Life Sciences
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.
Protein Pens: A New Diagnostic Instrument
Armitstead, Annie; Grether, Lara; Creech, Kealani (Brigham Young University)
Faculty Advisor: Watt, Richard (Brigham Young University, Biochemistry)
Lateral Flow Immunoassays (LFI) are simple tests that detect specific levels of antigens or antibodies in patient samples. Requiring only a few minutes, small sample sizes and no read-out equipment, LFI�s are an invaluable and time efficient testing technique. Made up of multiple layers they facilitate the capillary flow of a sample through protein detection zones and can be developed to detect virtually any disease or condition.
Despite the attractive attributes of these tests, the assembly of an LFI strip requires expensive machines, trained personnel, and materials not easily accessible to low-resourced labs or clinics. Developing an innovative point-of-care platform designed to be streamlined, low-cost, and intelligible to the unskilled would open the door of medicine to even the most underprivileged clinics in the world.
We are currently developing a paper LFI that uses a single sheet of copy paper with the ability to filter whole blood as well as replacing high-priced machines with stencils and pens which can still deliver detection proteins to the designated test zones. This avenue of testing is supported by previous experiments we have done with protein pens and tagged antibodies. Using anti-mouse and anti-hCG antibodies as our control and test lines respectively, we spike our sample with hCG mouse antibodies tagged with nanoparticles, and we are able to see binding of both proteins with their respective antibodies. We have seen results in our new testing technique that is easily comparable with currently commercialized LFI's: visual results of binding within 1 min, successful transformation of printer paper into a functional binding platform, and consistent protein binding at a 1/10^5 concentration. Once this concept can be translated to different inks in order to diagnose a plethora of varying conditions, we'll be able to detect diseases and other important biomarkers no matter the limiting low-resource circumstances.
Faculty Advisor: Watt, Richard (Brigham Young University, Biochemistry)
Lateral Flow Immunoassays (LFI) are simple tests that detect specific levels of antigens or antibodies in patient samples. Requiring only a few minutes, small sample sizes and no read-out equipment, LFI�s are an invaluable and time efficient testing technique. Made up of multiple layers they facilitate the capillary flow of a sample through protein detection zones and can be developed to detect virtually any disease or condition.
Despite the attractive attributes of these tests, the assembly of an LFI strip requires expensive machines, trained personnel, and materials not easily accessible to low-resourced labs or clinics. Developing an innovative point-of-care platform designed to be streamlined, low-cost, and intelligible to the unskilled would open the door of medicine to even the most underprivileged clinics in the world.
We are currently developing a paper LFI that uses a single sheet of copy paper with the ability to filter whole blood as well as replacing high-priced machines with stencils and pens which can still deliver detection proteins to the designated test zones. This avenue of testing is supported by previous experiments we have done with protein pens and tagged antibodies. Using anti-mouse and anti-hCG antibodies as our control and test lines respectively, we spike our sample with hCG mouse antibodies tagged with nanoparticles, and we are able to see binding of both proteins with their respective antibodies. We have seen results in our new testing technique that is easily comparable with currently commercialized LFI's: visual results of binding within 1 min, successful transformation of printer paper into a functional binding platform, and consistent protein binding at a 1/10^5 concentration. Once this concept can be translated to different inks in order to diagnose a plethora of varying conditions, we'll be able to detect diseases and other important biomarkers no matter the limiting low-resource circumstances.
Role of novel receptor GPR171 in chemotherapy-induced neuropathic pain
Edwards, Taylor; Ram, Akila; McCarty, Ashley; Bobeck, Erin N (Utah State University)
Faculty Advisor: Bobeck, Erin (College of Science, Biology Department)
First-line chemotherapies against solid tumors are highly efficacious in reducing the tumor burden, but have many adverse side-effects including nerve damage, leading to chronic pain. Non-addictive, efficacious pain relievers are an area of active interest, and we propose a novel target to address this pressing issue. GPR171 is a G-Protein Coupled Receptor that was recently deorphanized and was identified to be expressed in the brain in regions that regulate reward, anxiety, and pain. Within the pain circuit, it was shown previously that systemic administration of the GPR171 agonist enhances morphine antinociception in acute pain tests. Preliminary data from our lab has shown that GPR171 activation can also alleviate persistent inflammatory pain. However, the role of this receptor has not been investigated in other chronic pain models. Given these findings in acute and inflammatory pain, we hypothesize that GPR171 can reduce neuropathic pain. To test this hypothesis, we investigate the role of GPR171 in chronic neuropathic pain. We tested the efficacy of a GPR171 agonist in a chemotherapy-induced neuropathy mouse model. Neuropathic pain was induced by injecting paclitaxel (16 mg/kg) followed by assessment of the pain-relieving effects of activating GPR171 receptors. Mechanical pain thresholds were assessed using Von Frey filaments. We observed an increase in mechanical thresholds following GPR171 agonist treatment. Further, using immunofluorescence we observed that there is a decrease in GPR171 receptors in the periaqueductal gray (PAG) of these mice that have neuropathic pain, indicating that the agonist can bind to the available receptors to produce pain relief. Overall, this study proposes that GPR171 may be a novel target for the treatment of neuropathic pain.
Faculty Advisor: Bobeck, Erin (College of Science, Biology Department)
First-line chemotherapies against solid tumors are highly efficacious in reducing the tumor burden, but have many adverse side-effects including nerve damage, leading to chronic pain. Non-addictive, efficacious pain relievers are an area of active interest, and we propose a novel target to address this pressing issue. GPR171 is a G-Protein Coupled Receptor that was recently deorphanized and was identified to be expressed in the brain in regions that regulate reward, anxiety, and pain. Within the pain circuit, it was shown previously that systemic administration of the GPR171 agonist enhances morphine antinociception in acute pain tests. Preliminary data from our lab has shown that GPR171 activation can also alleviate persistent inflammatory pain. However, the role of this receptor has not been investigated in other chronic pain models. Given these findings in acute and inflammatory pain, we hypothesize that GPR171 can reduce neuropathic pain. To test this hypothesis, we investigate the role of GPR171 in chronic neuropathic pain. We tested the efficacy of a GPR171 agonist in a chemotherapy-induced neuropathy mouse model. Neuropathic pain was induced by injecting paclitaxel (16 mg/kg) followed by assessment of the pain-relieving effects of activating GPR171 receptors. Mechanical pain thresholds were assessed using Von Frey filaments. We observed an increase in mechanical thresholds following GPR171 agonist treatment. Further, using immunofluorescence we observed that there is a decrease in GPR171 receptors in the periaqueductal gray (PAG) of these mice that have neuropathic pain, indicating that the agonist can bind to the available receptors to produce pain relief. Overall, this study proposes that GPR171 may be a novel target for the treatment of neuropathic pain.
Sexual dimorphism and sexual selection in Alfaro cultratus and the effects of predation on these attributes
Bonnett, Kelsie; Golden, Kaitlyn; Johnson, Jerry (Brigham Young University)
Faculty Advisor: Johnson, Jerald (Brigham Young University, Biology)
Understanding life-history strategies allows us to know how a changing environment affects species and communities. Livebearing Poeciliid fish are commonly used as models to gain a better understanding of these strategies, but some species like Alfaro cultratus have been neglected in this process. A. cultratus is a freshwater fish with a unique keel-shaped anal fin commonly found along the eastern coast of Central America. To understand the life-history strategies of this species and use it as a future model, I am performing an experiment to: 1) determine if there is sexual selection in Alfaro cultratus considering both body size and anal fin length; 2) determine whether A. cultratus displays sexual selection; and 3) understand how predation influences both dimorphism and selection. To do this I will be performing a two-part experiment in which I will first analyze previously collected samples for morphological differences, and second perform a live experiment to test Alfaro female preference. By doing so I will be able to not only advance our understanding of A. cultratus, but of life-history theory and conservation strategies.
Faculty Advisor: Johnson, Jerald (Brigham Young University, Biology)
Understanding life-history strategies allows us to know how a changing environment affects species and communities. Livebearing Poeciliid fish are commonly used as models to gain a better understanding of these strategies, but some species like Alfaro cultratus have been neglected in this process. A. cultratus is a freshwater fish with a unique keel-shaped anal fin commonly found along the eastern coast of Central America. To understand the life-history strategies of this species and use it as a future model, I am performing an experiment to: 1) determine if there is sexual selection in Alfaro cultratus considering both body size and anal fin length; 2) determine whether A. cultratus displays sexual selection; and 3) understand how predation influences both dimorphism and selection. To do this I will be performing a two-part experiment in which I will first analyze previously collected samples for morphological differences, and second perform a live experiment to test Alfaro female preference. By doing so I will be able to not only advance our understanding of A. cultratus, but of life-history theory and conservation 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.
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.
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.
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.
Prophylactic Treatment of Post-Traumatic Stress Disorder with Mifepristone and Propranolol
Boyce, Zach; Smith, Calvin; Martin, Ashlyn; Ketch, Yuko; Dugan, James; Wright, Cole (Brigham Young University)
Faculty Advisor: Jeffrey, Edwards (Brigham Young University, Physiology and Developmental Biology)
Post-traumatic stress disorder (PTSD) is a complex psychological disorder that affects about 1 of 4 individuals after a stressful/traumatic experience. One common model to induce PTSD in rats is social defeat (SD) combined with chronic light exposure. First, we screened rats for natural anxiety to use in the SD protocol. Next, elevated plus maze (EPM) and light-dark transition (LDT) tests were used to detect anxious behavior after SD. The SD protocol induced significant anxious behavior when compared to controls. Next, we performed long-term potentiation (LTP) field electrophysiology synaptic plasticity physiology experiments in brain slices of the ventral hippocampus (VH) and basolateral amygdala (BLA), regions known to have altered enhanced plasticity in PTSD. SD significantly increased LTP in the VH (~25% greater than control) and BLA (~35% greater than control). To determine whether a prophylactic treatment could prevent the physiological changes of PTSD, we simultaneously administered two drugs at 10 mg/kg doses by intraperitoneal injection one week prior to and for the duration of SD. The first, propranolol, is a beta-adrenergic receptor antagonist, and the second, mifepristone, is a glucocorticoid receptor antagonist; thus, treatment would target the action of stress hormones altered in PTSD. To determine whether a prophylactic treatment could prevent the physiological changes of PTSD, propranolol and mifepristone, antagonists of two stress receptors, were simultaneously administered at 10 mg/kg doses by intraperitoneal (IP) injection one week prior to and for the duration of SThese drugs significantly decreased LTP in the VH and BLA back to near-control levels while SD rats with vehicle injections still had elevated LTP. However, SD drug-treated rats did not show significant reductions in anxious behavior compared to non-injected SD rats and also exhibited significantly more anxious behavior than control rats, suggesting the IP injection induced added stress. Next, we used rtPCR to examine gene expression of drug targets and plasticity markers to determine potential mechanisms for observed LTP changes. In both the VH and BLA, SD was associated with a significant decrease in glucocorticoid and mineralocorticoid receptor expression, which was restored to control levels under drug treatment. Overall, our data suggest that propranolol and mifepristone together may be a potential prophylactic treatment for preventing PTSD through a mechanism likely mediated by glucocorticoid/mineralocorticoid receptors.
Faculty Advisor: Jeffrey, Edwards (Brigham Young University, Physiology and Developmental Biology)
Post-traumatic stress disorder (PTSD) is a complex psychological disorder that affects about 1 of 4 individuals after a stressful/traumatic experience. One common model to induce PTSD in rats is social defeat (SD) combined with chronic light exposure. First, we screened rats for natural anxiety to use in the SD protocol. Next, elevated plus maze (EPM) and light-dark transition (LDT) tests were used to detect anxious behavior after SD. The SD protocol induced significant anxious behavior when compared to controls. Next, we performed long-term potentiation (LTP) field electrophysiology synaptic plasticity physiology experiments in brain slices of the ventral hippocampus (VH) and basolateral amygdala (BLA), regions known to have altered enhanced plasticity in PTSD. SD significantly increased LTP in the VH (~25% greater than control) and BLA (~35% greater than control). To determine whether a prophylactic treatment could prevent the physiological changes of PTSD, we simultaneously administered two drugs at 10 mg/kg doses by intraperitoneal injection one week prior to and for the duration of SD. The first, propranolol, is a beta-adrenergic receptor antagonist, and the second, mifepristone, is a glucocorticoid receptor antagonist; thus, treatment would target the action of stress hormones altered in PTSD. To determine whether a prophylactic treatment could prevent the physiological changes of PTSD, propranolol and mifepristone, antagonists of two stress receptors, were simultaneously administered at 10 mg/kg doses by intraperitoneal (IP) injection one week prior to and for the duration of SThese drugs significantly decreased LTP in the VH and BLA back to near-control levels while SD rats with vehicle injections still had elevated LTP. However, SD drug-treated rats did not show significant reductions in anxious behavior compared to non-injected SD rats and also exhibited significantly more anxious behavior than control rats, suggesting the IP injection induced added stress. Next, we used rtPCR to examine gene expression of drug targets and plasticity markers to determine potential mechanisms for observed LTP changes. In both the VH and BLA, SD was associated with a significant decrease in glucocorticoid and mineralocorticoid receptor expression, which was restored to control levels under drug treatment. Overall, our data suggest that propranolol and mifepristone together may be a potential prophylactic treatment for preventing PTSD through a mechanism likely mediated by glucocorticoid/mineralocorticoid receptors.
Positioning Nucleosomes with 601 DNA Sequence to Restore GFP Expression
Hales, Emily; Lundgren, Jane; Carter, John; Kempton, Colton; Johnson, Steven (Brigham Young University)
Faculty Advisor: Johnson, Steven (Brigham Young University, Molecular and Microbiology)
The mechanisms of transgene silencing in C. elegans are poorly understood, despite the importance of the nematode as a model for genetic research. Insertion of a transgene led to the expression of GFP in both the body wall and pharyngeal muscle cells of C. elegans as expected. However, subsequent generations stopped expressing body wall GFP. To reverse silencing, we have flanked the enhancers responsible for GFP expression with 601 sequences. The 601 sequence strongly positions nucleosomes. We hypothesize that this positioning will eliminate transgenerational gene silencing of body wall GFP.
Faculty Advisor: Johnson, Steven (Brigham Young University, Molecular and Microbiology)
The mechanisms of transgene silencing in C. elegans are poorly understood, despite the importance of the nematode as a model for genetic research. Insertion of a transgene led to the expression of GFP in both the body wall and pharyngeal muscle cells of C. elegans as expected. However, subsequent generations stopped expressing body wall GFP. To reverse silencing, we have flanked the enhancers responsible for GFP expression with 601 sequences. The 601 sequence strongly positions nucleosomes. We hypothesize that this positioning will eliminate transgenerational gene silencing of body wall GFP.
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
Supplemental treatment options for diabetes: how DHE induces Nr4a1 expression and subsequent β-cell function
Brown, Nathan; Herring, Jacob; Tessem, Jeffery (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. 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.
It is shown that β-cell mitochondrial respiration is dependent on the nuclear receptor Nr4a1. Respiration rates of cells lacking Nr4a1 in the presence of 16 mM glucose resulted in a significant decrease in glucose-stimulated insulin secretion by impeding the production of ATP. It was also found that knockdown of Nr4a1 results in decreased expression of mitochondrial dehydrogenase subunits Idh3g and Sdhb. Thus, the orphan nuclear receptor Nr4a1 is critical for β-cell mitochondrial function and insulin secretion.
In subsequent studies it was shown that dihydroergotamine (DHE) induces Nr4a1 expression via recruitment of the super elongation complex to enable elongation of Nr4a1 promoter paused RNA polymerase II. While these experiments have been shown in cancer cells, I hypothesize that DHE will up-regulate Nr4a1 and other downstream targets. To test this I will use an in-vitro model to culture INS-1 832/3 rat insulinoma cell lines as a useful model for insulin secretion regulation and pancreatic islet beta-cell function studies. This study will shed further light on the regulation of the Nr4a1 nuclear receptor in pancreatic β-cells.
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. 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.
It is shown that β-cell mitochondrial respiration is dependent on the nuclear receptor Nr4a1. Respiration rates of cells lacking Nr4a1 in the presence of 16 mM glucose resulted in a significant decrease in glucose-stimulated insulin secretion by impeding the production of ATP. It was also found that knockdown of Nr4a1 results in decreased expression of mitochondrial dehydrogenase subunits Idh3g and Sdhb. Thus, the orphan nuclear receptor Nr4a1 is critical for β-cell mitochondrial function and insulin secretion.
In subsequent studies it was shown that dihydroergotamine (DHE) induces Nr4a1 expression via recruitment of the super elongation complex to enable elongation of Nr4a1 promoter paused RNA polymerase II. While these experiments have been shown in cancer cells, I hypothesize that DHE will up-regulate Nr4a1 and other downstream targets. To test this I will use an in-vitro model to culture INS-1 832/3 rat insulinoma cell lines as a useful model for insulin secretion regulation and pancreatic islet beta-cell function studies. This study will shed further light on the regulation of the Nr4a1 nuclear receptor in pancreatic β-cells.
Understand whether folic acid can rescue fumonisin, ceramide, and valproic acid induced NTDs
Park, Yeram; Lin, Jade; Ross, Micah; Stark, Michael; (Brigham Young University)
Faculty Advisor: Stark, Michael (Life Sciences, Physiological and Developmental Biology); Hansen, Marc (Life Sciences, Physiological and Developmental Biology)
Neural tube defects (NTDs), which result from failure to close the neural tube during embryonic development, are one of the most widespread and common congenital malformations. Variance in these malformations can range from anencephaly (failure of the neural tube to close on the cranial end) to spina bifida (failure of closure on the posterior/dorsal end). Over the years, scientists have explored this field and have found different environmental factors that may attribute to the likelihood of NTDs. Some of these include fumonisin, valproic acid and more recently discovered, ceramide. To help counter NTDs, studies have shown that folic acid supplementation given to pregnant women has reduced the risk of NTDs and this has become a recommended suggestion by doctors. With its known preventative effects, this study aims to determine whether the preventative effects of folic acid can counter the harmful effects of fumonisin, valproic acid, or ceramide.
Faculty Advisor: Stark, Michael (Life Sciences, Physiological and Developmental Biology); Hansen, Marc (Life Sciences, Physiological and Developmental Biology)
Neural tube defects (NTDs), which result from failure to close the neural tube during embryonic development, are one of the most widespread and common congenital malformations. Variance in these malformations can range from anencephaly (failure of the neural tube to close on the cranial end) to spina bifida (failure of closure on the posterior/dorsal end). Over the years, scientists have explored this field and have found different environmental factors that may attribute to the likelihood of NTDs. Some of these include fumonisin, valproic acid and more recently discovered, ceramide. To help counter NTDs, studies have shown that folic acid supplementation given to pregnant women has reduced the risk of NTDs and this has become a recommended suggestion by doctors. With its known preventative effects, this study aims to determine whether the preventative effects of folic acid can counter the harmful effects of fumonisin, valproic acid, or ceramide.
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.
Uncovering Data for Susceptible Populations: STR DNA findings on Male Rape Victims
Pugh, Sam; Valentine, Julie; Miles, Leslie (Brigham Young University)
Faculty Advisor: Valentine, Julie (Brigham Young University, College Of Nursing); Miles, Leslie (Brigham Young University, College of Nursing)
Rape is generally recognized as a sexual assault by a male perpetrator to a female victim. However, sexual assault is a crime that affects all genders. Although the majority of rapes are male to female, current findings indicate that one in seventy-one men will be raped in their lifetime. Over time, research has recognized the psychological effects and underreporting that ail male rape victims. However, very little has been reported regarding short tandem repeat (STR) DNA findings from sexual assault kits of male victim rapes. These STR DNA profiles prove to be highly influential in the detainment and prosecution of perpetrators. After an extensive search for earlier publications concerning the topic, only three articles were found to have relative correlation to this topic. Current best practice is to obtain STR DNA profiles from sexual assault kit samples to enter into the FBI Combined DNA Index System (CODIS). The purpose of this study was to evaluate DNA analysis findings from 266 sexual assault kits collected from male sexual assault victims and compare predictors for the development of CODIS-eligible STR DNA profiles of male victims to female victims. Our study methodology is an exploratory, retrospective design to identify male rape victims from a sample size of 5,758 victims who received sexual assault forensic examinations with sexual assault kit evidence collection. Approximately 5% of the victims in our study were male (N=266). Male victims were found to have more physical or mental impairments. Male victim cases revealed significantly less development of STR DNA profiles and CODIS-eligible DNA profiles of the perpetrator (p=.007). Due to low STR DNA profile yields and increased targeting of mentally impaired or otherwise vulnerable male victims, we must improve our response to male victims to ensure justice to all victims of sexual assault.
Faculty Advisor: Valentine, Julie (Brigham Young University, College Of Nursing); Miles, Leslie (Brigham Young University, College of Nursing)
Rape is generally recognized as a sexual assault by a male perpetrator to a female victim. However, sexual assault is a crime that affects all genders. Although the majority of rapes are male to female, current findings indicate that one in seventy-one men will be raped in their lifetime. Over time, research has recognized the psychological effects and underreporting that ail male rape victims. However, very little has been reported regarding short tandem repeat (STR) DNA findings from sexual assault kits of male victim rapes. These STR DNA profiles prove to be highly influential in the detainment and prosecution of perpetrators. After an extensive search for earlier publications concerning the topic, only three articles were found to have relative correlation to this topic. Current best practice is to obtain STR DNA profiles from sexual assault kit samples to enter into the FBI Combined DNA Index System (CODIS). The purpose of this study was to evaluate DNA analysis findings from 266 sexual assault kits collected from male sexual assault victims and compare predictors for the development of CODIS-eligible STR DNA profiles of male victims to female victims. Our study methodology is an exploratory, retrospective design to identify male rape victims from a sample size of 5,758 victims who received sexual assault forensic examinations with sexual assault kit evidence collection. Approximately 5% of the victims in our study were male (N=266). Male victims were found to have more physical or mental impairments. Male victim cases revealed significantly less development of STR DNA profiles and CODIS-eligible DNA profiles of the perpetrator (p=.007). Due to low STR DNA profile yields and increased targeting of mentally impaired or otherwise vulnerable male victims, we must improve our response to male victims to ensure justice to all victims of sexual assault.
The Effects of Glucolipotoxicity on Nkx6.1 Expression and Insulin Secretion in the Beta Cell
Elison, Weston; Bauchle, Casey; Bunker, Libby; Stephens, Samuel; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics and Food Science)
Type 2 Diabetes (T2D) effects hundreds of millions of people worldwide, with that number increasing rapidly. It is characterized by increased insulin resistance and dysfunctional insulin secretion. The beta cell of the pancreas is the primary insulin secreting tissue, found in the endocrine tissue of the pancreas called islets of Langerhans. In T2D beta cells become glucose intolerant and disease progression is characterized by loss of functional beta cell mass. Previous studies have shown that the transcription factor Nkx6.1 is vital for beta cell differentiation, identity, and insulin secretion. Research has indicated that Nkx6.1 expression and protein levels decrease in pancreatic islets from human donors with T2D. Our data indicates that glucolipotoxicity, a common model for obesity and diabetes in cell culture, leads to decreased Nkx6.1 mRNA expression, protein levels and nuclear localization in Ins-1 832/13 cells. Nkx6.1 regulates genes in the nucleus , and its loss inhibits proper insulin secretion. We propose that reactive oxygen species created by metabolism of excess fuel decreases Nkx6.1 expression and Nkx6.1 target gene expression, as measured by quantitative polymerase chain reaction (qPCR). Also, increased glucose concentrations causes increased Nkx6.1 protein degradation and translocation out of the nucleus. Protein levels will be measured by western blot and localization by confocal microscopy. In order to understand how these changes effect beta cell function, we will measure glucose stimulated insulin secretion by sandwich Enzyme Linked Immunosorbent Assay (ELISA). We further propose that Nkx6.1 overexpression will restore beta cell function. These results will assist in unraveling the cause of beta cell dysfunction in T2D.
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics and Food Science)
Type 2 Diabetes (T2D) effects hundreds of millions of people worldwide, with that number increasing rapidly. It is characterized by increased insulin resistance and dysfunctional insulin secretion. The beta cell of the pancreas is the primary insulin secreting tissue, found in the endocrine tissue of the pancreas called islets of Langerhans. In T2D beta cells become glucose intolerant and disease progression is characterized by loss of functional beta cell mass. Previous studies have shown that the transcription factor Nkx6.1 is vital for beta cell differentiation, identity, and insulin secretion. Research has indicated that Nkx6.1 expression and protein levels decrease in pancreatic islets from human donors with T2D. Our data indicates that glucolipotoxicity, a common model for obesity and diabetes in cell culture, leads to decreased Nkx6.1 mRNA expression, protein levels and nuclear localization in Ins-1 832/13 cells. Nkx6.1 regulates genes in the nucleus , and its loss inhibits proper insulin secretion. We propose that reactive oxygen species created by metabolism of excess fuel decreases Nkx6.1 expression and Nkx6.1 target gene expression, as measured by quantitative polymerase chain reaction (qPCR). Also, increased glucose concentrations causes increased Nkx6.1 protein degradation and translocation out of the nucleus. Protein levels will be measured by western blot and localization by confocal microscopy. In order to understand how these changes effect beta cell function, we will measure glucose stimulated insulin secretion by sandwich Enzyme Linked Immunosorbent Assay (ELISA). We further propose that Nkx6.1 overexpression will restore beta cell function. These results will assist in unraveling the cause of beta cell dysfunction in T2D.
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 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.
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.
Temperature Effects on D. melanogaster Microbiota Content
Not yet published (Brigham Young University)
Faculty Advisor: Chaston, John (Brigham young University, Life Sciences)
Within an organism's gut are many strains of bacteria that are constantly interacting with their host. Microbiota composition has been shown to impact many aspects of host health such as metabolism, fat-storage, starvation resistance, and reproduction. Certain behaviors and outcomes have been correlated with certain microbial taxa present in the host gut.
D. melanogaster serves as a useful tool for studying this relationship because its microbiota contains relatively few bacterial strains and is both widely studied and largely understood. Previous research within our lab involving D. melanogaster has found trends in many life-history strategies (ie. reproduction, fecundity, lifespan) that correlate with the presence of certain gut bacteria. While there are many aspects of health that microbiota composition affects, there are also a variety of factors that impact microbiota composition thus leading to these end results.
This experiment seeks to further understand the role that environment has in determining microbiota composition. By rearing gnotobiotic flies in environments that differ in temperature, we can then analyze microbiota content to see if any fluctuations occur due to environmental temperature. If temperature is found to have an effect on the taxa present in fully developed D. melanogaster, we can then seek to determine whether or not there are evolutions taking place in host genotype that yield differing microbiota phenotypically.
Faculty Advisor: Chaston, John (Brigham young University, Life Sciences)
Within an organism's gut are many strains of bacteria that are constantly interacting with their host. Microbiota composition has been shown to impact many aspects of host health such as metabolism, fat-storage, starvation resistance, and reproduction. Certain behaviors and outcomes have been correlated with certain microbial taxa present in the host gut.
D. melanogaster serves as a useful tool for studying this relationship because its microbiota contains relatively few bacterial strains and is both widely studied and largely understood. Previous research within our lab involving D. melanogaster has found trends in many life-history strategies (ie. reproduction, fecundity, lifespan) that correlate with the presence of certain gut bacteria. While there are many aspects of health that microbiota composition affects, there are also a variety of factors that impact microbiota composition thus leading to these end results.
This experiment seeks to further understand the role that environment has in determining microbiota composition. By rearing gnotobiotic flies in environments that differ in temperature, we can then analyze microbiota content to see if any fluctuations occur due to environmental temperature. If temperature is found to have an effect on the taxa present in fully developed D. melanogaster, we can then seek to determine whether or not there are evolutions taking place in host genotype that yield differing microbiota phenotypically.
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.
The effect of IL-1β on Nf-_B and ICAM-1 mechanism
Hendricks, Kyle; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
Over 30 million Americans suffer from type 1 (T1D) or type 2 diabetes (T2D), the seventh leading cause of death in the US. T1D and T2D is caused by a significant decrease in pancreatic β-cell mass, resulting in the body's inability to regulate blood glucose. Specifically, T1D is classified as an autoimmune disease due to pancreatic β-cell death by the body's T cells. Nf-κB is required for T cell mediated β-cell destruction. Nf-κB interacts with ICAM-1 on the T cell and acts in conjunction with IL-1β which acts as a T cell activator. This pathway is part of the mechanism that contributes to T cell mediated cell destruction. Here we hypothesize that IL-1β is involved in the mechanism that contributes to Nf-κB and ICAM-1 binding. We will begin with an electrophoretic mobility shift assay to identify the interactions between the ICAM-1 site on IL-1β treated cells and the Nf-κB binding complex. A better understanding of this pathology can, in the future, lead to a treatment that could regulate T cell mediated death of β-cells.
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
Over 30 million Americans suffer from type 1 (T1D) or type 2 diabetes (T2D), the seventh leading cause of death in the US. T1D and T2D is caused by a significant decrease in pancreatic β-cell mass, resulting in the body's inability to regulate blood glucose. Specifically, T1D is classified as an autoimmune disease due to pancreatic β-cell death by the body's T cells. Nf-κB is required for T cell mediated β-cell destruction. Nf-κB interacts with ICAM-1 on the T cell and acts in conjunction with IL-1β which acts as a T cell activator. This pathway is part of the mechanism that contributes to T cell mediated cell destruction. Here we hypothesize that IL-1β is involved in the mechanism that contributes to Nf-κB and ICAM-1 binding. We will begin with an electrophoretic mobility shift assay to identify the interactions between the ICAM-1 site on IL-1β treated cells and the Nf-κB binding complex. A better understanding of this pathology can, in the future, lead to a treatment that could regulate T cell mediated death of β-cells.
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 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.
Understanding Defecation Patterns of Alouatta palliata in Costa Rica
Lengele, Alexius (Weber State University)
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
Primate defecation behaviors contribute notably to forest growth and diversity because primates are important seed dispersers in their environment. Understanding these defecation patterns is important to understand how primate populations impact tropical dry forests. Preexisting research on Allouatta seniculous (red howler monkeys) avoided defecating near their foraging and resting areas, potentially to avoid parasitic infection from contaminated feces. The goal of my research is to test whether the same pattern is found in A. palliata, the mantled howler monkey. I predicted that A. palliata would not defecate near their food resources as a parasite-avoidance behavior. La Selva Biological Station, where this research was conducted, is located in Sarapiqui, in northeastern Costa Rica. Data were collected in May 2017 at the beginning of the rainy season. Using all-occurrence sampling, I recorded all defecation events for A. palliata and whether any group members were feeding. I recorded 15 instances of defecation. Howler monkeys defecated in the same area where they had been feeding 46.7% of the time (n=7) and they defecated in an area where feeding did not occur 53.3% of the time (n=8). My hypothesis, that A. palliata would not defecate near food resources, was not supported as the data showed no bias toward non-feeding areas versus feeding areas. These results differ from prior research of Alouatta in the Amazon Basin, Venezuela, and Brazil, which all reported evidence of parasite-avoidance in defecation behavior. My sample size is small, and my data were collected in a short time span, likely contributing to this discrepancy. Additionally, the parasites infecting those species in South America may not be present in this Central American location, suggesting a lack of the need for this adaptive behavior.
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
Primate defecation behaviors contribute notably to forest growth and diversity because primates are important seed dispersers in their environment. Understanding these defecation patterns is important to understand how primate populations impact tropical dry forests. Preexisting research on Allouatta seniculous (red howler monkeys) avoided defecating near their foraging and resting areas, potentially to avoid parasitic infection from contaminated feces. The goal of my research is to test whether the same pattern is found in A. palliata, the mantled howler monkey. I predicted that A. palliata would not defecate near their food resources as a parasite-avoidance behavior. La Selva Biological Station, where this research was conducted, is located in Sarapiqui, in northeastern Costa Rica. Data were collected in May 2017 at the beginning of the rainy season. Using all-occurrence sampling, I recorded all defecation events for A. palliata and whether any group members were feeding. I recorded 15 instances of defecation. Howler monkeys defecated in the same area where they had been feeding 46.7% of the time (n=7) and they defecated in an area where feeding did not occur 53.3% of the time (n=8). My hypothesis, that A. palliata would not defecate near food resources, was not supported as the data showed no bias toward non-feeding areas versus feeding areas. These results differ from prior research of Alouatta in the Amazon Basin, Venezuela, and Brazil, which all reported evidence of parasite-avoidance in defecation behavior. My sample size is small, and my data were collected in a short time span, likely contributing to this discrepancy. Additionally, the parasites infecting those species in South America may not be present in this Central American location, suggesting a lack of the need for this adaptive behavior.
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.
Tbx2 Pigment Study by CRISPR/CAS9 Mutation
Bell, McKenzie; Porter, Tyrel; Naylor, Emily; Domyan, Eric (Utah Valley University)
Faculty Advisor: Domyan, Eric (Utah Valley University, Biology)
The domestic rock pigeon has been the subject of selective breeding for over a hundred years and so displays an immense variety of phenotypes. This variety provides opportunities to further understand the genetic basis of phenotypic evolution. Pigmentation of pigeon feathers is controlled by multiple alleles at different loci, which influences the type and amount of melanin deposited in the feathers. A specific phenotype, known as "recessive red", consists of distinctly red plumage and is caused by a mutation that greatly reduces the expression of the gene SOX10. This gene encodes a transcription factor, known to play a key role in melanocyte maturation and proliferation. SOX10 likely regulates the transcription of multiple downstream genes but the identities of these genes are largely unknown. To identify downstream targets of SOX10, we compared the transcriptomes of regenerating feathers from wild-type and recessive red birds to identify genes that had different expression levels between the two groups. We identified 46 genes that are expressed at different levels between wild-type and recessive red birds, and thus potential targets of SOX101. Of the 46 genes, Tbx2 was selected as a starter because it is one of the only transcription factors regulated by Sox10 in melanocytes. This mechanism makes it a plausible candidate given the critical role proteins play in phenotypic expression ("TBX2 T-box transcription factor 2—Gene—NCBI," n.d.).
Faculty Advisor: Domyan, Eric (Utah Valley University, Biology)
The domestic rock pigeon has been the subject of selective breeding for over a hundred years and so displays an immense variety of phenotypes. This variety provides opportunities to further understand the genetic basis of phenotypic evolution. Pigmentation of pigeon feathers is controlled by multiple alleles at different loci, which influences the type and amount of melanin deposited in the feathers. A specific phenotype, known as "recessive red", consists of distinctly red plumage and is caused by a mutation that greatly reduces the expression of the gene SOX10. This gene encodes a transcription factor, known to play a key role in melanocyte maturation and proliferation. SOX10 likely regulates the transcription of multiple downstream genes but the identities of these genes are largely unknown. To identify downstream targets of SOX10, we compared the transcriptomes of regenerating feathers from wild-type and recessive red birds to identify genes that had different expression levels between the two groups. We identified 46 genes that are expressed at different levels between wild-type and recessive red birds, and thus potential targets of SOX101. Of the 46 genes, Tbx2 was selected as a starter because it is one of the only transcription factors regulated by Sox10 in melanocytes. This mechanism makes it a plausible candidate given the critical role proteins play in phenotypic expression ("TBX2 T-box transcription factor 2—Gene—NCBI," n.d.).
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 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 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.
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.