2013 Abstracts

Introduction:

The central research question of this project was to determine if high-frequency ultrasound is sensitive to tissue pathology at the microscopic level. Previous studies on surgical specimens have shown that high-frequency ultrasound may be sensitive to a range of breast pathologies including fibroadenomas, atypical ductal hyperplasia, fibrocystic changes, and carcinomas. The ultrasonic parameters that were sensitive to pathology were the number of peaks (the peak density) of the first-order spectra of the waveforms (one forward Fourier transform), and the slope of the second-order spectra of the waveforms (two consecutive forward Fourier transforms). The ability to determine pathology rapidly and with minimal specimen preparation would make high-frequency ultrasound particularly well-suited for real-time use during cancer surgery to ensure all of the malignant tissue has been removed. The purpose of this research was to determine the sensitivity of the peak density and spectral slope to tissue microstructures other than those found in breast cancer. The results of this study would not only support the results from the breast cancer studies, but also extend those results to the detection of cancer and other diseases in a range of organs and tissues. The research methodology included the following steps. (1) Freshly excised bovine organs were obtained from a meat packaging facility, including the heart, liver, and kidney. (2) Specimens approximately 3x3x1 cm in size were dissected from the organs and tested immediately with ultrasound. (3) Both pitch-catch and pulse-echo waveforms were acquired from the samples. (4) The data were analyzed by determining the peak densities and spectral slopes. The results showed that the more heterogeneous tissues of the heart, the vascular structures (aorta, vena cava, etc.), displayed significantly higher peak densities than the muscle tissues. Similarly, the ureter, which has greater heterogeneities in its structure (larger and more varied), displayed significantly higher peak densities than the cortex and medulla tissues. No significant trends were observed for the liver tissue, or for the spectral slopes except for kidney medulla tissue. Heterogeneity and peak density in high-frequency ultrasonic spectra that may be useful for performing real-time pathology during cancer surgery.

Introduction:

A new N-oxide was produced from 3-(N-Boc-aminomethyl)-5-bromopyridine by bioconversion with Cunninghamella echinulata var. elegans ATCC 9245, and its structure was established based on spectral data. The microbial N-oxidation is efficient and highly selective. The substrate was transformed into the product in 7 days.

Most organisms exhibit latitudinal gradients in diversity (i.e., taxonomic richness decreases as latitude increases). Few studies have sought latitudinal gradients in lichens, especially in the midlatitudes. Our primary questions were: 1) do lichens along the west coast of the United States show latitudinal gradients? 2) If so, what is the rate of change and does the level of taxonomic richness affect this rate? We hypothesized that lichens would show a reverse latitudinal gradient in the region, as has been documented for lichens elsewhere in the northern hemisphere, but at a considerably smaller scale. This study fills in the gap in our understanding of lichen latitudinal gradients over large areas of North America. It also functions as a baseline for future climate change and conservation efforts. Our study area is bound at the south by the California-Mexico border (32.331° N) and at the north by the Washington-Canada border (47.178° N), and extends inland from the coastline to the crest of the Sierra Nevada Mountains (116.083° W, at its eastern -most point). We divided the region into 218 roughly equal-area (cite) grid cells using GIS, each bordered with latitudinal and longitudinal lines. We derived a list of all vouchered lichen specimens in each grid cell using Consortium of North American Lichen Herbaria, an online database. The data were synonymized, and species, generic, and familial richness were calculated for each grid cell. We found no correlation (R2 = 0.2306) between latitude and species richness, using the raw vouchered data. What we did find was a strong correlation (R2=0.9069) between sample density and species richness. These results are biased by sample density and do not reflect what is naturally occurring. We hypothesize that we can get an unbiased estimate of richness with MaxEnt models. Using the georeferenced lichen distributions and related climate data, we constructed species distribution models of all species with five or more occurrences (990 species). In GIS, we projected all 990 distribution models and our 218 grid cells together to calculate species richness for each cell at various thresholds (i.e. likelihood of occurrence at 10%, 20%, 30%, etc.).

The use of metallic nanoparticles (NPs) in various applications presents the possibility for environmental contamination by these NPs. CuO and ZnO NPs have dose-dependent phytotoxicity. However, NPs are modified by different environmental factors such as salts: chlorides of Na, K, and Ca are used on roads for de-icing, dust control, irrigation, water treatments, and fertilizer production. At low concentrations these salts promote plant growth, but elevated concentrations become phytotoxic. This study examines the impact of Na, K and Ca chlorides on the phytotoxicity of CuO and ZnO NPs for wheat grown in sand. Atomic force microscopy and dynamic light scattering showed dose-dependent aggregation of the NPs in the wheat rhizosphere. The salts also caused NP-aggregation, but the degree of dissolution of the NPs was salt specific. Release of Cu from CuO NPs in the planted sand was reduced by all salt treatments whereas Zn release from ZnO NPs was reduced by Na and K but not Ca. Overall, a phytotoxic synergistic effect was seen at lower salt concentrations, but at higher salt levels, toxicity was mediated mainly by the salts. Therefore, effects upon plant growth of NPs and salts will vary according to concentrations.

NANOGP8 is a human (Homo sapiens) retrogene, expressed predominantly in cancer cells where its protein product is tumorigenic. It arose through retrotransposition from its parent gene, NANOG, which is expressed predominantly in embryonic stem cells. Based on identification of fixed and polymorphic variants in a genetically diverse set of human NANOG and NANOGP8 sequences, we estimated the evolutionary origin of NANOGP8 at approximately 0.9 to 2.5 million years ago, more recent than previously estimated. We also discovered that NANOGP8 arose from a derived variant allele of NANOG containing a 22-nucleotide pair deletion in the 3? UTR, which has remained polymorphic in modern humans. Evidence from our experiments indicates that NANOGP8 is fixed in modern humans even though its parent allele is polymorphic. The presence of NANOGP8-specific sequences in Neanderthal reads provided definitive evidence that NANOGP8 is also present in the Neanderthal genome. Some variants between the reference sequences of NANOG and NANOGP8 utilized in cancer research to distinguish RT-PCR products are polymorphic within NANOG or NANOGP8 and thus are not universally reliable as distinguishing features. NANOGP8 was inserted in reverse orientation into the LTR region of an SVA retroelement that arose in a human-chimpanzee-gorilla common ancestor after divergence of the orangutan ancestral lineage. Transcription factor binding sites within and beyond this LTR may promote expression of NANOGP8 in cancer cells, although current evidence is inferential. The fact that NANOGP8 is a human-specific retro-oncogene may partially explain the higher genetic predisposition for cancer in humans compared with other primates.

Although the insect fauna of the Colorado Plateau region are somewhat well known, our specific understanding of the diversity in Capitol Reef National Park is sparse. Here we present an overview of the insect diversity of the park based on a number of undergraduate student-led excursions to collect specimens. The collections will contribute to an increased understanding of the parks insect diversity and will result in an appropriately curated natural history museum collection. Furthermore, an insect field guide for the park will be created in order to serve as an educational tool for visitors to the park. Field collecting efforts, consisting of traditional insect collecting methods (aerial nets, aquatic nets, light traps, etc.) will be used. The collected material will be curated, photographed, and stored in a natural history museum (Brigham Young University ML Bean Museum). This presentation will be a summary of material gathered in the last three years.

The long term objective of this research is to explore oil-based materials that can be used to replace trans-fats in the food industry. Vegetable oils such as olive (OO), corn (CO), soybean (SBO), sunflower (SFO), safflower (SAFO) and canola (CAO) were mixed with sunflower wax (SFW), paraffin wax (PW), and beeswax (BW) at different concentration levels (0.1%-100%). Crystal Morphology, oil stability, thermal behavior and viscoelastic properties were studied in this research. Results in our laboratory showed that the crystallization occurred sooner when the concentration of waxes increased. For example, crystallization in 0.1% of SFW in SFO occurred after 24 hours of incubation at 25°C, but when concentration increased to 0.25% the crystallization was observed at the first 15 minutes of measurement. In addition, a change in the thermal behavior of wax/oil soft materials was observed when the concentration of waxes increased. For example, the enthalpy change (H) in SFW/ SFO materials increased from 0.87 J/g at 1% of SFW to 21J/g at 10% of SFW. This increase in enthalpy indicated that the amount of crystal increased when the concentration increased. Overall, the crystallization behavior in wax/oil materials was affected by wax concentration and chemical compositions of oils and waxes.

Arctic lakes are very sensitive to the effects of climate change. It is important to understand the current food web dynamic and energy flow within these lakes to better understand how they will change in the future due to the effects of a rapidly changing climate. In order to understand the current conditions in arctic lakes, this project consists of an analysis of stable isotopes of carbon (13C) and nitrogen (15N) from invertebrates among fish and fishless lakes in arctic Alaska to compare their trophic level positions and primary energetic sources. In order to carry out this analysis, I collected pelagic invertebrates were collected from 6 different lakes, 3 of which have resident fish populations and 3 of which are fishless. Samples collected in 2011 were analyzed for stable isotope composition by a mass spectrometer at University of California Davis. Results from samples collected in 2012 are pending at Washington State University. I will analyze and correlate the stable isotope results with isotopic data collected from other related projects. With this analysis, I will create food webs to 1) assign trophic positions to each species in each lake and compare those positions across lakes, 2) assess the potential effect fish predation has on pelagic invertebrate community structure. I hypothesize that fish predation will determine zooplankton community structure (e.g., dominant taxa) and alter trophic linkages (e.g., secondary trophic level predation rates).

Ovarian cancer is the leading cause of death from gynecologic malignancies in the United States and is the fifth leading cause of cancer death among American women. It is estimated that over 22,000 women in 2012 will be diagnosed with ovarian cancer in the United States and approximately 15,500 women will succumb to the disease. This is due to the fact that only 20 percent of cases are diagnosed before the cancer has spread to the peritoneal cavity. Currently, there are no reliable, standard screening tests; the only diagnostic test currently available is the CA125 tumor antigen blood test. This test is inadequate and not available as a general screening tool; additional diagnostics are required to effectively diagnose this disease. It has been previously shown that the tight junction protein Claudin-16, found only in normal kidney, is aberrantly expressed in epithelial ovarian tumors. Therefore, this protein is a good candidate for ovarian cancer diagnostics and targeted therapy. By identifying the promoter region that controls cldn-16 gene expression in ovarian tumors, we can create a luciferase reporter assay to identify cells that express Claudin-16 in culture. To do this, PCR-amplify of various upstream regulatory elements previously identified in kidney cell lines were subcloned into the pGL3 luciferase reporter vector. A higher amount of luminescence is present if the promoter sequence successfully up-regulates the luciferase gene in the vector. This is measured using a Dual Luciferase Assay to determine which promoter region is responsible for the over-expression of Claudin 16. Promoter activity was verified in kidney cell lines that normally express Claudin-16. Next, the assays will be repeated in ovarian cancer cell lines known to express Claudin-16 compared to cell lines that do not express the protein. The promoter assay will then be tested on a collection of ovarian cancer cell lines to determine if luciferase activity correlates with Claudin-16 expression. Once validated, we can test our construct as a cell based assay for identifying therapeutics that can lower Claudin-16 expression in ovarian cancer cells.

Nearly half of Americans reject evolution as an explanation of the origin of the diversity of life on Earth and rejection of evolutionary science has been shown to be correlated with a belief in a Christian God-creator (Gallup 2007; NSB 2010). The goal of this study is to explore the student perceptions and knowledge of Evolution at UVU in the non-majors and Biology majors introductory courses. Our study is designed to examine three hypotheses: Hypothesis 1 (Modern secularist): high levels of religiosity and fatalism should correlate with low levels of interest in science and scientific understandings of the world; Hypothesis 2 (Postmodern, postsecularist): no correlation between high levels of religiosity and fatalism, and interest in science and scientific understandings of the world; Hypothesis 3 (Social Psychology): that identifiable structural, contextual features will correlate with identifiable behaviors, irrespective of preexisting beliefs or previous behaviors. Due to the high % of LDS students that attend UVU, we are also interested in looking at the acceptance of Evolution among LDS before learning about the theory of Evolution (Pre-test) and after taking the introductory courses (Post-test). Preliminary results suggest that those students who had a strong sense of God-mediated control also had, in general, a negative attitude towards evolution.

The Baetidae are an important group of Mayflies because of their position on the mayfly tree of life in that they are a key to understanding key evolutionary trends, such as the origin of wings and flight. Phylogenetic relationships of mayflies are still not very well known, however molecular and morphological data have begun to shed light on the relationships of these insects (Ogden et al. 2009). Recent molecular and morphological analyses have examined relationships of some of the lineages of Baetidae distributed in different regions of the world (Gattolliat et al., 2008; Kluge, 1997; Monaghan et al., 2005; Nieto, 2010;). Most of these studies suggest two subfamilies, Baetinae and Cloeoninae. This study represents one of the largest (in terms of data) and most diverse (in terms of taxa) phylogenetic analysis performed on the family Baetidae to date. The dataset consists of more than 50 taxa sequenced for six genes (Nuclear 18S rDNA; Nuclear 28S rDNA; Histone 3, Mitochondrial COI, Mitochondrial 16S rDNA; and Mitochondrial 12S rDNA). DNA sequences were aligned in Muscle (Edgar 2004) and tree reconstruction and nodal support was performed under Maximum Parsimony, Maximum Likelihood, and Baysian frameworks. The results indicate that the subfamilies were not strongly supported as monophyletic, contradicting conclusions from morphological data. Missing data in the data set might be contributing to low support across the tree.

Nanotechnology is revolutionizing imaging techniques, antibiotic therapy and cancer treatments. Nanoparticles (NPs) are also utilized in many commercial products such as sunscreens, paints, ceramics and semiconductors. Consequently, it is inevitable that NPs find their way into the environment. The effects that NPs have on agriculture and soil ecosystems are the focus of this project. Metal-oxide NPs such as ZnO are toxic to many bacterial pathogens but the beneficial root-colonizing isolate, Pseudomonas chlororaphis O6 (PcO6), has high tolerance. This bacterium represents an important group of micro-organisms that colonize plant roots improving their resilience to both abiotic and biotic stress. At sublethal dose, the ZnO NPs remodel the secondary metabolism of PcO6 in ways that could have an impact on agricultural ecosystems. Formation of antibiotic phenazines produced by PcO6 is strongly inhibited by ZnO NPs. Phenazines are important for PcO6 growth in biofilms, induction of tolerance mechanisms in the colonized plant, and antagonism of other rhizosphere pathogens. A role of NPs as a point source for soluble metal release is involved in these processes but does not explain the total effect of the NPs. The sublethal effects of the ZnO NPs on bacteria are similar to changes reported by sublethal doses of traditional antibiotics on human pathogens. These changes have a strong impact on fitness of bacterial pathogens, the most significant of which is induction of antibiotic resistance. Although many metal-containing NPs are being used in antimicrobial formulations, these alternative antibiotics too may be inducers of antibiotic resistance.

Nematodes are microscopic roundworms and are known to be some of the most difficult agricultural pests to control. Most of the 700 different plant parasitic nematodes species infect the root system of their host plant, causing galling, stunting and allow secondary pathogens to infect the host. For diagnostic purposes, in order to identify species of root-knot nematodes, the use of species specific primers can speed up the identification; many farmers need fast and reliable identification to determine which crops can still be planted in root-knot nematode infested fields. The purpose of my project was to test species-specific primers developed in Europe and Asia for Meloidogyne incognita and Meloidogyne arenaria on North American populations of both species using PCR. The results showed that primers developed in Asia and Europe for M. incognita amplified North American populations but primers developed in Europe for M. arenaria did not amplify North American M. arenaria populations. Based on results, M. incognita specific primers are useful for diagnostic purposes in the United States.

Spider silk is a biomaterial with extraordinary physical properties. It has a unique combination of tensile strength, elasticity, and even biocompatibility that has sparked interest in a wide range of disciplines. Potential implementations of spider silk include: medical advances (skin grafts, biomedical sutures, and artificial tendons/ligaments), automotive safety (seat belts, airbags), and military applications (parachutes, body armor). Although spider silk has a great variety of possible uses, collecting this product is not as easy as merely farming spiders and harvesting the silk. Spiders are naturally territorial and cannibalistic, thus alternative means of production are necessary to generate enough spider silk for realistic use. A non-pathogenic laboratory strain of Escherichia coli commonly used for research purposes will be modified to produce synthetic spider silk. Through the use of synthetic biology and molecular cloning techniques, recombinant DNA is inserted with the genetic code for dragline silk of the Argiope aurantia spider; once constructed, this DNA is transformed into E. coli. The goal is to take advantage of E. coli’s ability to be used as a “factory” for creating silk in a controllable and cost efficient system. Supplementation of additional tRNAs will be employed as a strategy to extend cell life and boost the overall spider silk protein yield. Preliminary results have been obtained that show the production of synthetic spider silk by the engineered E. coli. More research is being conducted to increase yields so that we can one day take advantage of this amazing biomaterial.

The presence of mercury in a food chain can have harmful effects, including altering behavior, on organisms. Mercury is typically found in aquatic ecosystems, however recently is also recognized as a potential problem in terrestrial ecosystems. High levels of mercury have been found in the Great Salt Lake in a form able to bioaccumulate up the food chain. If the aquatic ecosystem is linked to the terrestrial ecosystem, then some of the highest predators included in this food chain may be songbirds that eat spiders. We quantified mercury in the blood of the Loggerhead Shrike, Lanius ludoviciaus, on Antelope Island. Upon being caught, the birds were measured, color banded, and a blood and feather sample taken from each of the 15 shrikes to be tested for mercury. Mercury concentrations ranged from 0.96 to 4.00 ppm, with a mean 1.14+/0.31 ppm. Sub-lethal effects in songbirds from another study were seen with concentrations ranging from 2.0-3.5 ppm. Two shrikes we tested exceeded that range. There was high variability in concentrations of mercury, which might be related to distance from shore. These preliminary data demonstrate that some mercury from the aquatic ecosystem is bioaccumulating in the terrestrial ecosystem on Antelope Island, and there may be harmful levels in the birds which could lead to change in behavior, and eventually decline in population. Future studies will investigate the costs of mercury contamination.

Obesity predisposes individuals with Type II Diabetes to cardiovascular complications such as impaired blood vessel function. Due to the elevation of free fatty acids (FFAs) in obese individuals, ceramide, a lipid metabolite, accumulates and might contribute to the inability of a blood vessel to constrict or relax appropriately. Vessel dysfunction is partly caused by the inability of the endothelium, the innermost protective lining of blood vessels, to synthesize and release nitric oxide (NO). Our data indicate that ceramide impairs endothelial NO synthase (eNOS), the enzyme that synthesizes NO. In order to study mechanisms by which ceramide might impair eNOS, it is important to measure cellular ceramide production in response to pharmacological and genetic manipulations. Previously we used P-32 radioactive assays to measure ceramide accumulation. However, the use of radioactivity is expensive, potentially hazardous, and waste disposal is an environmental concern. Therefore, I sought to import a less harmful, more cost effective, yet accurate technique of measuring ceramide production by immunofluorescence (IF). IF allows ceramide to be tagged with a primary antibody which can be detected by a secondary antibody conjugated with a fluorescent dye. I have observed that 250, 500, and 750 uM palmitate (pal) incubation for 3 h increases (p<0.05) endothelial cell ceramide accrual in a dose-dependent manner. Further, a FFA-independent method to alter ceramide accrual i.e., 3 h incubation of cells with N-oleoylethanolamine, also elevates (p<0.05) ceramide production. Importantly, I have shown that 500 uM palmitate-induced ceramide accrual can be prevented (p<0.05) by two structurally dissimilar inhibitors (10 uM myriocin, 1mM L-cycloserine) of the rate-limiting enzyme responsible for ceramide biosynthesis i.e., serine palmitoyl transferase (SPT). None of these inhibitors impairs cell viability. These data indicate that IF is an accurate and reproducible method whereby ceramide accrual can be quantified in endothelial cell systems.

Both human language and birdsong are learned sensorimotor processes that require exposure to adult communication signals in order to be reproduced. The social aspect of the development of these vocal signals is distinctive because interaction is not required for the development of vocal signals in other organisms. Human infants isolated from interaction will not learn to speak normally. The need for interaction is also characteristic of the ontogeny of vocal signals in the zebra finch (Taeniopygia guttata), but only zebra finch males develop vocal signals, in the form of song. Juveniles raised in isolation will develop highly atypical song characterized by prolonged, monotonic syllables and irregular rhythms. When juveniles are tutored by these isolated birds in one-on-one conditions their song drifts towards normally raised birds. It is unclear how tutoring, as a social interaction, influences this observation. We hypothesized that different social experience influences birdsong. We predicted the song of juveniles raised in isolation with an experienced female to be closer to normal song than that of those raised in isolated sibling groups, or those raised in isolation. Current data suggest that the syntax and temporal features of song from juveniles raised in isolation with experienced females is indeed closer to normal song than that of isolated juveniles.

Many zygotes and spores of brown algae are photosensitive and establish a developmental axis in accordance with directional light cues. Ectocarpus siliculosus is being advanced as a genetic and genomic model organism for investigating brown alga development, and this report investigates photopolarization of the growth axis of mitospores. When exposed to unidirectional light, mitospores photopolarized and established a growth axis such that germination was preferentially localized to the shaded hemisphere of the spore body. The roles of the microtubule cytoskeleton and endomembrane cycling in the photopolarization process were investigated using pharmacological agents. Disruption of microtubule dynamics progressively reduced the percentage of mitospores that photopolarized, while inhibition of vesicle secretion blocked photopolarization nearly completely. Chronic treatment with these pharmacological agents severely affected algal morphogenesis. Microtubules in mitospores and algal filaments were imaged by confocal microscopy. Mitospores contained a radial microtubule array emanating from a centrosome associated with the nuclear envelope. At germination, the radial array gradually transitioned into a longitudinal array with microtubules extending into the emerging apex. At mitosis, spindles were aligned with the growth axis of cylindrical cells in the filament, and the division plane bisected the spindle axis. These studies demonstrate that dynamic membrane cycling and microtubule assembly play fundamental roles in photopolarization, and provide a foundation for future genetic and genomic investigations of this important developmental process.

Speckled dace, Rhinichthys osculus, is a small minnow that occupies river systems in Utah, Arizona, and Nevada. These minnows prefer clean, faster-moving water with plenty of cover such as rocks or undercuts. The invasive crayfish Orconectes virilis inhabits areas where the Virgin spinedace is found and has a similar diet being an opportunistic feeder. I tested the hypothesis that the natural behavior of Virgin spinedace will be altered due to the presence of crayfish. I predicted that the spinedace would avoid the crayfish and stay out in the open exposing them to a higher risk of predation, since crayfish are a more aggressive organism and will try and eat virtually anything they come into contact with. The experimental trials were conducted (N=24) in Quail Creek, Utah. Large plastic bins with the front and back replaced with screen were used to allow water flow through our experimental chambers. Each bin was placed in the creek and had rocks, and an undercut bank to resemble the native habitat. Controls consisted of placing isolated Virgin spinedace in one of the bins to observe its behavior. In both controls and experimental trials, either 2 spinedace, or 5+ spinedace were added. In experimental trials, I introduced 2 crayfish into the experimental chambers with the Virgin spinedace and allowed them to acclimate for 10 minutes. After the 10 minute acclimation period, behavioral observations were made. Results indicated that when 2 spinedace were present along with the crayfish, they would stay out in the open and the crayfish would occupy the cover or undercut. When 5+ spinedace were present with crayfish, the spinedace displayed a mobbing or “strength in numbers” behavior where they would occupy the undercut and either clog it up with their presence so the crayfish would leave, or mob the crayfish to make them leave. This behavior was opposite of what I predicted would happen. Since this mobbing or “strength in numbers” behavior was not observed with the samples of just 2 spinedace, this may be a behavioral adaptation resulting from the presence crayfish. Further research can be done on the behavioral effects of various fish species in the presence of introduced or invasive species to determine if this behavior is seen in other fishes.

Pradimicin, a small molecule produced by the soil bacterium Actinomadura hibisica, is a promising candidate as a combined antifungal/antiviral therapeutic. It is active against a broad-spectrum of opportunistic, pathogenic fungi, interferes with the replication of influenza virus, and inhibits the reproduction of HIV-1. Toxicity and solubility problems have hindered past efforts to develop pradimicin as a therapeutic. Our research focuses on elucidating the bio-synthetic pathway of pradimicin in order to design and chemoenzymatically create pradimicin structural analogs with improved solubility and activity, and less toxicity. Several enzymes in pradimicin biosynthesis have been identified. We intend to characterize one of the key enzymes, PdmS, a putative glycosyltransferase, and to manipulate its gene to create novel, more efficacious pradimicin analogs. This project is funded for three years by the NIH NIAID (3 years). Methods: A gene knockout experiment was used to determine the role of PdmS in pradimicin production. Bio-synthetic precursors of pradimicin were subjected to bio-transformations in E. coli with recombinant genes for PdmS and another glycosyltransferase, OleD, to generate analogs with new sugar attachments. Analogs of pradimicin will be screened for bio-activity using standard microbroth dilution assay techniques. Confirmed results: The enzyme PdmS was identified and characterized as a glycosyltransferase. Expected results: newly created analogs of pradimicin exhibited minimal inhibitory concentrations of 10μg/mL against Candida albicans. Conclusion: Knockout of pdmS yielded the pradimicin aglycon, which confirmed the function of this glycosyltransferase and provides a start molecule for further structural modification to yield new analogs for bioactivity studies.

There are numerous arresting differences between the hands of humans and those of chimps, principally among them the shortening of metacarpals 2-5, the relative increase in length of the thumb, and the development of separate musculature controlling flexion of the thumb. In addition to improving manual dexterity, these changes allow humans, in contrast to chimps and other apes, to make a buttressed fist with the phalanges pressed against the central palm and the thumb adducted onto the dorsal surface of the phalanges. We believe this hand posture greatly reduces the strain experienced by a fully formed fist when striking with force, thereby rendering the human hand as a more effective weapon. We tested this idea by manipulating the tendons in a cadaver arm to induce the hand to form into buttressed and non-buttressed conformations, and then tested each conformation by having the hand strike a hanging instrumented mass to measure the force of each strike. Additionally, the 2nd metacarpal of the hand was fitted with a strain gauge so that strain in the bone could be compared to the force of the strikes. The results of these tests showed that there was a significant drop in strain in the buttressed fist (in comparison to the non-buttressed) for a given force.

Impaired endothelium-dependent vasodilation has been associated with various cardiovascular diseases, including heart failure, is linked to oxidative stress, and persists despite heart transplantation (HTx). PURPOSE: This study aimed to determine if changes in vascular function following HTx can be assessed using incremental handgrip (HG) exercises to induce nitric oxide-dependent vasodilation of the brachial artery. Furthermore, the efficacy of an acute oral antioxidant cocktail (AOC) to reduce oxidative stress and improve vascular function in this population will be assessed. METHODS: 31 HTx recipients (14 years post-HTx) and 10 healthy age-matched controls were given either and AOC (Vitamin C, E, and alpha-lipoic acid) or placebo (PL; randomized for the subjects’ two visits. Measurements of the brachial arterial blood velocity and vessel diameter were performed during three absolute workloads (4, 8, and 12 kg) of HG exercise using ultrasound Doppler. RESULTS: Maximal vasodilation during PL HG exercise was not different between the controls (8.2 ± 1.5%), and recent (< 3 years post) HTx group (8.5 ± 1.2%), but the 5-10 years post-HTx groups had a tendency to be lower (6.5 ± 1.9%). The > 14 years post-HTx group recipients (5.2 ± 1.9%) were significantly attenuated compared to both the controls and the recent (< 3 years post) HTx recipients. CONCLUSION: The study supports the use of HG exercise as an assessment of vascular function in a patient population with known cardiovascular risk. Additionally, these results suggest that vascular function is similar between controls and early HTx recipients but declines as time passes following surgery despite normalized cardiac function.

Kentucky bluegrass (Poa pratensis L.) is a commonly used turfgrass species with many varieties being sold around the world. However, those varieties are very difficult to tell apart morphologically. Our objective was to use genetic markers (primers) to identify varieties, even if they are visually similar. This is valuable to the turfgrass industry for plant variety protection. We also wanted to use this data to explore the apomictic tendency (clonal reproduction) of the varieties. We collected leaf tissue from 24 Kentucky bluegrass varieties, extracted DNA, and sequenced portions using 29 EST and 21 genomic primers. This data was used to determine genetic relationships using a neighbor-joining dendrogram. Similarities of the genetic sequences from the varieties were estimated using the DICE coefficient. We found more polymorphisms in genomic primers than in EST primers with high variability between the varieties. Both types of primers were robust enough to distinguish varieties and that each variety was unique and genetically distinguishable. In addition, we discovered some varieties had large amounts of variation within a variety. This was unexpected due to the usual apomictic nature of the species. The markers resulting from our research will be available to the turfgrass industry.

All cells, including retinal neurons and glia, must sense and adapt to physical changes in their local environment (e.g. changes in osmotic pressure). Osmotic water flux can cause aberrant cell volume changes, which can contribute to tissue damage, edema, and neuronal hyperexcitability and excitotoxicity. We hypothesized that force-sensitive proteins enable retinal cells to monitor their physical form (e.g. volume) and help maintain homeostasis by regulating cell volume. To test this, we first investigated the properties of cell morphology when cells were bathed in solutions with different tonicities. Under these conditions, we measured changes in cell volume due to osmosis in retinal ganglion cells (RGCs) and Müller glia. We found RGCs were unable to actively adjust their volume, whereas Müller glia reduced their swollen volume in the continued presence of hypotonicity. The regulation of cellular volume often involves calcium signals. We tested whether calcium plays a role in the regulation of retinal cell volume. Free calcium within the cells was sequestered using cytosolic BAPTA, which decreased the extent of hypotonic swelling. This demonstrates that calcium elevations increase the extent of cell swelling. Because cell volume was dependent on calcium, which was elevated by membrane stretch, we hypothesized that the osmosensitive cation channel TRPV4 would transduce osmotic pressure and contribute to cell volume regulation. In an experiment using a selective TRPV4 inhibitor, the extent of hypotonic-induced swelling was reduced. Thus, the opening of TRPV4 leads to a calcium influx that exacerbated cellular swelling. In addition, we tested the idea that TRPV1 cation channel is involved in responses to osmotic stimuli. In an experiment using a selective TRPV1 antagonist, preliminary results show that the extent of hypotonic-induced swelling decreased. This signals that TRPV1 may have a role in volume regulation in retinal neurons and glia. Thus, inhibition of these force-sensitive protein channels might alleviate the deleterious effects of volume changes in pathological contexts. Our findings therefore have implications for our understanding of retinal mechanotransduction and osmoregulation as well as provide a mechanistic framework for developing new therapeutic strategies aimed at blinding conditions that involve mechanical stress and cellular morphology.

Cardiovascular disease (CVD) is the leading cause of death in the United States. Gamma-tocopherol (GTC) has been studied in its relation to hypertension and CVD. GTC has protective effects on the endothelium of arteries; the endothelium is in part responsible for regulating arterial relaxation and reducing blood pressure (BP). Diabetic mice (DB) were chosen for this study because of the associated co-morbidities of hypertension and excess body weight. Eight-week-old mice were divided into 3 separate groups: wild-type fed control (CON) diets (n=12), DB fed CON diets (n=12), and DB fed GTC supplemented diets (0.1%, n=12). BP was measured every week for 8 wk. Body weights were measured every wk and morphometric values were measured at the end of the 8-wk period. There was no statistically significant (P<0.05) difference in systolic and diastolic BP between DB-GTC and DB-CON. There was also no statistically significant (P<0.05) difference in heart rate between DB-GTC and DB-CON. After 8 wk, DB-GTC mice had lower (P<0.05) body weight versus the DB-CON group. In conclusion, a GTC supplemented diet did not result in a change in BP. However, GTC may have blunted weight gain and adipose tissue mass in DB-GTC compared to DB-CON. We speculate this may be due to a reduction of food consumption during the experimental period.

The nervous system is responsible for cognition, memory, and motor function. Neurons communicate with each other at intercellular connections called synapses. It is at these locations that vesicles fuse with the plasma membrane and release their contents into the space between the neurons (synaptic cleft). SNARE proteins (synaptobrevin, syntaxin and SNAP-25) facilitate synaptic vesicle fusion by winding together in a four-helix bundle forcing the mixture of opposing membranes. After exocytosis, the cell regenerates vesicles by a process called endocytosis. This process involves the budding of membrane back into the cytoplasm re-forming a functional synaptic vesicle. This progression from vesicle fusion to internalization is termed the “synaptic vesicle cycle” and is coordinated by a long list of molecular players. The SNARE proteins are classically considered to be unique to exocytosis. However, preliminary results from Erik Jorgensen’s lab have implicated SNAP-25 in endocytosis. Although we have evidence that SNAP-25 is required for endocytosis, the molecular mechanism is completely unknown. In this proposal we use forward genetics in the model organism C. elegans to identify novel protein interactions required for SNAP-25 mediated endocytosis. We have designed three suppressor screens that are predicted to target the role of SNAP-25 in exocytosis, endocytosis and general function. The first two screens use crippled forms of SNAP-25 (hypomorphs) while the last screen uses a deletion allele of the SNAP-25 gene (null mutation). With these screens we hope to precisely identify the molecular players involved in SNAP-25 mediated endocytosis versus exocytosis.

Glossiphoniids are jawless leeches that feed using a proboscis, which they use to impale prey so that they can feed on their prey’s blood and tissues. This study focuses on two different species, Glossiphonia complanata and Helobdella stagnalis. Helobdella stagnalis feed on a variety of prey, including bloodworms and freshwater snails, often feeding together in large groups. When a prey item is placed in the container, H. stagnalis immediately begin smelling the water and moving toward the prey. After the first H. stagnalis makes contact and secures prey, other individuals within the group will move to that area, often passing prey along the way, to attach to the same prey item. Once a number of individuals have started feeding, the prey is often broken into smaller pieces and individuals can then begin feeding on various sized pieces. This is in contrast to members of the species Glossiphonia complanata, who appear to be selective with their prey, and have been observed going up to 4 weeks without feeding, even in the presence of their non-preferred prey (bloodworms). When preferred prey (freshwater snails) were placed in the container, unfed G. complanata did not immediately begin smelling the water or initiate hunting. Unlike H. stagnalis which feed in groups, G. complanata appear to prefer feeding alone, and have even exhibited competitive behaviors.

The tumor suppressor p53 is mutated in more than 50% of all cancers, while the majority of remaining cancers contain mislocalized p53(1). p53 is part of a network of cellular pathways that regulate growth, cell cycle arrest, and clearly delineated apoptotic pathways(2). Restoring the function of p53 can be seen as an ultimate cancer treatment. Restoring p53 would activate an already existing cell pathway that prevents cells from becoming cancerous and cause cancer cells to go through apoptosis. Playing a role in tumor suppression has made p53 an attractive target for gene therapy. However, despite the potential of p53 as a powerful treatment, it is limited by the dominant-negative effect of dysfunctional mutant p53. This effect imposes one of the greatest limits on the effectiveness of a p53 based treatment. This project is focused on bypassing the dominant-negative effect of dysfunctional p53 over exogenous functional p53. The attempted solution was substitution of the p53 binding domain with a different but structurally analogous coiled-coil, based on a modified Breakpoint cluster region (Bcr) protein. By doing this, the dominant-negative effect of mutant p53 may be bypassed. The purpose of this project has been to synthesis and test rationally modified forms of p53 with modified Bcr coil that are introduced into cells via a plasmid to restore cellular p53 activity. In vivo cell tests have already shown the effectiveness of these constructs at causing higher rates of cell death in cancer cells and constructs are currently being refined to carry forward to xenograft model animal trails. The ultimate goal is to develop a treatment for human cancer patients where modified p53 will selectively cause apoptosis in cancer cells.

Fire induced soil water repellency has been characterized across a variety of soils and landscapes as being a cause of watershed degradation and surface water pollution. The repellency is due to the condensation of volatile polyaromatic hydrocarbons onto soil particles. Although repellency is known to reverse, in some locations the reversal takes months; whereas, in other locations it may take years. Little is known about the reversal mechanisms and how they lead to such a large range of reversal times. Access to untreated fired-affected sites, can be unpredictable and samples vary greatly from site to site. Therefore, a model compound that mimics fire-induced soil hydrophobicity is needed to be able to systematically investigate soil hydrophobilicty reversal mechanisms. Fluorene is a relatively non-toxic, hydrophobic polyaromatic hydrocarbon. The behavior of fluorene coated sand grains was investigated under laboratory conditions using quartz sand. Moreover, its fluorescent properties could be used to non-destructively monitor its degradation over time. In this context fluorene was studied as a possible model compound for the study of mechanisms involved in the reversal of fire-induced soil hydrophobilicy. The compound was subjected to conditions known to degrade or reverse water repellency including temperature, UV-light, and moisture content. Changes in hydrophobicity were monitored using the ethanol drop test and fluorescent imaging. Digital image processing techniques with the public domain software, ImageJ, produced by the National Institute of Health, were used to analyze the images and generate spatial maps of treatment effects on fluorene degradation and hyrdophobicity reversal. Our results indicate that the hydrophobic fluorene-coated sand layers were degraded by treatments such UV light known to reverse hydrophobicity in fire-affected soils, thus suggesting that fluorene may serve as a suitable model compound for producing hydrophobic layers on course grained material.

In C. elegans, sperm activation is the process through which sperm become motile. Genes that regulate sperm motility are being identified through a genetic screen. In this screen, swm-1 mutants that have sperm that activate too early are mutagenized, and genes regulating sperm motility can be found when the sperm of mutant worms no longer activated too early. Two genes have already been identified, try-5 and snf-10, and these can help identify new mutations. By selecting specific strains with sperm activation defects, the mutations causing the defect can be characterized. To sort through the many mutants, complementation tests to identify them as a new gene, or a new allele of try-5, snf-10 are being done. Selectively crossing an unknown mutant strain to a strain with a known mutation in try-5 or snf-10 will help identify the unknown mutation. If the progeny of the cross have activated sperm then the two strains complement each other. In this case the alleles must be in different genes and the unknown mutation from that cross has not been identified. If the progeny instead has non-activated sperm then the mutations failed to complement and the unknown mutation is allelic to the mutation it was crossed with. One new allele of try-5 and of snf-10 has been found. It is suspected that the other strains contain alleles of new genes not previously identified. Further complementation tests will be done to test this theory.

Agricultural herbivores, such as cattle, often encounter plants containing toxins. One class of toxins, tannins, bind to proteins and inhibit digestive enzymes from acting to liberate nutrients. In this way, tannins prevent optimal absorption of nutrients. To overcome this challenge, some herbivores host bacteria with the ability to degrade tannins. Currently, there is a push to find novel microbes capable of aiding animals in detoxifying these compounds. Therefore, we aimed to isolate tannin-degrading bacteria from the desert woodrat (Neotoma lepida), a wild, herbivorous rodent that feeds largely on a tannin-rich shrub, creosote bush (Larrea tridentata). Woodrat feces were cultured on tannin-treated media, and bacteria capable of degrading tannins were further characterized with DNA sequencing. Results revealed that from 9 isolates, three species of tannin-degrading bacteria were present: Enterococcus faecalis, Bacillus subtilis, and Escherichia coli. Further characterization was performed through measurement of tannase activity. The activity of each isolate varied significantly between bacterial species as well as within a species. Ultimately, I hypothesize that these tannin-degrading bacteria facilitate the ingestion of tannin-rich plants by woodrats. Additionally, this research exemplifies how wild herbivores, such as the desert woodrat, provide an essential source of tannin-degrading microbes that could be introduced into domestic herbivores in order to improve agricultural practices.

Chronic methamphetamine (METH) abuse leads to structural and functional damage in the brain, which likely contributes to cognitive and behavioral dysfunction. Recent data suggest an association between METH abuse and impaired inhibitory control over behavior; that is an impaired ability to inhibit inappropriate actions or thoughts. However, the extent to which METH-induced neurotoxicity is responsible for such impairment remains to be determined. Previously, we reported that rats with METH-induced partial dopamine (DA) and serotonin (5-HT) loss in striatum and prefrontal cortex (PFC) exhibited impaired response inhibition in the form of perseverative responding. Furthermore, levels of serotonin transporter (SERT) binding in PFC correlated with perseverative responding. Herein, we have examined another dimension of response inhibition impulsive action, which is an inability to withhold a “pre-potent” response in rats with METH-induced neurotoxicity. Rats were trained to perform a stop-signal task (SST). Once rats achieved stable responding (>80% correct response) on both “Go” and “Stop” trials, they were treated with saline (0.9%), a neurotoxic regimen of METH (4 x 10 mg/kg, 2-hr intervals, s.c.) under normal ambient temperature conditions (“neurotoxic” METH group) or the METH regimen under cooling conditions (“normothermic” METH group). One week after the treatment, rats were again tested on the SST. After behavioral tests were done, animals were sacrificed and brains removed for determination of monoamine loss. The results showed: 1) the “neurotoxic” METH-, but not the “normothermic” METH or saline-treated rats, showed a 40-60% loss of SERT and dopamine transporter (DAT) binding in PFC and striatum; 2) the “neurotoxic” METH group showed normal behavioral performance in “Go” trials of the SST relative to the “normothermic” METH group or saline controls; 3) the “neurotoxic” METH group exhibited impaired withholding of a “pre-potent” response, as reflected by increased numbers of errors on the “Stop” trials of the SST. Overall, these data suggest that impaired inhibitory control over behavior (i.e., increased impulsive action) can arise as a consequence of METH-induced neurotoxicity to central dopamine and serotonin systems. Supported by NIH grant DA 024036

Photosynthesis is driven primarily by radiation between 400 and 700 nm; however, not all wavelengths are equally efficient. Red light (600 to 700 nm) is 25 to 35% more efficient than blue light (400 to 500 nm) and 5 to 30% more efficient than green light (500 to 600 nm). Although blue light is less efficient than red light, it has been shown to be necessary for normal plant development in all tested crop species. The mixture of pigments in plant leaves allows them to absorb all colors of light. Both red and blue light are absorbed primarily in the upper leaf. Green light penetrates deep into the lower leaf and transmits to leaf layers below the upper leaf canopy. Accordingly, once the upper canopy is saturated with red and blue light, supplemental green light is beneficial in increasing whole plant photosynthesis. Although the effect of spectral quality on single leaves is well characterized, the effect on whole plant growth and development is poorly understood. The narrow spectral output of LEDs makes them particularly effective for photobiological studies. From our initial studies with radish, soybean, and wheat, we determined that blue light levels can be used to better predict plant development than red:far-red light ratios. We also found that plants require at least 80 μmol of blue photons m-2 s-1 in order to develop normally. Further studies are currently being conducted to determine the optimal ratio of red and green light for maximizing whole plant photosynthesis in lettuce and radish.

Differences in the genomes of organisms control an organism’s ability to deal with and adapt to environmental stresses. In this project, two strain isolates of the nematode Caenorhabditis elegans were analyzed using high-throughput assays measuring growth and offspring production to determine the genes that confer a greater resistance to the herbicide paraquat. Paraquat acts by interfering with electron transport mechanisms within the cells of living organisms. This mechanism not only allows it to act as an effective herbicide, but also causes it to pose a considerable risk to the health of animals, including livestock and humans. Using statistical genetics, regions of the genome were identified that are likely responsible for differences in growth rate and fecundity observed in the two strain isolates when grown in paraquat. Near-isogenic lines and extra-chromosomal arrays were then created to isolate these portions of the genome in a control genetic background. This approach allowed us to be able to attribute any differences in the two traits to the genes contained within the intervals. After analyzing the body size data, representing the growth of the animals over 72 hours, we were able to draw a preliminary conclusion that an interval on chromosome V may have a small but significant effect on growth determination. As a result of this project, a specific interval was identified that may be responsible for a greater growth rate, three near-isogenic lines were created, and 34 extra-chromosomal arrays were generated. This work will be used in the future to identify the gene(s) responsible for the greater growth rate and fecundity observed in some animals exposed to the herbicide paraquat. These results will allow us to draw conclusions about the roles that these genes, and others like them, play in an organism’s ability to cope with environmental stresses.

Parasites are arguably the most diverse group of organisms on the planet. One key factor in determining patterns of parasite biodiversity is the number of host species a parasite can infect, or its host specificity. However, surprisingly very little is known about the evolution of host specificity. Here I use domesticated pigeons (Columba livia) and their ectoparasitic feather lice (Columbicola columbae) to demonstrate how parasite body size influences host use. The current study has two main goals: 1) to use different sized hosts to experimentally evolve louse body size, and 2) to determine how parasite body size effects host specificity. Four breeds of domesticated pigeons were infested with parasitic feather lice. After roughly six generations, mean louse body size changed with respect to host body size, with the largest pigeons having the largest lice. Interestingly, the smallest pigeons had larger lice as well. However, this increase in louse size appears to be the result of a change in microhabitat use, with lice shifting from predominantly the wings and tail to the head. This study is one of the first to experimentally demonstrate how changes in parasite morphology and behavior influence the evolution of host specificity.

There is growing concern regarding the contribution of infilled turf fields on increased athlete infections. Abrasions that occur on these fields create a portal of entry for pathogens such as Staphylococcus aureus. This study compares microbial populations on two infilled turf fields (year old vs. 6 year old turf). Infill material from both fields was sampled at three sites for 5 months during the football season (sidelines, middle of the field, and end of the field). Tryptic Soy Agar was used to determine total microbial load, MSA for S. aureus, and EMB Agar for coliforms such as Escherichia coli. Much higher microbial populations were found on the older turf field, a 1E+04 increase over similar locations on the new turf; suggesting microbial populations can accumulate in synthetic turf infill over time. On the older field the sideline has the highest counts with an average of 1.60E+08 CFUs/g of rubber infill. On the new synthetic turf, the area with the highest number of microorganisms was the end of the field rather than the sideline. This is probably due to where practices are held on the new turf field rather than actual game play. A high number of salt-tolerant, mannitol-fermenting bacteria (indicators of S. aureus) were also found (3.25E+02 CFUs/g on the new turf vs. 2.73E+03 CFUs/g on the old turf). Escherichia coli was isolated from the new turf, in an area of high usage. These results indicate that infill material can serve as a source for pathogens among athletes and that organisms accumulate over time posing a greater risk if proper cleaning is not routinely performed.

Hydrocarbon metabolizing microbes play a critical role in bioremediation and bio-augmentation projects. In 1978, Brock et al. identified microbes in the Great Salt Lake (GSL) capable of metabolizing hydrocarbons, but since then these microbes have gone unstudied. This research has isolated and identified bacterial species capable of hydrocarbon metabolism from GSL. Though capable of survival in the high salt concentration of the Great Salt North Arm, we have demonstrated that these species are also capable of thriving in low salt concentrations. This research will continue with the characterization process to identify what hydrocarbon sources each species is capable of degrading.

Nuclear Pore Complexes (NPC) create aqueous channels embedded in the nuclear envelope and are made from a network of proteins called nucleoporins (nups). The nucleoporin Nup153 has previously been found to be required for proper assembly of the nuclear lamina. In this study, a fragment of Nup153 was overexpressed in order to impede Nup153 function in T-Rex HeLa cells. After inducing the expression of the dominant negative fragment, I examined the localization of lamins A, B1 and B2, as well as SUN1, Emerin, and BiP, three proteins that mark cell membrane compartments. Lamins B1, B2, SUN1, and Emerin were found to mislocalize to the cytoplasm of the cells, and colocalization among the proteins was observed. Lamin A also had an abnormal phenotype unlike that seen with the B lamins, but indicative of a problem with integration of Lamin A into the nuclear lamina. Colocalization of the various lamin isoforms with membrane proteins such as SUN1 and Emerin indicates a problem with membrane assembly. The distinct localization of BiP, however, suggests that there is a “nuclear-like” membrane in the cytoplasm that either does not incorporate normally into the nuclear envelope as it forms or is newly-recruited to mislocalize the lamin proteins.

Arnica is a common form of complementary medicine used to treat bruises and sore muscles. Most arnica preparations are made as an oil or gel by soaking the flowers of Arnica montana (a member of the Asteraceae) in an oil, such as olive oil, to extract helenalin, the presumed active component. Helenalin, a sesquiterpene lactone, has been shown to inhibit transcription factor NF-?B, a factor which controls the expression of dozens of genes involved in inflammation responses (Rungeler et al. 1999). Due to arnica’s popularity, A. montana is becoming scarce in Europe (its native range) and is sometimes substituted commercially with A. chamissonis (Cassells et al. 1999). Therefore, this study focused on finding other sources of helenalin among the Asteraceae. A. chamissonis, A. cordifolia, A. latifolia, A. longifolia, A. mollis and Helianthella uniflora were collected from Alta, UT, and Helenium autumnale from Salt Lake City, UT, at elevations of approximately 10,500 feet and 4,200 feet, respectively. Sesquiterpenes were extracted from the flowers by dipping them in methylene chloride and separated by thin-layer chromatography. Helenalin and possible helenalin derivatives were identified by their reaction with vanillin and comparison to a pure helenalin standard. Flowers from all seven of the species contained helenalin, with Helenium autumnale containing the highest concentration and A. latifolia the lowest. Therefore, all of the species tested in this study have the potential to be used commercially to prepare arnica salves and thus allow the A. montana population to re-establish itself. An additional finding was that A. chamissonis grew the best under greenhouse conditions, which may explain its substitution in some commercial salves.

Little is known about the details regarding gene expression which accounts for the light harvesting pigments in bacteria; specifically in the cyanobacteria Fremyella diplosiphon. The purpose of our experiment is to identify the upstream sequence that controls the expression of the scytonemin light harvesting pigment in Fremyella diplosiphon. This experiment will help us better understand the regulation and expression of genes that control light absorption. To test our hypothesis, that there is a regulating upstream promoter sequence for light sensitivity in Fremyella diplosiphon, we will culture the bacteria and extract its DNA. PCR will be performed to isolate the upstream sequence of the gene from Fremyella. Each plasmid was designed by adding restriction sites that will allow the proper cloning of the PCR fragment. This sequence will be introduced into the pSUN 199 and pSUN202 plasmids. These plasmids contain the GFP gene that will be activated by the promoter. The plasmids will be transformed onto Fremyella and analysis of gene expression will be done under different light conditions. This experiment will be able to identify the upstream regulatory sequence of the Scytonemin gene in Fremyella.

Batrachochytrium dendrobatidis belongs to the fungal phylum Chytridiomycota (chytrids), and in 1999, was both identified as a new species and associated with causing chytridiomycosis in amphibians. Since its identification, this fungus has been implicated in rapid population declines in multiple amphibian species in North and Central America, Europe, and Australia. In adult amphibians, the fungus colonizes only the keratinized skin cells on the ventral and dorsal surfaces as well as on the webbing between the toes. Infection is associated with hyperkeratosis resulting in disruption of the skin’s osmoregulatory function, dehydration, electrolyte imbalance and death. There is data that indicates some bacteria that live on the epidermis of amphibians produce chemicals that inhibit the growth of B. dendrobatidis. However, this data is associated only with salamanders. There is no similar inhibitor data published for frog and toad species, and no data published on the identification of normal skin biota of frogs and toads. We have isolated over 175 bacterial isolates from 67 toads and frogs, extracted DNA, and used PCR to amplify the 16S ribosomal DNA gene. Amplicons have been sequenced to determine speciation based on genetics. This is the first report of normal bacterial skin biota in frogs. Further, a skin swab was also taken from these frogs and used to analyze which are infected with B. dendrobatidis via quantitative PCR screening. While the prevalence of fungal infection appears to be low, we have been able to correlate fungal-positive hosts with number and/or species of normal bacterial skin biota.

Introduction:

The invertebrate ribbon worm, Paranemertes peregrina, serves as a unique model in that it thrives in an environment exposed to drastic fluctuations in salinity during tidal interchange. Its ability to sustain homeostatic integrity is not well understood. With an aim to clarify this phenomenon, total RNA was isolated, and reverse transcription with polymerase chain reaction allowed us to serendipitously clone and elucidate a 488 base pair region of a gene coding for a Tachykinin Receptor (TKR), a subtype of G-protein coupled receptor (GPCR). This region shows 79% homology to the mouse TKR-2 mRNA sequence, and 44% homology to human Neuromedin-K receptor’s amino acid sequence. Specifically, we have isolated a portion containing a cytosolic carboxy-terminus that has classically been associated with palmitoylation or otherwise hydrophibicity-enhancing interactions. This process facilitates the docking of cytosolic subunits to the membrane in the assembly of GPCRs, acting as a regulatory component. In vertebrate models, much of these proteins are conserved. Given that only a few of these genes have been reported for invertebrates, this suggests a critical need for investigation of the evolution of TKRs as they relate to stress response. Additionally, this receptor poses questions about its potential role in pain, with the prospect of revealing insight about the long-elusive pain perception in invertebrates.

Emigration Creek, part of the Salt Lake City watershed, runs through Westminster campus at two points: by Garfield school and through main campus. We assessed several measures of water quality, to understand the impacts of anthropogenic activities and urbanization on this stream ecosystem. Additionally, we identified areas for restoration with the ultimate goal of reintroducing the endangered native Bonneville Cutthroat trout. Beginning in June 2012, we monitored the creek monthly, and will continue for a full year’s time. We assessed the structure and stability of the stream along with dissolved oxygen, pH, temperature, and turbidity. The macroinvertebrate community composition was analyzed also as a biological indicator of stream health. Along both reaches lack of vegetative cover, lack of large rocks to support the lower banks, and scouring and deposition along the steam bottom were common. We found that for all water quality measures monthly average levels were in acceptable limits for trout populations except for dissolved oxygen. Dissolved oxygen dropped below optimal levels in July and August, most likely due to an increase in water temperature. We found that both reaches have moderate species diversity of macroinvertebrates, though the community is dominated by species that thrive in water highly polluted with organic waste. Preliminary results suggest that the current condition of Emigration Creek is not optimal for cutthroat trout, largely due to seasonally low dissolved oxygen levels and lack of preferred food. Continued monitoring, along with a comparative study of nearby creeks, will further our understanding of the impacts of urbanization and the restoration efforts needed to support native fish populations in the Salt Lake Valley.

One of the major challenges facing U.S. military caregivers is the presence of multidrug resistant organisms in extremity wounds. The most frequently identified drug resistant strains of bacteria found in these wounds are Acinetobacter baumanni, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. Due to these organisms rapid increase in resistance to the commonly used drugs, it is crucial to discover and establish alternative methods for treating these microbial infections. Antibiotics are currently the most common treatment for infections by these pathogens, and there is little data on the evaluation of phytochemicals as potential chemotherapeutic agents that could take their place. We have screened 24 individual compounds from 9 major compound families to determine if plant-based phytochemicals could be explored further for use in treating bacterial infections in patients with military wounds. All compounds were tested to determine the minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC). Of the 24 compounds tested, 21 (87.5%) inhibited at least one strain used in this study, with only 1 of the 24 (4.2%) inhibiting all strains. There were 10 (41.7%) of the compounds that displayed MIC values less than 100 _g/ml. For compounds displaying MLCs, they ranged from 2.5 mg/ml to 78.13 g/ml. While there is much more research that needs to be done with each of these compounds, this work is a crucial first step in the drug discovery process. We believe that several of these may serve as potential novel inhibitors of these drug-resistant bacteria.

According to the National Substance Abuse Index (2006), Utah women account for 31.7 % of the entire drug abusing population. Mind-altering substance abuse among women creates many unique problems in their lives according to Wesa and Culliton (2004). Additional research by Wesa and Cullliton shows this can affect their futures, finances, families, health, and relationships. Many of the women have other underlying problems that lead them to use mind altering substances to cope with these problems as researched by James (2011). In the Ogden, Utah area, mind-altering substances have created an epidemic that needs to be helped. Serenity, A Drug Abuse Recovery Center, will be an 18,000 square foot building located in Ogden, Utah. Serenity helps facilitate mental and physical healing to create an overall wellbeing for the women. Research has said that women are more likely to receive help for mental conditions than the substance according to Greenfield (2006). Mental disorders and substance abuse will be addressed at the recovery center. Individual and group therapy methods will be used because of their effectiveness shown by Greenfield. Research by Greenfield has shown that women in treatment centers have a greater chance of retention if dependent children are present. Serenity will provide a private room for a woman and two dependent children to stay at the center. The center will include nutrition education and dining area, and a children’s area. The children’s area will have reading, playing, learning, and therapy treating areas.

Re-feeding syndrome is the result of the rapid re-introduction of nutrients following a period of severe malnourishment or starvation. During starvation, critical nutrient absorption and anabolic pathways are shifted to conserve energy and maintain cellular function in intestinal enterocytes. Rapid reintroduction of nutrients can often overwhelm the remnant absorptive ability of the intestinal tract, leading to complications such as, failure to thrive and intestinal failure. We hypothesized that a polymeric diet would improve refeeding response compared to an elemental diet. We utilized the nematode Caenorhabditis elegans (C.elegans) as a minimalist model of refeeding. We induced a developmental state of starvation known as, dauer, and then transferred animals into either a polymeric diet or elemental diet. We measured body length daily for 5 days until the worms reached adulthood. Data were statistically analyzed using a general linear model of analysis of variance and expressed as mean+/-SEM. Our results indicate that polymeric diet (695.6+/-25.2 ?m) demonstrated at least 10% (p<0.05) improved growth compared to elemental diet (646.8+/-21.7), with polymeric worms reaching adulthood by day 2 (960.2+/-26.9), whereas elemental animals required 3 days to reach adulthood (894.2+/-26.9 day 2, 972.6+/-27 day 3). Additionally, we assayed intestinal proteolysis by feeding the worms green fluorescent protein (GFP) and measuring fluorescence along the length of the intestinal tract. Our findings demonstrate a decreased rate (p<0.05) of intestinal proteolysis among worms fed exclusively a polymeric diet, despite no significant change in the rate of fluorescent protein intake. Our data confirm our hypothesis that a polymeric diet improves growth and development over an elemental diet. In addition, by requiring some hydrolytic digestion, growth is improved. These results might help to improve clinical management of refeeding syndrome as well as rehabilitate patients suffering intestinal failure.

Forensic researchers as well as law enforcement officials rely heavily on herbarium type specimens for field investigation. An electronic catalog could greatly assist forensic professionals by providing easy access to area specific specimens. The goal of this project was to compile a database and dichotomous key of native plants found in Weber County, Utah. Pollen and woody material resist degradation, making them perfect candidates for forensic analysis. Using the USDA plant database, all native flowering plants were identified (729 in total). Plant pressings, microscope slides, digital micrographs and line drawings were made of two-hundred native species to date. Pollen and woody tissue were gathered from representative plants and prepared for permanent herbarium collection. Woody tissue and pollen preparation had previously been optimized for best imaging results. Rudimentary dichotomous keys for both woody tissue and pollen were made using micrographs and line drawings to illustrate distinguishing features between plant species. As new species are prepared, they are included in the dichotomous key. The primary purpose of this collection is for it to be used by forensic investigators with limited botanical training. In the spring of 2013, field tests will be conducted to determine how effective the dichotomous key is when identifying unknown specimens. After the layout is finalized, the key will be presented to local law officials as an additional tool in the forensic repertoire.