Life Sciences
Improved Synthetic Bioengineering Production of Polysaccharides and size-specific Oligosaccharides of Low Molecular Weight Heparosan, a Heparin Precursor
Garg, Varun (University of Utah)
Faculty Advisor: Balagurunathan, Kuberan (University of Utah, College of Pharmacy; Medicinal Chemistry)
Heparin is a widely-used blood anticoagulant medicine that is used in surgeries and in treatment of circulatory system illnesses. Heparin is currently harvested from live pigs before they are slaughtered. There are many problems with the current production process of heparin. On a chemical level, the drug can be easily and accidentally altered due to its complex chemical nature. In addition, the environmental impact of the pigs that need to be raised on farms and fed is immense. These reasons and more are why bioengineering approaches towards the artificial synthesis of Heparin need to be researched. One of the major obstacles in the synthesis of heparin is the production of Low Molecular Weight (LMW) Heparosan, a chemical precursor. The research goal is to find methods to increase LMW Heparosan yields to a point where those same yields can be used to conduct further exploration into Heparosan oligosaccharide precursors. These oligosaccharides are useful in further understanding heparin structure-function relationships and will eventually help in the goal to find a commercially viable, cost and time effective method of heparin production. Using shake flask bacterial cultures, LMW Heparosan was produced while adjusting variables such as bacterial density at induction and incubation time to maximize yield. Once a sufficient yield was obtained, oligosaccharide structures spliced from the LMW Heparosan were classified for further research. The initial shake flask experiments showed increased yield and purity of Heparosan for each successive trial. Further work is required to scale up production of LMW Heparosan to a mass-market synthetic Heparin but the research conducted shows a promising method to synthesize and improve yields to make that possible.
Faculty Advisor: Balagurunathan, Kuberan (University of Utah, College of Pharmacy; Medicinal Chemistry)
Heparin is a widely-used blood anticoagulant medicine that is used in surgeries and in treatment of circulatory system illnesses. Heparin is currently harvested from live pigs before they are slaughtered. There are many problems with the current production process of heparin. On a chemical level, the drug can be easily and accidentally altered due to its complex chemical nature. In addition, the environmental impact of the pigs that need to be raised on farms and fed is immense. These reasons and more are why bioengineering approaches towards the artificial synthesis of Heparin need to be researched. One of the major obstacles in the synthesis of heparin is the production of Low Molecular Weight (LMW) Heparosan, a chemical precursor. The research goal is to find methods to increase LMW Heparosan yields to a point where those same yields can be used to conduct further exploration into Heparosan oligosaccharide precursors. These oligosaccharides are useful in further understanding heparin structure-function relationships and will eventually help in the goal to find a commercially viable, cost and time effective method of heparin production. Using shake flask bacterial cultures, LMW Heparosan was produced while adjusting variables such as bacterial density at induction and incubation time to maximize yield. Once a sufficient yield was obtained, oligosaccharide structures spliced from the LMW Heparosan were classified for further research. The initial shake flask experiments showed increased yield and purity of Heparosan for each successive trial. Further work is required to scale up production of LMW Heparosan to a mass-market synthetic Heparin but the research conducted shows a promising method to synthesize and improve yields to make that possible.
Great Salt Lake Halophilic Archaea: A model for mineral-entrapment of life
Bonnie K. Baxter (Westminster College)
Faculty Advisor: Baxter, Bonnie (Westminster College, Biology)
Gypsum is a calcium sulfate mineral in a hydrated form. NASA's Mars Exploration Rover, Opportunity, found veins of gypsum deposited by water in 2011(Figure 7), and gypsum has been detected on Mars as early as 2005 by the ESA's Mars Express Orbiter. On Earth, gypsum is formed in hypersaline environments, in minerals left behind when water evaporates and it can trap microorganisms in fluid inclusions. Gypsum obtained from Great Salt Lake was used to develop a method to extract halophilic archaea and culture it in the lab. Our studies show that the mineral was difficult to dissolve in aqueous microbiological media. We tested various methods of dissolution involving mechanical crushing and tested solvents including microbiological media. We also employed a variety of cultivation methods. We will present data on best practices for obtaining halophilic microorganisms from gypsum samples. The method obtained could be used to isolate potential microorganisms present in gypsum samples from Mars.
Faculty Advisor: Baxter, Bonnie (Westminster College, Biology)
Gypsum is a calcium sulfate mineral in a hydrated form. NASA's Mars Exploration Rover, Opportunity, found veins of gypsum deposited by water in 2011(Figure 7), and gypsum has been detected on Mars as early as 2005 by the ESA's Mars Express Orbiter. On Earth, gypsum is formed in hypersaline environments, in minerals left behind when water evaporates and it can trap microorganisms in fluid inclusions. Gypsum obtained from Great Salt Lake was used to develop a method to extract halophilic archaea and culture it in the lab. Our studies show that the mineral was difficult to dissolve in aqueous microbiological media. We tested various methods of dissolution involving mechanical crushing and tested solvents including microbiological media. We also employed a variety of cultivation methods. We will present data on best practices for obtaining halophilic microorganisms from gypsum samples. The method obtained could be used to isolate potential microorganisms present in gypsum samples from Mars.
Finding the interactors of Cbf1 and its role in the regulation of respiration and lipid biosynthesis
Sirrine, Michael; Grose, Julianne (Brigham Young University)
Faculty Advisor: Grose, Julianne (Brigham Young University, Microbiology and Molecular Biology)
PAS kinase is a serine/threonine protein kinase known to regulate the pivotal switch between cellular respiration and lipid biosynthesis. One substrate of PAS kinase is Cbf1, a known transcription factor which regulates lipid biosynthesis in yeast and mammalian cells (human homolog USF1). USF1 is associated with hyperlipidemia and hypercholesterolemia in several GWAS studies. We have recently identified and characterized a role for Cbf1 in the regulation of respiration as well, making it a key player in partitioning cellular resources towards respiration versus lipid metabolism. The goal of this proposal is to use the powerful tools of yeast genetics to identify physical interactors of Cbf1 in order to characterize the molecular mechanisms of its action.
Faculty Advisor: Grose, Julianne (Brigham Young University, Microbiology and Molecular Biology)
PAS kinase is a serine/threonine protein kinase known to regulate the pivotal switch between cellular respiration and lipid biosynthesis. One substrate of PAS kinase is Cbf1, a known transcription factor which regulates lipid biosynthesis in yeast and mammalian cells (human homolog USF1). USF1 is associated with hyperlipidemia and hypercholesterolemia in several GWAS studies. We have recently identified and characterized a role for Cbf1 in the regulation of respiration as well, making it a key player in partitioning cellular resources towards respiration versus lipid metabolism. The goal of this proposal is to use the powerful tools of yeast genetics to identify physical interactors of Cbf1 in order to characterize the molecular mechanisms of its action.
Genetic influences on the microbiome of Drosophila melanogaster using CRISPR/Cas9
Lemmon, Skyler; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)
Over the course of the last year, I have dedicated most of my time in the lab to learning about CRISPR/Cas9 and practicing the laboratory techniques that are necessary to make genetic changes in Drosophila melanogaster. Here I aim to expand on that expertise by applying CRISPR to study a genetic question: how the microbial composition of the D. melanogaster microbiome is affected by the modification of 4 specifically selected genes in flies from Florida and Maine. For each of the selected genes, the Florida fly allele will be put into the Maine fly genome and the Maine fly allele will be put into the Florida fly genome. The microbiome composition of these two new flies will be compared against the original lines in a factorial design. Embryos will be injected with the necessary plasmids for a double-stranded cut to take place. After injection, homology dependent repair that will incorporate the new allele. Sanger sequencing will be used to screen for successful knock-in of the allele. Finally, the concentrations of each type of bacteria found in the microbiota of the flies will be measured and compared against the flies from which the allele came from.
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)
Over the course of the last year, I have dedicated most of my time in the lab to learning about CRISPR/Cas9 and practicing the laboratory techniques that are necessary to make genetic changes in Drosophila melanogaster. Here I aim to expand on that expertise by applying CRISPR to study a genetic question: how the microbial composition of the D. melanogaster microbiome is affected by the modification of 4 specifically selected genes in flies from Florida and Maine. For each of the selected genes, the Florida fly allele will be put into the Maine fly genome and the Maine fly allele will be put into the Florida fly genome. The microbiome composition of these two new flies will be compared against the original lines in a factorial design. Embryos will be injected with the necessary plasmids for a double-stranded cut to take place. After injection, homology dependent repair that will incorporate the new allele. Sanger sequencing will be used to screen for successful knock-in of the allele. Finally, the concentrations of each type of bacteria found in the microbiota of the flies will be measured and compared against the flies from which the allele came from.
High-Fat Diet Induces Nr4a3-Dependent Decrease in Respiratory Capacity of Mouse Soleus Muscle
Marchant, Nathan; Marchant Erik; Elison, Weston; Herring, Jacob; Yang, Haokun; Tessem, Jeffrey; Hancock, Chad (Brigham Young University)
Faculty Advisor: Hancock, Chad (Brigham Young University; Nutrition, Dietetics, and Food Science); Tessem, Jeffrey (Brigham Young University; Nutrition, Dietetics, and Food Science)
Purpose: To analyze the effect of the Nr4a3 gene on respiratory capacity of mitochondria in skeletal muscle of mice on a normal or high fat diet.
Methods: Nr4a3-/- and WT mice were fed a normal chow (NC) or high fat diet (HF) for at least 20 weeks. After euthanasia, soleus muscle was harvested and wet weight was measured and prepared for respirometry. Mitochondrial respiration was evaluated using an Oroboros Oxygraph Respirometer. Respiratory capacity comparisons were made with a two-way ANOVA and Tukey multiple comparison test.
Results: Oxygen consumption is reported as pmol/(s*mg wet tissue) and statistics are represented as mean ± SEM. In the WT male mice there was a decrease in coupled complex I supported respiration in HF vs. NC diet (25.9 ± 7.3 vs. 64.5 ± 5.0, p=0.004). In the HF WT group there was also a decrease in uncoupled respiration (61.4 ± 15.0 vs. 107.8 ± 7.1, p=0.0004) compared to NC WT. In female mice there was also a decrease between HF WT and NC WT in complex I (28.2 ± 3.7 vs. 57.4 ± 5.7, p=0.0005) and uncoupled respiration (87.1 ± 7.1 vs. 119.4 ± 8.9, p=0.0001). However, there was no significant difference between the WT NC mice and either of the Nr4a3-/- groups. Coupled complex I and uncoupled respiration states in both Nr4a3-/- groups were not significantly different from WT.
Conclusion: Feeding mice a high fat diet impairs proper mitochondrial function in muscle when compared to a normal chow diet. The decrease in respiration from the HF diet is dependent upon the function of the Nr4a3 gene, as no decrease was observed in Nr4a3-/- mice.
Faculty Advisor: Hancock, Chad (Brigham Young University; Nutrition, Dietetics, and Food Science); Tessem, Jeffrey (Brigham Young University; Nutrition, Dietetics, and Food Science)
Purpose: To analyze the effect of the Nr4a3 gene on respiratory capacity of mitochondria in skeletal muscle of mice on a normal or high fat diet.
Methods: Nr4a3-/- and WT mice were fed a normal chow (NC) or high fat diet (HF) for at least 20 weeks. After euthanasia, soleus muscle was harvested and wet weight was measured and prepared for respirometry. Mitochondrial respiration was evaluated using an Oroboros Oxygraph Respirometer. Respiratory capacity comparisons were made with a two-way ANOVA and Tukey multiple comparison test.
Results: Oxygen consumption is reported as pmol/(s*mg wet tissue) and statistics are represented as mean ± SEM. In the WT male mice there was a decrease in coupled complex I supported respiration in HF vs. NC diet (25.9 ± 7.3 vs. 64.5 ± 5.0, p=0.004). In the HF WT group there was also a decrease in uncoupled respiration (61.4 ± 15.0 vs. 107.8 ± 7.1, p=0.0004) compared to NC WT. In female mice there was also a decrease between HF WT and NC WT in complex I (28.2 ± 3.7 vs. 57.4 ± 5.7, p=0.0005) and uncoupled respiration (87.1 ± 7.1 vs. 119.4 ± 8.9, p=0.0001). However, there was no significant difference between the WT NC mice and either of the Nr4a3-/- groups. Coupled complex I and uncoupled respiration states in both Nr4a3-/- groups were not significantly different from WT.
Conclusion: Feeding mice a high fat diet impairs proper mitochondrial function in muscle when compared to a normal chow diet. The decrease in respiration from the HF diet is dependent upon the function of the Nr4a3 gene, as no decrease was observed in Nr4a3-/- mice.
Genomic Assembly Targeting Repetitive Regions in the Caddisfly Genome
Wilson, Seth; Taylor Adam; Bursell, Madeline; Frandsen, Paul; Stewart, Russell; Steeneck, Amy (Brigham Young University)
Faculty Advisor: Frandsen, Paul (Brigham Young University, Plant and Wildlife Sciences)
Caddisflies (Insecta: Trichoptera) have evolved to produce silk with adhesive and elastic properties in aqueous environments. The silk is used in several ways by different species within the order such as case making, retreat making and using the silk as an anchor in the stream. Previous research on caddisfly silk has focused on understanding the evolutionary changes in the H-fibroin gene, the main protein found in caddisfly silk, which underlies the structural transformation behind these phenotypic properties that allow for diverse usage of the silk across the order (Ashton et al. 2013). Understanding the genetic foundation of the silk is crucial to understanding the phenotypic interactions that determine the unique qualities of caddisfly silk. An accurate assembly of the caddisfly genome will allow us to resolve the H-fibroin gene that plays an integral role in the formation of the caddisfly silk. Next-generation sequencing, Oxford Nanopore, and PacBio will allow us to sequence long reads that can span repetitive regions of the genome. These regions have made it difficult to resolve the H-fibroin gene as there are many repetitive motifs found in the gene. We will combine this next-generation sequencing with second-generation sequencing, Illumina and Sanger Sequencing to optimize the assembly. In this study, we used a combination of next-generation sequencing technologies to assemble the complex H-Fibroin gene in order to look at the underlying genetic structure of the silk protein. We identified unique repetitive motifs in the gene that contribute to the silk's adhesive strength and elasticity when in aqueous environments.
Faculty Advisor: Frandsen, Paul (Brigham Young University, Plant and Wildlife Sciences)
Caddisflies (Insecta: Trichoptera) have evolved to produce silk with adhesive and elastic properties in aqueous environments. The silk is used in several ways by different species within the order such as case making, retreat making and using the silk as an anchor in the stream. Previous research on caddisfly silk has focused on understanding the evolutionary changes in the H-fibroin gene, the main protein found in caddisfly silk, which underlies the structural transformation behind these phenotypic properties that allow for diverse usage of the silk across the order (Ashton et al. 2013). Understanding the genetic foundation of the silk is crucial to understanding the phenotypic interactions that determine the unique qualities of caddisfly silk. An accurate assembly of the caddisfly genome will allow us to resolve the H-fibroin gene that plays an integral role in the formation of the caddisfly silk. Next-generation sequencing, Oxford Nanopore, and PacBio will allow us to sequence long reads that can span repetitive regions of the genome. These regions have made it difficult to resolve the H-fibroin gene as there are many repetitive motifs found in the gene. We will combine this next-generation sequencing with second-generation sequencing, Illumina and Sanger Sequencing to optimize the assembly. In this study, we used a combination of next-generation sequencing technologies to assemble the complex H-Fibroin gene in order to look at the underlying genetic structure of the silk protein. We identified unique repetitive motifs in the gene that contribute to the silk's adhesive strength and elasticity when in aqueous environments.
Human health and economic costs of air pollution in Utah: an expert assessment
Errigo, Isabella; Abbott, Benjamin; Reimer, Jessica; Glenn, Jeff; Chaney, Robert; Freeman, Andrew; Frei, Rebecca; Howe, Peter, Mendoza, Daniel; Kelly, Kerry; Summers, Laura; Johnston, James; Carter, Thom; Bratsman, Samuel; Stacey, Audrey; Wilson, Derrek; Lange, Leslie (Brigham Young University)
Faculty Advisor: Abbott, Benjamin (Life Sciences, Plant and Wildlife Sciences)
Recent medical and economic research has found that air pollution causes much more damage to our health and economy than previously understood. Globally, 16% of all deaths are attributable to air pollution—15 times more than from all wars and other forms of violence. However, translating these research findings into policy and behavior change at local levels remains a major challenge, partially because of mismatch between the spatial scale of the air pollution research (often national or global) and governance frameworks (typically multi-scale from local to regional). Here, we tested the effectiveness of expert assessment as a tool to resolve research-policy mismatch. We distributed a questionnaire to over 80 researchers living in Utah, asking for quantitative estimates of human health and economic costs of air pollution, and recommendations for what policy actions would be most effective at reducing those costs. Expert responses of air pollution costs varied widely, but were consistently higher than recent public health studies, indicating that experts were including a more complete suite of factors, or that they were biased. We discuss the response of Utah policymakers to these results and present a framework of involving local researchers to increase the assimilation of data into decision making.
Faculty Advisor: Abbott, Benjamin (Life Sciences, Plant and Wildlife Sciences)
Recent medical and economic research has found that air pollution causes much more damage to our health and economy than previously understood. Globally, 16% of all deaths are attributable to air pollution—15 times more than from all wars and other forms of violence. However, translating these research findings into policy and behavior change at local levels remains a major challenge, partially because of mismatch between the spatial scale of the air pollution research (often national or global) and governance frameworks (typically multi-scale from local to regional). Here, we tested the effectiveness of expert assessment as a tool to resolve research-policy mismatch. We distributed a questionnaire to over 80 researchers living in Utah, asking for quantitative estimates of human health and economic costs of air pollution, and recommendations for what policy actions would be most effective at reducing those costs. Expert responses of air pollution costs varied widely, but were consistently higher than recent public health studies, indicating that experts were including a more complete suite of factors, or that they were biased. We discuss the response of Utah policymakers to these results and present a framework of involving local researchers to increase the assimilation of data into decision making.
Exploring the Synergies Between Vancomycin and Electrohydraulic Shockwave in Treating Staphylococcus Aureus Biofilms
Escarate, Ashley; Brunetti, Bryce; Conway, Matthew (Utah Valley University)
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology); Slezak, Cyrill (Utah Valley University, Physics)
Medical device-associated infections can lead to serious complications affecting the health of patients. Electrohydraulic shockwave treatments have shown bactericidal activity in some microorganisms. Biofilms are structures formed by microorganisms enclosed in an extracellular matrix. They form on a variety of surfaces protecting the microorganisms from antibiotics and facilitating their growth. This can result in a high rate of drug resistance and in many cases result in chronic bacterial infections.
Previously determined MIC50 concentrations of vancomycin had little effect on biofilms at twelve hours of treatment when not paired with shockwave therapy. This research evaluates the synergistic effect of different concentrations of vancomycin and shockwaves after twelve and twenty four hours of treatment given that vancomycin has shown time-dependent activity. Biofilms were grown in 96 well plates and the corresponding treatments were applied. XTT and Crystal Violet assays were used to quantify and qualify the presence of the biofilm and the antibiosis effect. The results of this experiment will be discussed in detail. A better understanding of the synergistic effects of antibiotics and shockwave therapy may lead to a more effective treatment of biofilm and device-associated infections.
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology); Slezak, Cyrill (Utah Valley University, Physics)
Medical device-associated infections can lead to serious complications affecting the health of patients. Electrohydraulic shockwave treatments have shown bactericidal activity in some microorganisms. Biofilms are structures formed by microorganisms enclosed in an extracellular matrix. They form on a variety of surfaces protecting the microorganisms from antibiotics and facilitating their growth. This can result in a high rate of drug resistance and in many cases result in chronic bacterial infections.
Previously determined MIC50 concentrations of vancomycin had little effect on biofilms at twelve hours of treatment when not paired with shockwave therapy. This research evaluates the synergistic effect of different concentrations of vancomycin and shockwaves after twelve and twenty four hours of treatment given that vancomycin has shown time-dependent activity. Biofilms were grown in 96 well plates and the corresponding treatments were applied. XTT and Crystal Violet assays were used to quantify and qualify the presence of the biofilm and the antibiosis effect. The results of this experiment will be discussed in detail. A better understanding of the synergistic effects of antibiotics and shockwave therapy may lead to a more effective treatment of biofilm and device-associated infections.
Habitat Accessibility May Alter Mule Deer Activity Schedule in Response to Hunting Pressure
Stoner, David; Clark, Debbie; Bufton, Ali (Utah State University)
Faculty Advisor: Stoner, David (S.J. & Jessie E. Quinney College of Natural Resources, Wildland Resources Department)
A challenge facing Utah mule deer is habitats with older shrubs and little to no regeneration of young plants, or habitats being replaced with cheatgrass (Bromus tectorum)—both in critical winter ranges. Forested habitats provide cover but when there is little understory, there isn't enough forage. Hunting funds the management of big game animals. If there is inadequate funding, there will not be funding for habitat. Thus population objectives will not be met. Mule deer (Odocoileus hemionus) may alter activity schedule during the hunting season by selecting restricted hunter access areas, including agricultural lands. Here we test the hypothesis that deer alter activity schedules in space (accessible and WILD sites) and time in response to human presence. We defined WILD as being further than 100 meters away from a human or OHV trail, or road. Our research question asks if accessible roads have an effect on mule deer activity schedule in response to hunting pressure, as indexed by access differences. We predicted that increased human activity during the hunting season would cause reduced deer activity during daylight. We expect more deer detections in September, less in October (hunting season), and an increase in November during the rut. Spatially we expect higher buck detection rates in remote areas, with no change in activity schedule, or are active in the day. The study area is in the Bear River Range east of Logan, Utah. We are using Before-After-Control-Impact (BACI) Sampling Design. We used camera trap data from October 2017 through December 2017 to measure (1) detection rates of bucks (photos/camera-day), and (2) activity times (diurnal, crepuscular, or nocturnal) by sex-age class. We controlled for habitat type by grouping cameras within common elevation bands and plant communities. Reduced hunter success may result in decreased hunting interest with economic implications for mule deer conservation.
Faculty Advisor: Stoner, David (S.J. & Jessie E. Quinney College of Natural Resources, Wildland Resources Department)
A challenge facing Utah mule deer is habitats with older shrubs and little to no regeneration of young plants, or habitats being replaced with cheatgrass (Bromus tectorum)—both in critical winter ranges. Forested habitats provide cover but when there is little understory, there isn't enough forage. Hunting funds the management of big game animals. If there is inadequate funding, there will not be funding for habitat. Thus population objectives will not be met. Mule deer (Odocoileus hemionus) may alter activity schedule during the hunting season by selecting restricted hunter access areas, including agricultural lands. Here we test the hypothesis that deer alter activity schedules in space (accessible and WILD sites) and time in response to human presence. We defined WILD as being further than 100 meters away from a human or OHV trail, or road. Our research question asks if accessible roads have an effect on mule deer activity schedule in response to hunting pressure, as indexed by access differences. We predicted that increased human activity during the hunting season would cause reduced deer activity during daylight. We expect more deer detections in September, less in October (hunting season), and an increase in November during the rut. Spatially we expect higher buck detection rates in remote areas, with no change in activity schedule, or are active in the day. The study area is in the Bear River Range east of Logan, Utah. We are using Before-After-Control-Impact (BACI) Sampling Design. We used camera trap data from October 2017 through December 2017 to measure (1) detection rates of bucks (photos/camera-day), and (2) activity times (diurnal, crepuscular, or nocturnal) by sex-age class. We controlled for habitat type by grouping cameras within common elevation bands and plant communities. Reduced hunter success may result in decreased hunting interest with economic implications for mule deer conservation.
Experimental adaptation of Influenza A Virus to specific host genotypes
Kelleher, Justin; Costa, Rodrigo; Potts, Wayne (University of Utah)
Faculty Advisor: Potts, Wayne (University of Utah, School of Biological Sciences)
Influenza A Virus (IAV) is a highly adaptable pathogen with the ability to cross over into different host species. Theory predicts that when a pathogen adapts to specific host genotypes, it loses virulence when encountering novel genotypes. This study focuses on whether influenza virulence is lost when infecting novel host genotypes. To test whether influenza adaptation to different genotypes leads to viral fitness and virulence tradeoffs, IAV was adapted to 2 strains of mouse via serial passage and subsequently tested against the host of passage (familiar) and in the novel host (unfamiliar). After 10 rounds of passage, IAV virulence increased in the familiar host. However, when adapted IAV strains were used to infect unfamiliar hosts, influenza virulence effects were mitigated, but not to a statistically significant degree. This study helps elucidate why different barriers to infection, including novel host genotypes, affect IAV virulence and fitness. Studying genotype-dependent virulence tradeoffs focuses can further research on more effective Influenza control in epidemiological, agricultural and conservation settings.
Faculty Advisor: Potts, Wayne (University of Utah, School of Biological Sciences)
Influenza A Virus (IAV) is a highly adaptable pathogen with the ability to cross over into different host species. Theory predicts that when a pathogen adapts to specific host genotypes, it loses virulence when encountering novel genotypes. This study focuses on whether influenza virulence is lost when infecting novel host genotypes. To test whether influenza adaptation to different genotypes leads to viral fitness and virulence tradeoffs, IAV was adapted to 2 strains of mouse via serial passage and subsequently tested against the host of passage (familiar) and in the novel host (unfamiliar). After 10 rounds of passage, IAV virulence increased in the familiar host. However, when adapted IAV strains were used to infect unfamiliar hosts, influenza virulence effects were mitigated, but not to a statistically significant degree. This study helps elucidate why different barriers to infection, including novel host genotypes, affect IAV virulence and fitness. Studying genotype-dependent virulence tradeoffs focuses can further research on more effective Influenza control in epidemiological, agricultural and conservation settings.
Genomic Analysis Between Trichoptera and Lepidoptera Show Evolutionary Innovations Allowing Trichoptera to Adapt to an Aquatic Environment
Olsen, Lindsey; Frandsen, Paul (Brigham Young University)
Faculty Advisor: Frandsen, Paul (Life Science, Plant and Wildlife)
Trichoptera (caddisflies) have evolved to become the most diverse, exclusively aquatic insects, yet many of the genomic changes that contribute to Trichoptera's the success of this order of insects are still unknown. Trichoptera and Lepidoptera (moths and butterflies) are reciprocally monophyletic meaning that they both share their most recent common ancestor. Despite being closely related, Trichoptera have evolved into the most diverse, exclusively aquatic insects, whereas, Lepidoptera have evolved to become a diverse, almost exclusively terrestrial insect (Holzenthal et al. 2007). Trichoptera and Lepidoptera are the subjects of scientific inquiry because they are both capable of spinning silk. Trichoptera produce silk as larvae and use it to make cases or fixed retreats. Trichoptera silk is of particular interest because its properties allow for it to be an underwater adhesive. While other research has focused primarily on the evolution of Trichoptera silk, little research has been done to identify the evolutionary innovations that allowed Trichoptera to adapt and diversify in an aquatic environment. Our research focuses on identifying the genomic basis of their evolutionary innovations. We report the genome annotation of four newly sequenced Trichoptera species Hesperophylax magnus, Parapsyche elsis, Philanisus plebeius, and Rhyacophila brunnea. These annotations will reveal levels of homozygosity, conserved elements, and gene duplications. We then conducted a genome-wide search for gene family expansions and retractions using CAFE, in order to identify genomic regions that could contribute to Trichoptera's unique qualities and evolutionary history.
Holzenthal R. W., R. J. Blahnik, A. L. Prather, and K. M. Kjer, 2007 Order Trichoptera Kirby, 1813 (Insecta), Caddisflies*. Zootaxa 1668: 639—698. https://doi.org/10.11646/zootaxa.1668.1.29
Faculty Advisor: Frandsen, Paul (Life Science, Plant and Wildlife)
Trichoptera (caddisflies) have evolved to become the most diverse, exclusively aquatic insects, yet many of the genomic changes that contribute to Trichoptera's the success of this order of insects are still unknown. Trichoptera and Lepidoptera (moths and butterflies) are reciprocally monophyletic meaning that they both share their most recent common ancestor. Despite being closely related, Trichoptera have evolved into the most diverse, exclusively aquatic insects, whereas, Lepidoptera have evolved to become a diverse, almost exclusively terrestrial insect (Holzenthal et al. 2007). Trichoptera and Lepidoptera are the subjects of scientific inquiry because they are both capable of spinning silk. Trichoptera produce silk as larvae and use it to make cases or fixed retreats. Trichoptera silk is of particular interest because its properties allow for it to be an underwater adhesive. While other research has focused primarily on the evolution of Trichoptera silk, little research has been done to identify the evolutionary innovations that allowed Trichoptera to adapt and diversify in an aquatic environment. Our research focuses on identifying the genomic basis of their evolutionary innovations. We report the genome annotation of four newly sequenced Trichoptera species Hesperophylax magnus, Parapsyche elsis, Philanisus plebeius, and Rhyacophila brunnea. These annotations will reveal levels of homozygosity, conserved elements, and gene duplications. We then conducted a genome-wide search for gene family expansions and retractions using CAFE, in order to identify genomic regions that could contribute to Trichoptera's unique qualities and evolutionary history.
Holzenthal R. W., R. J. Blahnik, A. L. Prather, and K. M. Kjer, 2007 Order Trichoptera Kirby, 1813 (Insecta), Caddisflies*. Zootaxa 1668: 639—698. https://doi.org/10.11646/zootaxa.1668.1.29
Identifying Conservation Needs of Dwarf Bear-Claw Poppy Populations
Thatcher, Spencer; Eyere, Gloria (Dixie State University)
Faculty Advisor: O'Brien, Erin (College of Science, Engineering, and Sciences; Biological Sciences Department)
Dwarf bear-claw poppies (Arctomecon humilis Coville) are an endemic species of wildflower in Washington county Utah. These poppies only flower for about a month during the summer. There are currently seven known locations where these poppies are found. Conservation efforts to protect these populations includes research to understand the reproductive success of each site. Populations with low reproductive success may indicate the need for additional efforts or interventions to save this unique species. Four specific locations were monitored over a three-year period. Poppies in these areas were tagged and studied during their flowering seasons and individual inflorescences were bagged and collected to determine struggling locations and their correlating issues related to population decline. Seeds were split into two groups: immature and mature and were weighed and counted. A large percentage of immature seeds may indicate a resource or pollinator issue for the population. A low overall seed production may indicate that a resource limitation or other stressor is reducing the population viability.
Faculty Advisor: O'Brien, Erin (College of Science, Engineering, and Sciences; Biological Sciences Department)
Dwarf bear-claw poppies (Arctomecon humilis Coville) are an endemic species of wildflower in Washington county Utah. These poppies only flower for about a month during the summer. There are currently seven known locations where these poppies are found. Conservation efforts to protect these populations includes research to understand the reproductive success of each site. Populations with low reproductive success may indicate the need for additional efforts or interventions to save this unique species. Four specific locations were monitored over a three-year period. Poppies in these areas were tagged and studied during their flowering seasons and individual inflorescences were bagged and collected to determine struggling locations and their correlating issues related to population decline. Seeds were split into two groups: immature and mature and were weighed and counted. A large percentage of immature seeds may indicate a resource or pollinator issue for the population. A low overall seed production may indicate that a resource limitation or other stressor is reducing the population viability.
Genome-wide CRISPR-Cas9 Screen Identifies Genes Required for Ꞵ-cell Survival of Metabolic Stressors.
Ekpo, Idongesit; Yates, Joshua; Tessem, Jeffery; Hill, Jonathan (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutrition, Dietetics, and Food Science); Hill, Jonathan (Life Sciences, Physiology and Developmental Biology)
By the year 2040, an estimated 642 million people are expected to have diabetes globally. Diabetes results from an elevation of metabolic stressors, such as glucotoxicity, lipotoxicity and oxidative stress induced by reactive oxygen and nitrogen species. Current treatment methods for diabetes are not curative and do not help us understand its pathogenesis. A more effective method involves exploring the pathogenesis of diabetes by probing the genetic variation involved in diabetes so that we can understand the disease better and develop curative methods to combat it. Gene therapy is a method for determining genetic variation in disease and CRISPR-Cas9 is a gene-editing tool that can be used. Because of its convenience, CRISPR-Cas9 has been used to create many forward genetic screens. We use the CRISPR-Cas9 tool to create a knockout forward genetic screen of all the genes in the INS-1 Ꞵ-cell line that are required for _-cell survival of metabolic stressors. We hypothesize that the gene knockouts generated by the CRISPR-Cas9 system will help us identify genes that are involved in the mechanistic pathways of these metabolic stressors. Here we present the results of our forward genetic screen.
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutrition, Dietetics, and Food Science); Hill, Jonathan (Life Sciences, Physiology and Developmental Biology)
By the year 2040, an estimated 642 million people are expected to have diabetes globally. Diabetes results from an elevation of metabolic stressors, such as glucotoxicity, lipotoxicity and oxidative stress induced by reactive oxygen and nitrogen species. Current treatment methods for diabetes are not curative and do not help us understand its pathogenesis. A more effective method involves exploring the pathogenesis of diabetes by probing the genetic variation involved in diabetes so that we can understand the disease better and develop curative methods to combat it. Gene therapy is a method for determining genetic variation in disease and CRISPR-Cas9 is a gene-editing tool that can be used. Because of its convenience, CRISPR-Cas9 has been used to create many forward genetic screens. We use the CRISPR-Cas9 tool to create a knockout forward genetic screen of all the genes in the INS-1 Ꞵ-cell line that are required for _-cell survival of metabolic stressors. We hypothesize that the gene knockouts generated by the CRISPR-Cas9 system will help us identify genes that are involved in the mechanistic pathways of these metabolic stressors. Here we present the results of our forward genetic screen.
Implications of Testing Sexual Assault Kits: Justice for Suspects and Victims
Caten, Reilly; Valentine, Julie; Miles, Leslie (Brigham Young University)
Faculty Advisor: Valentine, Julie (Brigham Young University, Nursing); Leslie, Miles (Brigham Young University, Nursing)
In the United States, there is a push to mandate submission and testing of all sexual assault kits. A typically-overlooked benefit of testing sexual assault kits is the exoneration of wrongfully accused suspects. Sexual assault kits include DNA samples from the victim of sexual assault, and occasionally are submitted with DNA samples collected from the identified suspect for comparison. Inclusion of suspect samples is beneficial because it can lead to a DNA match with samples collected from the victim or exclude the suspect as the source of DNA. For a sexual assault kit to meet the criteria for "excluded the suspect" a DNA profile had to be developed from analysis of the sexual assault kit evidence which excluded the named suspect with submitted DNA sample.
A large retrospective study of 2,727 sexual assault kits with completed DNA analysis found 66 cases in which DNA findings excluded the suspect. Findings will be presented on descriptive data on cases in which the named suspect was excluded: relationship between victim and suspect, victim loss of consciousness/awareness at time of assault, alcohol/drug use during the assault, suspected drug-facilitated sexual assault, and multiple perpetrator sexual assault. Each of these cases excluding a suspect represents an individual who was erroneously identified, but DNA analysis findings excluded the identified suspect.
Testing sexual assault kits uses science to aid in the establishment of justice. Sexual assault kit testing transcends accusations and labeling to provide factual evidence supporting the claims of the innocent, whether they are identified as victim or suspect. Additionally, the liberation of an unjustly accused suspect promotes the correct identification and conviction of the responsible perpetrator. Thus, mandated testing of sexual assault kit promotes justice for victims of sexual assault and innocent suspects alike.
Faculty Advisor: Valentine, Julie (Brigham Young University, Nursing); Leslie, Miles (Brigham Young University, Nursing)
In the United States, there is a push to mandate submission and testing of all sexual assault kits. A typically-overlooked benefit of testing sexual assault kits is the exoneration of wrongfully accused suspects. Sexual assault kits include DNA samples from the victim of sexual assault, and occasionally are submitted with DNA samples collected from the identified suspect for comparison. Inclusion of suspect samples is beneficial because it can lead to a DNA match with samples collected from the victim or exclude the suspect as the source of DNA. For a sexual assault kit to meet the criteria for "excluded the suspect" a DNA profile had to be developed from analysis of the sexual assault kit evidence which excluded the named suspect with submitted DNA sample.
A large retrospective study of 2,727 sexual assault kits with completed DNA analysis found 66 cases in which DNA findings excluded the suspect. Findings will be presented on descriptive data on cases in which the named suspect was excluded: relationship between victim and suspect, victim loss of consciousness/awareness at time of assault, alcohol/drug use during the assault, suspected drug-facilitated sexual assault, and multiple perpetrator sexual assault. Each of these cases excluding a suspect represents an individual who was erroneously identified, but DNA analysis findings excluded the identified suspect.
Testing sexual assault kits uses science to aid in the establishment of justice. Sexual assault kit testing transcends accusations and labeling to provide factual evidence supporting the claims of the innocent, whether they are identified as victim or suspect. Additionally, the liberation of an unjustly accused suspect promotes the correct identification and conviction of the responsible perpetrator. Thus, mandated testing of sexual assault kit promotes justice for victims of sexual assault and innocent suspects alike.
Field based real time sequencing of microbial samples via nanopore technology
Jackson, Ryan; Miller, Charles (Utah State University)
Faculty Advisor: Jackson, Ryan (College of Science, Chemistry and Biochemistry Department); Miller, Charles (College of Engineering, Biological Engineering Department)
The concept of real time species identification in situ is a long time researchers dream. This dream now lies within reach due to the recent innovation of nanopore sequencing technology. These machines, with their small size and powerful computing capability, have made it possible to preform 16s and whole genome sequencing, with a setup that can fit in a backpack. Not only will this increase convenience of sampling for researchers, but a recent study in Wales has shown that sampling on site may help to identify closely related organisms at a greater level of accuracy (Parker, 2017). If sampling in the field really can give more accurate results, field sequencing may help to identify an extraordinarily large amount of biodiversity and genetic pathways.
One obstacle that stands in the way of this technology becoming more accessible across the globe is the lack of scientific literature on how to build the infrastructure necessary to sample on site. This study aims to construct a complete, self-contained kit with which you could field sequence. I have, currently at my disposal, a portable thermocycler, a nanopore sequencer, and computer designed with a workflow to do real time sequencing analysis. Using this technology already available, we aim to round out the kit with the necessary reagents, and structure to house the equipment. We will provide in depth analysis of the equipment, reagents, and all other materials provided to sequence a sample in any given location.
Faculty Advisor: Jackson, Ryan (College of Science, Chemistry and Biochemistry Department); Miller, Charles (College of Engineering, Biological Engineering Department)
The concept of real time species identification in situ is a long time researchers dream. This dream now lies within reach due to the recent innovation of nanopore sequencing technology. These machines, with their small size and powerful computing capability, have made it possible to preform 16s and whole genome sequencing, with a setup that can fit in a backpack. Not only will this increase convenience of sampling for researchers, but a recent study in Wales has shown that sampling on site may help to identify closely related organisms at a greater level of accuracy (Parker, 2017). If sampling in the field really can give more accurate results, field sequencing may help to identify an extraordinarily large amount of biodiversity and genetic pathways.
One obstacle that stands in the way of this technology becoming more accessible across the globe is the lack of scientific literature on how to build the infrastructure necessary to sample on site. This study aims to construct a complete, self-contained kit with which you could field sequence. I have, currently at my disposal, a portable thermocycler, a nanopore sequencer, and computer designed with a workflow to do real time sequencing analysis. Using this technology already available, we aim to round out the kit with the necessary reagents, and structure to house the equipment. We will provide in depth analysis of the equipment, reagents, and all other materials provided to sequence a sample in any given location.
How long is too long? Developing CODIS eligible profiles from sexual assault evidence collection
Lauren Schagel; Julie Valentine; Leslie Miles (Brigham Young University)
Faculty Advisor: Valentine, Julie (Brigham Young University, Nursing); Miles, Leslie (Brigham Young University, Nursing)
Question: What is the impact of time between assault and evidence collection on the development of CODIS eligible DNA profiles?
Learning Overview/Synopsis: After attending this presentation, attendees will understand the length of time between sexual assault and evidence collection can be extended to five to six days post-assault and develop an eligible STR DNA profile.
Impact on the Forensic Science Community: This presentation will impact the forensic community by providing individuals with knowledge about the length of time in which victims can receive a sexual assault forensic examination (SAFE) to obtain a DNA profile of their perpetrator.
Synopsis/Abstract: The methodology of the study is an exploratory, retrospective design of over 2,700 submitted and analyzed sexual assault kits from a Mountain West state in the United States. Review of the current literature on time between assault and evidence collection indicate male DNA (Y-STR DNA) can be recovered up to 7 days post-coitus from a cervico-vaginal swab and develop a Y-STR profile.
Findings: In our large-scale, retrospective study of 2,727 sexual assault kits, probative STR DNA CODIS eligible profiles were developed in 39% of kits. Generalized estimating equation (GEE) logistic regression analysis found that for every 24 hours that passes between assault and SAFE, there is a 10% reduction in the development of a CODIS eligible DNA profile. The longest length of time between assault and exam and development of CODIS eligible profile in our study was 122.5 hours, over 5 days. This finding supports the testing of DNA samples collected five to six days post sexual assault due to the possibility of developing a CODIS eligible profile.
Conclusion: This retrospective study is impactful due to large-scale of the sample size. The 2,727 sexual assault kits used in this study are representative of what sexual assault looks like in modern society.
Faculty Advisor: Valentine, Julie (Brigham Young University, Nursing); Miles, Leslie (Brigham Young University, Nursing)
Question: What is the impact of time between assault and evidence collection on the development of CODIS eligible DNA profiles?
Learning Overview/Synopsis: After attending this presentation, attendees will understand the length of time between sexual assault and evidence collection can be extended to five to six days post-assault and develop an eligible STR DNA profile.
Impact on the Forensic Science Community: This presentation will impact the forensic community by providing individuals with knowledge about the length of time in which victims can receive a sexual assault forensic examination (SAFE) to obtain a DNA profile of their perpetrator.
Synopsis/Abstract: The methodology of the study is an exploratory, retrospective design of over 2,700 submitted and analyzed sexual assault kits from a Mountain West state in the United States. Review of the current literature on time between assault and evidence collection indicate male DNA (Y-STR DNA) can be recovered up to 7 days post-coitus from a cervico-vaginal swab and develop a Y-STR profile.
Findings: In our large-scale, retrospective study of 2,727 sexual assault kits, probative STR DNA CODIS eligible profiles were developed in 39% of kits. Generalized estimating equation (GEE) logistic regression analysis found that for every 24 hours that passes between assault and SAFE, there is a 10% reduction in the development of a CODIS eligible DNA profile. The longest length of time between assault and exam and development of CODIS eligible profile in our study was 122.5 hours, over 5 days. This finding supports the testing of DNA samples collected five to six days post sexual assault due to the possibility of developing a CODIS eligible profile.
Conclusion: This retrospective study is impactful due to large-scale of the sample size. The 2,727 sexual assault kits used in this study are representative of what sexual assault looks like in modern society.
Effects of TrkB activation by alcohol withdrawal-mediated secretions of BDNF following chronic alcohol exposure
Murley, Jordan; Stockard, Alyssa; Payne, Andrew; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
Brain-derived neurotrophic factor (BDNF) is implicated in varied physiological processes. Its main receptor in the central nervous system is tyrosine receptor kinase B (TrkB), and the main ligand for TrkB in the central nervous system is BDNF. It has been shown previously that activating TrkB can cause a downregulation of the chloride-exporting potassium chloride cotransporter 2 (KCC2), presumably resulting in a decreased chloride gradient. It is hypothesized that this down-regulation, caused by an increase in BDNF levels, creates hyperexcitable GABA neurons in the ventral tegmental area (VTA) due to a reduced efficacy of inhibitory currents. Here we investigate this mechanism of adaptation in the context of chronic alcohol exposure. We demonstrate that BDNF levels are elevated in the VTA during withdrawal from chronic alcohol exposure. We also observe that blocking TrkB activity decreases alcohol seeking behavior. Further, we investigate the expression patterns of KCC2 in connection with chronic alcohol administration. Additional work is underway to validate this mechanism and further elucidate its putative role in alcohol dependence.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
Brain-derived neurotrophic factor (BDNF) is implicated in varied physiological processes. Its main receptor in the central nervous system is tyrosine receptor kinase B (TrkB), and the main ligand for TrkB in the central nervous system is BDNF. It has been shown previously that activating TrkB can cause a downregulation of the chloride-exporting potassium chloride cotransporter 2 (KCC2), presumably resulting in a decreased chloride gradient. It is hypothesized that this down-regulation, caused by an increase in BDNF levels, creates hyperexcitable GABA neurons in the ventral tegmental area (VTA) due to a reduced efficacy of inhibitory currents. Here we investigate this mechanism of adaptation in the context of chronic alcohol exposure. We demonstrate that BDNF levels are elevated in the VTA during withdrawal from chronic alcohol exposure. We also observe that blocking TrkB activity decreases alcohol seeking behavior. Further, we investigate the expression patterns of KCC2 in connection with chronic alcohol administration. Additional work is underway to validate this mechanism and further elucidate its putative role in alcohol dependence.
Ethanol enhancement of dopamine release in the nucleus accumbens and ethanol reward is mediated by peripheral dopamine 2 receptors
Diaz, Paula; Baptista, Gabriella; Obray, J Daniel; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
The prevailing view is that enhancement of dopamine (DA) transmission in the mesolimbic DA system underlies the rewarding properties of alcohol. The aim of the present study was to help reconcile the relative insensitivity of DA neurons to ethanol in vitro (EC50 = 96 mM) with the sensitivity of DA neurons in vivo (EC50 = 3 mM). To do this we investigated the role of peripheral dopamine 2 receptor (D2R) mediated neuroimmune responses in ethanol enhancement of DA release in the NAc and ethanol reward. We found that systemic administration of ethanol (0.5-4.0 g/kg) markedly enhanced DA release in the NAc while pretreatment with a peripheral-only D2 receptor (D2R) antagonist blocked these effects. A place conditioning paradigm was used to test rats for ethanol preference. Administration of a peripheral D2R antagonist before ethanol conditioning trials was found to prevent acquisition of ethanol conditioned place preference. Finally, ethanol suppression of locomotor activity in rats was attenuated by domperidone pre-administration. Domperidone pre-administration did not affect ethanol impairment of motor coordination. These findings suggest that ethanol enhancement of DA release, intoxication, and ethanol reward are all mediated, at least in part, by a peripheral mechanism involving D2Rs. These results challenge the dogma regarding direct ethanol actions on mesolimbic DA transmission and potentially provides novel pharmacological targets for the treatment of alcohol use disorder.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
The prevailing view is that enhancement of dopamine (DA) transmission in the mesolimbic DA system underlies the rewarding properties of alcohol. The aim of the present study was to help reconcile the relative insensitivity of DA neurons to ethanol in vitro (EC50 = 96 mM) with the sensitivity of DA neurons in vivo (EC50 = 3 mM). To do this we investigated the role of peripheral dopamine 2 receptor (D2R) mediated neuroimmune responses in ethanol enhancement of DA release in the NAc and ethanol reward. We found that systemic administration of ethanol (0.5-4.0 g/kg) markedly enhanced DA release in the NAc while pretreatment with a peripheral-only D2 receptor (D2R) antagonist blocked these effects. A place conditioning paradigm was used to test rats for ethanol preference. Administration of a peripheral D2R antagonist before ethanol conditioning trials was found to prevent acquisition of ethanol conditioned place preference. Finally, ethanol suppression of locomotor activity in rats was attenuated by domperidone pre-administration. Domperidone pre-administration did not affect ethanol impairment of motor coordination. These findings suggest that ethanol enhancement of DA release, intoxication, and ethanol reward are all mediated, at least in part, by a peripheral mechanism involving D2Rs. These results challenge the dogma regarding direct ethanol actions on mesolimbic DA transmission and potentially provides novel pharmacological targets for the treatment of alcohol use disorder.
Effect of Mercury Chloride and Methyl Mercury
Villanueva,Xitlalli; Berthelemy Nicole (Weber State University)
Faculty Advisor: Berthelemy, Nicole (Weber State University, Zoology)
Introduction: Mercury, originating from mining and industrial activity, has been accumulating in the Great Salt Lake water and has contaminated its food web. Some of the mercury chloride (HgCl) is transformed by benthic bacteria to the more toxic monomethyl mercury (MeHg). While the effect of HgCl on the brine shrimp Artemia franciscana has been described, little is known about the effects of MeHG on this shrimp, except that it is more toxic. We correlated the tissue mercury concentrations to the amount of mercury present in the sea-water (SW). We also tested the ability of the shrimp to depurate mercury accumulated in the tissues.
Results: - Adult Artemia survived at least one week when exposed to concentrations ranging from 10-9 to 10-5g/L HgCl or MeHg. Shrimp exposed to 10-4g/L HgCl or MeHg died within two days.
Fertility was influenced by mercury exposure. The number of offspring per brood produced by females exposed to 10-6g/L or less HgCl was around 150, significantly higher than the average 80 offspring per brood produced by females exposed to 10-6g/L MeHg. When exposed to 10-5g/l HgCl and MeHg, the females brood size was 47 and 24, respectively.
- Artemia is able to maintain low level of tissue mercury, up to exposure to 10-6g/L HgCl and 10-7 g/L for MeHg. Exposure to higher mercury concentrations leads to much higher mercury levels, about 35 ppm HgCl for SW reaching 10-5g/L and 300 ppm with exposure to 10-4 MeHg.
Discussion: While both HgCl and MeHg are lethal to the shrimp, after a few days exposure, at concentration higher than 10-6 g/L, MeHg has a stronger effect on fertility, with brood size near half the size. Also, MeHg, the organic form of mercury, is accumulated in much higher level than the inorganic form. The organic form has a better ability to cross cell membranes. Since the mercury levels in the Great Salt Lake reach 55 ng/L (= 0.055 10-6g/L) and 30 ng/L (0.03 10-6 g/L) for HgCl and MeHG, respectively, the brine shrimp population is likely not affected by this pollutant.
Conclusion: Mercury has a strong effect on Artemia metabolism. We will study, next, the shrimp response to the oxidative stress induced by the mercury exposure.
Faculty Advisor: Berthelemy, Nicole (Weber State University, Zoology)
Introduction: Mercury, originating from mining and industrial activity, has been accumulating in the Great Salt Lake water and has contaminated its food web. Some of the mercury chloride (HgCl) is transformed by benthic bacteria to the more toxic monomethyl mercury (MeHg). While the effect of HgCl on the brine shrimp Artemia franciscana has been described, little is known about the effects of MeHG on this shrimp, except that it is more toxic. We correlated the tissue mercury concentrations to the amount of mercury present in the sea-water (SW). We also tested the ability of the shrimp to depurate mercury accumulated in the tissues.
Results: - Adult Artemia survived at least one week when exposed to concentrations ranging from 10-9 to 10-5g/L HgCl or MeHg. Shrimp exposed to 10-4g/L HgCl or MeHg died within two days.
Fertility was influenced by mercury exposure. The number of offspring per brood produced by females exposed to 10-6g/L or less HgCl was around 150, significantly higher than the average 80 offspring per brood produced by females exposed to 10-6g/L MeHg. When exposed to 10-5g/l HgCl and MeHg, the females brood size was 47 and 24, respectively.
- Artemia is able to maintain low level of tissue mercury, up to exposure to 10-6g/L HgCl and 10-7 g/L for MeHg. Exposure to higher mercury concentrations leads to much higher mercury levels, about 35 ppm HgCl for SW reaching 10-5g/L and 300 ppm with exposure to 10-4 MeHg.
Discussion: While both HgCl and MeHg are lethal to the shrimp, after a few days exposure, at concentration higher than 10-6 g/L, MeHg has a stronger effect on fertility, with brood size near half the size. Also, MeHg, the organic form of mercury, is accumulated in much higher level than the inorganic form. The organic form has a better ability to cross cell membranes. Since the mercury levels in the Great Salt Lake reach 55 ng/L (= 0.055 10-6g/L) and 30 ng/L (0.03 10-6 g/L) for HgCl and MeHG, respectively, the brine shrimp population is likely not affected by this pollutant.
Conclusion: Mercury has a strong effect on Artemia metabolism. We will study, next, the shrimp response to the oxidative stress induced by the mercury exposure.
Effects of Exercise on Subjective Feelings of Vitality: A Randomized Crossover Study
Stevens, Andrew; Hicks, Joseph; Savage, Ryland; Bailey, Bruce (Brigham Young University)
Faculty Advisor: Bailey, Bruce (Life Sciences, Exercise Science)
Purpose: This study examined the effect that differing exercise intensities have on subjective ratings of vitality both immediately following the condition and after 60 minutes of intense cognitive testing.
Methods: These findings come as a secondary analysis of a larger randomized crossover study that examined the effects of differing exercise intensities on cognitive performance. We recruited 228 men and women in roughly equal numbers to participate in the study. Each participant reported to the study lab four times one week apart. During the first day the participants ran a VO2 max test to establish pacing and baseline values. During the next three visits the participant completed a subjective vitality questionnaire before exercise, after 40 minutes of exercise or watching a video, and after 60 minutes of cognitive testing. The exercise conditions of vigorous (70% VO2 Max), moderate (35% VO2 Max) and sedentary activity (watching a video) were randomly assigned each week.
Results: The vitality rating before exercise was not significantly different between the three conditions with a mean rating of 19.9 ± 8.6 cm. After the sedentary condition, subjective ratings of vitality decreased from 19.9 cm to 12.5 cm after the condition and increased to 15.0 cm after the cognitive testing, remaining below pre-exercise values. After the moderate exercise condition, vitality did not change significantly after exercise (20.2 cm to 21.0 cm) but was lower following the cognitive tasks (15.9 cm). Vitality was slightly elevated following the vigorous exercise condition (19.8 cm to 21.3 cm) but went down after the cognitive tasks (17.1 cm). Vitality ratings did not differ significantly between exercise conditions at any time. The sedentary condition was lower than both exercise conditions after the 40-min intervention (p<0.001) and was lower than the vigorous exercise condition after the cognitive tasks (p<0.001).
Conclusion: Sedentary tasks may reduce subjective feelings of energy and vitality and this subjective evaluation remains suppressed after a period of cognitive exertion. Feelings of vitality and energy remain similar before and after exercise (vigorous or moderate) but then reduce after cognitive exertion. Vigorous exercise may be slightly beneficial for subjective feeling of vitality after exercise and following extended cognitive exertion.
Faculty Advisor: Bailey, Bruce (Life Sciences, Exercise Science)
Purpose: This study examined the effect that differing exercise intensities have on subjective ratings of vitality both immediately following the condition and after 60 minutes of intense cognitive testing.
Methods: These findings come as a secondary analysis of a larger randomized crossover study that examined the effects of differing exercise intensities on cognitive performance. We recruited 228 men and women in roughly equal numbers to participate in the study. Each participant reported to the study lab four times one week apart. During the first day the participants ran a VO2 max test to establish pacing and baseline values. During the next three visits the participant completed a subjective vitality questionnaire before exercise, after 40 minutes of exercise or watching a video, and after 60 minutes of cognitive testing. The exercise conditions of vigorous (70% VO2 Max), moderate (35% VO2 Max) and sedentary activity (watching a video) were randomly assigned each week.
Results: The vitality rating before exercise was not significantly different between the three conditions with a mean rating of 19.9 ± 8.6 cm. After the sedentary condition, subjective ratings of vitality decreased from 19.9 cm to 12.5 cm after the condition and increased to 15.0 cm after the cognitive testing, remaining below pre-exercise values. After the moderate exercise condition, vitality did not change significantly after exercise (20.2 cm to 21.0 cm) but was lower following the cognitive tasks (15.9 cm). Vitality was slightly elevated following the vigorous exercise condition (19.8 cm to 21.3 cm) but went down after the cognitive tasks (17.1 cm). Vitality ratings did not differ significantly between exercise conditions at any time. The sedentary condition was lower than both exercise conditions after the 40-min intervention (p<0.001) and was lower than the vigorous exercise condition after the cognitive tasks (p<0.001).
Conclusion: Sedentary tasks may reduce subjective feelings of energy and vitality and this subjective evaluation remains suppressed after a period of cognitive exertion. Feelings of vitality and energy remain similar before and after exercise (vigorous or moderate) but then reduce after cognitive exertion. Vigorous exercise may be slightly beneficial for subjective feeling of vitality after exercise and following extended cognitive exertion.
Ecology of Male Mating in Alfaro cultratus: Is morphology a response to the environment?
Mueller, Kaeli; Kaitlyn, Golden; Johnson, J.B. (Brigham Young University)
Faculty Advisor: Johnson, Jerry (Brigham Young University, College of Life Sciences);
Human-induced changes to the environment are causing declines in many of the world's species. Studying model organisms, such as Poeciilidae fishes, can provide crucial information about a variety of other species (Johnson & Bagley, 2011). Alfaro, a monophyletic genus within the family Poeciilidae (Bagley & Johnson, 2014), has the potential to become an exemplary model species. There is currently much scientific knowledge about the two distinct reproductive tactics of Poeciliid males. However, studies about the presence of this trait are lacking for Alfaro. Therefore, I propose adding to the current knowledge about Alfaro cultratus by researching the presence of two distinct sets of traits that accompany different male reproductive tactics. The hypothesis is that male Alfaro will show similar physical characteristics to other Poeciliids, but that these traits will not differ across varying habitats. Measurements of deceased Alfaro specimens will be analyzed to identify differences in physical traits. This will be followed by a live experiment, in which the male size differential is expected to mirror that of the deceased specimens. This research will produce novel information about Alfaro cultratus because its life history strategies have yet to be described. Many intriguing evolution questions could follow.
References:
Bagley, J. C., & Johnson, J. B. (2014). Testing for shared biogeographic history in the lower Central American freshwater fish assemblage using comparative phylogeography: Concerted, independent, or multiple evolutionary responses? Ecology and Evolution, 4(9), 1686—1705.
Johnson, J. B., & Bagley, J. C. (2011). Ecology and Evolution of Poeciliid Fishes, Chapter 4, Ecological drivers of life-history divergence.
Faculty Advisor: Johnson, Jerry (Brigham Young University, College of Life Sciences);
Human-induced changes to the environment are causing declines in many of the world's species. Studying model organisms, such as Poeciilidae fishes, can provide crucial information about a variety of other species (Johnson & Bagley, 2011). Alfaro, a monophyletic genus within the family Poeciilidae (Bagley & Johnson, 2014), has the potential to become an exemplary model species. There is currently much scientific knowledge about the two distinct reproductive tactics of Poeciliid males. However, studies about the presence of this trait are lacking for Alfaro. Therefore, I propose adding to the current knowledge about Alfaro cultratus by researching the presence of two distinct sets of traits that accompany different male reproductive tactics. The hypothesis is that male Alfaro will show similar physical characteristics to other Poeciliids, but that these traits will not differ across varying habitats. Measurements of deceased Alfaro specimens will be analyzed to identify differences in physical traits. This will be followed by a live experiment, in which the male size differential is expected to mirror that of the deceased specimens. This research will produce novel information about Alfaro cultratus because its life history strategies have yet to be described. Many intriguing evolution questions could follow.
References:
Bagley, J. C., & Johnson, J. B. (2014). Testing for shared biogeographic history in the lower Central American freshwater fish assemblage using comparative phylogeography: Concerted, independent, or multiple evolutionary responses? Ecology and Evolution, 4(9), 1686—1705.
Johnson, J. B., & Bagley, J. C. (2011). Ecology and Evolution of Poeciliid Fishes, Chapter 4, Ecological drivers of life-history divergence.
Does phylogenetic relatedness influence response to heterospecific alarm cues in Brachyrhaphis rhabdophora?
Chou, Audrey; Duffy, Alexandra; Johnson, Jerald (Brigham Young University)
Faculty Advisor: Johnson, Jerald (Brigham Young University, Biology)
Upon injury, fish release chemicals that cause fear and a dramatic change in behavior in nearby members of the species. Chemical alarm cues are released from injured epidermal tissue and provide an early warning signal for individuals to detect the presence of an active predator, and potentially increase the chances of individual survival. This research project investigates the conservation of alarm cue recognition and response across species boundaries by measuring the behavioral response of Brachyrhaphis rhabdophora to the alarm cues from five different species. Since these species will be of known phylogenetic relatedness, we will determine conservation of alarm cue response as a function of phylogenetic distance. I will also discuss the method of quantifying behavioral response through the use of a two-choice flume tank to measure both changes in activity and spatial avoidance upon alarm cue exposure.
Faculty Advisor: Johnson, Jerald (Brigham Young University, Biology)
Upon injury, fish release chemicals that cause fear and a dramatic change in behavior in nearby members of the species. Chemical alarm cues are released from injured epidermal tissue and provide an early warning signal for individuals to detect the presence of an active predator, and potentially increase the chances of individual survival. This research project investigates the conservation of alarm cue recognition and response across species boundaries by measuring the behavioral response of Brachyrhaphis rhabdophora to the alarm cues from five different species. Since these species will be of known phylogenetic relatedness, we will determine conservation of alarm cue response as a function of phylogenetic distance. I will also discuss the method of quantifying behavioral response through the use of a two-choice flume tank to measure both changes in activity and spatial avoidance upon alarm cue exposure.
Effects of Mycorrhizal Associations on Tomato Defensive Compounds
Jones, Dalton (Weber State University)
Faculty Advisor: Schramm, Katharina (Science, Botany)
Tomatoes (Solanum lycopersicum) are an important agricultural crop around the world. In order to produce food with less impact on the environment, many researchers are looking to utilize natural systems to maximize production with minimal inputs. Maximizing tomato defenses is one possible way to increase productivity. Tomatoes produce both physical and chemical defenses in response to the stress. Increasing the number of trichomes on the plant is a physical means of deterring insects from eating the plant. Trichomes come in two forms glandular trichomes and non-glandular trichomes. The non-glandular trichomes are strictly a physical defense while the glandular trichomes produce chemical defensive compounds against a wide variety of insects. Most land plants can form a symbiosis with arbuscular mycorrhizal fungi (AMF). This symbiosis has been shown to increase nutrient supply, decrease drought stress, and prime plant defenses, all increasing a plant's ability to withstand herbivory stress better. This study examines the response of trichomes and the number of chemical defenses after insect herbivory has occurred with the additional support of the AMF symbiosis. Treatments were exposed to Manduca sexta to stimulate trichome and chemical production. The extracted leaves were analyzed via gas chromatography to examine the make-up of the tomato's chemical defenses. The growth rates of M. sexta were recorded to examine the effects of tomato's chemical defenses after feeding the insects the induced tomato leaves. Measuring trichome density quantifies the change in physical defenses. The addition of the AMF increased the plant defenses, both the number of trichomes present on the plants and the quantity of the chemical defenses. Insects feeding on plants with increased defenses were also shown to have decreased growth. This study shows an alternative strategy for the use of commercial pesticides, lessening the impact of tomato crops on the ecosystem.
Faculty Advisor: Schramm, Katharina (Science, Botany)
Tomatoes (Solanum lycopersicum) are an important agricultural crop around the world. In order to produce food with less impact on the environment, many researchers are looking to utilize natural systems to maximize production with minimal inputs. Maximizing tomato defenses is one possible way to increase productivity. Tomatoes produce both physical and chemical defenses in response to the stress. Increasing the number of trichomes on the plant is a physical means of deterring insects from eating the plant. Trichomes come in two forms glandular trichomes and non-glandular trichomes. The non-glandular trichomes are strictly a physical defense while the glandular trichomes produce chemical defensive compounds against a wide variety of insects. Most land plants can form a symbiosis with arbuscular mycorrhizal fungi (AMF). This symbiosis has been shown to increase nutrient supply, decrease drought stress, and prime plant defenses, all increasing a plant's ability to withstand herbivory stress better. This study examines the response of trichomes and the number of chemical defenses after insect herbivory has occurred with the additional support of the AMF symbiosis. Treatments were exposed to Manduca sexta to stimulate trichome and chemical production. The extracted leaves were analyzed via gas chromatography to examine the make-up of the tomato's chemical defenses. The growth rates of M. sexta were recorded to examine the effects of tomato's chemical defenses after feeding the insects the induced tomato leaves. Measuring trichome density quantifies the change in physical defenses. The addition of the AMF increased the plant defenses, both the number of trichomes present on the plants and the quantity of the chemical defenses. Insects feeding on plants with increased defenses were also shown to have decreased growth. This study shows an alternative strategy for the use of commercial pesticides, lessening the impact of tomato crops on the ecosystem.
Electrohydraulic Shockwaves as a Possible Treatment for Bacterial Biofilms
Brunetti, Bryce; Escarate, Ashley; Conway, Matthew; Slezak, Cyrill; Kopp, Olga (Utah Valley University)
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology); Slezak, Cyrill (Utah Valley University, Physics)
Purpose:
This study evaluates the effect of electrohydraulic shockwaves on Staphylococcus aureus biofilms. This system could be a great alternative to the use of antibiotics, and potentially life-saving technology that could save billions of dollars.
Background:
The rise of antibiotic-resistant bacteria is a global threat. Staphylococcus aureus is typically harmless, but this gram-positive species has become highly resistant and extremely pathogenic. Strains like MRSA and VRSA have the highest rate of drug resistance and are the leading cause of chronic bacterial infections via bacterial biofilms on medical devices. Biofilms are an aggregation of microbes that excrete an extracellular matrix providing an ideal environment for gene exchange and quorum sensing. Their complexity hinders the diffusion of antimicrobials. A proposed method to prevent device-associated infection is shockwave sterilization and therapy. A shockwave is a high-energy wave causing a sudden change in temperature, pressure and density in the medium. This study investigates the potential disruption of bacterial biofilms by electrohydraulic shockwaves.
Methods:
E. coli and S. aureus biofilms were grown on polystyrene plates. Biofilms were treated with shockwaves (0.19mJ/mm2, 300 pulses, 3 Hz) in a water bath and compared with those treated with Vancomycin. Cell viability was determined through XTT/menadione absorbance and specific biofilm formation through crystal violet absorbance.
Results:
Current testing has shown that electrohydraulic shockwaves have a bacteriostatic effect on biofilms. Other finding show potential for shockwaves to increase bacterial susceptibility to lower levels of antibiotics.
Conclusions:
Device-associated infections are a serious threat to patients' health. The diminishing effectiveness of antibiotics in treating and preventing infections along with evolution of mass resistance in bacteria have given rise to the term "post-antibiotic era." The better understanding of electrohydraulic shockwaves bacteriostatic effect could lead to more effective treatments for antibiotic resistant bacteria such as S. aureus.
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology); Slezak, Cyrill (Utah Valley University, Physics)
Purpose:
This study evaluates the effect of electrohydraulic shockwaves on Staphylococcus aureus biofilms. This system could be a great alternative to the use of antibiotics, and potentially life-saving technology that could save billions of dollars.
Background:
The rise of antibiotic-resistant bacteria is a global threat. Staphylococcus aureus is typically harmless, but this gram-positive species has become highly resistant and extremely pathogenic. Strains like MRSA and VRSA have the highest rate of drug resistance and are the leading cause of chronic bacterial infections via bacterial biofilms on medical devices. Biofilms are an aggregation of microbes that excrete an extracellular matrix providing an ideal environment for gene exchange and quorum sensing. Their complexity hinders the diffusion of antimicrobials. A proposed method to prevent device-associated infection is shockwave sterilization and therapy. A shockwave is a high-energy wave causing a sudden change in temperature, pressure and density in the medium. This study investigates the potential disruption of bacterial biofilms by electrohydraulic shockwaves.
Methods:
E. coli and S. aureus biofilms were grown on polystyrene plates. Biofilms were treated with shockwaves (0.19mJ/mm2, 300 pulses, 3 Hz) in a water bath and compared with those treated with Vancomycin. Cell viability was determined through XTT/menadione absorbance and specific biofilm formation through crystal violet absorbance.
Results:
Current testing has shown that electrohydraulic shockwaves have a bacteriostatic effect on biofilms. Other finding show potential for shockwaves to increase bacterial susceptibility to lower levels of antibiotics.
Conclusions:
Device-associated infections are a serious threat to patients' health. The diminishing effectiveness of antibiotics in treating and preventing infections along with evolution of mass resistance in bacteria have given rise to the term "post-antibiotic era." The better understanding of electrohydraulic shockwaves bacteriostatic effect could lead to more effective treatments for antibiotic resistant bacteria such as S. aureus.
Effects of Exercise on Subjective Feelings of Fatigue: A Randomized Crossover Study
Savage, Ryland; Stevens, Andrew; Hicks, Joseph (Brigham Young University)
Faculty Advisor: Bailey, Bruce (Brigham Young University, Exercise Science)
Purpose: This study examined the effect of different intensities of exercise on subjective feelings of fatigue both immediately following exercise and after 60 minutes of intense cognitive testing in men and woman.
Methods: These results come from a secondary analysis of a larger randomized crossover study examining the different levels of exercise intensity on cognitive performance. We recruited 228 men and woman to participate in the study. Each participant was tested after three exercise conditions: moderate (35% VO2 max), vigorous (70% VO2 max), and sedentary (no exercise). Each exercise condition was randomized and each test was separated by one week. After each 40-minute exercise condition, we administered a visual analog scale to evaluate subjective feelings of fatigue. This assessment of fatigue was delivered before treatment, immediately after treatment and then again after a variety of cognitive tasks were performed for the duration of one hour.
Results: For the sedentary condition, feelings of fatigue increased after the 40-minute intervention (p<0.001) and decreased after 60 minutes of cognitive work but remaining elevated compared to baseline (p<0.01). For the moderate condition, feelings of fatigue did not change immediately after exercise (p=0.063) but then increased after 60 minutes of cognitive work (p<0.001). For vigorous condition, feelings of fatigue increased after exercise(p<0.001) and remained elevated after performing the cognitive tasks (p=0.163). Following the 40-minute intervention, fatigue was highest after the sedentary condition, followed by the vigorous condition and did not change after completion of the moderate condition. Following 60 minutes of cognitive work, fatigue was highest in the vigorous condition (p<0.001) and did not differ between the sedentary and moderate conditions (p=0.063).
Conclusion: The results of this study show that moderate exercise has no significant impact on feelings of fatigue but exercise at a higher intensity and being sedentary increase fatigue. Doing cognitive work increases fatigue if exercise precedes the tasks but being sedentary beforehand decreases feelings of fatigue during cognitive tasks. This suggests cognitive work is usually draining, but if performed after sedentary or low activity states such as watching TV, cognitive work will be invigorating and help feel more alert.
Faculty Advisor: Bailey, Bruce (Brigham Young University, Exercise Science)
Purpose: This study examined the effect of different intensities of exercise on subjective feelings of fatigue both immediately following exercise and after 60 minutes of intense cognitive testing in men and woman.
Methods: These results come from a secondary analysis of a larger randomized crossover study examining the different levels of exercise intensity on cognitive performance. We recruited 228 men and woman to participate in the study. Each participant was tested after three exercise conditions: moderate (35% VO2 max), vigorous (70% VO2 max), and sedentary (no exercise). Each exercise condition was randomized and each test was separated by one week. After each 40-minute exercise condition, we administered a visual analog scale to evaluate subjective feelings of fatigue. This assessment of fatigue was delivered before treatment, immediately after treatment and then again after a variety of cognitive tasks were performed for the duration of one hour.
Results: For the sedentary condition, feelings of fatigue increased after the 40-minute intervention (p<0.001) and decreased after 60 minutes of cognitive work but remaining elevated compared to baseline (p<0.01). For the moderate condition, feelings of fatigue did not change immediately after exercise (p=0.063) but then increased after 60 minutes of cognitive work (p<0.001). For vigorous condition, feelings of fatigue increased after exercise(p<0.001) and remained elevated after performing the cognitive tasks (p=0.163). Following the 40-minute intervention, fatigue was highest after the sedentary condition, followed by the vigorous condition and did not change after completion of the moderate condition. Following 60 minutes of cognitive work, fatigue was highest in the vigorous condition (p<0.001) and did not differ between the sedentary and moderate conditions (p=0.063).
Conclusion: The results of this study show that moderate exercise has no significant impact on feelings of fatigue but exercise at a higher intensity and being sedentary increase fatigue. Doing cognitive work increases fatigue if exercise precedes the tasks but being sedentary beforehand decreases feelings of fatigue during cognitive tasks. This suggests cognitive work is usually draining, but if performed after sedentary or low activity states such as watching TV, cognitive work will be invigorating and help feel more alert.
Evaluating the Knock Out Effects of Nr4a1 and Nr4a3 on Delta cells of the Pancreas
Hess, Kavan; Herring, Jacob; Yang, Haokun; Tessem, Jeff (Brigham Young University)
Faculty Advisor: Tessem, Jeff (Brigham Young University; Department of Nutrition, Dietetics, and Food Science)
Diabetes is the seventh leading cause of death in the United States, and often accompanies other life-threating complications. There are two main types of diabetes that are both characterized by disfunction or destruction of insulin producing beta cells found in the islets of Langerhans. Islets of Langerhans are composed of endocrine hormone secreting cells, including alpha cells (glucagon), beta cells (insulin) delta cells (somatostatin), epsilon cells (ghrelin) and PP cells (pancreatic polypeptide). While alpha and beta cells make up ~90% of all the cells in the islet, delta cells comprise only ~10% and are responsible for cross talk in the islet. Delta cells regulate intra-islet cross talk through the secretion of somatostatin-14. It has been shown that Nr4a1 and Nr4a3 overexpression induces beta cell proliferation, while Nr4a1 or Nr4a3 deletion inhibits insulin secretion when challenged with glucose. Delta cells contain three times the amount of Nr4a1 mRNA than beta cells. However, no research has been done on the role of either of these transcription factors in the cross talk between the different cell types of the islet. Here we aim to show how a lack of Nr4a1 and Nr4a3 affects delta cell somatostatin release when challenged with glucose.
Faculty Advisor: Tessem, Jeff (Brigham Young University; Department of Nutrition, Dietetics, and Food Science)
Diabetes is the seventh leading cause of death in the United States, and often accompanies other life-threating complications. There are two main types of diabetes that are both characterized by disfunction or destruction of insulin producing beta cells found in the islets of Langerhans. Islets of Langerhans are composed of endocrine hormone secreting cells, including alpha cells (glucagon), beta cells (insulin) delta cells (somatostatin), epsilon cells (ghrelin) and PP cells (pancreatic polypeptide). While alpha and beta cells make up ~90% of all the cells in the islet, delta cells comprise only ~10% and are responsible for cross talk in the islet. Delta cells regulate intra-islet cross talk through the secretion of somatostatin-14. It has been shown that Nr4a1 and Nr4a3 overexpression induces beta cell proliferation, while Nr4a1 or Nr4a3 deletion inhibits insulin secretion when challenged with glucose. Delta cells contain three times the amount of Nr4a1 mRNA than beta cells. However, no research has been done on the role of either of these transcription factors in the cross talk between the different cell types of the islet. Here we aim to show how a lack of Nr4a1 and Nr4a3 affects delta cell somatostatin release when challenged with glucose.
Examining the Trafficking of Normal and TYRP1 Ash-Red Proteins' in Melanocytes
Domyan, Eric; Godoy, Daniela; Gardiner, Kylan (Utah Valley University)
Faculty Advisor: Domyan, Eric (Science, Biology)
Pigmentation is one of the main traits we notice when we look at something, whether it be a flower, an animal, or another human. Variation in pigmentation arises when random mutations affect the function of a gene involved in pigmentation production. In this research our goal is to understand a specific mutation that happens in Pigeons which involves the TYRP1 gene.
The TYRP1 gene instructs the making of the tyrosinase-related protein. This enzyme is located in melanocytes, which are cells that produce melanin. Studies suggest that this enzyme may help stabilize tyrosinase, which is responsible for the first step in melanin production.
TYRP1 has a signal peptide which directs the protein to the ER (endoplasmic reticulum) where the signal peptide is removed before the mature protein is trafficked to melanosomes to perform its normal function. The Ash-red mutation, however, prevents removal of the signal peptide, which somehow results in pheomelanin synthesis (red) instead of eumelanin synthesis (dark blue). These findings suggest that the Ash-red mutation is causing the TYRP1 protein to perform a new function. The goal of this project is to better understand the synthesis and trafficking of TYRP1 throughout the cell organelles.
To study this, we plan to use transgenesis to express normal, or Ash-red versions of TYRP1 protein in melanocytes, label the different intracellular compartments using an immunostain, and determine whether normal or Ash-red TYRP1 proteins are being sent to the same, or different compartments of the cell.
Faculty Advisor: Domyan, Eric (Science, Biology)
Pigmentation is one of the main traits we notice when we look at something, whether it be a flower, an animal, or another human. Variation in pigmentation arises when random mutations affect the function of a gene involved in pigmentation production. In this research our goal is to understand a specific mutation that happens in Pigeons which involves the TYRP1 gene.
The TYRP1 gene instructs the making of the tyrosinase-related protein. This enzyme is located in melanocytes, which are cells that produce melanin. Studies suggest that this enzyme may help stabilize tyrosinase, which is responsible for the first step in melanin production.
TYRP1 has a signal peptide which directs the protein to the ER (endoplasmic reticulum) where the signal peptide is removed before the mature protein is trafficked to melanosomes to perform its normal function. The Ash-red mutation, however, prevents removal of the signal peptide, which somehow results in pheomelanin synthesis (red) instead of eumelanin synthesis (dark blue). These findings suggest that the Ash-red mutation is causing the TYRP1 protein to perform a new function. The goal of this project is to better understand the synthesis and trafficking of TYRP1 throughout the cell organelles.
To study this, we plan to use transgenesis to express normal, or Ash-red versions of TYRP1 protein in melanocytes, label the different intracellular compartments using an immunostain, and determine whether normal or Ash-red TYRP1 proteins are being sent to the same, or different compartments of the cell.
Effects of Parenting Styles on Child Delinquency and Bullying
Leavitt, Amanda; Harris, Heather; Szanter, Kathryn; Stokes, Alexis (Weber State University)
Faculty Advisor: Dunn, Charles (Weber State University, Child and Family Studies); Hubler, Daniel (Weber State University, Child and Family Studies); Osai, Keith (Weber State University, Child and Family Studies)
How a child is parented can influence their success socially, mentally, physically, and emotionally. Agarwal (2017) states that "Parenting plays the most important part in any child's life, it can help a child by teaching them to deal with people, situations and adapt to a better living standard" (p.1335). Parents help form worldviews, shape a child's attitude towards personal achievement, teach how to approach adversity in life, and satisfy their needs whether it be psychological and/or physiological. Much is known about how the base of one's childhood is built upon one's primary caregivers' parenting style, however, less is known about the intergenerational transmission of parenting philosophies.
The current study was designed to assess how parenting styles affect child delinquency and bullying. Through social media invitations and snowball sampling, 200 people accepted invitations to participate in an online survey incorporating both open-ended and quantitative items. Participants were asked to reflect on how they were parented and then state what they have maintained or changed in their own parenting styles. Considering the rich nature of the responses to the qualitative items, a corroborative narrative came together through personal stories.
Through thematic analysis, several themes emerged. When participants reported being parented with: open communication, flexibility, love, trust, and higher expectations they wanted to keep those traits. When asked what participants carried on from their parents one participant stated, "Cherish the family and the moments we have together." Additionally, 28% (n = 113) of our participants shared the importance of spending quality time together. However, when participants' parents did not display those traits, most participants indicated that similar traits, namely communication, flexibility, love, quality time, trust, and higher expectations were desired in their homes. The findings from the current study can be used to inform future parenting research assessing influence on subsequent generations.
Faculty Advisor: Dunn, Charles (Weber State University, Child and Family Studies); Hubler, Daniel (Weber State University, Child and Family Studies); Osai, Keith (Weber State University, Child and Family Studies)
How a child is parented can influence their success socially, mentally, physically, and emotionally. Agarwal (2017) states that "Parenting plays the most important part in any child's life, it can help a child by teaching them to deal with people, situations and adapt to a better living standard" (p.1335). Parents help form worldviews, shape a child's attitude towards personal achievement, teach how to approach adversity in life, and satisfy their needs whether it be psychological and/or physiological. Much is known about how the base of one's childhood is built upon one's primary caregivers' parenting style, however, less is known about the intergenerational transmission of parenting philosophies.
The current study was designed to assess how parenting styles affect child delinquency and bullying. Through social media invitations and snowball sampling, 200 people accepted invitations to participate in an online survey incorporating both open-ended and quantitative items. Participants were asked to reflect on how they were parented and then state what they have maintained or changed in their own parenting styles. Considering the rich nature of the responses to the qualitative items, a corroborative narrative came together through personal stories.
Through thematic analysis, several themes emerged. When participants reported being parented with: open communication, flexibility, love, trust, and higher expectations they wanted to keep those traits. When asked what participants carried on from their parents one participant stated, "Cherish the family and the moments we have together." Additionally, 28% (n = 113) of our participants shared the importance of spending quality time together. However, when participants' parents did not display those traits, most participants indicated that similar traits, namely communication, flexibility, love, quality time, trust, and higher expectations were desired in their homes. The findings from the current study can be used to inform future parenting research assessing influence on subsequent generations.
Differences in anthropometric characteristics between intermediate and high level climbers
Miriam Reber; Ethan Del Toro; Holden Lyman; Weston Hargis; Travis Ficklin (Dixie State University)
Faculty Advisor: Ficklin, Travis (Dixie State University, Health and Human Performance)
In recent years sport climbing has increased in popularity, and with that have come questions regarding what factors improve performance. While multiple studies have examined the effect of anthropometric factors on injury in climbing, few have examined their relationship with climbing performance. Therefore, the purpose of this study is to compare the anthropometric measures of high-level to intermediate level climbers to determine if certain body types, or dimensions, lend themselves to better performance in climbing.
Twenty-one subjects were divided into an intermediate (INT) or high-level (HL) group based on highest graded climb achieved. Various measures including height, limb lengths, and experience. These were compared between the two groups using t-tests.
Significant differences were found in the number of days climbed per week (HL 2.9±1.1, INT 2.17±0.6, p = 0.03), and the ratios of upper arm to full arm length (HL 0.44±0.05, INT 0.41±0.02, p=0.03), and trunk to height (HL 0.26±0.03, INT 0.23±0.01, p=0.01). There were also statistical trends towards years of experience (HL 5.46±3.8, INT 3.39±1.8, p = 0.06), and trunk length (HL 44.96±7.1cm, INT 41.28±2.7cm, p=0.06).
The current data suggest that for the levels of climbing examined (5.9-5.13b on the Yosemite Decimal Scale) a greater trunk to height ratio, greater upper arm to arm length ratio, and increased frequency of practice (a variable within the climber's control) may positively impact climbing ability.
Faculty Advisor: Ficklin, Travis (Dixie State University, Health and Human Performance)
In recent years sport climbing has increased in popularity, and with that have come questions regarding what factors improve performance. While multiple studies have examined the effect of anthropometric factors on injury in climbing, few have examined their relationship with climbing performance. Therefore, the purpose of this study is to compare the anthropometric measures of high-level to intermediate level climbers to determine if certain body types, or dimensions, lend themselves to better performance in climbing.
Twenty-one subjects were divided into an intermediate (INT) or high-level (HL) group based on highest graded climb achieved. Various measures including height, limb lengths, and experience. These were compared between the two groups using t-tests.
Significant differences were found in the number of days climbed per week (HL 2.9±1.1, INT 2.17±0.6, p = 0.03), and the ratios of upper arm to full arm length (HL 0.44±0.05, INT 0.41±0.02, p=0.03), and trunk to height (HL 0.26±0.03, INT 0.23±0.01, p=0.01). There were also statistical trends towards years of experience (HL 5.46±3.8, INT 3.39±1.8, p = 0.06), and trunk length (HL 44.96±7.1cm, INT 41.28±2.7cm, p=0.06).
The current data suggest that for the levels of climbing examined (5.9-5.13b on the Yosemite Decimal Scale) a greater trunk to height ratio, greater upper arm to arm length ratio, and increased frequency of practice (a variable within the climber's control) may positively impact climbing ability.
Dopamine 2 receptors display rapid adaptation in response to acute ethanol administration
LeBaron, Josh; Obray, J Daniel; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences, Psychology)
Dopamine neurons in the substancia nigra (SN) and ventral tegmental area (VTA) are inhibited by dopamine (DA) via dopamine 2 receptor (D2R) activation. D2R expression in the striatum is a well-known biomarker for brain DA levels, drug abuse, and dependence. Markers of D2R expression are not only detectable in the brain but are also expressed in peripheral tissues, including the blood, where DA appears to play a pivotal role in mediating communication between the nervous and immune systems. Alteration in lympocytic D2Rs are seen in chronic psychostimulant use (Ersche et al., 2011). For the last two decades it has been generally accepted that D2R expression in the striatum is reduced by chronic ethanol use. Additionally, research has suggested that these changes mirror changes in DA levels in the striatum and predict risk of relapse. Despite this, the timecourse over which these changes occur has not been demonstrated. Further, recent research has challenged both the reduction in D2R expression produced by chronic ethanol and the mechanism whereby it was believed to be produced (reductions in striatal DA levels). This research has suggested that alterations in D2R levels may be due to disruption of sleep in individuals with substance use disorders. Here we demonstrate that dopamine 2 receptor expression in the brain and the blood follows brain and blood dopamine levels on a timescale of minutes to hours following an acute dose of ethanol. This research provides evidence for transient changes in D2R expression following a single dose of ethanol.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences, Psychology)
Dopamine neurons in the substancia nigra (SN) and ventral tegmental area (VTA) are inhibited by dopamine (DA) via dopamine 2 receptor (D2R) activation. D2R expression in the striatum is a well-known biomarker for brain DA levels, drug abuse, and dependence. Markers of D2R expression are not only detectable in the brain but are also expressed in peripheral tissues, including the blood, where DA appears to play a pivotal role in mediating communication between the nervous and immune systems. Alteration in lympocytic D2Rs are seen in chronic psychostimulant use (Ersche et al., 2011). For the last two decades it has been generally accepted that D2R expression in the striatum is reduced by chronic ethanol use. Additionally, research has suggested that these changes mirror changes in DA levels in the striatum and predict risk of relapse. Despite this, the timecourse over which these changes occur has not been demonstrated. Further, recent research has challenged both the reduction in D2R expression produced by chronic ethanol and the mechanism whereby it was believed to be produced (reductions in striatal DA levels). This research has suggested that alterations in D2R levels may be due to disruption of sleep in individuals with substance use disorders. Here we demonstrate that dopamine 2 receptor expression in the brain and the blood follows brain and blood dopamine levels on a timescale of minutes to hours following an acute dose of ethanol. This research provides evidence for transient changes in D2R expression following a single dose of ethanol.
Effect of Gender on Shear Wave Elastography and Cross Sectional Area of the Gastrocnemius in Senior Athletes.
Hutchison, Cortland; Preece, J. Caleb; Seibold, Tanner; Feland, J. Brent (Brigham Young University)
Faculty Advisor: Feland, Brent (Life Sciences, Exercise Science)
BACKGROUND: Muscle morphology changes with age and the fibrous/fatty infiltration should affect the overall stiffness of aging muscle. However, little info intrinsic stiffness as measured by SWE is available on aging muscle and the effect of gender using this measurement is not clear.
PURPOSE: This study aimed to determine how gender affects cross sectional area (CSA) and SWE of both the medial and lateral heads of the gastrocnemius muscle of the dominant leg in active older athletes participating in the HuntsmanWorld Senior Games.
METHODS: Data was collected from 116 volunteers (participants in the Huntsman World Senior Games) in St. George, Utah, 2019. Subjects (62 males: mean age = 68.9 ± 7.8 yrs, Ht = 177.4 ± 8.1 cm, Wt= 85.4±20 kg; 54 females: mean age = 66.9 ± 8.5 yrs, Ht =164.4 ± 7.2 cm, Wt= 69.8±18 kg) signed an approved consent form and then lay prone on a treatment table for ultrasonic measurement of both CSA and SWE of both heads of the gastrocnemius.
ANALYSIS: All data were analyzed a generalized linear model analysis using SPSS ver25 comparing CSA and SWE of both medial and lateral gastrocnemius heads by gender with age, height and weight as covariates.
RESULTS & CONCLUSION: We hypothesized that gender would affect both CSA and SWE with males exhibiting increases in both CSA and SWE values. We found a significant difference in CSA of both the medial and lateral gastroc (p=.000 for both) between genders, but no significant difference in lateral gastroc SWE (p=.337) or medial gastroc SWE (p=.320). Both age (p=.004) and weight (p=.000) were also found to significantly affect CSA and SWE between genders. The larger CSA values for men in this study may be more of a function of overall body size and weight since both populations are active athletes.
Faculty Advisor: Feland, Brent (Life Sciences, Exercise Science)
BACKGROUND: Muscle morphology changes with age and the fibrous/fatty infiltration should affect the overall stiffness of aging muscle. However, little info intrinsic stiffness as measured by SWE is available on aging muscle and the effect of gender using this measurement is not clear.
PURPOSE: This study aimed to determine how gender affects cross sectional area (CSA) and SWE of both the medial and lateral heads of the gastrocnemius muscle of the dominant leg in active older athletes participating in the HuntsmanWorld Senior Games.
METHODS: Data was collected from 116 volunteers (participants in the Huntsman World Senior Games) in St. George, Utah, 2019. Subjects (62 males: mean age = 68.9 ± 7.8 yrs, Ht = 177.4 ± 8.1 cm, Wt= 85.4±20 kg; 54 females: mean age = 66.9 ± 8.5 yrs, Ht =164.4 ± 7.2 cm, Wt= 69.8±18 kg) signed an approved consent form and then lay prone on a treatment table for ultrasonic measurement of both CSA and SWE of both heads of the gastrocnemius.
ANALYSIS: All data were analyzed a generalized linear model analysis using SPSS ver25 comparing CSA and SWE of both medial and lateral gastrocnemius heads by gender with age, height and weight as covariates.
RESULTS & CONCLUSION: We hypothesized that gender would affect both CSA and SWE with males exhibiting increases in both CSA and SWE values. We found a significant difference in CSA of both the medial and lateral gastroc (p=.000 for both) between genders, but no significant difference in lateral gastroc SWE (p=.337) or medial gastroc SWE (p=.320). Both age (p=.004) and weight (p=.000) were also found to significantly affect CSA and SWE between genders. The larger CSA values for men in this study may be more of a function of overall body size and weight since both populations are active athletes.
Effects of Flavanols on β-cell proliferation.
Tessem, Jeffery; Lloyd, Trevor; Brown, Nathan (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics and Food Science)
Diabetes is a global epidemic affecting millions of people. The total estimated cost of diabetes in the U.S. during 2017 was 327 billion dollars [1]. Diabetes is characterized by the loss of pancreatic β-cell function which is caused by an autoimmune disorder in Type 1 diabetes or insulin resistance and β-cell exhaustion in Type 2 (T2D) diabetes. Lifestyle changes in diet are beneficial in treating T2D. Phytochemicals are commonly utilized in these diets, and recent studies show diets high in flavanols exert beneficial bioactivity for β-cells. While flavanols demonstrate beneficial effects on β-cells, these flavanols are rarely observed in circulation, suggesting a necessary intermediate step. Flavanols are metabolized by gut bacteria to smaller metabolites that are absorbable. We hypothesize that these gut bacteria derived flavanol metabolites cross the gut and affect β-cell function. We have fed rats catechin supplemented or unsupplemented diets and collected urine as a means to isolate all absorbable gut flavanol metabolites. Here we present the effects of these absorbed metabolites on β-cell proliferation. This study begins to explain the mechanism by which flavanols exert their beneficial effect on glucose metabolism through the β-cell.
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics and Food Science)
Diabetes is a global epidemic affecting millions of people. The total estimated cost of diabetes in the U.S. during 2017 was 327 billion dollars [1]. Diabetes is characterized by the loss of pancreatic β-cell function which is caused by an autoimmune disorder in Type 1 diabetes or insulin resistance and β-cell exhaustion in Type 2 (T2D) diabetes. Lifestyle changes in diet are beneficial in treating T2D. Phytochemicals are commonly utilized in these diets, and recent studies show diets high in flavanols exert beneficial bioactivity for β-cells. While flavanols demonstrate beneficial effects on β-cells, these flavanols are rarely observed in circulation, suggesting a necessary intermediate step. Flavanols are metabolized by gut bacteria to smaller metabolites that are absorbable. We hypothesize that these gut bacteria derived flavanol metabolites cross the gut and affect β-cell function. We have fed rats catechin supplemented or unsupplemented diets and collected urine as a means to isolate all absorbable gut flavanol metabolites. Here we present the effects of these absorbed metabolites on β-cell proliferation. This study begins to explain the mechanism by which flavanols exert their beneficial effect on glucose metabolism through the β-cell.
Engineered bacterial tight junctions: a high-throughput method to characterize claudins and identify epithelial modulators
Rollins, Jay; Whitney, Jordan; Hope, Sandra; Mizrachi, Dario (Brigham Young University)
Faculty Advisor: Mizrachi, Dario (Brigham Young University, Physiology and Developmental Biology)
Epithelial and endothelial tissues form selectively permeable barriers, with the permeability largely controlled by intercellular tight junctions. Claudin (CLDN) proteins are critical components of these tight junctions, making them the gatekeepers that control the paracellular space in multicellular organisms. CLDN proteins are thus targets for studies on epithelial and endothelial absorption, to therefore learn how to regulate them for potential drug delivery or therapeutics. CLDN characterization is still in progress. Previously, the relative strength of each member of the CLDN family was unknown. Additionally, no high-throughput method to study absorption enhancers or inhibitors had been found.
Through CLDN expression in Escherichia coli, we determined the relative strength of each CLDN protein and confirmed the effects of various absorption enhancers from previous studies. Therefore, we propose that CLDN expression in Escherichia coli is a valid model for the study of tight junctions and that, through flow cytometry, it is a high-throughput method for interrogating large libraries of potential drug delivery compounds. Using CLDN 2 because of its role in cancer-metastasis prevention and its measured sensitivity towards epithelial modulators, we studied a fifty thousand compound library (DIVERSet-CL Library) to identify absorption moderators, drug delivery compounds, and possible cancer-metastasis prevention.
Faculty Advisor: Mizrachi, Dario (Brigham Young University, Physiology and Developmental Biology)
Epithelial and endothelial tissues form selectively permeable barriers, with the permeability largely controlled by intercellular tight junctions. Claudin (CLDN) proteins are critical components of these tight junctions, making them the gatekeepers that control the paracellular space in multicellular organisms. CLDN proteins are thus targets for studies on epithelial and endothelial absorption, to therefore learn how to regulate them for potential drug delivery or therapeutics. CLDN characterization is still in progress. Previously, the relative strength of each member of the CLDN family was unknown. Additionally, no high-throughput method to study absorption enhancers or inhibitors had been found.
Through CLDN expression in Escherichia coli, we determined the relative strength of each CLDN protein and confirmed the effects of various absorption enhancers from previous studies. Therefore, we propose that CLDN expression in Escherichia coli is a valid model for the study of tight junctions and that, through flow cytometry, it is a high-throughput method for interrogating large libraries of potential drug delivery compounds. Using CLDN 2 because of its role in cancer-metastasis prevention and its measured sensitivity towards epithelial modulators, we studied a fifty thousand compound library (DIVERSet-CL Library) to identify absorption moderators, drug delivery compounds, and possible cancer-metastasis prevention.
Effects of Grape Seed Extract Metabolites on ß-cell Proliferation and Function
Beales, Joseph; Lloyd, Trevor; Krueger, Emily; Barlow, Andrew (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutritional, Dietetics, and Food Science)
Worldwide, an estimated 415 million people suffer from diabetes.1 Diabetes is characterized by chronic dysfunction of the pancreatic ß-cell, which leads to unregulated insulin secretion and abnormal blood glucose levels. Therefore, methods which increase the number of ß-cells or improve their function have potential for complementary treatment of type 2 diabetes. Compounds such as antioxidants and their gut metabolites have received attention in literature as having potential ß-cell-regulating properties.2,3 Therefore, we hypothesize that supplementation of grape seed extract (GSE), which is rich in antioxidants, will enhance ß-cell proliferation and insulin secretion. Accordingly, we obtained metabolites, derived from rats on either a control or grape seed extract diet, to measure the metabolites' impact on ß-cell function through in vitro assays such as glucose stimulated insulin secretion (GSIS) and 3H-thymidine incorporation. Discoveries regarding GSE metabolites' effects on ß-cell function could be fundamental to understanding ß-cell regulation and potential pharmaceutical or dietary treatments for diabetes.
1 Ogurtsova, K., et al. "IDF Diabetes Atlas: Global Estimates for the Prevalence of Diabetes for 2015 and 2040." Diabetes Research and Clinical Practice, Elsevier, 31 Mar. 2017, www.sciencedirect.com/science/article/pii/S0168822717303753?via%3Dihub.
2 Bajaj, Sarita, and Afreen Khan. "Antioxidants and diabetes." Indian journal of endocrinology and metabolism vol. 16,Suppl 2 (2012): S267-71. Doi:10.4103/2230-8210.104057
3 Tsuda, Takanori. "Recent Progress in Anti-Obesity and Anti-Diabetes Effect of Berries." MDPI, Multidisciplinary Digital Publishing Institute, 6 Apr. 2016, www.mdpi.com/2076-3921/5/2/13.
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutritional, Dietetics, and Food Science)
Worldwide, an estimated 415 million people suffer from diabetes.1 Diabetes is characterized by chronic dysfunction of the pancreatic ß-cell, which leads to unregulated insulin secretion and abnormal blood glucose levels. Therefore, methods which increase the number of ß-cells or improve their function have potential for complementary treatment of type 2 diabetes. Compounds such as antioxidants and their gut metabolites have received attention in literature as having potential ß-cell-regulating properties.2,3 Therefore, we hypothesize that supplementation of grape seed extract (GSE), which is rich in antioxidants, will enhance ß-cell proliferation and insulin secretion. Accordingly, we obtained metabolites, derived from rats on either a control or grape seed extract diet, to measure the metabolites' impact on ß-cell function through in vitro assays such as glucose stimulated insulin secretion (GSIS) and 3H-thymidine incorporation. Discoveries regarding GSE metabolites' effects on ß-cell function could be fundamental to understanding ß-cell regulation and potential pharmaceutical or dietary treatments for diabetes.
1 Ogurtsova, K., et al. "IDF Diabetes Atlas: Global Estimates for the Prevalence of Diabetes for 2015 and 2040." Diabetes Research and Clinical Practice, Elsevier, 31 Mar. 2017, www.sciencedirect.com/science/article/pii/S0168822717303753?via%3Dihub.
2 Bajaj, Sarita, and Afreen Khan. "Antioxidants and diabetes." Indian journal of endocrinology and metabolism vol. 16,Suppl 2 (2012): S267-71. Doi:10.4103/2230-8210.104057
3 Tsuda, Takanori. "Recent Progress in Anti-Obesity and Anti-Diabetes Effect of Berries." MDPI, Multidisciplinary Digital Publishing Institute, 6 Apr. 2016, www.mdpi.com/2076-3921/5/2/13.
Evaluating the use of Drones for Yield Estimates, Disease Detection, and Other Problems in Agriculture
Nischwitz, Claudia; Compton, Tyson (Utah State University)
Faculty Advisor: Nischwitz, Claudia (College of Science, Biology Department)
This research evaluates the use of Unmanned Aerial Vehicles (UAVs) in agricultural applications. We center our research on early disease detection and yield estimation in vegetable crops using aerial imagery and computer software. Previous research on UAV use in agriculture has addressed topics such as soil and field analysis (Long, 2017), Precision Viticulture in Italy (Matese, et al., 2015), and other areas pertinent to agriculturists. Our research builds on previous studies and aims to provide Utah farmers with knowledge and tools to increase agricultural productivity. A DJI Inspire drone is used with both a traditional light camera and a Near-Infrared (NIR) camera. Normal and NIR images are taken at the USU Research Farm in Kaysville Utah, and over local farm fields in Utah throughout the growing season. Unhealthy plants, identified from the aerial images, are tested at the USU Plant Pathology lab to identify diseases. Computer software (ImageJ, Microsoft ICE, and MATLAB) is used to process the images and collect crop health and yield estimate data. At the end of the growing season, the yield for each crop is measured and correlated to the aerial image data to create a predictive model for yield. Some plant diseases including Beet curly top virus in tomato and powdery mildew in squash are readily identified. We find that yield estimation with aerial imagery works well for specific crops. Potato yield was correlated with plant size at different numbers of days after planting. Further tests in coming years will provide validation for these results. Our current data show that the use of an UAV can be a valuable tool for early disease detection and yield estimation in vegetable crops.
Faculty Advisor: Nischwitz, Claudia (College of Science, Biology Department)
This research evaluates the use of Unmanned Aerial Vehicles (UAVs) in agricultural applications. We center our research on early disease detection and yield estimation in vegetable crops using aerial imagery and computer software. Previous research on UAV use in agriculture has addressed topics such as soil and field analysis (Long, 2017), Precision Viticulture in Italy (Matese, et al., 2015), and other areas pertinent to agriculturists. Our research builds on previous studies and aims to provide Utah farmers with knowledge and tools to increase agricultural productivity. A DJI Inspire drone is used with both a traditional light camera and a Near-Infrared (NIR) camera. Normal and NIR images are taken at the USU Research Farm in Kaysville Utah, and over local farm fields in Utah throughout the growing season. Unhealthy plants, identified from the aerial images, are tested at the USU Plant Pathology lab to identify diseases. Computer software (ImageJ, Microsoft ICE, and MATLAB) is used to process the images and collect crop health and yield estimate data. At the end of the growing season, the yield for each crop is measured and correlated to the aerial image data to create a predictive model for yield. Some plant diseases including Beet curly top virus in tomato and powdery mildew in squash are readily identified. We find that yield estimation with aerial imagery works well for specific crops. Potato yield was correlated with plant size at different numbers of days after planting. Further tests in coming years will provide validation for these results. Our current data show that the use of an UAV can be a valuable tool for early disease detection and yield estimation in vegetable crops.
Drosophila melanogaster Determines Dietary Preference Through Volatile Detection
Walker, Carson; Burke, Tyler; Tanner, Call; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Brigham Young University, Plant and Wildlife Sciences)
Host-microbe interactions can dramatically influence Drosophila melanogaster phenotypes, but few studies have explained how these microbes are recognized by the host. For example, fruit flies from one area, Maine, prefer to consume diets inoculated with Lactic Acid Bacteria (LABs) over diets containing Acetic acid bacteria (AABs); whereas flies from another area, Florida, show no preference for either LAB or AAB. However, the bacterial mechanisms responsible for this preference are unknown. My follow-up analyses further suggest the hypothesis that Maine flies avoid AAB, rather than are attracted to LAB. Therefore, I propose a forward genetic approach to define the AAB processes that shape this fly preference. I will do this by comparing the feeding preferences of Maine and Florida flies to diets incorporated with LAB versus specific bacterial molecules, such as acetic acid, peptidoglycan, and lipopolysaccharides. Alternatively, if these molecules do not mediate the effects I will perform a metagenome wide association assay (MGWA) to identify genes linked to this preference. Together, these approaches will help to reveal the bacterial factors that influence fly feeding preferences.
Faculty Advisor: Chaston, John (Brigham Young University, Plant and Wildlife Sciences)
Host-microbe interactions can dramatically influence Drosophila melanogaster phenotypes, but few studies have explained how these microbes are recognized by the host. For example, fruit flies from one area, Maine, prefer to consume diets inoculated with Lactic Acid Bacteria (LABs) over diets containing Acetic acid bacteria (AABs); whereas flies from another area, Florida, show no preference for either LAB or AAB. However, the bacterial mechanisms responsible for this preference are unknown. My follow-up analyses further suggest the hypothesis that Maine flies avoid AAB, rather than are attracted to LAB. Therefore, I propose a forward genetic approach to define the AAB processes that shape this fly preference. I will do this by comparing the feeding preferences of Maine and Florida flies to diets incorporated with LAB versus specific bacterial molecules, such as acetic acid, peptidoglycan, and lipopolysaccharides. Alternatively, if these molecules do not mediate the effects I will perform a metagenome wide association assay (MGWA) to identify genes linked to this preference. Together, these approaches will help to reveal the bacterial factors that influence fly feeding preferences.
Age-Dependent Molecular Effects of Cyclin-Dependent Kinase Inhibitors on β-cell Proliferation
Jensen, Daelin; Aitken, Talon; Baxter, Melanie (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
As of 2017, over 400 million people were diagnosed with diabetes mellitus. This is becoming a worldwide concern as the number of people affected by diabetes is growing at an alarming rate. Functional pancreatic β-cell mass is responsible for maintaining healthy blood glucose levels through the production of insulin. A hallmark of both type 1 and type 2 diabetes is a relative or absolute loss of functional β-cell mass and, consequently, decreased insulin production. Two possible approaches for replenishing the β-cells are: 1) replacement through cadaveric donors and 2) regeneration of endogenous β-cells. A major impediment to these approaches is that aged β-cells are refractory to genes that are known to induce proliferation in young β-cells. It is currently unknown why aged β-cells are refractory. . We hypothesized that age-dependent changes to the proliferative capacity of β-cells are influenced by increasing levels of cyclin-dependent kinase inhibitors (CDKI). CDKI's bind to cyclin-dependent kinases, effectively halting the cell cycle and proliferation. Here, we demonstrate the expression of the Ink4 and Cip/Kip families of CDKI's by mRNA and protein expression in five week old and five month old Wistar rat β-cells. Greater understanding of the proliferative mechanisms of the β-cell will allow greater application of the aforementioned treatments.
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
As of 2017, over 400 million people were diagnosed with diabetes mellitus. This is becoming a worldwide concern as the number of people affected by diabetes is growing at an alarming rate. Functional pancreatic β-cell mass is responsible for maintaining healthy blood glucose levels through the production of insulin. A hallmark of both type 1 and type 2 diabetes is a relative or absolute loss of functional β-cell mass and, consequently, decreased insulin production. Two possible approaches for replenishing the β-cells are: 1) replacement through cadaveric donors and 2) regeneration of endogenous β-cells. A major impediment to these approaches is that aged β-cells are refractory to genes that are known to induce proliferation in young β-cells. It is currently unknown why aged β-cells are refractory. . We hypothesized that age-dependent changes to the proliferative capacity of β-cells are influenced by increasing levels of cyclin-dependent kinase inhibitors (CDKI). CDKI's bind to cyclin-dependent kinases, effectively halting the cell cycle and proliferation. Here, we demonstrate the expression of the Ink4 and Cip/Kip families of CDKI's by mRNA and protein expression in five week old and five month old Wistar rat β-cells. Greater understanding of the proliferative mechanisms of the β-cell will allow greater application of the aforementioned treatments.
Anti-Tumor Activity of Chalcone Derivatives
Allen, Brian; Covey, Tracy; Davies, Don; Eccles, Nick; Farnsworth, Brian; Ferguson, Parker; Hart, Sierra; Lowder, Jordan (Weber State University)
Faculty Advisor: Davies, Don (Weber State University, Chemistry and Biochemistry); Covey, Tracy (Weber State University, Chemistry and Biochemsitry)
Chalcones refer to biological molecules with the structure trans 1,3-diphenylprop-2-en-1-one. Biological chalcones and chalcone derivatives display anti-tumor, anti-fungal, anti-inflammatory and antibiotic properties. To understand the role of the chalcone structure in tumor cessation, derivatives to the original chalcone were synthesized using aldol condensation reactions. HeLa and HEK-293 cells were treated with the synthesized chalcone and an LD50, or the concentration of chalcone required to kill half of the cells, was calculated. The LD50 was then used to determine the efficiency of the chalcone derivative. Correlations between the structure and activity suggest that a Michael reaction occurs at the cell and indicate that that an aromatic ring at C3 is likely necessary. Further research will help determine the structures of more cytotoxic compounds.
Faculty Advisor: Davies, Don (Weber State University, Chemistry and Biochemistry); Covey, Tracy (Weber State University, Chemistry and Biochemsitry)
Chalcones refer to biological molecules with the structure trans 1,3-diphenylprop-2-en-1-one. Biological chalcones and chalcone derivatives display anti-tumor, anti-fungal, anti-inflammatory and antibiotic properties. To understand the role of the chalcone structure in tumor cessation, derivatives to the original chalcone were synthesized using aldol condensation reactions. HeLa and HEK-293 cells were treated with the synthesized chalcone and an LD50, or the concentration of chalcone required to kill half of the cells, was calculated. The LD50 was then used to determine the efficiency of the chalcone derivative. Correlations between the structure and activity suggest that a Michael reaction occurs at the cell and indicate that that an aromatic ring at C3 is likely necessary. Further research will help determine the structures of more cytotoxic compounds.
Antimicrobial Activity of Artemisia tridentata
Wasden, Kayla; Suisse, David; Kaundal, Amita (faculty mentor) (Utah State University)
Faculty Advisor: Kaundal, Amita (College of Agriculture and Applied Sciences; Plants, Soils, and Climate Department)
Many plants secrete substances to create a more favorable environment, including chemicals that kill pathogenic microbes or competing plants. Artemisia tridentata, also known as "Big Sagebrush," is prevalent in the Rocky Mountain region of the United States and is known to have antimicrobial capabilities. We will study the potential antimicrobial activity of Artemisia tridentata.
Studies report that chemicals released by the leaves and branches of A. tridentata affect bacteria native to deer rumen. Another study showed that 27 actinomycetes (anaerobic bacteria that form colonies) strains found in the rhizosphere of A. tridentata demonstrated antibacterial activities when tested on E. coli, Bacillus subtilis and Staphylococcus aureus. Native Americans traditionally used A. tridentata to relieve stomach pain, colds, coughs, sore eyes, snake bites and as an insect repellent. Researchers found several compounds, including flavonoids, that can affect antimicrobial activity. Articles regarding antimicrobial activities in A. tridentata were published between 1967 and 2004. With the chronological gaps and considering the progress that biological and molecular technology has made in recent years, knowledge of the chemicals released by A. tridentata lies largely untapped. In this study, we will investigate the antimicrobial activities of the leaves, stem, roots, and flowers of A. tridentata initially by the agar well diffusion method and followed by validating with the agar disk diffusion method. We will check the antimicrobial activity of the extract from different plant parts of A. tridentata on common bacteria such as E. coli, Bacillus subtilis, and some Pseudomonas spp. of plant pathogens.
The knowledge obtained from this research will further help in the identification and characterization of the secondary metabolites or chemicals involved in antimicrobial activity of sagebrush. Medicinal plants provide a healthy, natural alternative to conventional medication, and may lead to new insights on antibiotics and pharmaceuticals. Besides, Artemisia tridentata is a plant native to Utah and Idaho. It grows everywhere in the surrounding area, making it inexpensive (free) to produce.
Faculty Advisor: Kaundal, Amita (College of Agriculture and Applied Sciences; Plants, Soils, and Climate Department)
Many plants secrete substances to create a more favorable environment, including chemicals that kill pathogenic microbes or competing plants. Artemisia tridentata, also known as "Big Sagebrush," is prevalent in the Rocky Mountain region of the United States and is known to have antimicrobial capabilities. We will study the potential antimicrobial activity of Artemisia tridentata.
Studies report that chemicals released by the leaves and branches of A. tridentata affect bacteria native to deer rumen. Another study showed that 27 actinomycetes (anaerobic bacteria that form colonies) strains found in the rhizosphere of A. tridentata demonstrated antibacterial activities when tested on E. coli, Bacillus subtilis and Staphylococcus aureus. Native Americans traditionally used A. tridentata to relieve stomach pain, colds, coughs, sore eyes, snake bites and as an insect repellent. Researchers found several compounds, including flavonoids, that can affect antimicrobial activity. Articles regarding antimicrobial activities in A. tridentata were published between 1967 and 2004. With the chronological gaps and considering the progress that biological and molecular technology has made in recent years, knowledge of the chemicals released by A. tridentata lies largely untapped. In this study, we will investigate the antimicrobial activities of the leaves, stem, roots, and flowers of A. tridentata initially by the agar well diffusion method and followed by validating with the agar disk diffusion method. We will check the antimicrobial activity of the extract from different plant parts of A. tridentata on common bacteria such as E. coli, Bacillus subtilis, and some Pseudomonas spp. of plant pathogens.
The knowledge obtained from this research will further help in the identification and characterization of the secondary metabolites or chemicals involved in antimicrobial activity of sagebrush. Medicinal plants provide a healthy, natural alternative to conventional medication, and may lead to new insights on antibiotics and pharmaceuticals. Besides, Artemisia tridentata is a plant native to Utah and Idaho. It grows everywhere in the surrounding area, making it inexpensive (free) to produce.
Akt and Inflammatory Pathways Activation by Cache Valley Particulate Air Pollution
Sagers, Rachel; Nguyen, Andy; Weston, Jake; Grooms, Nicholas; Eggleston, Morgan; Martin, Randy; Coulombe, Roger (Utah State University)
Faculty Advisor: Coulombe, Roger (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)
The scenic mountain views of Cache Valley in Northern Utah stand in stark contrast with the valley's high concentrations of fine particulate air pollution (PM2.5), some of the worst reported in the United States. The unique geography promotes formation of ammonium nitrate (NH4NO3) from nitrogen oxides produced by motor vehicles and ammonia from dairy cow excreta. Winter atmospheric inversions, exacerbated by the mountainous terrain, trap and concentrate air pollutants. Epidemiological studies have revealed an association between PM exposure and early all-cause mortality. Exposure to PM2.5 is also associated with a variety of cardiovascular, cardiopulmonary, and neurodegenerative diseases, including myocardial infarction, stroke, COPD, lung cancer, Alzheimer's disease, and Parkinson's disease. Previous studies have shown that Cache Valley PM (CVPM) has pro-inflammatory effects, which has been linked to enhanced activation of Akt in human pulmonary epithelial cells. This research examined the cellular responses of human lung (BEAS-2B) cells exposed to CVPM and diesel exhaust particles (DEP), at 1 and 12 µg/ml concentrations of each particle type for a 24 hour exposure period. The CVPM used was collected onto stainless steel plates by a Tisch impactor. Assessment by the comet assay reveal genetic damage to CVPM exposed cells with equal potency to DEP exposed cells. Flow cytometry (p < 0.05) showed CVPM exposed cells had a significant increase in the number of actively-dividing cells compared to control cells. Whole-genome microarray identified affected genes related to inflammatory pathways, as well as activated Akt-dependent pathways. Subsequent qRT-PCR showed that CVPM exposure significantly increased expression of inflammatory markers, including IL-6, CD40LG, PLAG27, and cytochrome P450 (CYP) 1A1 (p < 0.05). Immunoblotting confirmed activation of Akt by phosphorylation of Thr308 in both CVPM and DEP exposed cells. This data supports the hypothesis that CVPM may induce pro-carcinogenic pathways with potency similar to DEP.
Faculty Advisor: Coulombe, Roger (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)
The scenic mountain views of Cache Valley in Northern Utah stand in stark contrast with the valley's high concentrations of fine particulate air pollution (PM2.5), some of the worst reported in the United States. The unique geography promotes formation of ammonium nitrate (NH4NO3) from nitrogen oxides produced by motor vehicles and ammonia from dairy cow excreta. Winter atmospheric inversions, exacerbated by the mountainous terrain, trap and concentrate air pollutants. Epidemiological studies have revealed an association between PM exposure and early all-cause mortality. Exposure to PM2.5 is also associated with a variety of cardiovascular, cardiopulmonary, and neurodegenerative diseases, including myocardial infarction, stroke, COPD, lung cancer, Alzheimer's disease, and Parkinson's disease. Previous studies have shown that Cache Valley PM (CVPM) has pro-inflammatory effects, which has been linked to enhanced activation of Akt in human pulmonary epithelial cells. This research examined the cellular responses of human lung (BEAS-2B) cells exposed to CVPM and diesel exhaust particles (DEP), at 1 and 12 µg/ml concentrations of each particle type for a 24 hour exposure period. The CVPM used was collected onto stainless steel plates by a Tisch impactor. Assessment by the comet assay reveal genetic damage to CVPM exposed cells with equal potency to DEP exposed cells. Flow cytometry (p < 0.05) showed CVPM exposed cells had a significant increase in the number of actively-dividing cells compared to control cells. Whole-genome microarray identified affected genes related to inflammatory pathways, as well as activated Akt-dependent pathways. Subsequent qRT-PCR showed that CVPM exposure significantly increased expression of inflammatory markers, including IL-6, CD40LG, PLAG27, and cytochrome P450 (CYP) 1A1 (p < 0.05). Immunoblotting confirmed activation of Akt by phosphorylation of Thr308 in both CVPM and DEP exposed cells. This data supports the hypothesis that CVPM may induce pro-carcinogenic pathways with potency similar to DEP.
A Proposal to Investigate Protein Expression of Rhizopus oryzae Biofilms Upon Treatment with Extracorporeal Shockwaves and Amphotericin B
Nanasi Sekona, Ashley Balderrama, Carlos Nunez, Kyle Hendricks, Tyson Hillock, and Dr. Olga Kopp (Utah Valley University)
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology)
Over the last 30 years, the incidence of fungal infections has gradually increased. Mucormycosis is a fungal infection primarily caused by Rhizopus Oryzae. The majority of patients who develop invasive mucormycosis die within 12 weeks of diagnosis. Mucormycosis is commonly treated with an antifungal agent called Amphotericin B (AMB). When used in high concentrations, AMB causes severe side effects such as nephrotoxicity. It has been reported that 99% of microbes exist as biofilm: thus, there is a direct association between mucormycosis and biofilms. Shockwave has been shown to inhibit living bacteria in biofilm, but few studies have focused on the effects of shockwave on fungal biofilm. Previous work in our lab showed that shockwaves were effective in damaging biofilms of R. oryzae; but at the same time helped promote the metabolism of surviving R. oryzae. This study aims to investigate the proteins expressed in fungal biofilms when introduced to different intensities of shockwave coupled with the treatment of AMB. This will be accomplished by culturing sporangiospores and propagating R. oryzae biofilms. Standardized biofilm will be treated with 0.5 µg/mL AMB in 1% DMSO, and/or shockwave treatment of 300 pulses at 0.19 mJ/mm2 energy density to be measured against a control group. The proteins will be extracted, determined by 2D gel electrophoresis, and identified by mass spectrometry. Studying protein expression resulting from combination therapy of extracorporeal shockwave and AMB on R. oryzae biofilm could progress research surrounding the difficulties of mucormycosis treatments. Particularly, research aimed at counteracting the antifungal and antimicrobial resistance contributed by proteins in the fungi's biofilm.
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology)
Over the last 30 years, the incidence of fungal infections has gradually increased. Mucormycosis is a fungal infection primarily caused by Rhizopus Oryzae. The majority of patients who develop invasive mucormycosis die within 12 weeks of diagnosis. Mucormycosis is commonly treated with an antifungal agent called Amphotericin B (AMB). When used in high concentrations, AMB causes severe side effects such as nephrotoxicity. It has been reported that 99% of microbes exist as biofilm: thus, there is a direct association between mucormycosis and biofilms. Shockwave has been shown to inhibit living bacteria in biofilm, but few studies have focused on the effects of shockwave on fungal biofilm. Previous work in our lab showed that shockwaves were effective in damaging biofilms of R. oryzae; but at the same time helped promote the metabolism of surviving R. oryzae. This study aims to investigate the proteins expressed in fungal biofilms when introduced to different intensities of shockwave coupled with the treatment of AMB. This will be accomplished by culturing sporangiospores and propagating R. oryzae biofilms. Standardized biofilm will be treated with 0.5 µg/mL AMB in 1% DMSO, and/or shockwave treatment of 300 pulses at 0.19 mJ/mm2 energy density to be measured against a control group. The proteins will be extracted, determined by 2D gel electrophoresis, and identified by mass spectrometry. Studying protein expression resulting from combination therapy of extracorporeal shockwave and AMB on R. oryzae biofilm could progress research surrounding the difficulties of mucormycosis treatments. Particularly, research aimed at counteracting the antifungal and antimicrobial resistance contributed by proteins in the fungi's biofilm.
Analysis of binding affinity and molecular cloning of two PRMT1 substrates
Jones, Abigail; Hevel, Joan (Utah State University)
Faculty Advisor: Hevel, Joan (College of Science, Chemistry and Biochemistry Department)
PRMT1 is one of nine known mammalian Protein Arginine Methyltransferases (PRMTs) whose function are to transfer methyl groups from S-adenosyl methionine (SAM) to arginine residues of specific proteins. PRMT1 is known to methylate many different proteins in cells, but the mechanism of target recognition and binding is still unknown. Correct regulation of PRMT1 is critical to proper cellular function; thus, the action of PRMT1 is important to understand. In this study, we seek to elucidate how PRMT1 recognizes and binds its targets by identifying protein substrates of PRMT1 that form a stable complex with the enzyme. Such a protein would allow for additional studies (e.g. crystallographic or cryo-EM studies) to help visualize PRMT1-substrate interactions. Two substrates of PRMT1, TWIST1 and Smad6, have been purified, and the binding affinity of each to PRMT1 has been qualitatively assessed via pull-down assay and Western blot. Ligation-independent-cloning has been used to clone each substrate gene out of a GST-tagged vector and into a His-tagged vector, which will allow for further experiments assessing the stoichiometry of PRMT1-substrate binding.
Faculty Advisor: Hevel, Joan (College of Science, Chemistry and Biochemistry Department)
PRMT1 is one of nine known mammalian Protein Arginine Methyltransferases (PRMTs) whose function are to transfer methyl groups from S-adenosyl methionine (SAM) to arginine residues of specific proteins. PRMT1 is known to methylate many different proteins in cells, but the mechanism of target recognition and binding is still unknown. Correct regulation of PRMT1 is critical to proper cellular function; thus, the action of PRMT1 is important to understand. In this study, we seek to elucidate how PRMT1 recognizes and binds its targets by identifying protein substrates of PRMT1 that form a stable complex with the enzyme. Such a protein would allow for additional studies (e.g. crystallographic or cryo-EM studies) to help visualize PRMT1-substrate interactions. Two substrates of PRMT1, TWIST1 and Smad6, have been purified, and the binding affinity of each to PRMT1 has been qualitatively assessed via pull-down assay and Western blot. Ligation-independent-cloning has been used to clone each substrate gene out of a GST-tagged vector and into a His-tagged vector, which will allow for further experiments assessing the stoichiometry of PRMT1-substrate binding.
Antibiotic Resistance in Staphylococcus aureus: Effects of Biofilm Synthesis in Gene Transfer
Hirschi, Blake; Pickett, Brad; Thompson, Jared; Telford, Mady; Berges, Bradford (Brigham Young University)
Faculty Advisor: Berges, Brad (Life Sciences, Microbiology and Molecular Biology)
Staphylococcus aureus (S. aureus) is a commensal bacterium commonly found amongst livestock and near 30% of humans' nostrils. However, through acquisition of certain genes S. aureus may develop antibiotic resistance such as in methicillin-resistant Staphylococcus aureus (MRSA). One hypothesized component lending to acquisition of genetic resistance in S. aureus is the synthesis of colony biofilms. Biofilms are comprised of a variety of substances including secreted polysaccharides, protein and even extracellular DNA. Our work postulates that extracellular DNA-based biofilms will transfer genes for antibiotic resistance at a higher rate than in polysaccharide/protein biofilms. Through employment of polymerase chain reaction (PCR), we aim to characterize a wide sample of methicillin-susceptible S. aureus (MSSA) human associated strains and MRSA livestock associated strains for multiple antibiotic resistances. Co-inoculating pairs of human associated and livestock associated strains, each lacking the other's resistance genes, will provide an environment wherein biofilm-mediated gene transfer may occur. Further pairing based on biofilm composition (DNA or polysaccharide/protein) will yield data concerning which biofilm facilitates gene transfer more efficiently. Subsequent genotyping will confirm whether resulting isolates acquired new antibiotic resistance through biofilm-mediated transfer, thus increasing pathogenicity.
Faculty Advisor: Berges, Brad (Life Sciences, Microbiology and Molecular Biology)
Staphylococcus aureus (S. aureus) is a commensal bacterium commonly found amongst livestock and near 30% of humans' nostrils. However, through acquisition of certain genes S. aureus may develop antibiotic resistance such as in methicillin-resistant Staphylococcus aureus (MRSA). One hypothesized component lending to acquisition of genetic resistance in S. aureus is the synthesis of colony biofilms. Biofilms are comprised of a variety of substances including secreted polysaccharides, protein and even extracellular DNA. Our work postulates that extracellular DNA-based biofilms will transfer genes for antibiotic resistance at a higher rate than in polysaccharide/protein biofilms. Through employment of polymerase chain reaction (PCR), we aim to characterize a wide sample of methicillin-susceptible S. aureus (MSSA) human associated strains and MRSA livestock associated strains for multiple antibiotic resistances. Co-inoculating pairs of human associated and livestock associated strains, each lacking the other's resistance genes, will provide an environment wherein biofilm-mediated gene transfer may occur. Further pairing based on biofilm composition (DNA or polysaccharide/protein) will yield data concerning which biofilm facilitates gene transfer more efficiently. Subsequent genotyping will confirm whether resulting isolates acquired new antibiotic resistance through biofilm-mediated transfer, thus increasing pathogenicity.
Transcriptomics of Ephemeroptera (Mayflies): Generation of New Data and Bioinformatics Workflow
Vilela, Ernie; Seal, Isaac; Ogden, Heath T. (Utah Valley University)
Faculty Advisor: Ogden, Thomas (College of Science, Biology Deapartment)
We are interested in using transcriptome data, generated with next generation sequencing technology, to investigate the evolutionary trends of specific genes and their associated expression in mayflies. We generated an additional transcriptome for mayflies. RNA was extracted from a freshly frozen specimen preserved in RNAlater® (Ambion) using TRIzol® Reagent (Ambion) and cDNA libraries were prepared from mRNA. RNA-seq data was generated using a paired-end protocol (PE100) on Illumina HiSeq2000 with an expected 60 million reads. In order to effectively investigate the large amount of sequences, we created a bioinformatics workflow to analyze the newly generated transcriptome data along with previous data for mayflies. The workflow consists of these main steps: Trinity (Assemblying the transcripts), Transdecoder (Identifying candidate coding regions), HMMER (Searching biological sequence databases for homologous sequences). We tested the workflow looking at opsin genes.
Faculty Advisor: Ogden, Thomas (College of Science, Biology Deapartment)
We are interested in using transcriptome data, generated with next generation sequencing technology, to investigate the evolutionary trends of specific genes and their associated expression in mayflies. We generated an additional transcriptome for mayflies. RNA was extracted from a freshly frozen specimen preserved in RNAlater® (Ambion) using TRIzol® Reagent (Ambion) and cDNA libraries were prepared from mRNA. RNA-seq data was generated using a paired-end protocol (PE100) on Illumina HiSeq2000 with an expected 60 million reads. In order to effectively investigate the large amount of sequences, we created a bioinformatics workflow to analyze the newly generated transcriptome data along with previous data for mayflies. The workflow consists of these main steps: Trinity (Assemblying the transcripts), Transdecoder (Identifying candidate coding regions), HMMER (Searching biological sequence databases for homologous sequences). We tested the workflow looking at opsin genes.
A Study of the Changes in the Oral Microbiome of Horses by Age Group
Forbush, Micah; Gold, Roger (Southern Utah University)
Faculty Advisor: Gold, Roger (Southern Utah University, Biology)
The health of a horse's teeth and oral cavity is strongly correlated with the horse's overall health and well-being. As a horse ages the oral cavity undergoes many distinct physical changes, which may lead to changes in the composition of the oral microbiome as well. While there has been extensive research performed on the oral microbiomes of healthy horses compared to horses with various oral diseases, very little is known about normal changes to the horse oral microbiome as they age. The aim of this study was to use high-throughput sequencing to compare the oral microbiomes of horses in different age categories. Total genomic DNA was isolated from oral swabs taken from horses in the 7 day-old, 1-7 years old, 8-15 years old and 16+ years old age categories. The V3-V4- region of the 16S rRNA gene was amplified by PCR and amplicons were submitted for paired-end sequencing on the Illumina HiSeq system. Sample reads were analyzed using the QIIME 2.0 microbiome bioinformatics platform and overall bacterial diversity was compared among age classes.
Faculty Advisor: Gold, Roger (Southern Utah University, Biology)
The health of a horse's teeth and oral cavity is strongly correlated with the horse's overall health and well-being. As a horse ages the oral cavity undergoes many distinct physical changes, which may lead to changes in the composition of the oral microbiome as well. While there has been extensive research performed on the oral microbiomes of healthy horses compared to horses with various oral diseases, very little is known about normal changes to the horse oral microbiome as they age. The aim of this study was to use high-throughput sequencing to compare the oral microbiomes of horses in different age categories. Total genomic DNA was isolated from oral swabs taken from horses in the 7 day-old, 1-7 years old, 8-15 years old and 16+ years old age categories. The V3-V4- region of the 16S rRNA gene was amplified by PCR and amplicons were submitted for paired-end sequencing on the Illumina HiSeq system. Sample reads were analyzed using the QIIME 2.0 microbiome bioinformatics platform and overall bacterial diversity was compared among age classes.
A Novel Natural Product PIM3 Inhibitor with Anti-Cancer Activity
Fazio, Nicholas; Russell, Michael; Krapohl, John; Andrus, Brayden; Hansen, Marc (Brigham Young University)
Faculty Advisor: Hansen, Marc (Brigham Young University, Physiology and Developmental Biology)
PIM 3 is a proto-oncogene with serine/threonine kinase activity that can prevent apoptosis, promote cell survival and protein translation. Abnormal PIM3 activity contributes to tumorigenesis by phosphorylation of targets that release anti-apoptotic proteins. Pathological PIM3 expression is common in pancreatic and prostate cancer. Inhibiting this kinase activity can be used to therapeutically suppress uncontrolled cell growth in cancerous tissues. Synthetic inhibitors are being developed as therapeutics to treat PIM3 related disorders. Compounds derived from plants and natural sources have therapeutically-relevant biological activity. Additionally, they often well tolerated, making them important starting points for drug discovery efforts. A less widely used approach to discover the biological activity of molecules is built around using a large scale in-silico molecular screening, which has emerged as a critical drug discovery tool. Here, we screen a large (>100,000 compound) virtual library of natural product compounds for binding in the PIM3 ATP binding site, then validate compounds with using cell-based and immuno-based assays. This approach reveals PIM3 inhibition by a saponin scaffold, which suggests potential utility as a therapeutic or as a lead for further optimization.
Faculty Advisor: Hansen, Marc (Brigham Young University, Physiology and Developmental Biology)
PIM 3 is a proto-oncogene with serine/threonine kinase activity that can prevent apoptosis, promote cell survival and protein translation. Abnormal PIM3 activity contributes to tumorigenesis by phosphorylation of targets that release anti-apoptotic proteins. Pathological PIM3 expression is common in pancreatic and prostate cancer. Inhibiting this kinase activity can be used to therapeutically suppress uncontrolled cell growth in cancerous tissues. Synthetic inhibitors are being developed as therapeutics to treat PIM3 related disorders. Compounds derived from plants and natural sources have therapeutically-relevant biological activity. Additionally, they often well tolerated, making them important starting points for drug discovery efforts. A less widely used approach to discover the biological activity of molecules is built around using a large scale in-silico molecular screening, which has emerged as a critical drug discovery tool. Here, we screen a large (>100,000 compound) virtual library of natural product compounds for binding in the PIM3 ATP binding site, then validate compounds with using cell-based and immuno-based assays. This approach reveals PIM3 inhibition by a saponin scaffold, which suggests potential utility as a therapeutic or as a lead for further optimization.
Analysis of the Gut Microbiome of Drosophila melanogaster Models of Parkinson's Disease
Marshman, Evan; Peterson, Samara; Call, Gerald; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Science, Plant and Wildlife Science)
In recent years the association between the human gut microbiome and the brain has become a promising field of study. Often referred to as the "gut-brain axis", this connection has greatly enriched our scientific understanding of many disorders that affect the brain and nervous system. A recent study showed the differing richness of bacteria in the microbiota of Parkinson's patients and healthy control subjects. Because recent research shows this connection, we predicted that we would detect variation in the microbiota of D. melanogaster (fruit flies) models of Parkinson's disease, relative to wild type flies. To test this hypothesis, I analyzed 16s rRNA sequence data, reporting the microbiota composition in flies that are a model of Parkinson's Disease, as well as wild type flies. I found one strain of the genus Acetobacter that was differentially abundant between the two fly types. Therefore, for my CURA I will extend my analysis by performing similar analyses by taking a larger set of Parkinson's fly models. Once they are sequenced, I will use QIIME, the same software I used in my preliminary analyses to further our understanding of the taxonomic differences between the gut bacteria of Parkinson's models and wild type flies.
Faculty Advisor: Chaston, John (Life Science, Plant and Wildlife Science)
In recent years the association between the human gut microbiome and the brain has become a promising field of study. Often referred to as the "gut-brain axis", this connection has greatly enriched our scientific understanding of many disorders that affect the brain and nervous system. A recent study showed the differing richness of bacteria in the microbiota of Parkinson's patients and healthy control subjects. Because recent research shows this connection, we predicted that we would detect variation in the microbiota of D. melanogaster (fruit flies) models of Parkinson's disease, relative to wild type flies. To test this hypothesis, I analyzed 16s rRNA sequence data, reporting the microbiota composition in flies that are a model of Parkinson's Disease, as well as wild type flies. I found one strain of the genus Acetobacter that was differentially abundant between the two fly types. Therefore, for my CURA I will extend my analysis by performing similar analyses by taking a larger set of Parkinson's fly models. Once they are sequenced, I will use QIIME, the same software I used in my preliminary analyses to further our understanding of the taxonomic differences between the gut bacteria of Parkinson's models and wild type flies.
Census and Distribution Analysis of Alouatta palliata (mantled howler monkey) in La Selva Biological Station
Smith, Mick; Desdames, Chloe (University of Utah)
Faculty Advisor: Seaboch, Melissa (University of Utah, Anthropology)
Census and sampling work are important because they provide critical information on population size, distribution, and habitat preference — all important factors in conservation. La Selva Biological station is located on the north eastern side of Costa Rica and is a protected biological reserve. It is home to three species of primates including Alouatta palliata, commonly known as the mantled howler monkey. It is comprised of primary forest that has been undisturbed by human forces and secondary forest that is regenerating from past disturbances, such as deforestation. Past research conducted on howler monkeys shows they prefer primary forest over secondary because primary forests have higher species richness and a wider range of trees which provides food for howler monkey's selective diet. I predict that the majority of A. palliata in Costa Rica will be located in primary forest because it offers more resources. I censused 11 established trails at La Selva (two trails in primary forest, seven trails in secondary forest, and two trails crossing both forest types). For each howler monkey encountered, I recorded the location, forest type, and group size. I identified four different groups of A. palliata with group size ranging between 6-10 individuals. One group was located in primary forest and three groups in secondary forest; thus, my hypothesis that A. palliata would prefer primary forests was not supported. Anecdotally, A. palliata were observed more frequently around rivers or streams. This could indicate that rather than being concerned with primary versus secondary forest, they prefer riparian habitats found along the banks of rivers or other actively moving sources of water. A study conducted by Stoner found howler monkeys in riparian habits supporting this hypothesis. This type of research continues to provide critical information for understanding primate's habitats which helps with conservation of species.
Faculty Advisor: Seaboch, Melissa (University of Utah, Anthropology)
Census and sampling work are important because they provide critical information on population size, distribution, and habitat preference — all important factors in conservation. La Selva Biological station is located on the north eastern side of Costa Rica and is a protected biological reserve. It is home to three species of primates including Alouatta palliata, commonly known as the mantled howler monkey. It is comprised of primary forest that has been undisturbed by human forces and secondary forest that is regenerating from past disturbances, such as deforestation. Past research conducted on howler monkeys shows they prefer primary forest over secondary because primary forests have higher species richness and a wider range of trees which provides food for howler monkey's selective diet. I predict that the majority of A. palliata in Costa Rica will be located in primary forest because it offers more resources. I censused 11 established trails at La Selva (two trails in primary forest, seven trails in secondary forest, and two trails crossing both forest types). For each howler monkey encountered, I recorded the location, forest type, and group size. I identified four different groups of A. palliata with group size ranging between 6-10 individuals. One group was located in primary forest and three groups in secondary forest; thus, my hypothesis that A. palliata would prefer primary forests was not supported. Anecdotally, A. palliata were observed more frequently around rivers or streams. This could indicate that rather than being concerned with primary versus secondary forest, they prefer riparian habitats found along the banks of rivers or other actively moving sources of water. A study conducted by Stoner found howler monkeys in riparian habits supporting this hypothesis. This type of research continues to provide critical information for understanding primate's habitats which helps with conservation of species.
Deep learning for image segmentation
Jenkins, Abigail; Baugh, Makinnon; Frandsen, Paul; White, Alexander; Dikow, Rebecca (Brigham Young University)
Faculty Advisor: Frandsen, Paul (Life Sciences, Plant and Wildlife Sciences)
Historically, physical plant specimens have been preserved and mounted on paper sheets and stored in plant collections, or herbaria. Herbarium collections are used for a wide variety of research purposes, including plant taxonomy, ecology, and evolutionary biology. The process of digitizing the herbarium sheets is simple and involves taking high resolution photos of each sheet and recording the corresponding metadata and attributes of the particular sample. Digitized herbarium sheets can be useful for a variety of purposes, and, by making images freely available online, they become immediately accessible to the scientific community, facilitating remote analysis. In addition, in a digital format, the images become computable and usable for purposes such as training deep learning models for classification or analysis of morphology.
While the process of digitizing is simple, herbarium sheets contain other features not directly representative of the plant, such as annotations, labels, museum stamps, color palettes, and rulers. There are additional inconsistencies in the herbarium sheets that are introduced through staining, record keeping, and natural degradation. Taken together, this information can contribute a substantial amount of noise if one is to use the image for downstream research analysis concerning the pattern, shape, or color of the specimen. We have developed a pipeline to filter this extraneous information, using image segmentation (whereby the specimen material is partitioned from the background) and deep learning.
We present this pipeline for generating training data for image segmentation tasks along with a novel dataset of highly resolved image masks segmenting plant material from background noise. We used this dataset to train a neural network to segment plant material in herbarium sheets more generally, and our method is applicable to other museum data sources where masking may be useful for quantitative analysis of patterns and shapes.
Faculty Advisor: Frandsen, Paul (Life Sciences, Plant and Wildlife Sciences)
Historically, physical plant specimens have been preserved and mounted on paper sheets and stored in plant collections, or herbaria. Herbarium collections are used for a wide variety of research purposes, including plant taxonomy, ecology, and evolutionary biology. The process of digitizing the herbarium sheets is simple and involves taking high resolution photos of each sheet and recording the corresponding metadata and attributes of the particular sample. Digitized herbarium sheets can be useful for a variety of purposes, and, by making images freely available online, they become immediately accessible to the scientific community, facilitating remote analysis. In addition, in a digital format, the images become computable and usable for purposes such as training deep learning models for classification or analysis of morphology.
While the process of digitizing is simple, herbarium sheets contain other features not directly representative of the plant, such as annotations, labels, museum stamps, color palettes, and rulers. There are additional inconsistencies in the herbarium sheets that are introduced through staining, record keeping, and natural degradation. Taken together, this information can contribute a substantial amount of noise if one is to use the image for downstream research analysis concerning the pattern, shape, or color of the specimen. We have developed a pipeline to filter this extraneous information, using image segmentation (whereby the specimen material is partitioned from the background) and deep learning.
We present this pipeline for generating training data for image segmentation tasks along with a novel dataset of highly resolved image masks segmenting plant material from background noise. We used this dataset to train a neural network to segment plant material in herbarium sheets more generally, and our method is applicable to other museum data sources where masking may be useful for quantitative analysis of patterns and shapes.
Cell Adhesion and Morphology in Relation to Neural Tube Defects
Lin, Jade; Park, Yeram; Ross, Micah; Stark, Michael; Hansen, Marc (Brigham Young University)
Faculty Advisor: Stark, Michael (Brigham Young University, Physiology and Developmental Biology); Hansen, Marc (Brigham Young University, Physiology and Developmental Biology)
Neural Tube Defects (NTDs) such as spina bifida and anencephaly are due to incomplete closure of neural tubes in developing embryos. While the etiology is still unknown, environmental and genetic factors, toxicants, and maternal health are implicated as potential causes. Previous research shows that NTDs are associated with increased levels of ceramide (C2) and possible exposure to fumonisin (FB1), and valproic acid (VPA). We hypothesize that these molecules may interfere with cell-to-cell interactions which are important for neural tube formation. To investigate the potential mechanisms by which these toxicants can induce NTDs, I will use a well-characterized MDCK cell model treated with C2, FB1, and VPA to assess their impact on cell adhesion.
Faculty Advisor: Stark, Michael (Brigham Young University, Physiology and Developmental Biology); Hansen, Marc (Brigham Young University, Physiology and Developmental Biology)
Neural Tube Defects (NTDs) such as spina bifida and anencephaly are due to incomplete closure of neural tubes in developing embryos. While the etiology is still unknown, environmental and genetic factors, toxicants, and maternal health are implicated as potential causes. Previous research shows that NTDs are associated with increased levels of ceramide (C2) and possible exposure to fumonisin (FB1), and valproic acid (VPA). We hypothesize that these molecules may interfere with cell-to-cell interactions which are important for neural tube formation. To investigate the potential mechanisms by which these toxicants can induce NTDs, I will use a well-characterized MDCK cell model treated with C2, FB1, and VPA to assess their impact on cell adhesion.