Life Sciences
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.
Intravenous dopamine enhancement of dopamine release in the nucleus accumbens is peripheral dopamine 2 receptor dependent
Small, Christina; Obray, J Daniel; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
Recent studies have shown that administration of dopamine in the periphery (outside of the brain) produces a robust enhancement of dopamine release in the nucleus accumbens and alleviates cognitive deficits associated with schizophrenia-like and attention deficit hyperactivity disorder (ADHD)-like phenotypes in rodent models. Despite this, the mechanism whereby peripheral administration of dopamine produces these effects is unknown as dopamine does not cross the blood brain barrier. Activation of dopamine 2 receptors on circulating leukocytes encourages extravasation and can trigger production and release of cytokines such as TNF-_ and IL-10 as well as IL-1_. IL-1_ and IL-10 are both known to enhance dopamine release. In this study we demonstrate that the effects of intravenous dopamine on dopamine release in the NAc are mediated by peripheral dopamine 2 receptors. Additionally, we show that intravenous dopamine is rewarding and that these rewarding effects can be blocked by antagonism of peripheral dopamine 2 receptors. As many drugs of abuse enhance plasma dopamine levels this research elucidates a secondary pathway which may play a role in the development of substance use disorders.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
Recent studies have shown that administration of dopamine in the periphery (outside of the brain) produces a robust enhancement of dopamine release in the nucleus accumbens and alleviates cognitive deficits associated with schizophrenia-like and attention deficit hyperactivity disorder (ADHD)-like phenotypes in rodent models. Despite this, the mechanism whereby peripheral administration of dopamine produces these effects is unknown as dopamine does not cross the blood brain barrier. Activation of dopamine 2 receptors on circulating leukocytes encourages extravasation and can trigger production and release of cytokines such as TNF-_ and IL-10 as well as IL-1_. IL-1_ and IL-10 are both known to enhance dopamine release. In this study we demonstrate that the effects of intravenous dopamine on dopamine release in the NAc are mediated by peripheral dopamine 2 receptors. Additionally, we show that intravenous dopamine is rewarding and that these rewarding effects can be blocked by antagonism of peripheral dopamine 2 receptors. As many drugs of abuse enhance plasma dopamine levels this research elucidates a secondary pathway which may play a role in the development of substance use disorders.
Mapping the Potential Distribution of an Invasive Plant, Lythrum salicaria, using Crowd-Sourced Survey Data.
Wertz, Parker (Weber State University)
Faculty Advisor: Dorsey, Bryan (Weber State University, Geography)
Prevention and predicting spread is the best method of control against invasive species. Land managers require accurate and reliable methods for containment and eradication to prevent land cover change and loss of biodiversity. Ecological niche models exist and are used by ecologists to map habitat suitability, but many rely on presence-absence samples which are difficult to obtain. Maximum entropy species distribution modeling (Maxent) is a popular model that has been increasingly used since it can make valid predictions using presence-only data. Many studies have used Maxent to model species distributions, but few have done so with crowdsourced data since it is more likely to be bias and unreliable. The purpose of this study is to test the robustness of Maxent using crowdsourced presence-only data on Lyrthum salicaria, a perennial herb that invades wetlands and pushes out native flora. The study is set in northern and central Utah, and uses environmental variables in climate, landcover, and topography, with landcover being the most contributive factor to the model. Model performance was very good, even with species data being bias towards areas of higher population, proving Maxent as a worthy method to use in species distribution modeling with crowdsourced species presence data. This results of this study show promise for use in modeling other invasive plants in the future.
Faculty Advisor: Dorsey, Bryan (Weber State University, Geography)
Prevention and predicting spread is the best method of control against invasive species. Land managers require accurate and reliable methods for containment and eradication to prevent land cover change and loss of biodiversity. Ecological niche models exist and are used by ecologists to map habitat suitability, but many rely on presence-absence samples which are difficult to obtain. Maximum entropy species distribution modeling (Maxent) is a popular model that has been increasingly used since it can make valid predictions using presence-only data. Many studies have used Maxent to model species distributions, but few have done so with crowdsourced data since it is more likely to be bias and unreliable. The purpose of this study is to test the robustness of Maxent using crowdsourced presence-only data on Lyrthum salicaria, a perennial herb that invades wetlands and pushes out native flora. The study is set in northern and central Utah, and uses environmental variables in climate, landcover, and topography, with landcover being the most contributive factor to the model. Model performance was very good, even with species data being bias towards areas of higher population, proving Maxent as a worthy method to use in species distribution modeling with crowdsourced species presence data. This results of this study show promise for use in modeling other invasive plants in the future.
Patients' Perceptions of Stress During Hospitalization
Larson, Rebecca; Jimenez, Misty (Utah Valley University)
Faculty Advisor: Jensen, Francine (Utah Valley University, Nursing)
Stress is a known barrier to patient recovery. Patients experience increased emotions, such as stress, while hospitalized due to high stakes from risks to life, health and well-being. Patients' emotions can affect their perceptions, future intentions, and behaviors. In pediatrics, the way parents react to their child's illness may affect the children's compliance, emotional response to medical treatment, and even some development processes, demonstrating the premise that there are many possible stressors that can have significant impacts on patients. Hospitals have taken several measures to evaluate patient stress, such as encouraging hospital staff to discuss patient satisfaction surveys with their patient. However, not all patients recognize their own stressors, and some patients may not initially feel comfortable sharing them. For example, a study showed specific stressors that may experienced by patients of different demographics. These stressors may not always be apparent to nurses. Patients' stress can be reduced if the hospital environment fosters perceptions of control, social support and positive distraction. A change in patient environment can promote healing, as evidenced by a hospital, Navicent Health, that demonstrated in their neonatal intensive care unit that reducing stress and anxiety for both newborns and their parents facilitated healing growth and bonding. Nurses can improve the care they provide to patients by learning how to recognize and reduce stressors during the hospital stay.
Faculty Advisor: Jensen, Francine (Utah Valley University, Nursing)
Stress is a known barrier to patient recovery. Patients experience increased emotions, such as stress, while hospitalized due to high stakes from risks to life, health and well-being. Patients' emotions can affect their perceptions, future intentions, and behaviors. In pediatrics, the way parents react to their child's illness may affect the children's compliance, emotional response to medical treatment, and even some development processes, demonstrating the premise that there are many possible stressors that can have significant impacts on patients. Hospitals have taken several measures to evaluate patient stress, such as encouraging hospital staff to discuss patient satisfaction surveys with their patient. However, not all patients recognize their own stressors, and some patients may not initially feel comfortable sharing them. For example, a study showed specific stressors that may experienced by patients of different demographics. These stressors may not always be apparent to nurses. Patients' stress can be reduced if the hospital environment fosters perceptions of control, social support and positive distraction. A change in patient environment can promote healing, as evidenced by a hospital, Navicent Health, that demonstrated in their neonatal intensive care unit that reducing stress and anxiety for both newborns and their parents facilitated healing growth and bonding. Nurses can improve the care they provide to patients by learning how to recognize and reduce stressors during the hospital stay.
Probiotic Survival in Non-Dairy Fermentation
Smith, June; Mishra, Niharika (Weber State University)
Faculty Advisor: Oberg, Craig (Weber State University, Microbiology); Culumber, Michele (Weber State University, Microbiology)
Non-dairy food options have become a growing cultural necessity, however, providing fermented or probiotic supplemented non-dairy alternatives is difficult. Little is known about the activity and survival of probiotic cultures in dairy alternatives. We evaluated the activities of several probiotics at various concentrations and in different combinations in oat, almond, and coconut beverages. Probiotic culture strains of Streptococcus thermophilus (YFLO1), Lactobacillus rhamnose (LGG), L. casei (Casei 431), and Bifidobacterium animalis subsp. lactis (BB12), and commercial probiotic mixtures, YFLO2, and Fresh Q, were inoculated in MRS broth, transferred to MRS agar plates, and incubated anaerobically for 24 hours at 37_. BB12 was grown anaerobically in MRS + cystine broth and agar. Isolated colonies were assayed on API 50 CH panels, and a carbohydrate use panel was developed for each organism. Oat, almond, and coconut beverages were inoculated in duplicate with the isolated strains and incubated in a water bath at 40_. The pH was recorded at regular intervals for up to 41 hours. The oat beverage had the most rapid and significant pH change, when incubated with either YFLO1, casei431, and LGG, dropping between 1.5 to 3 pH units over 3 hours depending on the culture. The almond and coconut beverages did not show rapid pH change with the organisms tested. Due to the quick decrease in pH change, further tests on the oat beverage. It was inoculated with Lactobacillus casei 431, LGG, and YFLO1. Organisms were tested at 0.5%, 1.0%, and 2.0% concentrations in oat beverage in triplicate. These inoculations were again incubated at 40°C and pH monitored after 5 hours, then plated on MRS agar plates after 24 hours. Final ranged between 1.0 x 109 - 1.8 x 109 for the 1% inoculum. It appears that these organisms survive, and may even grow in the oat beverage. This research demonstrates that probiotic cultures can grow in non-dairy beverages and can ferment the available carbohydrates and decrease pH. These results provide insights that can be used for beverages, yogurt, ice cream, and other fermented food production.
Faculty Advisor: Oberg, Craig (Weber State University, Microbiology); Culumber, Michele (Weber State University, Microbiology)
Non-dairy food options have become a growing cultural necessity, however, providing fermented or probiotic supplemented non-dairy alternatives is difficult. Little is known about the activity and survival of probiotic cultures in dairy alternatives. We evaluated the activities of several probiotics at various concentrations and in different combinations in oat, almond, and coconut beverages. Probiotic culture strains of Streptococcus thermophilus (YFLO1), Lactobacillus rhamnose (LGG), L. casei (Casei 431), and Bifidobacterium animalis subsp. lactis (BB12), and commercial probiotic mixtures, YFLO2, and Fresh Q, were inoculated in MRS broth, transferred to MRS agar plates, and incubated anaerobically for 24 hours at 37_. BB12 was grown anaerobically in MRS + cystine broth and agar. Isolated colonies were assayed on API 50 CH panels, and a carbohydrate use panel was developed for each organism. Oat, almond, and coconut beverages were inoculated in duplicate with the isolated strains and incubated in a water bath at 40_. The pH was recorded at regular intervals for up to 41 hours. The oat beverage had the most rapid and significant pH change, when incubated with either YFLO1, casei431, and LGG, dropping between 1.5 to 3 pH units over 3 hours depending on the culture. The almond and coconut beverages did not show rapid pH change with the organisms tested. Due to the quick decrease in pH change, further tests on the oat beverage. It was inoculated with Lactobacillus casei 431, LGG, and YFLO1. Organisms were tested at 0.5%, 1.0%, and 2.0% concentrations in oat beverage in triplicate. These inoculations were again incubated at 40°C and pH monitored after 5 hours, then plated on MRS agar plates after 24 hours. Final ranged between 1.0 x 109 - 1.8 x 109 for the 1% inoculum. It appears that these organisms survive, and may even grow in the oat beverage. This research demonstrates that probiotic cultures can grow in non-dairy beverages and can ferment the available carbohydrates and decrease pH. These results provide insights that can be used for beverages, yogurt, ice cream, and other fermented food production.
Proteomic Analysis of Trichopteran Silk Fibre
Frandsen, Paul; Bursell, Madeline; Taylor, Adam; Wilson, Seth; Steeneck, Amy; Stewart, Russell (Brigham Young University)
Faculty Advisor: Frandsen, Paul (Life Sciences, Plant and Wildlife Sciences)
Caddisfly (Insecta: Trichoptera) silk is unique from other insect's silk in that it retains its adhesive capabilities, strength and viscoelasticity when submerged in water. To understand how caddisfly silk is capable of possessing these characteristics, it is essential to understand the protein foundation of the silk proteins. Caddisfly silk is complex and made up of different structures generated by processes that are unique to caddisfly silk. H-Fibroin and L-Fibroin have been identified as two of the major protein components within caddisfly silk (Hatano & Nagashima, 2015). The caddisfly silk fibre experiences unique structures not typically seen in nature. An understanding of the primary structure of the silk fibre is essential in understanding the complexity of the silk's capabilities. In this study, we used proteomic techniques to analyze the complex H-Fibroin protein and the silk fibre in order to look at the underlying structural features of the protein. In doing so, we identified post-translational phosphorylation, metal cation incorporation, and other structural features which contributes to Caddisfly silk's adhesive capabilities, strength and viscoelasticity when submerged in water.
Faculty Advisor: Frandsen, Paul (Life Sciences, Plant and Wildlife Sciences)
Caddisfly (Insecta: Trichoptera) silk is unique from other insect's silk in that it retains its adhesive capabilities, strength and viscoelasticity when submerged in water. To understand how caddisfly silk is capable of possessing these characteristics, it is essential to understand the protein foundation of the silk proteins. Caddisfly silk is complex and made up of different structures generated by processes that are unique to caddisfly silk. H-Fibroin and L-Fibroin have been identified as two of the major protein components within caddisfly silk (Hatano & Nagashima, 2015). The caddisfly silk fibre experiences unique structures not typically seen in nature. An understanding of the primary structure of the silk fibre is essential in understanding the complexity of the silk's capabilities. In this study, we used proteomic techniques to analyze the complex H-Fibroin protein and the silk fibre in order to look at the underlying structural features of the protein. In doing so, we identified post-translational phosphorylation, metal cation incorporation, and other structural features which contributes to Caddisfly silk's adhesive capabilities, strength and viscoelasticity when submerged in water.
Small Mammal Communities of the Darhad Valley, Mongolia
Smith, Chyanne; Jal, Tumursukh; Duuji, Nyam-Ochir; Tumur, Battogtokh; Mull, John (Weber State University)
Faculty Advisor: Mull, John (Weber State University, College of Science; Zoology)
The Darhad Valley, Mongolia, is a sparsely populated area with abundant wildlife and numerous livestock, including: goats, yaks, horses, and sheep. Few studies completed in this location have placed an importance on obtaining baseline species data. To our knowledge, no data have been collected on small mammal diversity, density, and distribution. This study focused on live-trapping small mammals, with an emphasis on rodents, in six locations throughout the Darhad. We aimed to identify species currently present and develop protocols for future work. Captured rodents represented four families: Sciuridae, Arvicolinae, Cricetidae, and Muridae. Common species included striped dwarf hamsters (Cricetulus barabensis), Mongolian silver voles (Alticola semicanus), and Korean field mice (Apodemus peninsulae). Challenges encountered, which must be mitigated in future studies, include: curious humans, resource and waste management, grazing animals, and novel food sources. These studies should also emphasize community composition, range, and presence of ectoparasites, which could transfer zoonotic diseases.
Faculty Advisor: Mull, John (Weber State University, College of Science; Zoology)
The Darhad Valley, Mongolia, is a sparsely populated area with abundant wildlife and numerous livestock, including: goats, yaks, horses, and sheep. Few studies completed in this location have placed an importance on obtaining baseline species data. To our knowledge, no data have been collected on small mammal diversity, density, and distribution. This study focused on live-trapping small mammals, with an emphasis on rodents, in six locations throughout the Darhad. We aimed to identify species currently present and develop protocols for future work. Captured rodents represented four families: Sciuridae, Arvicolinae, Cricetidae, and Muridae. Common species included striped dwarf hamsters (Cricetulus barabensis), Mongolian silver voles (Alticola semicanus), and Korean field mice (Apodemus peninsulae). Challenges encountered, which must be mitigated in future studies, include: curious humans, resource and waste management, grazing animals, and novel food sources. These studies should also emphasize community composition, range, and presence of ectoparasites, which could transfer zoonotic diseases.
The Effects of High Glucose and Pterostilbene on the Oxidant Status of the Red Blood Cell
Garcia, Jessica; Hanks, Hana; Kist, Taylor; Suman, Tanner (Dixie State University)
Faculty Advisor: Meyer, Jennifer (Dixie State University, Physical Sciences)
Antioxidants in the human body regulate reactive oxygen species (ROS). If ROS are increased within the body it can potentially lead to oxidative stress and cell injury. Glucose-6-phosphate dehydrogenase (G6PD) is an enzyme found in the pentose phosphate pathway that assists in reducing the amount of ROS in the bloodstream. Increased concentrations of glucose, commonly found in patients that suffer from type 1 and type 2 diabetes mellitus have been shown to impair G6PD activity, thereby causing damage to erythrocytes (RBCs)2. It has been shown that hyperglycemic conditions decrease the activity of G6PD in RBCs, which is improved by the addition of pterostilbene, a potent antioxidant1. Glutathione, another antioxidant found in the pentose phosphate pathway, is decreased in RBCs within hyperglycemic conditions. The addition of pterostilbene is predicted to increase levels of glutathione under high glucose conditions.
References:
1. Richins, M., & Meyer, J. (2018). Pterostilbene Ameliorates Lipid Peroxidation and Increases Glucose-6-Phosphate Dehydrogenase Activity in Erythrocytes Subjected to High Glucose Conditions. American Heart Association Journals, 138.
2. Zang, Z., Apse, K., Pang, J., & Stanton, R. C. (2000). High glucose inhibits glucose-6-phosphate dehydrogenase via cAMP in aortic endothelial cells. The Journal of Biological Chemistry, 275(51), 40042-40047. Doi: 10.1074/jbc.M007505200
Faculty Advisor: Meyer, Jennifer (Dixie State University, Physical Sciences)
Antioxidants in the human body regulate reactive oxygen species (ROS). If ROS are increased within the body it can potentially lead to oxidative stress and cell injury. Glucose-6-phosphate dehydrogenase (G6PD) is an enzyme found in the pentose phosphate pathway that assists in reducing the amount of ROS in the bloodstream. Increased concentrations of glucose, commonly found in patients that suffer from type 1 and type 2 diabetes mellitus have been shown to impair G6PD activity, thereby causing damage to erythrocytes (RBCs)2. It has been shown that hyperglycemic conditions decrease the activity of G6PD in RBCs, which is improved by the addition of pterostilbene, a potent antioxidant1. Glutathione, another antioxidant found in the pentose phosphate pathway, is decreased in RBCs within hyperglycemic conditions. The addition of pterostilbene is predicted to increase levels of glutathione under high glucose conditions.
References:
1. Richins, M., & Meyer, J. (2018). Pterostilbene Ameliorates Lipid Peroxidation and Increases Glucose-6-Phosphate Dehydrogenase Activity in Erythrocytes Subjected to High Glucose Conditions. American Heart Association Journals, 138.
2. Zang, Z., Apse, K., Pang, J., & Stanton, R. C. (2000). High glucose inhibits glucose-6-phosphate dehydrogenase via cAMP in aortic endothelial cells. The Journal of Biological Chemistry, 275(51), 40042-40047. Doi: 10.1074/jbc.M007505200
The role of Fibroblast Growth Factor 21 (FGF21) in Mitochondrial Disorders (MDs)
Almaw, Naredos; Chaudhuri, Dipayan (University of Utah)
Faculty Advisor: Chaudhuri, Dipayan (School of Medicine, Internal Medicine)
Fibroblast Growth Factor 21 (FGF21), a regulator of metabolism that is typically expressed in the liver, has recently been shown to be induced by other tissues in the body as a response to mitochondrial stress. Elevated levels of serum FGF21 was exhibited in children with mitochondrial mutation-induced mitochondrial dysfunctions. Similarly, in dilated cardiomyopathy, a common type of heart failure (HF) mitochondrial dysfunction is associated with mitochondrial DNA damage. This study aims to determine the signaling pathway that leads to the production and effects of FGF21 during mitochondrial dysfunction associated HF. We hypothesize that in left ventricular failure, cardiomyocytes experience oxidative stress, which initiates signaling pathways that leads to the production of FGF21 by other organs.
To test this hypothesis, HF was induced in four mice models via Transverse Aortic Constriction (TAC), and tissue samples were collected. Messenger RNA (mRNA) was extracted, and quantitative Polymerase Chain Reaction (qPCR) was performed to examine the FGF21 gene expression in control and experiment mice models. The qPCR data showed an upregulation of FGF21 in the heart, liver, and pancreas of experiment mice. qPCR results were confirmed through FGF21 protein expression via western blot. Our preliminary results appear to support our hypothesis that during heart failure, the heart sends stress signals to other organs to produce FGF21. Understanding the origin of FGF21 production could help better understand the critical role it plays in preventing disease progression in HF patients.
Faculty Advisor: Chaudhuri, Dipayan (School of Medicine, Internal Medicine)
Fibroblast Growth Factor 21 (FGF21), a regulator of metabolism that is typically expressed in the liver, has recently been shown to be induced by other tissues in the body as a response to mitochondrial stress. Elevated levels of serum FGF21 was exhibited in children with mitochondrial mutation-induced mitochondrial dysfunctions. Similarly, in dilated cardiomyopathy, a common type of heart failure (HF) mitochondrial dysfunction is associated with mitochondrial DNA damage. This study aims to determine the signaling pathway that leads to the production and effects of FGF21 during mitochondrial dysfunction associated HF. We hypothesize that in left ventricular failure, cardiomyocytes experience oxidative stress, which initiates signaling pathways that leads to the production of FGF21 by other organs.
To test this hypothesis, HF was induced in four mice models via Transverse Aortic Constriction (TAC), and tissue samples were collected. Messenger RNA (mRNA) was extracted, and quantitative Polymerase Chain Reaction (qPCR) was performed to examine the FGF21 gene expression in control and experiment mice models. The qPCR data showed an upregulation of FGF21 in the heart, liver, and pancreas of experiment mice. qPCR results were confirmed through FGF21 protein expression via western blot. Our preliminary results appear to support our hypothesis that during heart failure, the heart sends stress signals to other organs to produce FGF21. Understanding the origin of FGF21 production could help better understand the critical role it plays in preventing disease progression in HF patients.
The Synthesis and Characterization of Liposomes for Future Delivery of Gentamicin to Biofilms
Buehler, Nate; Hoehn, Nick; Stokes, Britt; Tyler, Areiann; Dr. Kopp, Olga (Utah Valley University)
Faculty Advisor: Kopp, Dr.Olga (Utah Valley University, Biology)
Bacterial infections are difficult to treat with antibiotics because of the protective nature of the biofilms produced by bacteria. Biofilms are a common cause of nosocomial and medical devices-related infections. The current treatments for biofilms include mechanically removing the biofilm itself or by treatments with antibiotics. Biofilms usually become resistant to drugs because of the higher frequency of mutation and horizontal gene transfer compared to planktonic cells. Liposomes are promising delivery systems because of their small size, surface characteristics and ability to encapsulate drugs and other molecules. Liposomal particles can slowly release the encapsulated drugs, increasing their distribution in targeted areas. Studies have shown that the fusion between liposomes and bacterial cells enhances the penetration of antibiotics. The purpose of this study is to form liposomes to encapsulate Gentamicin and characterize the formation and characteristics of these liposomes. Liposomes will be formed using the thin film hydration method and characterized using a scanning electron microscope. This project will present an analysis of the use of different ratios of phospholipids and cholesterol to evaluate the stability and ability to carry Gentamicin.
Faculty Advisor: Kopp, Dr.Olga (Utah Valley University, Biology)
Bacterial infections are difficult to treat with antibiotics because of the protective nature of the biofilms produced by bacteria. Biofilms are a common cause of nosocomial and medical devices-related infections. The current treatments for biofilms include mechanically removing the biofilm itself or by treatments with antibiotics. Biofilms usually become resistant to drugs because of the higher frequency of mutation and horizontal gene transfer compared to planktonic cells. Liposomes are promising delivery systems because of their small size, surface characteristics and ability to encapsulate drugs and other molecules. Liposomal particles can slowly release the encapsulated drugs, increasing their distribution in targeted areas. Studies have shown that the fusion between liposomes and bacterial cells enhances the penetration of antibiotics. The purpose of this study is to form liposomes to encapsulate Gentamicin and characterize the formation and characteristics of these liposomes. Liposomes will be formed using the thin film hydration method and characterized using a scanning electron microscope. This project will present an analysis of the use of different ratios of phospholipids and cholesterol to evaluate the stability and ability to carry Gentamicin.
Tree of Trees
Lahy, Neve (Westminster College)
Faculty Advisor: Kruback, Matt (Westminster College, Art)
Tree of Trees
Neve Lahy:
The Uinta-Wasatch-Cache National Forest is a group of national forests within our Wasatch Mountains. The forest is a host to many species of trees, both coniferous and flowering. I have chosen seven of each of the most prominent local cone bearing and flowering trees to focus on.
The trees are a crucial part of the ecosystem and participate in many symbiotic relationships. Without the trees the ecosystem wouldn't survive. It is important to acknowledge the vitality of our local forests not only for us, but for all other organisms dependent on the trees.
A phylogenetic tree is a branching diagram showing the evolutionary relationships among various biological species. Phylogeny is based upon similarities and differences in their physical or genetic characteristics. The branches indicate closeness in relation to one another. The species nearest to each other on the phylogeny are the most closely related organisms based on special derived traits.
Learning how to identify the trees that I am so often surrounded by has brought me a deeper appreciation and love for the organisms around me. Now that I can spend time in trees and know exactly what I am looking at has given me a broader understanding of how the trees not only interact with me but the other ecological factors present in the area. I feel more keenly aware of what it takes to protect these trees and enjoy them more respectfully.
Faculty Advisor: Kruback, Matt (Westminster College, Art)
Tree of Trees
Neve Lahy:
The Uinta-Wasatch-Cache National Forest is a group of national forests within our Wasatch Mountains. The forest is a host to many species of trees, both coniferous and flowering. I have chosen seven of each of the most prominent local cone bearing and flowering trees to focus on.
The trees are a crucial part of the ecosystem and participate in many symbiotic relationships. Without the trees the ecosystem wouldn't survive. It is important to acknowledge the vitality of our local forests not only for us, but for all other organisms dependent on the trees.
A phylogenetic tree is a branching diagram showing the evolutionary relationships among various biological species. Phylogeny is based upon similarities and differences in their physical or genetic characteristics. The branches indicate closeness in relation to one another. The species nearest to each other on the phylogeny are the most closely related organisms based on special derived traits.
Learning how to identify the trees that I am so often surrounded by has brought me a deeper appreciation and love for the organisms around me. Now that I can spend time in trees and know exactly what I am looking at has given me a broader understanding of how the trees not only interact with me but the other ecological factors present in the area. I feel more keenly aware of what it takes to protect these trees and enjoy them more respectfully.
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.
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.
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.
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.
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.
Habitat Preference of Ateles geoffroyi at La Selva Biological Station, Costa Rica
Desdames, Chloe; Smith, Mick (Salt Lake Community College)
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
Spider monkeys (genus Ateles) is one of the many endangered species in Costa Rica and they can now only be found in very few places due to deforestation. It is important for conservation reasons to know what forest types spider monkeys prefer. According to prior research in Surinam, Mexico, and Bolivia, Ateles prefers primary forests (undisturbed, high canopy) over secondary forests (degraded, lower canopy). Ateles are frugivores and their habitat preference depends highly on fruit availability. Fruit is more abundant in primary over secondary forests. This goal of this study was to determine whether spider monkeys in Costa Rica have a similar habitat preference. I hypothesize that spider monkeys will be found more often in high canopy primary forests compared to secondary forests. This research was conducted at La Selva Biological Station in northeastern Costa Rica. La Selva is a protected lowland tropical rainforest consisting of both primary forest (55%) and secondary forest in various stages of regeneration. I conducted a census of Ateles geoffroyi by walking established trails, with markers every 50 meters, in May 2019. Whenever a spider monkey was spotted, the trail name, nearest meter marker, and forest type were recorded. On a census of 9 trails, five Ateles groups were located, with four of the sightings being in primary forests. My hypothesis, that Ateles geoffroyi would be found in primary over secondary forests, was supported because 80% of the sightings were in primary forests. This is especially significant because, of the 20.5 kilometers censused, only 4.6 km censused (22%) were in primary forests with the remaining 15.9 km (78%) in secondary forests. This preference for primary forests agrees with prior studies on Ateles in other Neotropical forests. This highlights the importance of conserving primary forest for the well-being of spider monkeys.
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
Spider monkeys (genus Ateles) is one of the many endangered species in Costa Rica and they can now only be found in very few places due to deforestation. It is important for conservation reasons to know what forest types spider monkeys prefer. According to prior research in Surinam, Mexico, and Bolivia, Ateles prefers primary forests (undisturbed, high canopy) over secondary forests (degraded, lower canopy). Ateles are frugivores and their habitat preference depends highly on fruit availability. Fruit is more abundant in primary over secondary forests. This goal of this study was to determine whether spider monkeys in Costa Rica have a similar habitat preference. I hypothesize that spider monkeys will be found more often in high canopy primary forests compared to secondary forests. This research was conducted at La Selva Biological Station in northeastern Costa Rica. La Selva is a protected lowland tropical rainforest consisting of both primary forest (55%) and secondary forest in various stages of regeneration. I conducted a census of Ateles geoffroyi by walking established trails, with markers every 50 meters, in May 2019. Whenever a spider monkey was spotted, the trail name, nearest meter marker, and forest type were recorded. On a census of 9 trails, five Ateles groups were located, with four of the sightings being in primary forests. My hypothesis, that Ateles geoffroyi would be found in primary over secondary forests, was supported because 80% of the sightings were in primary forests. This is especially significant because, of the 20.5 kilometers censused, only 4.6 km censused (22%) were in primary forests with the remaining 15.9 km (78%) in secondary forests. This preference for primary forests agrees with prior studies on Ateles in other Neotropical forests. This highlights the importance of conserving primary forest for the well-being of spider monkeys.
Impaired Glucose Metabolism in Niacin Deficient Transgenic Mice
Allen, Sierra; Meyer-Ficca, Mirella; Wandersee, Miles (Utah State University)
Faculty Advisor: Meyer-Ficca, Mirella (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)
Niacin, a component of vitamin B3, is necessary for the synthesis of nicotinamide adenine dinucleotide (NAD+). NAD+ is an essential coenzyme in several metabolic processes, including those that regulate glucose and fat homeostasis in the body. Niacin is available to humans in its dietary form through foods such as meat, various legumes, nuts and vegetables. Niacin deficiency has been linked to health problems including age-related decline of cognitive abilities, as seen in Alzheimer's disease, and impaired dermal repair. In cases of extreme niacin deficiency, individuals may even develop pellagra, a disease characterized by dermatitis, diarrhea and dementia. Recent studies in niacin deficient mice indicate that niacin deficiency impairs proper glucose metabolism. Compared to mice with adequate niacin levels, niacin deficient mice experience a significant loss of total body weight and body fat and have impaired glucose regulation in response to insulin. We hypothesize that these metabolic defects result from inadequate levels of NAD+ preventing the conversion of lactate to pyruvate in the gluconeogenesis pathway, which in turn results in decreased blood glucose levels. Another potential explanation is that niacin deficiency results in decreased glycogen stores in the liver, also impeding glucose production. To test these hypotheses, we are studying the relationship between dietary niacin and glucose metabolism in a transgenic mouse model with acquired niacin dependency that accurately represents human niacin metabolism. Results of this study will contribute to better understanding of niacin's role in proper glucose metabolism, with important implications for diabetes and other glucose-related diseases, particularly in populations with limited access to quality food.
Faculty Advisor: Meyer-Ficca, Mirella (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)
Niacin, a component of vitamin B3, is necessary for the synthesis of nicotinamide adenine dinucleotide (NAD+). NAD+ is an essential coenzyme in several metabolic processes, including those that regulate glucose and fat homeostasis in the body. Niacin is available to humans in its dietary form through foods such as meat, various legumes, nuts and vegetables. Niacin deficiency has been linked to health problems including age-related decline of cognitive abilities, as seen in Alzheimer's disease, and impaired dermal repair. In cases of extreme niacin deficiency, individuals may even develop pellagra, a disease characterized by dermatitis, diarrhea and dementia. Recent studies in niacin deficient mice indicate that niacin deficiency impairs proper glucose metabolism. Compared to mice with adequate niacin levels, niacin deficient mice experience a significant loss of total body weight and body fat and have impaired glucose regulation in response to insulin. We hypothesize that these metabolic defects result from inadequate levels of NAD+ preventing the conversion of lactate to pyruvate in the gluconeogenesis pathway, which in turn results in decreased blood glucose levels. Another potential explanation is that niacin deficiency results in decreased glycogen stores in the liver, also impeding glucose production. To test these hypotheses, we are studying the relationship between dietary niacin and glucose metabolism in a transgenic mouse model with acquired niacin dependency that accurately represents human niacin metabolism. Results of this study will contribute to better understanding of niacin's role in proper glucose metabolism, with important implications for diabetes and other glucose-related diseases, particularly in populations with limited access to quality food.
Is behavioral lateralization in the tropical fish Xenophallus umbratilis related to morphological asymmetry?
Johnson, Erik; Johnson, Ellie; Johnson; Jerald (Brigham Young University)
Faculty Advisor: Johnson, Jerald (Brigham Young University, Biology)
It seems counterintuitive that organisms should evolve handedness (or what we might more broadly refer to as "lateralization"). Individuals who can forage equally well with both hands, who can kick equally well with both feet, who can detect stimuli and orient equally well in both directions, and so on—these individuals should be favored relative to those who are either right handed or left handed. Yet in humans, and in several other species, handedness is common, but we still no very little about why. Here we explore this question using a tropical freshwater fish species with an unusual anatomy. Males have a modified fin—the gonopodium—that they use to internally inseminate females. Interestingly, males are either right or left handed for this structure, which terminates with either a dextral or sinistral twist. In this study, we ask a simple question: is there a link between male gonopodium morphology and male behavioral lateralization. We use a detour test approach to determine how males approach different stimuli, turning either to the left or right to more clearly see each type of stimulus. We focus on how males approach potential mates, predators, and novel items. We predict that males with a dextral gonopodium will orient differently than those with a sinistral gonopodium, consistent with the idea that there is link between behavioral lateralization and morphological handedness. If true, it would suggest that reproductive morphology could be linked to brain and behavioral lateralization in vertebrates.
Faculty Advisor: Johnson, Jerald (Brigham Young University, Biology)
It seems counterintuitive that organisms should evolve handedness (or what we might more broadly refer to as "lateralization"). Individuals who can forage equally well with both hands, who can kick equally well with both feet, who can detect stimuli and orient equally well in both directions, and so on—these individuals should be favored relative to those who are either right handed or left handed. Yet in humans, and in several other species, handedness is common, but we still no very little about why. Here we explore this question using a tropical freshwater fish species with an unusual anatomy. Males have a modified fin—the gonopodium—that they use to internally inseminate females. Interestingly, males are either right or left handed for this structure, which terminates with either a dextral or sinistral twist. In this study, we ask a simple question: is there a link between male gonopodium morphology and male behavioral lateralization. We use a detour test approach to determine how males approach different stimuli, turning either to the left or right to more clearly see each type of stimulus. We focus on how males approach potential mates, predators, and novel items. We predict that males with a dextral gonopodium will orient differently than those with a sinistral gonopodium, consistent with the idea that there is link between behavioral lateralization and morphological handedness. If true, it would suggest that reproductive morphology could be linked to brain and behavioral lateralization in vertebrates.
Low dose alcohol enhances dopamine release in the nucleus accumbens via alpha6-containing nicotinic receptors on GABAergic inputs from the ventral tegmental area
Hansen, Wade; Stockard, Alyssa; Anderson, Elizabeth; Yorgason, Jordan; Sudweeks, Sterling; Wu, Jie; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology); Yorgason, Jordan (Life Sciences, Physiology & Developmental Biology); Sudweeks, Sterling (Life Sciences, Physiology & Developmental Biology)
The prevailing view is that enhancement of dopamine (DA) transmission in the mesolimbic underlies the rewarding properties of ethanol (EtOH) and nicotine (NIC). Although the dogma is that EtOH enhancement of DA neural activity contributes to enhancement of DA transmission, DA neurons are not sensitive to rewarding levels of EtOH. However, VTA GABA neurons are sensitive to low-dose EtOH. We have shown previously that EtOH modulation of DA release in the NAc is mediated by α6-containing nicotinic receptors (α6*-nAChRs), that α6*-nAChRs mediate low-dose EtOH effects on VTA GABA neurons and EtOH preference, and α6*-nAChRs may be a molecular target for low-dose EtOH. The aim of this study was to evaluate EtOH effects on VTA GABAergic input to CINs and DA release in the NAc. Using DIO channel rhodopsin-2 (ChR2) viral injections into the VTA of VGAT Cre mice, we found that VTA GABA neurons send an inhibitory projection to CINs, replicating what has been demonstrated by others. Low-dose EtOH (IC50 = 10 mM) decreased optically-evoked IPSCs (oIPSCs) on CINs and enhanced (EC50 = 10 mM) CIN-mediated spontaneous DA release. Surprisingly, oIPSCs on CINs were not blocked by typical GABAA receptor (GABAAR) antagonists, but by GABAR rho-1 antagonists, suggesting involvement of atypical GABARs on CINs that are postsynaptic to VTA GABAergic input. The α6-conotoxin MII blocked the effects of EtOH on spontaneous DA release and optically-evoked DA release in choline acetyltransferase (ChAT) ChR2 mice. Chronic administration of NIC enhanced EtOH consumption in the drink-in-the-dark procedure and EtOH preference in the CPP procedure and concomitantly enhanced expression of α6*-nAChRs in VTA GABA neurons, without affecting other nAChR subunits. Taken together, these findings suggest that VTA GABA neuron inhibitory input to CINs is modulated by α6*-nAChRs and sensitive to low-dose EtOH, which may underlie the rewarding properties of EtOH.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology); Yorgason, Jordan (Life Sciences, Physiology & Developmental Biology); Sudweeks, Sterling (Life Sciences, Physiology & Developmental Biology)
The prevailing view is that enhancement of dopamine (DA) transmission in the mesolimbic underlies the rewarding properties of ethanol (EtOH) and nicotine (NIC). Although the dogma is that EtOH enhancement of DA neural activity contributes to enhancement of DA transmission, DA neurons are not sensitive to rewarding levels of EtOH. However, VTA GABA neurons are sensitive to low-dose EtOH. We have shown previously that EtOH modulation of DA release in the NAc is mediated by α6-containing nicotinic receptors (α6*-nAChRs), that α6*-nAChRs mediate low-dose EtOH effects on VTA GABA neurons and EtOH preference, and α6*-nAChRs may be a molecular target for low-dose EtOH. The aim of this study was to evaluate EtOH effects on VTA GABAergic input to CINs and DA release in the NAc. Using DIO channel rhodopsin-2 (ChR2) viral injections into the VTA of VGAT Cre mice, we found that VTA GABA neurons send an inhibitory projection to CINs, replicating what has been demonstrated by others. Low-dose EtOH (IC50 = 10 mM) decreased optically-evoked IPSCs (oIPSCs) on CINs and enhanced (EC50 = 10 mM) CIN-mediated spontaneous DA release. Surprisingly, oIPSCs on CINs were not blocked by typical GABAA receptor (GABAAR) antagonists, but by GABAR rho-1 antagonists, suggesting involvement of atypical GABARs on CINs that are postsynaptic to VTA GABAergic input. The α6-conotoxin MII blocked the effects of EtOH on spontaneous DA release and optically-evoked DA release in choline acetyltransferase (ChAT) ChR2 mice. Chronic administration of NIC enhanced EtOH consumption in the drink-in-the-dark procedure and EtOH preference in the CPP procedure and concomitantly enhanced expression of α6*-nAChRs in VTA GABA neurons, without affecting other nAChR subunits. Taken together, these findings suggest that VTA GABA neuron inhibitory input to CINs is modulated by α6*-nAChRs and sensitive to low-dose EtOH, which may underlie the rewarding properties of EtOH.
Loading a Novel Anti-biofilm Compound into Polyurethane Foam for Use in Negative Pressure Wound Therapy
Rawson, Kaden; Nueberger, Travis; Looper, Ryan; Sebahar, Paul; Williams, Dustin (University of Utah)
Faculty Advisor: Williams, Dustin (Engineering, Bioengineering)
Negative pressure wound therapy (NPWT) is commonly used to treat high energy, traumatic battlefield-related injuries, typically caused by an explosion. NPWT may be applied in the field at the time of injury or in the operating room as a therapeutic measure. Wounds are susceptible to contamination from the soil, which contains high amounts of bacteria (>10^9 colony forming units (CFU)/g of material). Greater than 99% of wild-type bacteria favor the biofilm phenotype in the natural world. Biofilms are aggregates of bacteria that are more resistant to traditional antibiotics due to their altered phenotypic and metabolic expressions. Thus, developed biofilms can potentially contaminate these wounds and lead to chronic infection. Furthermore, the lattice structure of polyurethane (PU) foam used in NPWT can potentially harbor and encourage increased biofilm growth. Since the introduction of NPWT as a standard of care for soldiers in 2004, "superficial and deep infections of soft tissue remain a clinical concern after sustaining combat-related trauma [while] using NPWT." To date, GRANUFOAM Silver by KCI is the only variation of PU foam for NPWT that possesses any degree of antimicrobial efficacy. However, silver nanoparticles are minimally effective against biofilms. Thus, the goal of this project is to develop a PU foam that is loaded with a biofilm-specific antimicrobial compound, CZ-01179 in order to decrease the rate of infection when NPWT is utilized in the field of battle.
To date, two prototypes have been developed: One prototype (V1) relies on THF and H2O to coat the Pu foam with CZ-01179 while the second prototype (V2) relies on a hydrogel scaffold to provide a sustained release of CZ-01179 over 24 hours. V1 has been shown to reduce MRSA AND A. baumanii by 7 Log10 CFU during in vitro dilution testing compared to a 1 Log10 reduction produced by GRANUFOAM Silver.
Faculty Advisor: Williams, Dustin (Engineering, Bioengineering)
Negative pressure wound therapy (NPWT) is commonly used to treat high energy, traumatic battlefield-related injuries, typically caused by an explosion. NPWT may be applied in the field at the time of injury or in the operating room as a therapeutic measure. Wounds are susceptible to contamination from the soil, which contains high amounts of bacteria (>10^9 colony forming units (CFU)/g of material). Greater than 99% of wild-type bacteria favor the biofilm phenotype in the natural world. Biofilms are aggregates of bacteria that are more resistant to traditional antibiotics due to their altered phenotypic and metabolic expressions. Thus, developed biofilms can potentially contaminate these wounds and lead to chronic infection. Furthermore, the lattice structure of polyurethane (PU) foam used in NPWT can potentially harbor and encourage increased biofilm growth. Since the introduction of NPWT as a standard of care for soldiers in 2004, "superficial and deep infections of soft tissue remain a clinical concern after sustaining combat-related trauma [while] using NPWT." To date, GRANUFOAM Silver by KCI is the only variation of PU foam for NPWT that possesses any degree of antimicrobial efficacy. However, silver nanoparticles are minimally effective against biofilms. Thus, the goal of this project is to develop a PU foam that is loaded with a biofilm-specific antimicrobial compound, CZ-01179 in order to decrease the rate of infection when NPWT is utilized in the field of battle.
To date, two prototypes have been developed: One prototype (V1) relies on THF and H2O to coat the Pu foam with CZ-01179 while the second prototype (V2) relies on a hydrogel scaffold to provide a sustained release of CZ-01179 over 24 hours. V1 has been shown to reduce MRSA AND A. baumanii by 7 Log10 CFU during in vitro dilution testing compared to a 1 Log10 reduction produced by GRANUFOAM Silver.
Non-occupational Crystalline Silica Exposure from Sand and Gravel Pits in Utah
Greenhalgh, Mitchell; Merrill, Alex; Lopez, David; Lefevre, Sam; Williams, Greg (Brigham Young University)
Faculty Advisor: Abbott, Ben (Brigham Young University, Plant and Wildlife Sciences)
The presence of sand and gravel pits around Utah are usually accompanied by public complaints of increased negative health outcomes. The primary risk from these areas is crystalline silica (CS)—the molecule released into the air as a result of crushing rocks and sand. Literature has given mixed results of the potentially harmful effects of crystalline silica. To address the potential health risk of Utah residents from living near sand pits, we performed a meta-analysis on CS-related literature to estimate the true effects of CS on human health. We then used GIS software to estimate the total population in Utah that lives within a 5000-meter buffer of the sand pits in Utah. Using Utah cancer data, birth data, and hospital emergency department data, we created risk ratios for residents within the buffer. The meta-analysis concluded that CS is a weak lung carcinogen. Other research suggests that air pollution leads to low birth weight and preterm births. In our study, lung cancer rates were significantly lower in populations within the 5000-meter buffer. We found no evidence of significant adverse birth outcomes as a result of living in close proximity to sand and gravel pits. Non-malignant respiratory disease also had significantly lower rates within the buffer. These findings are important in determining the role of sand/gravel pit operations in disease incidence in surrounding communities. More research is needed to evaluate confounding factors such as smoking prevalence and socioeconomic status and to investigate crystalline silica in non-occupational settings.
Faculty Advisor: Abbott, Ben (Brigham Young University, Plant and Wildlife Sciences)
The presence of sand and gravel pits around Utah are usually accompanied by public complaints of increased negative health outcomes. The primary risk from these areas is crystalline silica (CS)—the molecule released into the air as a result of crushing rocks and sand. Literature has given mixed results of the potentially harmful effects of crystalline silica. To address the potential health risk of Utah residents from living near sand pits, we performed a meta-analysis on CS-related literature to estimate the true effects of CS on human health. We then used GIS software to estimate the total population in Utah that lives within a 5000-meter buffer of the sand pits in Utah. Using Utah cancer data, birth data, and hospital emergency department data, we created risk ratios for residents within the buffer. The meta-analysis concluded that CS is a weak lung carcinogen. Other research suggests that air pollution leads to low birth weight and preterm births. In our study, lung cancer rates were significantly lower in populations within the 5000-meter buffer. We found no evidence of significant adverse birth outcomes as a result of living in close proximity to sand and gravel pits. Non-malignant respiratory disease also had significantly lower rates within the buffer. These findings are important in determining the role of sand/gravel pit operations in disease incidence in surrounding communities. More research is needed to evaluate confounding factors such as smoking prevalence and socioeconomic status and to investigate crystalline silica in non-occupational settings.
Optimal chemotherapeutic combination of 9 putative natural compounds
Berlin, Ian; Kenealey, Jason. (Brigham Young University)
Faculty Advisor: Kenealey, Jason (Life Science; Nutrition, Dietetics, and Food Science)
Prostate cancer accounts for 9.9% of all new cancer cases in the United States annually, and thought it has high 5-year survival rate of 98%, but its prognosis changes if the cancer becomes drug resistant or metastases. Natural compounds are often used and studied for their potential chemotherapeutic effects or their sensitizing effects which increases the cancer cells susceptibility to treatment. Traditional Chinese medicine is a common source for finding bioactive small molecules which may have chemotherapeutic effects. This study focused on 9 putative natural compounds and their effectiveness of treating PC-3 prostate cancer cells. First their IC50s were calculated and then used in Mixture Design Response Surface Methodology (MDRSM) to determine the optimal mixture ratio and used in Chou Talalay statistical analysis to determine if combination effects were synergistic, antagonistic or additive. The compounds used in ascending order starting at the most potent or lowest IC50 to highest; Triptolide, .01819uM (Ttd), Shikonin, .6002uM (Shk), Curcumin 20.83uM (Cur), Emodin, 57.38uM (Em), Wogonin, 97.87uM (Wo) Berberine, 101.4uM (BB), Silibinin, 106.2uM (or Silybin) (Sy), Epigallocatechin gallate, 272.6uM (EGCG), and beta Elemene, 304.3uM (beta-E). Emodin, Silibinin and EGCG all appeared to act primarily via cell cycle inhibition and their effectiveness was found to increase in combination with other small molecules. The ideal combination was provided a multi-faced approach reduce cell viability which suggests it may help treat prostate cancer cells in vivo either in tandem or alone.
Faculty Advisor: Kenealey, Jason (Life Science; Nutrition, Dietetics, and Food Science)
Prostate cancer accounts for 9.9% of all new cancer cases in the United States annually, and thought it has high 5-year survival rate of 98%, but its prognosis changes if the cancer becomes drug resistant or metastases. Natural compounds are often used and studied for their potential chemotherapeutic effects or their sensitizing effects which increases the cancer cells susceptibility to treatment. Traditional Chinese medicine is a common source for finding bioactive small molecules which may have chemotherapeutic effects. This study focused on 9 putative natural compounds and their effectiveness of treating PC-3 prostate cancer cells. First their IC50s were calculated and then used in Mixture Design Response Surface Methodology (MDRSM) to determine the optimal mixture ratio and used in Chou Talalay statistical analysis to determine if combination effects were synergistic, antagonistic or additive. The compounds used in ascending order starting at the most potent or lowest IC50 to highest; Triptolide, .01819uM (Ttd), Shikonin, .6002uM (Shk), Curcumin 20.83uM (Cur), Emodin, 57.38uM (Em), Wogonin, 97.87uM (Wo) Berberine, 101.4uM (BB), Silibinin, 106.2uM (or Silybin) (Sy), Epigallocatechin gallate, 272.6uM (EGCG), and beta Elemene, 304.3uM (beta-E). Emodin, Silibinin and EGCG all appeared to act primarily via cell cycle inhibition and their effectiveness was found to increase in combination with other small molecules. The ideal combination was provided a multi-faced approach reduce cell viability which suggests it may help treat prostate cancer cells in vivo either in tandem or alone.
Precipitation and Thunder Associated Vocalizations in Mantled Howler Monkeys (Alouatta palliata)
Pehkonen, Eliza (Salt Lake Community College)
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
Precipitation-associated behaviors have been observed in several species of primate including bonobos (e.g., building leafy shelters), chimpanzees (e.g., drinking, rain dancing displays), and mantled howler monkeys (e.g., licking rain from the air, altering typical behavior based on weather and season). The purpose of this study is to determine if mantled howler monkeys (Alouatta palliata) exhibit precipitation-associated vocalizations. A. palliata is well known for its vocalizations, which are the loudest sound made by any terrestrial mammal and are used for a wide variety of communicative purposes, such as attracting mates, defending territory, and deterring predation. Given the purpose with which A. palliata vocalizes and the existence of precipitation-associated behaviors within primate species, including A. palliata, it was hypothesized that A. palliata would vocalize in association with climatic events (precipitation and thunder). To test this hypothesis, 41.75 hours of data were collected on A. palliata over a two-week time period during the rainy season at La Selva Biological Station in Costa Rica. All-occurrence sampling was used to record the timing and duration of all A. palliata vocalizations, precipitation, and thunder events. Events were considered accompanied if they occurred within five minutes of one another. Of the 59 observed vocalization events 53% were associated with climatic events. Of the 20 observed precipitation events 90% were accompanied by vocalizations and of the 37 observed thunder events 57% were accompanied by vocalization. Associated vocalizations occurred before, during and after climatic events, however, during or after were most common. The data indicate an association between A. palliata vocalization and precipitation, confirming the hypothesis. Further research is warranted to investigate a possible purpose of precipitation-associated vocalizations, an understanding of which could provide further insight into A. palliata's behavioral interaction with climatic events.
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
Precipitation-associated behaviors have been observed in several species of primate including bonobos (e.g., building leafy shelters), chimpanzees (e.g., drinking, rain dancing displays), and mantled howler monkeys (e.g., licking rain from the air, altering typical behavior based on weather and season). The purpose of this study is to determine if mantled howler monkeys (Alouatta palliata) exhibit precipitation-associated vocalizations. A. palliata is well known for its vocalizations, which are the loudest sound made by any terrestrial mammal and are used for a wide variety of communicative purposes, such as attracting mates, defending territory, and deterring predation. Given the purpose with which A. palliata vocalizes and the existence of precipitation-associated behaviors within primate species, including A. palliata, it was hypothesized that A. palliata would vocalize in association with climatic events (precipitation and thunder). To test this hypothesis, 41.75 hours of data were collected on A. palliata over a two-week time period during the rainy season at La Selva Biological Station in Costa Rica. All-occurrence sampling was used to record the timing and duration of all A. palliata vocalizations, precipitation, and thunder events. Events were considered accompanied if they occurred within five minutes of one another. Of the 59 observed vocalization events 53% were associated with climatic events. Of the 20 observed precipitation events 90% were accompanied by vocalizations and of the 37 observed thunder events 57% were accompanied by vocalization. Associated vocalizations occurred before, during and after climatic events, however, during or after were most common. The data indicate an association between A. palliata vocalization and precipitation, confirming the hypothesis. Further research is warranted to investigate a possible purpose of precipitation-associated vocalizations, an understanding of which could provide further insight into A. palliata's behavioral interaction with climatic events.
Quantification of Staphylococcus Biofilm Clearance
Kaneshiro, Alma; Jordan, Adam; Crompton, Rhees; Brailsford, Samantha; Spencer, Jonathan (Weber State University)
Faculty Advisor: Clark, Daniel (Science, Microbiology Department and Neuroscience Center); Chaston, John (Life Sciences, Plant & Wildlife Sciences)
Antibiotic resistance is of great concern in the medical community, with bacterial resistance increasing proportional to their use. Staphylococcus aureus, such as methicillin resistant S. aureus (MRSA), can cause fatal infections. Problems due to this resistance are compounded when the infecting bacteria form a biofilm, thick sticky layers of bacterial secretions, which are difficult for antibiotics to penetrate. Biofilm formation is common in hospital settings on stents, catheters, and IV lines. Biofilms make antibiotic treatment risky due to incomplete killing—the most resistant survive exposure. There is evidence that bacteriophage can break up biofilms, possibly making them more susceptible to antibiotics. We induced a S. aureus biofilm formation using chemicals that mimic a skin wound. Using bacteriophage K, we inoculated the biofilm and observed clearance. Samples of cell pellets and liquid supernatant were collected, and DNA was extracted. Real-time PCR was used to quantify the levels of bacteriophage K replication, representing clearance of the bacteria. This research can be used to find efficient ways to treat an infection caused by a S. aureus biofilm. Bacteriophage used in combination with antibiotics may be able to better clear a biofilm infection and reduce antibiotic resistance risk due to more complete infection clearance.
Faculty Advisor: Clark, Daniel (Science, Microbiology Department and Neuroscience Center); Chaston, John (Life Sciences, Plant & Wildlife Sciences)
Antibiotic resistance is of great concern in the medical community, with bacterial resistance increasing proportional to their use. Staphylococcus aureus, such as methicillin resistant S. aureus (MRSA), can cause fatal infections. Problems due to this resistance are compounded when the infecting bacteria form a biofilm, thick sticky layers of bacterial secretions, which are difficult for antibiotics to penetrate. Biofilm formation is common in hospital settings on stents, catheters, and IV lines. Biofilms make antibiotic treatment risky due to incomplete killing—the most resistant survive exposure. There is evidence that bacteriophage can break up biofilms, possibly making them more susceptible to antibiotics. We induced a S. aureus biofilm formation using chemicals that mimic a skin wound. Using bacteriophage K, we inoculated the biofilm and observed clearance. Samples of cell pellets and liquid supernatant were collected, and DNA was extracted. Real-time PCR was used to quantify the levels of bacteriophage K replication, representing clearance of the bacteria. This research can be used to find efficient ways to treat an infection caused by a S. aureus biofilm. Bacteriophage used in combination with antibiotics may be able to better clear a biofilm infection and reduce antibiotic resistance risk due to more complete infection clearance.
Sex differences in MAP kinase activation in the periaqueductal gray after morphine treatment
Ashley McCarty, Akila Ram, Max V. McDermott, Erin N. Bobeck (Utah State University)
Faculty Advisor: Bobeck, Erin (College of Science, Biology Department)
Morphine is a potent opioid analgesic, but its long term use can lead to negative side effects, including tolerance, which is a decrease in the effectiveness of the opioid. An area of active interest is looking into the molecular effects of chronic morphine treatment in the Periaqueductal gray (PAG), a brain region that controls descending pain modulation. One such molecular target within the PAG is extracellular-signal regulated kinase 1/2 (ERK). Previous studies have shown that pharmacological inhibition of ERK enhanced morphine tolerance, indicating that ERK activity is associated with better responsiveness to morphine. The PAG is known to contain a heterogenous population of neurons including GABA and glutamate subtypes. However, which neurons ERK is activated in within the PAG following morphine tolerance is unknown. Further, there are known differences in PAG activity between male and female mice. However, these sex-differences have not been well studied after morphine tolerance using acute pain tests. The purpose of this research is to investigate differences in ERK activation following morphine tolerance in male and female mice. We treated wild-type male and female mice with morphine (10 mg/kg, i.p.) or saline for 5 days to induce morphine tolerance, following which both behavior and protein immunofluorescence were assessed. We observe sex-specific differences in ERK activation levels and morphine antinociceptive tolerance in mice. We also assessed co-localization of ERK with GABA and glutamate neurons after morphine tolerance. The study will help us understand the cell-type specificity of kinase activation following morphine tolerance. Further this will give us more information about the nature of neurons that are contributing to sex-differences in opioid functions within the PAG
Faculty Advisor: Bobeck, Erin (College of Science, Biology Department)
Morphine is a potent opioid analgesic, but its long term use can lead to negative side effects, including tolerance, which is a decrease in the effectiveness of the opioid. An area of active interest is looking into the molecular effects of chronic morphine treatment in the Periaqueductal gray (PAG), a brain region that controls descending pain modulation. One such molecular target within the PAG is extracellular-signal regulated kinase 1/2 (ERK). Previous studies have shown that pharmacological inhibition of ERK enhanced morphine tolerance, indicating that ERK activity is associated with better responsiveness to morphine. The PAG is known to contain a heterogenous population of neurons including GABA and glutamate subtypes. However, which neurons ERK is activated in within the PAG following morphine tolerance is unknown. Further, there are known differences in PAG activity between male and female mice. However, these sex-differences have not been well studied after morphine tolerance using acute pain tests. The purpose of this research is to investigate differences in ERK activation following morphine tolerance in male and female mice. We treated wild-type male and female mice with morphine (10 mg/kg, i.p.) or saline for 5 days to induce morphine tolerance, following which both behavior and protein immunofluorescence were assessed. We observe sex-specific differences in ERK activation levels and morphine antinociceptive tolerance in mice. We also assessed co-localization of ERK with GABA and glutamate neurons after morphine tolerance. The study will help us understand the cell-type specificity of kinase activation following morphine tolerance. Further this will give us more information about the nature of neurons that are contributing to sex-differences in opioid functions within the PAG
Role of the CD5 T cell co-receptor in T cell metabolism
Haynie, Christopher; Freitas, Claudia M. Tellez; Whitley, Kiara V.; Weber, K. Scott (Brigham Young University)
Faculty Advisor: Weber, K. Scott (Life Sciences, Microbiology and Molecular Biology)
T cells play a critical role in the adaptive immune response and undergo significant metabolic changes upon activation. T cell co-receptors influence T cell activation and function, yet their influence on T cell metabolism remains unclear. CD5, an inhibitory co-receptor expressed on the surface of T cells, is known to regulate thymocyte selection and T cell receptor (TCR) signaling. We previously observed that CD5 plays a critical role in calcium signaling in naïve helper T cells. As calcium signaling influences metabolic changes in cells, our current work focuses on understanding the role of CD5 in T cell metabolism. To understand how CD5 regulates metabolism in T cells, we used CD5 deficient T cells and compared them to wildtype CD5 sufficient T cells. We have characterized their metabolic activity using glycolytic and mitochondrial respiration assays. Interestingly, CD5 deficient naïve T cells have increased glycolysis, mitochondrial respiration, and spare respiratory capacity in comparison to wildtype T cells. We hypothesize that this is due to CD5 altering mitochondrial membrane potential and mass, gene regulation, and the influence of different cellular fuels. Understanding how CD5 regulates T cell metabolism will provide critical insights for improved immunotherapeutic strategies.
Faculty Advisor: Weber, K. Scott (Life Sciences, Microbiology and Molecular Biology)
T cells play a critical role in the adaptive immune response and undergo significant metabolic changes upon activation. T cell co-receptors influence T cell activation and function, yet their influence on T cell metabolism remains unclear. CD5, an inhibitory co-receptor expressed on the surface of T cells, is known to regulate thymocyte selection and T cell receptor (TCR) signaling. We previously observed that CD5 plays a critical role in calcium signaling in naïve helper T cells. As calcium signaling influences metabolic changes in cells, our current work focuses on understanding the role of CD5 in T cell metabolism. To understand how CD5 regulates metabolism in T cells, we used CD5 deficient T cells and compared them to wildtype CD5 sufficient T cells. We have characterized their metabolic activity using glycolytic and mitochondrial respiration assays. Interestingly, CD5 deficient naïve T cells have increased glycolysis, mitochondrial respiration, and spare respiratory capacity in comparison to wildtype T cells. We hypothesize that this is due to CD5 altering mitochondrial membrane potential and mass, gene regulation, and the influence of different cellular fuels. Understanding how CD5 regulates T cell metabolism will provide critical insights for improved immunotherapeutic strategies.
Smyd1 Histone Methyltransferase Activity in Heart Failure and Cardiac Hypertrophy Models
Szulik, Marta; Wang, Li; Franklin, Sarah. (University of Utah)
Faculty Advisor: Franklin, Sarah (Medicine, Internal Medicine)
Heart failure (HF) is a type of heart disease characterized by the structural and functional impairment of ventricular filling. In 2016, HF was the underlying cause of death in approximately 78,000 individuals and today more than 6.2 million Americans suffer from heart failure. HF is the final stage for many types of heart disease including cardiac hypertrophy. During hypertrophy, the ventricular walls thicken to help maintain the proper workload needed to continue supplying the body with oxygenated blood. In addition to increase in cell size, cardiac hypertrophy leads to cell death, fibrosis, metabolic reprogramming and reactivation of fetal gene expression. Gene expression is often modulated by changes in chromatin and histone structure via post-translational modifications (PTMs). Histone methylation, a covalent PTM, has been shown to play a significant role in cardiac development.
Smyd1 is a muscle specific lysine histone methyltransferase protein that has a role in early cardiac development and is known to methylate histone H3 on lysine-4. Additionally, loss of Smyd1 in adult mice models has been shown to induce heart failure and hypertrophy whereas overexpression of Smyd1 has been shown to restrict hypertrophic growth in cell model. Although Smyd1 knockdown experiments have been performed in vivo, the effects of knocking down Smyd1 in isolated cardiomyocytes has not been examined. Furthermore, the effects Smyd1 overexpression in adult mammalian heart failure is unknown.
This project seeks to characterize changes in global levels of histone PTM's as a result of either overexpressing or silencing Smyd1. Using proteomic analysis, we have identified the changes in histone methylation and consequently gene expression in the adult heart and isolated cells in response to Smyd1. Our results help us better understand Smyd1 role in the failing heart and help determine it therapeutic potential.
Faculty Advisor: Franklin, Sarah (Medicine, Internal Medicine)
Heart failure (HF) is a type of heart disease characterized by the structural and functional impairment of ventricular filling. In 2016, HF was the underlying cause of death in approximately 78,000 individuals and today more than 6.2 million Americans suffer from heart failure. HF is the final stage for many types of heart disease including cardiac hypertrophy. During hypertrophy, the ventricular walls thicken to help maintain the proper workload needed to continue supplying the body with oxygenated blood. In addition to increase in cell size, cardiac hypertrophy leads to cell death, fibrosis, metabolic reprogramming and reactivation of fetal gene expression. Gene expression is often modulated by changes in chromatin and histone structure via post-translational modifications (PTMs). Histone methylation, a covalent PTM, has been shown to play a significant role in cardiac development.
Smyd1 is a muscle specific lysine histone methyltransferase protein that has a role in early cardiac development and is known to methylate histone H3 on lysine-4. Additionally, loss of Smyd1 in adult mice models has been shown to induce heart failure and hypertrophy whereas overexpression of Smyd1 has been shown to restrict hypertrophic growth in cell model. Although Smyd1 knockdown experiments have been performed in vivo, the effects of knocking down Smyd1 in isolated cardiomyocytes has not been examined. Furthermore, the effects Smyd1 overexpression in adult mammalian heart failure is unknown.
This project seeks to characterize changes in global levels of histone PTM's as a result of either overexpressing or silencing Smyd1. Using proteomic analysis, we have identified the changes in histone methylation and consequently gene expression in the adult heart and isolated cells in response to Smyd1. Our results help us better understand Smyd1 role in the failing heart and help determine it therapeutic potential.
Spatial variation in mercury concentrations of flying insects at Antelope Island
Stoneham, Lisa; Brasso, Dr. Rebecka (Weber State University)
Faculty Advisor: Brasso, Rebecka (Weber State University, Zoology)
Mercury is a toxic heavy metal that poses significant health threats to people and wildlife. The organic form of mercury, methylmercury, is converted from its inorganic form via microbial methylation. Methylmercury is dangerous because it attaches to proteins in the blood, muscle, and other tissues and can cross the blood-brain and placental barriers. Microbial methylation is enhanced in anoxic environments such as wetlands, which are increasingly being classified as mercury hotspots where animals accumulate elevated concentrations relative to those in terrestrial systems. This is concerning for the wetlands of the Great Salt Lake due to its history of anthropogenic inputs of pollutants and its importance as a breeding ground and rest stop for migrating avian species. Previous research has shown significant mercury methylation occurring within the Deep Brine Layer of the GSL. The purpose of this project was to investigate potential spatial variation in mercury concentration in different portions of the GSL. With a focus on invertebrates, we collected insects including brine flies, midges, damselflies, and crane flies from three sites of varying salinity around Antelope Island State Park: Farmington Bay, White Rock Bay, and the Antelope Island Marina. Mercury concentrations in insects were determined using a Nippon MA-3000 Direct Mercury Analyzer. Our results will provide a preliminary assessment of mercury concentrations in flying insects from different habitats around the island. This will help in determining differential risk to insectivorous songbirds, waterfowl, and shorebirds foraging on these common prey species in the GSL.
Faculty Advisor: Brasso, Rebecka (Weber State University, Zoology)
Mercury is a toxic heavy metal that poses significant health threats to people and wildlife. The organic form of mercury, methylmercury, is converted from its inorganic form via microbial methylation. Methylmercury is dangerous because it attaches to proteins in the blood, muscle, and other tissues and can cross the blood-brain and placental barriers. Microbial methylation is enhanced in anoxic environments such as wetlands, which are increasingly being classified as mercury hotspots where animals accumulate elevated concentrations relative to those in terrestrial systems. This is concerning for the wetlands of the Great Salt Lake due to its history of anthropogenic inputs of pollutants and its importance as a breeding ground and rest stop for migrating avian species. Previous research has shown significant mercury methylation occurring within the Deep Brine Layer of the GSL. The purpose of this project was to investigate potential spatial variation in mercury concentration in different portions of the GSL. With a focus on invertebrates, we collected insects including brine flies, midges, damselflies, and crane flies from three sites of varying salinity around Antelope Island State Park: Farmington Bay, White Rock Bay, and the Antelope Island Marina. Mercury concentrations in insects were determined using a Nippon MA-3000 Direct Mercury Analyzer. Our results will provide a preliminary assessment of mercury concentrations in flying insects from different habitats around the island. This will help in determining differential risk to insectivorous songbirds, waterfowl, and shorebirds foraging on these common prey species in the GSL.
Substrate specificity in variants of an aldehyde oxidoreductase
Carter, Riley; Hertig, Jess; Durrant, Doran (Southern Utah University)
Faculty Advisor: Pierce, Elizabeth (Science and Engineering, Physical Science)
Aldehyde oxidoreductases (AOR) are enzymes used to catalyze the conversion between aldehydes and carboxylic acids. Certain bacteria use these enzymes as a source of metabolism or to detoxify aldehydes to less toxic carboxylic acids: Desulfovibrio gigas uses a highly efficient enzyme (DgAOR) to oxidize benzaldehyde in metabolism while E. coli uses a periplasmic AOR (PaoABC) to detoxify aldehydes. These AORs are members of the xanthine oxidase family, but they don't metabolize many of the normal substrates characteristic of this enzyme family, namely purines. Moreover, the active sites of these enzymes have very different environments. Correia, et al (2014) characterized the kinetics and structure of DgAOR with several substrates and found that the Phe425 and Tyr535 residues at the active site likely stabilize aromatic aldehydes by pi stacking. This active site was also buried away from solvent. The active site of PaoABC lacked any significant aromatic residues and was positioned at the surface of the protein. The substrate stabilizing elements at this active site are Leu246 and Pro352. We are interested in why these active sites both are unreactive towards purines given their different chemical and location compared to the solvent. We propose that by mutating PaoABC to have smaller, nonpolar residues at the 246 and 352 position, we may be able to change the specificity of PaoABC to include purines. We also will mutate these residues to aromatic groups to probe at the chemical environment of the active site and its similarities to DgAOR.
Faculty Advisor: Pierce, Elizabeth (Science and Engineering, Physical Science)
Aldehyde oxidoreductases (AOR) are enzymes used to catalyze the conversion between aldehydes and carboxylic acids. Certain bacteria use these enzymes as a source of metabolism or to detoxify aldehydes to less toxic carboxylic acids: Desulfovibrio gigas uses a highly efficient enzyme (DgAOR) to oxidize benzaldehyde in metabolism while E. coli uses a periplasmic AOR (PaoABC) to detoxify aldehydes. These AORs are members of the xanthine oxidase family, but they don't metabolize many of the normal substrates characteristic of this enzyme family, namely purines. Moreover, the active sites of these enzymes have very different environments. Correia, et al (2014) characterized the kinetics and structure of DgAOR with several substrates and found that the Phe425 and Tyr535 residues at the active site likely stabilize aromatic aldehydes by pi stacking. This active site was also buried away from solvent. The active site of PaoABC lacked any significant aromatic residues and was positioned at the surface of the protein. The substrate stabilizing elements at this active site are Leu246 and Pro352. We are interested in why these active sites both are unreactive towards purines given their different chemical and location compared to the solvent. We propose that by mutating PaoABC to have smaller, nonpolar residues at the 246 and 352 position, we may be able to change the specificity of PaoABC to include purines. We also will mutate these residues to aromatic groups to probe at the chemical environment of the active site and its similarities to DgAOR.
The Beta Cell Struggle: How CDKIs and Age Affect Cell Proliferation in Type 1 Diabetes
Jensen, Daelin; Baxter, Melanie (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Nutrition, Dietetics, and Food Science; Life Sciences)
Approximately 1.25 million people are currently living with type 1 diabetes. By 2050, 5 million people are expected to be diagnosed with the disease1. The insulin secreting pancreatic beta cells are essential to control proper glucose absorption and storage in insulin sensitive peripheral tissue. Both type 1 and type 2 diabetes are characterized by decreased functional beta cell mass and, consequently, decreased insulin production. One potential intervention is the use of beta cell transplantation from cadaveric donors. A major impediment to greater application of this treatment is the scarcity of transplant ready beta cells. Increasing the quantity of functional beta cells for transplantation will lead to increased insulin production and better management of the disease. Various genes have been defined that can induce beta cell replication. A major caveat of these findings, however, is that these factors induce replication in young beta cells but not in aged beta cells. Age-dependent morphological changes in the beta cell are poorly understood, despite its relevance to type 1 diabetes: here, we show that insulin-positive tissue area changes with age. Given that the majority of beta cells that will be used for transplant will come from aged donors, it is imperative to understand why aged beta cells are refractory to the aforementioned proliferative mechanisms. The cell cycle is tightly regulated by cyclin-dependent kinases. Cyclin-dependent kinase inhibitors (CDKI's) bind to cyclin dependent kinases, inhibiting cell proliferation. We hypothesized that these CDKIs are responsible for the observed lack of proliferation in aged animals. We demonstrate the expression of the Ink4 and Cip/Kip family of CDKI's by mRNA, protein and histological expression in 5 week and 5 month old primary rat beta cells. In addition, we show how size-related expression differences of CDKIs relate to beta cell proliferation.
Faculty Advisor: Tessem, Jeffery (Nutrition, Dietetics, and Food Science; Life Sciences)
Approximately 1.25 million people are currently living with type 1 diabetes. By 2050, 5 million people are expected to be diagnosed with the disease1. The insulin secreting pancreatic beta cells are essential to control proper glucose absorption and storage in insulin sensitive peripheral tissue. Both type 1 and type 2 diabetes are characterized by decreased functional beta cell mass and, consequently, decreased insulin production. One potential intervention is the use of beta cell transplantation from cadaveric donors. A major impediment to greater application of this treatment is the scarcity of transplant ready beta cells. Increasing the quantity of functional beta cells for transplantation will lead to increased insulin production and better management of the disease. Various genes have been defined that can induce beta cell replication. A major caveat of these findings, however, is that these factors induce replication in young beta cells but not in aged beta cells. Age-dependent morphological changes in the beta cell are poorly understood, despite its relevance to type 1 diabetes: here, we show that insulin-positive tissue area changes with age. Given that the majority of beta cells that will be used for transplant will come from aged donors, it is imperative to understand why aged beta cells are refractory to the aforementioned proliferative mechanisms. The cell cycle is tightly regulated by cyclin-dependent kinases. Cyclin-dependent kinase inhibitors (CDKI's) bind to cyclin dependent kinases, inhibiting cell proliferation. We hypothesized that these CDKIs are responsible for the observed lack of proliferation in aged animals. We demonstrate the expression of the Ink4 and Cip/Kip family of CDKI's by mRNA, protein and histological expression in 5 week and 5 month old primary rat beta cells. In addition, we show how size-related expression differences of CDKIs relate to beta cell proliferation.
The effect of Nr4a3 gene deletion on body weight, blood glucose levels, and glucose tolerance in mice
Yang, Haokun; Herring, Jacob; Elison, Weston; Wynn, Adam; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
Nearly 1 in 10 Americans have type 2 diabetes (T2D), a disease that is characterized by a loss of functional β-cell mass, resulting in decreased insulin secretion and glucose utilization. The pancreatic β-cell is responsible for producing and secreting insulin and monitoring blood glucose levels, and it is crucial to the understanding of T2D. The orphan nuclear receptor Nr4a3 (Nor1) has well-defined roles throughout the body, specifically with fuel utilization in the liver, muscle, and adipose tissues. Here we present data demonstrating that Nr4a3 KO mice have increased body weight, blood glucose levels (fasting and non-fasting), and impaired glucose tolerance when fed a standard diet. Respiration from adipose tissue is significantly impaired in male and female Nr4a3 KO animals. These data demonstrate that Nr4a3 is necessary for whole-body homeostasis. We believe that these data serve as a step toward understanding the pathway of T2D progression and finding a cure.
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
Nearly 1 in 10 Americans have type 2 diabetes (T2D), a disease that is characterized by a loss of functional β-cell mass, resulting in decreased insulin secretion and glucose utilization. The pancreatic β-cell is responsible for producing and secreting insulin and monitoring blood glucose levels, and it is crucial to the understanding of T2D. The orphan nuclear receptor Nr4a3 (Nor1) has well-defined roles throughout the body, specifically with fuel utilization in the liver, muscle, and adipose tissues. Here we present data demonstrating that Nr4a3 KO mice have increased body weight, blood glucose levels (fasting and non-fasting), and impaired glucose tolerance when fed a standard diet. Respiration from adipose tissue is significantly impaired in male and female Nr4a3 KO animals. These data demonstrate that Nr4a3 is necessary for whole-body homeostasis. We believe that these data serve as a step toward understanding the pathway of T2D progression and finding a cure.
The Effects Of Invasive Common Carp On Invertebrate Food Sources For Diving Ducks In Great Salt Lake Wetlands
Karin, Kettenring; Robison, Talin; Leonard, Emily (Utah State University)
Faculty Advisor: Kettering, Karin (S.J. & Jessie E. Quinney College of Natural Resource, Watershed Sciences Department)
The Great Salt Lake (GSL) and its wetlands are important habitat for migrating birds. The GSL wetlands provide crucial habitat for nesting, food, and areas to recover from migration. Common carp are a threat to GSL wetlands. Carp disturb sediments in the water, blocking some of the sunlight from entering the water, which is utilized by aquatic macrophytes and algae. Carp also may be affecting invertebrate populations, which are critical food resources for migrating birds, but these effects have not been well-documented. My research addressed the question: what are the effects of invasive common carp on invertebrate food sources for diving ducks in the Great Salt Lake wetlands? I answered my research question by addressing the following objectives: (1) to identify the benthic, epiphytic, and water-column dwelling invertebrates in Farmington Bay Waterfowl Management Area (WMA), and (2) to determine if common carp are having an impact on the overall density, diversity, and abundance of the invertebrate communities fed on by diving ducks. I compared invertebrate communities (diversity and abundance) between carp-excluded boxes and control boxes. I constructed my carp exclosures of wire mesh and t-posts to prohibit carp from entering while still allowing invertebrates and water to freely move in and out of the exclosure. The control boxes were constructed of t-posts and allowed carp to freely enter and exit the box. I used dipnet and substrate core samples to determine what invertebrates are living in the water column and substrates at Farmington Bay wetlands. Although sample processing is on-going, early results indicate that carp reduce water column invertebrate abundance while effects on invertebrate diversity are thus far inconclusive. Given the importance of GSL wetlands and their invertebrate food sources to migrating diving, my research findings underscore the importance of aggressive carp management.
Faculty Advisor: Kettering, Karin (S.J. & Jessie E. Quinney College of Natural Resource, Watershed Sciences Department)
The Great Salt Lake (GSL) and its wetlands are important habitat for migrating birds. The GSL wetlands provide crucial habitat for nesting, food, and areas to recover from migration. Common carp are a threat to GSL wetlands. Carp disturb sediments in the water, blocking some of the sunlight from entering the water, which is utilized by aquatic macrophytes and algae. Carp also may be affecting invertebrate populations, which are critical food resources for migrating birds, but these effects have not been well-documented. My research addressed the question: what are the effects of invasive common carp on invertebrate food sources for diving ducks in the Great Salt Lake wetlands? I answered my research question by addressing the following objectives: (1) to identify the benthic, epiphytic, and water-column dwelling invertebrates in Farmington Bay Waterfowl Management Area (WMA), and (2) to determine if common carp are having an impact on the overall density, diversity, and abundance of the invertebrate communities fed on by diving ducks. I compared invertebrate communities (diversity and abundance) between carp-excluded boxes and control boxes. I constructed my carp exclosures of wire mesh and t-posts to prohibit carp from entering while still allowing invertebrates and water to freely move in and out of the exclosure. The control boxes were constructed of t-posts and allowed carp to freely enter and exit the box. I used dipnet and substrate core samples to determine what invertebrates are living in the water column and substrates at Farmington Bay wetlands. Although sample processing is on-going, early results indicate that carp reduce water column invertebrate abundance while effects on invertebrate diversity are thus far inconclusive. Given the importance of GSL wetlands and their invertebrate food sources to migrating diving, my research findings underscore the importance of aggressive carp management.
The Effects of the Ketogenic Diet on Learning and Memory
Edwards, Jeffery; Saito, Erin; Blaylock, Tanner; Brantley, Adam; Winzenried, Eric (Brigham Young University)
Faculty Advisor: Edwards, Jeffrey (Life Sciences, Physiology and Developmental Biology)
The ketogenic diet initially began as a significant treatment to prevent epilepsy. More recently it has seen a rise in popularity again, with many attributing positive physiological and cognitive benefits. The purpose of this study is to assess the validity of those claims in an animal model in order to examine this at the cellular level as well as identify possible molecular mechanisms for the changes observed. To quantify this, mice will be fed a diet high in fats and low in carbohydrates. A Morris water maze, radial arm maze, and novel object recognition will then be used to assess the diets effect on behavioral memory. Field electrophysiology will then be performed in the CA1 region of the hippocampus, the region of the brain responsible for mediating memory, to measure two types of synaptic plasticity: long-term potentiation and long-term depression. It has been previously hypothesized that changes in BDNF concentration are a possible explanation for physiological changes caused by the keto diet. To assess this, ANA-12, a TrkB antagonist, will be used to block the effects caused by BDNF. Preliminary data gathered from bathed brain slices of both male and female animals have shown an enhancement of LTP, the cellular equivalent of learning and memory. These data lead us to our hypothesis that the ketogenic diet will cause significant changes in behavioral memory and CA1 synaptic plasticity through altered BDNF levels.
Faculty Advisor: Edwards, Jeffrey (Life Sciences, Physiology and Developmental Biology)
The ketogenic diet initially began as a significant treatment to prevent epilepsy. More recently it has seen a rise in popularity again, with many attributing positive physiological and cognitive benefits. The purpose of this study is to assess the validity of those claims in an animal model in order to examine this at the cellular level as well as identify possible molecular mechanisms for the changes observed. To quantify this, mice will be fed a diet high in fats and low in carbohydrates. A Morris water maze, radial arm maze, and novel object recognition will then be used to assess the diets effect on behavioral memory. Field electrophysiology will then be performed in the CA1 region of the hippocampus, the region of the brain responsible for mediating memory, to measure two types of synaptic plasticity: long-term potentiation and long-term depression. It has been previously hypothesized that changes in BDNF concentration are a possible explanation for physiological changes caused by the keto diet. To assess this, ANA-12, a TrkB antagonist, will be used to block the effects caused by BDNF. Preliminary data gathered from bathed brain slices of both male and female animals have shown an enhancement of LTP, the cellular equivalent of learning and memory. These data lead us to our hypothesis that the ketogenic diet will cause significant changes in behavioral memory and CA1 synaptic plasticity through altered BDNF levels.
The Influence of Staphylococcus Aureus Biofilm-associated Gene Mutations on Biofilm Composition
Berges, Bradford; Wienclaw, Trevor; Ball, Ashley; Richmond, Bradley (Brigham Young University)
Faculty Advisor: Berges, Bradford (Life Sciences, Microbiology and Molecular Biology)
Staphylococcus Aureus (SA) biofilms are serious impediments to immune defenses and antibiotics, making them a major factor in SA infection. Such infections can be highly lethal even using current treatments, representing a major challenge to the healthcare industry. Previous genetic screenings of SA have revealed several genes that may be associated with biofilm formation. While the roles of many of these genes have been studied, little research has been done on how mutations of these genes impact biofilm composition. As several therapeutic options for treating mature SA biofilms require understanding of biofilm composition, a better understanding of how genes influence that composition is critical to improving current treatments and developing new ones.
In this project, we will study the biofilm phenotypes of SA with mutations in common biofilm-associated genes. By comparing the biofilm mass and composition of the wild-type (wt) Je2 strain to strains containing mutated biofilm-associated genes, we hope to uncover the impact that each mutation has on the composition of the biofilm matrix. We will utilize crystal violet assays as well as extracellular DNA and protein quantifying procedures to determine biofilm composition, after which meaningful comparisons can be made between mutant biofilms and wt biofilms.
Faculty Advisor: Berges, Bradford (Life Sciences, Microbiology and Molecular Biology)
Staphylococcus Aureus (SA) biofilms are serious impediments to immune defenses and antibiotics, making them a major factor in SA infection. Such infections can be highly lethal even using current treatments, representing a major challenge to the healthcare industry. Previous genetic screenings of SA have revealed several genes that may be associated with biofilm formation. While the roles of many of these genes have been studied, little research has been done on how mutations of these genes impact biofilm composition. As several therapeutic options for treating mature SA biofilms require understanding of biofilm composition, a better understanding of how genes influence that composition is critical to improving current treatments and developing new ones.
In this project, we will study the biofilm phenotypes of SA with mutations in common biofilm-associated genes. By comparing the biofilm mass and composition of the wild-type (wt) Je2 strain to strains containing mutated biofilm-associated genes, we hope to uncover the impact that each mutation has on the composition of the biofilm matrix. We will utilize crystal violet assays as well as extracellular DNA and protein quantifying procedures to determine biofilm composition, after which meaningful comparisons can be made between mutant biofilms and wt biofilms.
The Role of Bacterial Genotype in Persistence of the Microbiota of Drosophila melanogaster
Gottfredson, Sarah; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)
The microbiome of Drosophila melanogaster can have significant effects on the host, and many of these have been studied. However, the reason why the bacterial species associate with and persist in D. melanogaster has not been studied in depth. Here we define persistence as how long a microbe associates with a host. The early assumption has been that the D. melanogaster gut microbiome is established solely through diet, but recent work suggests that other factors may be at play in the microbiome establishment. This experiment aims to study the correlation between bacterial genotype and persistence in the D. melanogaster microbiome. In this study, a metagenome wide association (MGWAS) was done using 40 different strains of bacteria to find distinct bacterial genes that are significantly correlated with persistence. To do this, each strain was mono-associated with twenty-four individual flies. The flies were reared for fourteen days, transferred onto new food three times a day for two days, homogenized, and plated. Using the significant genes found through the MGWAS, the same experiment protocol will be used to test mutants of these genes for their effect on persistence. These data will provide us with distinct genes that are necessary for effective bacterial persistence.
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)
The microbiome of Drosophila melanogaster can have significant effects on the host, and many of these have been studied. However, the reason why the bacterial species associate with and persist in D. melanogaster has not been studied in depth. Here we define persistence as how long a microbe associates with a host. The early assumption has been that the D. melanogaster gut microbiome is established solely through diet, but recent work suggests that other factors may be at play in the microbiome establishment. This experiment aims to study the correlation between bacterial genotype and persistence in the D. melanogaster microbiome. In this study, a metagenome wide association (MGWAS) was done using 40 different strains of bacteria to find distinct bacterial genes that are significantly correlated with persistence. To do this, each strain was mono-associated with twenty-four individual flies. The flies were reared for fourteen days, transferred onto new food three times a day for two days, homogenized, and plated. Using the significant genes found through the MGWAS, the same experiment protocol will be used to test mutants of these genes for their effect on persistence. These data will provide us with distinct genes that are necessary for effective bacterial persistence.
The search for Lactobacillus wasatchensis.
Thornton, Sherie; Cardona,Rebecca (Weber State University)
Faculty Advisor: Culumber, Michele (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology)
Lactobacillus wasatchensis was initially isolated from cheese produced at Utah State University and was found to be a Non-Starter Lactic Acid Bacteria (NSLAB) that causes late-gas production in cheese that can damage packaging and produce defects in the cheese. The goal of this project was to locate an environmental reservoir for Lactobacillus wasatchensis. Five samples of silage that were in different stages of fermentation and content and raw milk samples were obtained at the Utah State University dairy. Samples were serially diluted, plated on de Man, Rogosa and Sharpe agar supplemented with 1% D-Ribose (NRS-R) and incubated anaerobically for 5 days. Colonies that looked like potential Lb. wasatchensis were selected and regrown for isolation. All isolates were gram-positive rods. The isolates were further grown in broth for DNA extraction, sequencing, and analysis with API 50 carbohydrate panel (API 50CH). The API 50CH results were significantly different from Lb. wasatchensis, which only demonstrates use of ribose in this assay. Sequencing of the 16S rRNA gene, however, produced a match to three isolates from two different silage samples that had 99% sequence identity to Lb. wasatchensis. Further analysis of the isolates is being done to confirm this finding and describe the organism isolated from the soil. We hypothesize that these organisms are very closely related to Lb. wasatchensis and that silage could be an environmental source of contamination.
Faculty Advisor: Culumber, Michele (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology)
Lactobacillus wasatchensis was initially isolated from cheese produced at Utah State University and was found to be a Non-Starter Lactic Acid Bacteria (NSLAB) that causes late-gas production in cheese that can damage packaging and produce defects in the cheese. The goal of this project was to locate an environmental reservoir for Lactobacillus wasatchensis. Five samples of silage that were in different stages of fermentation and content and raw milk samples were obtained at the Utah State University dairy. Samples were serially diluted, plated on de Man, Rogosa and Sharpe agar supplemented with 1% D-Ribose (NRS-R) and incubated anaerobically for 5 days. Colonies that looked like potential Lb. wasatchensis were selected and regrown for isolation. All isolates were gram-positive rods. The isolates were further grown in broth for DNA extraction, sequencing, and analysis with API 50 carbohydrate panel (API 50CH). The API 50CH results were significantly different from Lb. wasatchensis, which only demonstrates use of ribose in this assay. Sequencing of the 16S rRNA gene, however, produced a match to three isolates from two different silage samples that had 99% sequence identity to Lb. wasatchensis. Further analysis of the isolates is being done to confirm this finding and describe the organism isolated from the soil. We hypothesize that these organisms are very closely related to Lb. wasatchensis and that silage could be an environmental source of contamination.
Transformation of Lactobacillus species
Evans, Justin; Murray, Cameron; Crowley, Bailey; Welker, Dennis; (Utah State University)
Faculty Advisor: Welker, Dennis (College of Science, Biology Department)
In previous experiments, we explored the abilities of a set of newly derived vectors to transform Lactobacillus casei, specifically, the 32G and the A2-362 strains. We have now expanded our research to study the abilities of these vectors to transform additional Lactobacillus species, Lactobacillus paracasei strain LPC-37 and Lactobacillus rhamnosus strain HN001. The vectors were transformed into the cells by electroporation, after which the cells were given a 4-hour incubation to allow expression of the erythromycin resistance gene carried on the vectors. The cells were then plated to MRS agar containing erythromycin and incubated for 2-3 days until colonies appeared. The colonies were counted and the transformation efficiencies for each vector tabulated as colony forming units per _g of vector DNA. These studies help us to understand how effective the vectors are at transforming different species of lactic acid bacteria. We can also start to ask why some vectors performed better in some bacterial strains than they did in other strains.
Faculty Advisor: Welker, Dennis (College of Science, Biology Department)
In previous experiments, we explored the abilities of a set of newly derived vectors to transform Lactobacillus casei, specifically, the 32G and the A2-362 strains. We have now expanded our research to study the abilities of these vectors to transform additional Lactobacillus species, Lactobacillus paracasei strain LPC-37 and Lactobacillus rhamnosus strain HN001. The vectors were transformed into the cells by electroporation, after which the cells were given a 4-hour incubation to allow expression of the erythromycin resistance gene carried on the vectors. The cells were then plated to MRS agar containing erythromycin and incubated for 2-3 days until colonies appeared. The colonies were counted and the transformation efficiencies for each vector tabulated as colony forming units per _g of vector DNA. These studies help us to understand how effective the vectors are at transforming different species of lactic acid bacteria. We can also start to ask why some vectors performed better in some bacterial strains than they did in other strains.
Characterizing Lampenflora Diversity in Great Basin National Park to Monitor Disturbances in Fragile Cave Ecosystems
Burgoyne, Jake; Leavitt, Steve (Brigham Young University)
Faculty Advisor: Leavitt, Steve (Life Sciences, Biology)
In show caves, artificially lighting is intended to highlight intricate cave formations for visitors. However, as an unintended consequence, artificial lighting promotes the growth of diverse biofilm communities termed Lampenflora that gain their energy from these novel light sources. Lampenflora, which generally consist of algae and cyanobacteria, discolor formations and introduce novel ecological interactions in simple cave ecosystems. Lampenflora communities have been understudied mainly due to technological limitations and difficult accessibility. However, by characterizing these communities, we can better monitor their impact and develop effective strategies for their removal. Using metagenomic high-throughput sequencing, this research provides the first molecular-based perspective into lampenflora diversity in cave systems in the Great Basin. The data collected, generated, and analyzed is vital in understanding Lampenflora biodiversity and how these communities develop. Furthermore, it offers ecologists a novel perspective on the use molecular detection to understand biodiversity within cave systems.
Faculty Advisor: Leavitt, Steve (Life Sciences, Biology)
In show caves, artificially lighting is intended to highlight intricate cave formations for visitors. However, as an unintended consequence, artificial lighting promotes the growth of diverse biofilm communities termed Lampenflora that gain their energy from these novel light sources. Lampenflora, which generally consist of algae and cyanobacteria, discolor formations and introduce novel ecological interactions in simple cave ecosystems. Lampenflora communities have been understudied mainly due to technological limitations and difficult accessibility. However, by characterizing these communities, we can better monitor their impact and develop effective strategies for their removal. Using metagenomic high-throughput sequencing, this research provides the first molecular-based perspective into lampenflora diversity in cave systems in the Great Basin. The data collected, generated, and analyzed is vital in understanding Lampenflora biodiversity and how these communities develop. Furthermore, it offers ecologists a novel perspective on the use molecular detection to understand biodiversity within cave systems.
Cocoa Epicatechin Metabolites' affect on β Cell Proliferation and Cell Cycle
Ross, Mimi; Tessem, Jeffery; Orton, Emily; Ekpo, Idongesit; Beales, Joseph (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutrition, Dietetics, & Food Science)
In 2015 there were over 30 million Americans with diabetes and over 84 million Americans ages 18 and older had pre-diabetes. With diabetes being the seventh leading cause of death in the United States and becoming more prevalent the race is on to find a cure. One of the main problems with this disease is the decrease in functional β-cell mass. β-cells produce insulin to maintain blood glucose levels at healthy levels. Thus, if we can increase β-cell proliferation we are one step closer to curing diabetes. Cocoa epicatechins have been shown to be beneficial in blocking diabetes progression. Studies have shown that oligomeric and polymeric cocoa epicatechin extracts improve diabetes onset in a mouse model of Type 2 diabetes. We have demonstrated that the oligomeric fraction of cocoa epicatechins enhances β-cell proliferation in an in vitro model. Absorption studies have shown that while the oligomeric and polymeric forms are not readily absorbed in the gut, they are metabolized by gut bacteria and that these metabolites can be observed in circulation. Using flow cytometry we have studied how these phytochemicals: epicatechin, 5-phenylvaleric acid, Homovanilic acid, and Hippuric acid. Here we present the data regarding the effect of microbial cocoa flavanol metabolites on β-cell cell cycle during proliferation.
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutrition, Dietetics, & Food Science)
In 2015 there were over 30 million Americans with diabetes and over 84 million Americans ages 18 and older had pre-diabetes. With diabetes being the seventh leading cause of death in the United States and becoming more prevalent the race is on to find a cure. One of the main problems with this disease is the decrease in functional β-cell mass. β-cells produce insulin to maintain blood glucose levels at healthy levels. Thus, if we can increase β-cell proliferation we are one step closer to curing diabetes. Cocoa epicatechins have been shown to be beneficial in blocking diabetes progression. Studies have shown that oligomeric and polymeric cocoa epicatechin extracts improve diabetes onset in a mouse model of Type 2 diabetes. We have demonstrated that the oligomeric fraction of cocoa epicatechins enhances β-cell proliferation in an in vitro model. Absorption studies have shown that while the oligomeric and polymeric forms are not readily absorbed in the gut, they are metabolized by gut bacteria and that these metabolites can be observed in circulation. Using flow cytometry we have studied how these phytochemicals: epicatechin, 5-phenylvaleric acid, Homovanilic acid, and Hippuric acid. Here we present the data regarding the effect of microbial cocoa flavanol metabolites on β-cell cell cycle during proliferation.
Collared Peccary (Pecari tajucu) Group Size at La Selva Biological Station, Costa Rica
Shin, Seungwon (Salt Lake Community College)
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
The abundance of collared peccaries (Pecari tajucu) is crucial to study because they are a keystone species that plays a large role in their ecosystems. They consume fallen fruits and nuts, disperse seeds, and provide food for predators. Additionally, they are ecosystem engineers altering the landscape for other species. Previous studies have shown that collared peccaries at La Selva Biological Station travel in smaller groups (averaging 10 individuals per group) compared to peccaries at other Neotropical sites. La Selva Biological Station is located in northeastern Costa Rica and it consists of both primary and secondary (i.e. degraded) forests surrounded on three sides by farmland. Due to the general decline of mammals in degraded habitats, I predicted that the average group size of collared peccaries at La Selva will be even smaller than previously reported. I collected data at La Selva Biological Station for two weeks in May 2019. I used three census methods: total count sampling (counting all the species in a certain area), line transect sampling (counting all the species I see when I walk through a trail), and point sampling (standing at selected viewpoints and recording the species visible from that location). I observed 39 peccaries in 17 separate sightings. Group size ranged from 1 to 7 peccaries with an average of 2.3 peccaries per group. Eight sightings (20%) were of single peccaries. My hypothesis that peccary group size would be smaller than 10 individuals was supported. Some limitations of the study were low visibility due to the dense forest and the dispersed social organization of peccary individuals within the group. Both of these factors would underestimate the actual group size of collared peccaries. Nevertheless, the results support previous findings that peccary group size at La Selva are smaller than at other Neotropical sites.
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
The abundance of collared peccaries (Pecari tajucu) is crucial to study because they are a keystone species that plays a large role in their ecosystems. They consume fallen fruits and nuts, disperse seeds, and provide food for predators. Additionally, they are ecosystem engineers altering the landscape for other species. Previous studies have shown that collared peccaries at La Selva Biological Station travel in smaller groups (averaging 10 individuals per group) compared to peccaries at other Neotropical sites. La Selva Biological Station is located in northeastern Costa Rica and it consists of both primary and secondary (i.e. degraded) forests surrounded on three sides by farmland. Due to the general decline of mammals in degraded habitats, I predicted that the average group size of collared peccaries at La Selva will be even smaller than previously reported. I collected data at La Selva Biological Station for two weeks in May 2019. I used three census methods: total count sampling (counting all the species in a certain area), line transect sampling (counting all the species I see when I walk through a trail), and point sampling (standing at selected viewpoints and recording the species visible from that location). I observed 39 peccaries in 17 separate sightings. Group size ranged from 1 to 7 peccaries with an average of 2.3 peccaries per group. Eight sightings (20%) were of single peccaries. My hypothesis that peccary group size would be smaller than 10 individuals was supported. Some limitations of the study were low visibility due to the dense forest and the dispersed social organization of peccary individuals within the group. Both of these factors would underestimate the actual group size of collared peccaries. Nevertheless, the results support previous findings that peccary group size at La Selva are smaller than at other Neotropical sites.
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.
Determining the Function and Structure of Cms1, A Type V CRISPR Effector Endonuclease
Tonks, Adam; Domgaard, Hannah; Crowley, Valerie; Neumann, Gina; Keiser, Dylan; Metcalf, Josie; Guo, Hongjie; Zhou, Yi; Begemann, Mathew; Taylor, David; Jackson, Ryan (Utah State University)
Faculty Advisor: Jackson, Ryan (College of Science, Chemistry and Biochemistry)
Cms1 is a Type V endonuclease that contains a novel domain, shares little sequence homology with other Type V endonucleases, and in some organisms, is found near genes coding for other single-subunit nucleases. Studies in rice (Oryza sativa) have shown Cms1 capable of RNA-directed DNA editing. However, the mechanism of DNA cleavage remains unknown.
Here we present biochemical data that demonstrate Cms1 from Sulfuricurvum processes an RNA guide and binds/cleaves single- and double-stranded DNA through RuvC nuclease motifs. 2-D classification of structures obtained by negative staining electron microscopy show a major conformational change between SuCms1 bound and unbound to an RNA guide. The predicted global structure appears to be different than those reported for other Type V effectors. These data provide for a greater understanding of Type V endonucleases and may provide an alternative tool for genome editing applications.
Faculty Advisor: Jackson, Ryan (College of Science, Chemistry and Biochemistry)
Cms1 is a Type V endonuclease that contains a novel domain, shares little sequence homology with other Type V endonucleases, and in some organisms, is found near genes coding for other single-subunit nucleases. Studies in rice (Oryza sativa) have shown Cms1 capable of RNA-directed DNA editing. However, the mechanism of DNA cleavage remains unknown.
Here we present biochemical data that demonstrate Cms1 from Sulfuricurvum processes an RNA guide and binds/cleaves single- and double-stranded DNA through RuvC nuclease motifs. 2-D classification of structures obtained by negative staining electron microscopy show a major conformational change between SuCms1 bound and unbound to an RNA guide. The predicted global structure appears to be different than those reported for other Type V effectors. These data provide for a greater understanding of Type V endonucleases and may provide an alternative tool for genome editing applications.
Development of a New Molecular Predictor for Risk of Melanoma Brain Metastases
Stehn, Christopher; Colman, Howard; Boucher, Kenneth; Grossman, Allie H; Holmen, Sheri L (University of Utah)
Faculty Advisor: Holmen, Sheri (University of Utah, Surgery)
Despite therapeutic advances in the treatment of melanoma, development of brain metastases continues to be a major cause of treatment failure. Prognosis for patients with brain metastases is exceedingly poor, therefore the development of sensitive and specific biomarkers to predict which melanoma patients are at highest risk for disease progression are needed. To accomplish this goal, we developed a novel combined molecular/clinical/pathologic predictor of brain metastasis risk. We first analyzed multiple gene expression datasets including The Cancer Genome Atlas (TCGA; n = 437) and an independent series from the European Genome-Phenome Archive (n = 183) and identified a list of 60 consensus genes that is robustly predictive of development of melanoma brain metastases (p < 0.05; FDR 5%). Next, we performed a similar analysis of association of miRNAs and melanoma brain metastasis risk which identified a set of miRNAs with significant predictive power. An optimized combined set of 15 mRNA and miRNA markers was a better predictor of brain metastasis risk than either mRNA or miRNA list alone when applied to the TCGA data set. The combined predictor was most sensitive in separating patients with no metastases from those with either brain metastases or systemic metastases. Current efforts are focused on optimizing miRNA and mRNA separation of patients specifically with brain metastases from those with other metastases using a machine learning linear classifier, and with integrating the expression classifier with other clinical and pathologic predictive factors including: age, stage, thickness, location, histology, ulceration, and gender. The sensitivity and specificity of the resulting clinical/molecular predictor will be validated in an independent retrospective patient dataset, and subsequently implemented in a prospective brain metastasis screening trial to determine real-world utility of this approach in preparation for prospective brain metastasis adjuvant/chemoprevention trials utilizing both immunotherapy and targeted therapy approaches.
Faculty Advisor: Holmen, Sheri (University of Utah, Surgery)
Despite therapeutic advances in the treatment of melanoma, development of brain metastases continues to be a major cause of treatment failure. Prognosis for patients with brain metastases is exceedingly poor, therefore the development of sensitive and specific biomarkers to predict which melanoma patients are at highest risk for disease progression are needed. To accomplish this goal, we developed a novel combined molecular/clinical/pathologic predictor of brain metastasis risk. We first analyzed multiple gene expression datasets including The Cancer Genome Atlas (TCGA; n = 437) and an independent series from the European Genome-Phenome Archive (n = 183) and identified a list of 60 consensus genes that is robustly predictive of development of melanoma brain metastases (p < 0.05; FDR 5%). Next, we performed a similar analysis of association of miRNAs and melanoma brain metastasis risk which identified a set of miRNAs with significant predictive power. An optimized combined set of 15 mRNA and miRNA markers was a better predictor of brain metastasis risk than either mRNA or miRNA list alone when applied to the TCGA data set. The combined predictor was most sensitive in separating patients with no metastases from those with either brain metastases or systemic metastases. Current efforts are focused on optimizing miRNA and mRNA separation of patients specifically with brain metastases from those with other metastases using a machine learning linear classifier, and with integrating the expression classifier with other clinical and pathologic predictive factors including: age, stage, thickness, location, histology, ulceration, and gender. The sensitivity and specificity of the resulting clinical/molecular predictor will be validated in an independent retrospective patient dataset, and subsequently implemented in a prospective brain metastasis screening trial to determine real-world utility of this approach in preparation for prospective brain metastasis adjuvant/chemoprevention trials utilizing both immunotherapy and targeted therapy approaches.
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.
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.
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.
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.
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.
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.