, these racers and carriers have been selected for their increased ability to home. While much research has been conducted on the mechanisms of homing, the heritability of homing is not very well understood. Furthermore, it is unclear if homing is predominantly learned or innate (Melhorn, Haastert, Rehkamper, 2010). The existence of homing breeds demonstrates that homing is heritable, but the degree to which this is the case or what traits in particular are improved with selective breeding are more uncertain. To attempt to better understand the heritability of homing, we will compare the homing ability of pigeons who are siblings to each other, and to unrelated pigeons. We first captured 90 wild pigeons from areas around Salt Lake City and allowed them to breed freely. We tracked the ancestry of all of the pigeons born from the feral birds and banded each of the birds in order to reliably distinguish which birds were related. We are now in the process of teaching the captive bred birds to home. Once this is completed, we will release the birds at varying distances from the loft. To quantify homing ability, we will measure angle of displacement from the loft when the birds first orient themselves and being homing, the time it takes to home, and which birds successfully make it home to the loft. This data will then be used to compare the homing ability of siblings to unrelated pigeons.

Stephen Tahan, Westminster College Life Sciences

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

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

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

Jacob Anderson, Utah Sate University Life Sciences Leafy spurge (LS) is an aggressive Eurasian forb that has been successfully reduced in many areas in western North America through the biological control releases of flea beetles. Long term studies of this phenomenon are sparse. Three flea beetle species were released in the mid-1990s at a site dominated by LS in Richmond, Utah. This study assessed the long term effects of LS biocontrol on an ecological community at this site by addressing five questions: (1) Is LS abundance significantly lower now than in the 1990s? (2) What plant species are replacing LS and are they native or non-native? (3) Have the flea beetle populations persisted since their initial release? (4) What part does soil type play in which flea beetle species now dominate at the site? (5) In response to their unexpected presence, what role may long-horned beetles contribute to the long-term reduction of LS? It was found that LS abundance has significantly decreased from the 1990s; the dominant plant species are those of non-native grasses; flea beetles have persisted in significantly smaller numbers, with Aphthona lacertosa being the most abundant; and long-horned beetles appear to play a significant role in the reduction of sexual success of LS. The results of this project have implications for land managers when considering the vegetative response to LS biological control and the importance of long-horned beetles for long-term in managed, LS-reduced habitats.

Michael Morgan, Utah State University Life Sciences The advancement of biologically derived alternatives to petroleum diesel fuel requires a multifaceted approach. At Utah State University we use an interdisciplinary team including the Colleges of Engineering, Agriculture & Applied Sciences, and Science in conjunction with industry partners to drive innovation in improving the science behind petroleum diesel alternatives. With increasing petroleum use, depleting reserves, increasing emissions standards, and other factors, there is need for petroleum diesel alternatives that are cost effective, offer improvement, and perform similarly to petroleum diesel. Our team has focused on the use of oleaginous microbes utilizing low value effluent and waste sources including sugars and CO2 to create biofuels. We have focused on a yeast, Cryptococcus curvatus, and a microalgae, Nannochloropsis salina which have shown high yields of fuel per cell mass. Using these microbes we have utilized USU’s own direct trans-esterification reaction to create sufficient quantities of biodiesel for engine performance and emissions testing, including a subset of ASTM tests characterizing the fuels from each organism. Our initial engine testing used petroleum diesel as a baseline in conjunction with commercial soybean biodiesel to establish the quality of our microbially derived biodiesel. Testing in stationary diesel engines and on the Bonneville Salt Flats has proven our microbial fuels perform similarly to soybean biodiesel and comparably to petroleum diesel. To further improve biological diesel replacements we have begun working to create green diesel, hydrocarbons from a biological source, using a novel method of hydrothermal liquefaction. Preliminary results of those tests are presented here. Through a multifaceted and interdisciplinary approach USU is successfully improving petroleum diesel alternatives from microbial sources including characterization of the properties of these fuels and is working to create the fuels at the scale necessary for exhaustive engine performance and emissions testing including ASTM testing of all important fuel properties.

Adam Olson, Brigham Young University Plant and Wildlife Sciences Forests systems of the Central Rocky Mountains rely on an intricate balance of natural disturbance cycles in order to develop properly. Forest fires are one such disturbance, however, certain fire characteristics, particularly fire severity, can vary widely across forest landscapes. In our study, we examined the influence of fire severity on aspen regeneration as well as aspen defense against wildlife and livestock browsing. Our results indicate that high or moderate burn severity is more favorable to aspen regeneration and survival. These areas of severe burn result in a higher density of aspen suckers, more vertical and lateral growth, greater chemical defense concentrations, and less browse damage than the neighboring plots of low or no burn. This data suggests that fire severity should be taken into account when considering plant regeneration and susceptibility to browse damage in burned landscapes.

Matthew Whited, Brigham Young University Biochemistry Several lines of evidence suggest that protein lysine acetylation pathways are deregulated in cancer (1). Moreover, deacetylase inhibitors are emerging as important anti-tumor therapeutics, suggesting that the forced reprogramming of protein-lysine acetylation is toxic to tumor cells. In this study we show that Sirt1, an NAD+-dependent Sirtuin deacetylase that promotes cancer cell survival, is aberrantly mislocalized to the cytoplasm of breast tumor cells. Moreover, the depletion of cytosolic Sirt1 by siRNA sensitizes breast tumor cells to paclitaxel-induced death. Previously, we developed a biotin-switch proteomics approach to identify cytosolic Sirt1 substrates (2). This approach yielded a variety of substrates with roles in metabolism, survival, and oxidative stress signaling. Our current work focuses on three of the proteins identified as Sirt1 substrates: SOD1, DJ-1, and 14-3-3z. SOD1 and DJ-1 both suppress oxidative stress-induced death, and high levels of 14-3-3z expression suppress chemotherapy-induced apoptosis and correlate with negative patient outcomes in breast cancer. Our preliminary results suggest that acetylation of DJ-1 and SOD1 suppress their anti-oxidant functions, while acetylation of 14-3-3z disrupts its binding to pro-survival proteins. Taken together, our data support a model in which cytosolic Sirt1 activates multiple pathways that work together to promote tumor cell survival.

Hyrum Shumway, Brigham Young University Microbiology and Molecular Biology Epigenetics is the study of factors of gene regulation that do not stem from the primary DNA sequence. One such example of epigenetic gene regulation is where DNA wraps around histone proteins to form nucleosomes. The positioning of nucleosomes is the first order of control for genic transcription. Wrapped DNA is less accessible for transcription compared to DNA that is nucleosome free. Model organisms for human epigenetics such as the nematode Caenorhabditis elegans are invaluable because of their ease in manipulation and because the components of nucleosomes (histone proteins and DNA) are highly conserved across phylogeny. The purpose of my particular research is to develop and validate a new protocol for tissue-specific isolation of nucleosomes through immunoprecipitation in young adult C. elegans. This protocol leverages green fluorescent protein fused to histones to research nucleosome positioning in the germ-line cells of C. elegans nematodes. When successful in our animal model, this procedure is the first step in allowing analysis of chromatin architecture of any tissue at any developmental or disease state including human cells. The widespread prevalence and implications of human disease is staggering in magnitude. Research is ongoing to decrease morbidity, increase prevention, and fight infection. With epigenetic tools, this vital research is benefitted and supplemented.

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

Ji Su Park, Brigham Young University Nutrition, Dietetics, and Food Science Selenium (Se) and soy have each been shown to reduce risk for prostate cancer when consumed at high levels. The purpose of this project was to define the molecular mechanisms of prostate cancer chemoprevention by Se and soy, and to describe how timing of dietary treatment modifies those effects. [C57BL/6 X FVB] F1 TRAMP (TRansgenic Adenocarcinoma of Mouse Prostate) male mice were fed stock diets high or low in soy, with or without a supplement of Se (4.0 mg Se/kg BW as Se-meth- ylselenocysteine) by gavage 5 d/wk in a 2 X 2 factorial design. Mice were exposed to different diets starting from conception, 6 weeks, or 12 weeks of age and were sacrificed at 18 weeks. Three-way ANOVA showed that supplemental Se increased serum and liver Se, with significant interactions with both time and soy intake. Selenium dosing decreased BW independent of soy intake and time of dietary intervention. Both Se and soy decreased epididymal fat pad weights, with Se’s effects being more pronounced in mice exposed to diets from conception than from 6 wk. Urogenital tract weights, a measure of prostate proliferation and tumor volume, were significantly reduced by Se supplementation (P<0.001) and soy (p=0.044), independent of time of dietary intervention. Histological examination of mouse prostates is in progress to determine dietary effects on disease progression. These data suggest that, in this model, chemopreventive efficacy of Se and soy does not differ between prenatal and early post-natal introduction.

Marcus Vranes, Brigham Young University Molecular Biology Recent studies have attempted to discover the correlation that exists between DNA methylation and nucleosome positioning, but none have explored the direct effect of DNA methylation on nucleosome formation and positioning. This proposed research will directly test the effects DNA methylation has on nucleosome positioning and whether the histone octamer has preferred sequences to which it binds, which will in turn add our understanding of gene expression and regulation. A better understanding of these concepts will help to aid efforts in gene therapy to better the quality of life of many who suffer from various genetic conditions.

Sarah Miller, Southern Utah University Agriculture and Nutrition Science The high intake of fat in the average American diet is a growing concern to the nutritional community. High levels of total fat intake increase cardiovascular disease (CVD) which is the number one killer in America. The successful development of low fat recipes that can be accessed and produced by the general population is valuable to the fight against obesity and CVD. If acceptable products can be made with lower fat content then it is possible to lower the fat intake in a person’s diet. This study is designed to test the acceptability of low-fat desserts using every day food products available for home use as fat substitutes. Four popular “name-brand” recipes were altered by removing 55-73% of the fat and replacing it with a common food item, either pureed white or pinto beans, applesauce or yogurt. Southern Utah University students were recruited via flyers, in class announcements, and word of mouth. Participants (n=56) were asked to taste and evaluate the four products. Afterwards they completed a short survey about their knowledge of low-fat food substitutes and acceptability of the products. Results and conclusions forthcoming.

Adrian Scottorn, Salt Lake Community College Natural Sciences Chocolate has long been the favorite snack of many, and recent studies have shown multiple positive health benefits of eating dark chocolate in particular, encouraging production and consumption of very high cacao content bars. We have quantified some of the changes that occur to the amount of known flavor compounds contained in a locally made artisan chocolate. The bars we chose were 70% minimum single origin cacao made with as few other ingredients as possible. By first measuring the amount of known flavor compounds, then stressing the bars in various ways, we looked in to what really happens when a quality chocolate bar is mistreated.

Michael Peterson, Brigham Young University Biology A person’s predisposition to Alzheimer’s Disease is known to be influenced by both genetic factors as well as environmental factors. One know environmental factor is that known to affect risk for disease is early parental death. The purpose of this research is to better understand the complex factors that influence the disease by analyzing the relationship between the environmental factor of early parental death with genetic variants known to influence the disease. We used extant data from the CCSMHA, an ongoing aging study including 89.7% (5092 of 5677) of all of the eligible residents of Cache County, Utah. This data includes information about environmental and psychosocial stressor of the subjects as well as information about physical examinations, metal screenings, and individuals’ genotypes at many loci that are known to be related to Alzheimers Disease. We used multivariate logistic regression to determine the effect of early parental death by SNP interactions on risk for AD. For the analysis we cleaned the data by removing SNPs less than a minor allele frequency of 0.01, a Hardy-Weinberg equilibrium value of 110-6, and a maximum missing snp call of 0.2. Individuals were also removed if genotyping rate was less than 0.2. After filtering we had 262 cases, 239 controls and 0 missing Final Results will be presented at the Conference.

Rachel Perry, Brigham Young University Life Sciences Previous studies have indicated that Visinin-like protein (VILIP) may be a powerful tool in predicting disease progression and guiding prognosis of Alzheimer’s disease (AD). Cerebral spinal fluid (CSF) was collected from hundreds of individuals with varying levels of AD. The CSF was then analyzed for levels of VILIP protein using Luminex technology. SNPs were genotyped using the Illumina OmniExpress chip. SNPs found to have a Hardy-Weinberg frequency less than 1×10-4 were not included, assuming that this variance was due to a genotyping error. SNPs and samples missing more than five percent of the data were also not included. Following the cleanup of the data, an association test using linear regression was performed. Covariates used in the analysis included age, gender, and covariates that accounted for population stratification (PC1 and PC2). Over one hundred SNPs were found with a p-value less than 1×10-5. The genomic inflation factor for the generated data was 1. One marker showed significance at the genome-wide level. We have identified a genetic marker that shows significant association with CSF VILIP levels. This finding may provide insight into genetic control of VILIP levels, which may be a useful in understanding the pathological processes involved in AD.

Chris Thurman, Salt Lake Community College Natural Sciences This project is an ongoing multifaceted investigation of the Jordan River. The study’s intent is to monitor and asses the overall condition of the river from Utah Lake to the Great Salt Lake. Samples of river and pond water, typical vegetation such as cattails, and some soils have been collected with reference to sites along the river that may serve as sites for the introduction of waste and other contaminants. These samples have so far been examined with respect to temperature and dissolved oxygen at the sampling point. Two river samples, one pond sample, and plant sample have been analyzed by ICP-MS for 20 different metals. The levels of Lead, Cadmium, and Copper have been determined by anodic stripping voltammetry at the ppb level in a variety of other water samples from the river. The results of these measurements are disused and presented Geospatially.

Matthew Schneider, Brigham Young University Physiology and Developmental Biology Alzheimer’s disease, the most common form of dementia, affects millions of people per year. Research has shown that Alzheimer’s affects the hippocampus brain region, which is involved in learning and memory. Understanding learning and memory functions is imperative to comprehending both healthy brain functions and Alzheimer’s disease. Many researchers seek to understand both the causes and treatments of the disease, but tangible information remains elusive. Studies thus far have shown that to encode memories, the brain must change neural synapses to either strengthen or weaken those pathways, a process known as synaptic plasticity. Using electrophysiology techniques on mouse hippocampal slices, this project will provide further insight on memory formation and regulation by imitating synaptic plasticity mechanisms. I will look at a specific cellular pathway involving the protein receptor GPR55, which has recently been shown to induce synaptic plasticity. By understanding how the GPR55 pathway functions, this research will contribute to the understanding and treatments of Alzheimer’s and other neurodegenerative diseases.

Lacie Cates, Salt Lake Community College Natural Sciences According to the Salt Lake Valley Health Department 1 the optimal level of fluoride is 0 .7 to 1.2 ppm in drinking water. Also acidity of many foods and drinks leads to tooth decay. In 2003 the State of Utah started adding fluoride to tap water. Then in 2008 Utah counties voted on addition of fluoride resulting in a variation of water treatment from county to county. At present the state has about 50% of the population receiving fluoride treated water with the aim of providing the 1 ppm fluoride level. It has also been reported that some counties are considering cutting back to about 0.7 ppm 2 . This study examined the pH and fluoride content of water samples from the major population counties in Utah. Fluoride levels ranged from 0.08 – 0.92 ppm. The pH values for these samples ranged from 6.26 -8.08. A further study of the fluoride levels and pH in bottled water and other drinks and foods such as fruit and cheese was conducted because many people in Utah do not drink tap water, particularly in regions of high water hardness or areas where taste and / or odor can be off putting. The pH values ranged from 2 to 8. The most acidic being colas and citrus based drinks and foods. The fluoride values varied from 0.03 to 0.47 ppm.

Kristian Johnson, Dixie State University Biology Antimicrobial methods, such antibiotics and Ultraviolet (UV) irradiation, have been a means of suppressing prokaryote proliferation for nearly a century. Over the last several years, scientists have found that numerous strains of prokaryotes have developed resistance to antibiotics. Concurrently, the process of bacterial irradiation using UV-C is common practice in a variety of sterilization applications. As revealed in the seminal work by Chang et al. inactivation curves for Microorganisms such as Staphylococcus aureus (Staph) were established in 1985. Their values indicate survival rates based on Intensity, which is defined as the time of UV irradiance per unit area. Similar to the evolutionary evidence of antibiotic resistance, we are interested in the selective pressure UV-C has on Staph. By recapitulating Chang’s experiment nearly 30 years later, our preliminary results indicate an increased resistance to UV-C in Staph. In this experiment, we determine a current UV-C dose-dependent kill rate function for Staph.

Chancen Hall and Nichkolas Hadley, Dixie State University Biological Sciences Batrachochytrium dendrobatidis (chytrid fungus) is prevalent worldwide, and the resulting chytridiomycosis has contributed to at least 168 amphibian species extinctions. In 2010, B. dendrobatidis was discovered in the greater Zion National Park area of southwestern Utah. Because few populations have shown resistance to chytridiomycosis, we decided to explore the effects of this disease on populations of Hyla arenicolor (canyon tree frog). We tracked the spread of B. dendrobatidis by testing skin samples taken annually from several different canyons and monitored population sizes. During the three years of our study, infected populations did not show subsequent population declines. This suggests that H. arenicolor population size in this region is unaffected by B. dendrobatidis. In the future, testing hypothesized explanations for surviving infection could help us identify populations not at risk and thus allocate conservation resources more efficiently.

Rachel Schneider, Brigham Young University Neuroscience The ability to create distinct memories for very similar stimuli and events is called pattern separation. Pattern separation is thought to be dependent on neurogenesis (the birth of new neurons) in the dentate gyrus, a subregion of the hippocampus. Neurogenesis is reduced in depression, as is overall memory performance. It has been proposed that depression negatively impacts pattern separation abilities, however a link between depression and performance in pattern separation memory tasks has yet to be identified. Accordingly, we designed a study to investigate the relationship between pattern separation performance and level of depression. Eighty-two participants completed a pattern separation memory test and a set of questionnaires to gauge their level of depression. During the task, participants were presented with 600 images one at a time on a computer screen in a continuous recognition paradigm. Participants were asked to determine whether each image was new, old, or similar. Images seen for the first time during the task qualified as “new”, images that were repeated following a variable delay qualified as “old”, and images that were similar to previously presented stimuli, but not exactly the same, qualified as “similar”. A pattern separation score was calculated based on the proportion of correctly identified similar stimuli. We found a negative correlation between depression scores and pattern separation scores (r(82) = – 0.301, p < 0.01). This relationship held constant even when we controlled for other factors known to affect neurogenesis, such as exercise and anxiety levels. These results provide support for the theory that depression is negatively related to pattern separation performance, possibly due to a decrease in neurogenesis in the hippocampus.

Michael Barney, Southern Utah University Biology Parkinson’s disease is caused by a decrease in dopaminergic neurons in the substantia nigra, which results in a loss of motor control. Although the exact causes of Parkinson’s disease are unclear, studies have shown that exposure to environmental contaminants causes death of cells in the substantia nigra (McCormack et al., 2002). The purpose of our study is to investigate the potential role of fire retardant as a risk factor for Parkinson’s disease. We subcutaneously injected 8 experimental mice with retardant solution (1 ml/60 g body weight) and 7 control mice with saline solution (1 ml/60 g body weight). Each mouse was given 8 injections over four weeks, after which the mice were sacrificed and brains were harvested. Frozen sections (40 mm thick) were mounted on slides and are being processed using anti-tyrosine hydroxylase, which will be visualized using a peroxidase reaction. The number of cells in the substantia nigra will be counted and compared between control and experimental groups. Our hypothesis is that exposure to fire retardant is a risk factor for Parkinson’s disease. If this hypothesis is supported, this would be the first study to show a link between fire retardants and Parkinson’s disease. This would have important implications for current forest fire fighting techniques.

Ryan Williamson, Brigham Young University Physiology and Developmental Biology The hippocampus is thought to mediate learning and memory by altering the strength of synapses within its circuitry. In many cases, this synaptic plasticity can be induced by intracellular signaling molecules. Lipid-based intracellular signaling molecules called endocannabinoids have been shown to modulate or mediate synaptic plasticity among hippocampal pyramidal cells and stratum radiatum interneurons; however, the role of endocannabinoids in mediating synaptic plasticity among interneurons in the stratum oriens is still unclear. Our goal was to determine whether stratum oriens interneurons have the machinery necessary for endocannabinoid production and, if so, whether this machinery is expressed in a sub-type specific manner. To do this, we used patch clamp electrodes to extract single cells from rat hippocampal slices and analyzed the expression of endocannabinoid biosynthetic enzyme mRNA using quantitative real-time PCR. In this analysis, we examined cellular expression of two interneuron markers, GAD65 and GAD67, as well as several calcium-binding proteins and neuropeptides to determine interneuron subtype. We also analyzed cellular expression of several endocannabinoid biosynthetic enzymes, including N-acyl phosphatidylethanolamine phospholipase D, diacylglycerol lipase alpha, and 12-lipoxygenase, as well as type I metabotropic glutamate receptors. Preliminary data suggests that stratum oriens interneurons express mRNA necessary for endocannabinoid biosynthetic enzymes. Additionally, we identified interneurons that coexpress mRNA for somatostatin and diacylglycerol lipase, suggesting that O-LM cells or another somatostatin-positive interneuron subtype may possess the enzymes necessary to produce the endocannabinoid 2-arachidonoylglycerol. Further work will allow us to examine how endocannabinoid biosynthetic enzyme expression correlates with other interneuron subtypes in the stratum oriens.

Aaron Sharp, Brigham Young University Biology The Cache County study on memory, health, and aging has played a significant role in several studies. However, there is some potential skepticism in the scientific community about its sample. The population in Cache County is derived from a diverse group of founders, but it is perceived by some to be an isolated population. If so, conclusions discovered there might not apply to other populations. Our objective is to compare the Cache County data to a panel of genetic data—provided by the International HapMap Project and the Alzheimer’s Disease Neuroimaging initiative—that is known to be representative of typical European-American populations. Doing so will indicate whether the genetic diversity in the Cache County sample is characteristic of an isolate or not. Analysis will be done using the open source “Plink” analysis toolset, including the –cluster and –mds-plot computational algorithms. Using –cluster groups individuals according to identity by state distances. The –mds-plot algorithm creates a scatter-plot of the individuals in 2-dimensional space, identifying any systematic difference between the Cache County data and the general population. We expect that the Cache County data will be representative of general European-American populations, because of its diverse group of founders.

Rachel Nettles, Brigham Young University Plant and Wildlife Sciences Background/Questions/Methods

Michael McEntire, Brigham Young University Biology Female mate choice (intersexual selection) and male dominance interactions (intrasexual selection) can each play important roles in sexual selection. These two mechanisms tend to be discussed in isolation. The goal of this study is to explore the interaction between these two forms of sexual selection. To test this idea, we focused on the livebearing tropical fish system Poecilia gillii. We grouped males into similarly sized pairs and observed them for a week to determine which male was dominant. These pairs were then presented to females in mate choice trials to ascertain female preference. We also photo- graphed the males to determine coloration. We found that females were unable to detect dominant males without viewing the physical contest and that carotenoid coloration bore no effect on female preference. Females tended to choose the male to their left, suggesting the preferential use of their right eyes in making decisions on mate choice.

Whitney Hoopes, Brigham Young University Microbiology and Molecular Biology HspB2 is one of eleven known small Heat Shock Proteins (sHSP) that is expressed in human heart and skeletal muscle. In response to cellular stress, heat shock proteins play a vital role to help misfolded proteins and proteins susceptible to denaturation maintain their structure. Two members of the sHSP family, CryAB and HspB2, are both required for normal heart function and cardiac muscle integrity. CryAB-deficient mice have defects in cardiac muscle structure whereas HspB2-deficient mice display energy deficits (Rajasekaran et al. 2007). The contrasting phenotypes of CryAB and HspB2 suggest differential roles for these molecular chaperones in the heart. HspB2 has been found to localize with the mitochondria in several different cell lines and overexpression of this sHSP has been shown to support survival of cells against heat stress (Nakagawa, 2001). To understand the role and mechanism of HspB2 in cardiac muscle energy regulation, we have used a yeast two-hybrid (Y2H) system to uncover the novel protein binding partners specific to HspB2. From screening a human heart cDNA library, HspB2 interacted with approximately 10,000 out of 20 million plasmids. We have sequenced more than 1000 of these putative interactors and have identified over 100 unique proteins. Over 40% of these protein partners are involved in mitochondrial energy production and another 25% in cardiac muscle structure maintenance. In addition, we have identified an interaction between HspB2 and the related sHSP CryAB. We then compared this data with mitochondrial HspB2 binding partners identified by mass spectroscopy (MS) through a large-scale bioinformatics analysis and constructed a protein-protein network. Y2H dependency tests were conducted to verify interactions identified by both Y2H and MS. Following yeast verification, a subset of the interactions were confirmed in C9H2 cardiac cells through coimmunopurification. Our research describes the first protein-protein interaction network for any sHSP, supports a role for HspB2 in mitochondrial energy production and suggests a link between mitochondrial energy production/redox stasis and stressed cardiac muscle maintenance.

Christian Boekweg, Brigham Young University Plant and Wildlife Sciences It has been shown that the population densities of deer and elk in the Rocky Mountains are at an unprecedented high. The heavy browsing of said species (ungulates) on regenerating aspen suckers can be devastating, leading to homogenously aged aspen stands that now have reduced resilience to drought, fire or logging disturbance, and an increased susceptibility to pathogens. Lastly, the extensive, highly selective herbivory of the aspen suckers may cause a shift in forest composition away from the aspen tree to an increase in other, less palatable species. We selected 186 sites across the 3 national forests of Central and Southern Utah, and characterized stand composition using the point quarter method. Stands were defined by successional stages from early to late; aspen, mixed or conifer. We then used pellet counts to estimate animal density and evaluated the effect of animal density and stand type on the regeneration success of aspen suckers. The key result of our study is that high ungulate density is highly correlated with reduced sucker heights (p<0.001). This indicates that high ungulate density impedes aspen regeneration by preventing aspen from recruiting into the overstory. Our study suggests that closer monitoring of the long term effects of herbivory on aspen development and regeneration is necessary to ensure vigorous aspen forests.

Jesse Cobell, Brigham Young University Biology Alzheimer’s disease (AD) is the sixth major cause of death in the U.S. However, at present, no diagnostically useful serum markers for AD have been identified. Hence, we used a novel serum proteomic approach to interrogate the low molecular weight proteome for serum biomarkers. This allowed for survey of around 5000 low MW species. To reduce ion suppression, an acetonitrile precipitation step was used to remove high abundance serum proteins. Protein-depleted sera from 58 cases and 55 controls were analyzed by cLC-ESI-QTOFMS/MS using reverse phase chromatography. Data were reviewed using Applied Biosystem’s Analyst-QS software to compile spectra. Differentially expressed peptides (cases vs. controls) were analyzed statistically using the Student’s t-test. This led to discovery of 36 candidate biomarkers. Additionally, we compared AD subjects with more severe disease (Clinical Dementia Rating (CDR) =3) with non-demented individuals (CDR=0) and found 23 biomarkers. Furthermore, on comparison of mild and moderate stage AD individuals (CDR = 0.5, 1, 2) with those with severe disease (CDR = 3), we found 24 biomarkers. Some of these biomarkers appeared more prominent in one gender. We then fragmented several of these biomarkers on an LTQ-Orbitrap XL hybrid mass spectrometer and cLC-ESI-QTOF-MS/ MS system using collision-induced dissociation to determine amino acid sequence analysis. We have identified 5 biomarkers and are in the process of identifying the remaining biomarker species. This serum proteomics approach found statistically different peptide abundances in subjects with AD. Additional biostatistical evaluations are underway to determine sensitivity and specificity of individual biomarkers and their combinations. Future studies will assess biomarkers according to disease stage and validate current biomarkers in blinded comparisons of other AD sera. This serum proteomics approach appears promising in locating and identifying clinically useful serum biomarkers of AD.