Life Sciences
Development of a New Molecular Predictor for Risk of Melanoma Brain Metastases
Stehn, Christopher; Colman, Howard; Boucher, Kenneth; Grossman, Allie H; Holmen, Sheri L (University of Utah)
Faculty Advisor: Holmen, Sheri (University of Utah, Surgery)
Despite therapeutic advances in the treatment of melanoma, development of brain metastases continues to be a major cause of treatment failure. Prognosis for patients with brain metastases is exceedingly poor, therefore the development of sensitive and specific biomarkers to predict which melanoma patients are at highest risk for disease progression are needed. To accomplish this goal, we developed a novel combined molecular/clinical/pathologic predictor of brain metastasis risk. We first analyzed multiple gene expression datasets including The Cancer Genome Atlas (TCGA; n = 437) and an independent series from the European Genome-Phenome Archive (n = 183) and identified a list of 60 consensus genes that is robustly predictive of development of melanoma brain metastases (p < 0.05; FDR 5%). Next, we performed a similar analysis of association of miRNAs and melanoma brain metastasis risk which identified a set of miRNAs with significant predictive power. An optimized combined set of 15 mRNA and miRNA markers was a better predictor of brain metastasis risk than either mRNA or miRNA list alone when applied to the TCGA data set. The combined predictor was most sensitive in separating patients with no metastases from those with either brain metastases or systemic metastases. Current efforts are focused on optimizing miRNA and mRNA separation of patients specifically with brain metastases from those with other metastases using a machine learning linear classifier, and with integrating the expression classifier with other clinical and pathologic predictive factors including: age, stage, thickness, location, histology, ulceration, and gender. The sensitivity and specificity of the resulting clinical/molecular predictor will be validated in an independent retrospective patient dataset, and subsequently implemented in a prospective brain metastasis screening trial to determine real-world utility of this approach in preparation for prospective brain metastasis adjuvant/chemoprevention trials utilizing both immunotherapy and targeted therapy approaches.
Faculty Advisor: Holmen, Sheri (University of Utah, Surgery)
Despite therapeutic advances in the treatment of melanoma, development of brain metastases continues to be a major cause of treatment failure. Prognosis for patients with brain metastases is exceedingly poor, therefore the development of sensitive and specific biomarkers to predict which melanoma patients are at highest risk for disease progression are needed. To accomplish this goal, we developed a novel combined molecular/clinical/pathologic predictor of brain metastasis risk. We first analyzed multiple gene expression datasets including The Cancer Genome Atlas (TCGA; n = 437) and an independent series from the European Genome-Phenome Archive (n = 183) and identified a list of 60 consensus genes that is robustly predictive of development of melanoma brain metastases (p < 0.05; FDR 5%). Next, we performed a similar analysis of association of miRNAs and melanoma brain metastasis risk which identified a set of miRNAs with significant predictive power. An optimized combined set of 15 mRNA and miRNA markers was a better predictor of brain metastasis risk than either mRNA or miRNA list alone when applied to the TCGA data set. The combined predictor was most sensitive in separating patients with no metastases from those with either brain metastases or systemic metastases. Current efforts are focused on optimizing miRNA and mRNA separation of patients specifically with brain metastases from those with other metastases using a machine learning linear classifier, and with integrating the expression classifier with other clinical and pathologic predictive factors including: age, stage, thickness, location, histology, ulceration, and gender. The sensitivity and specificity of the resulting clinical/molecular predictor will be validated in an independent retrospective patient dataset, and subsequently implemented in a prospective brain metastasis screening trial to determine real-world utility of this approach in preparation for prospective brain metastasis adjuvant/chemoprevention trials utilizing both immunotherapy and targeted therapy approaches.
Comparative anatomy and connectivity of the Aii amacrine cell in mouse and rabbit retina
Sigulinsky, Crystal; Anderson, James; Emrich, Daniel; Rapp, Christopher; Dahal, Jeebika; Pfeiffer, Rebecca; Rapp, Kevin; Yang, Jia-Hui; Watt, Carl; Marc, Robert; Jones, Bryan (University of Utah)
Faculty Advisor: Jones, Bryan (University of Utah, Ophthalmology/Visual Sciences)
Purpose: Mouse retina differs structurally from rabbit retina, as it is thicker and vascular, while rabbit retina is thinner and avascular. The implications of these differences on neuronal morphology and connectivity is unclear. We compare the morphology and connectivity of the Aii amacrine cell (AC) at ultrastructural precision in connectomes of mouse (RC2) and rabbit (RC1) retina.
Methods: RC1 and RC2 are 0.25 mm diameter volumes built by automated transmission electron microscopy at 2 nm/pixel resolution. RC1 is from a 13 month old, female Dutch Belted rabbit. RC2 is from a 5 month old female C57BL/6J mouse. The Viking application was used to annotate Aii ACs.
Results: Mouse Aii ACs are elongated with a prominent neck region. Lobular appendages in both species extend from the soma, neck and proximal arboreal dendrites in the OFF sublamina, forming reciprocal synapses with OFF cone bipolar cells (BCs). In rabbits, multiple arboreal dendrites emerge from the base of the neck, branch and travel obliquely through the ON sublamina, forming gap junctions with ON cone BCs, neighboring Aii ACs, and itself. They extend laterally at the base of the IPL, collecting ribbon input from rod BCs. In contrast, mouse arboreal dendrites branch from a single primary dendrite, travel vertically through the IPL with limited self-interaction, and terminate at variable depths that align with the more broadly ramified axon terminals of mouse rod BCs. Synaptology reveals greater output in the OFF vs ON layer in mouse versus rabbit. Uniquely, descending axons of mouse ON cone BCs form gap junctions with Aii AC somas.
Conclusions: Lateral expansion of rabbit Aii ACs may be attributable to eccentricity. However, morphological differences correlate with connectivity differences. Comparative anatomy connectomics is essential for understanding implications of retinal structure on neuronal morphology and connectivity underlying network differences between the mouse and rabbit.
Faculty Advisor: Jones, Bryan (University of Utah, Ophthalmology/Visual Sciences)
Purpose: Mouse retina differs structurally from rabbit retina, as it is thicker and vascular, while rabbit retina is thinner and avascular. The implications of these differences on neuronal morphology and connectivity is unclear. We compare the morphology and connectivity of the Aii amacrine cell (AC) at ultrastructural precision in connectomes of mouse (RC2) and rabbit (RC1) retina.
Methods: RC1 and RC2 are 0.25 mm diameter volumes built by automated transmission electron microscopy at 2 nm/pixel resolution. RC1 is from a 13 month old, female Dutch Belted rabbit. RC2 is from a 5 month old female C57BL/6J mouse. The Viking application was used to annotate Aii ACs.
Results: Mouse Aii ACs are elongated with a prominent neck region. Lobular appendages in both species extend from the soma, neck and proximal arboreal dendrites in the OFF sublamina, forming reciprocal synapses with OFF cone bipolar cells (BCs). In rabbits, multiple arboreal dendrites emerge from the base of the neck, branch and travel obliquely through the ON sublamina, forming gap junctions with ON cone BCs, neighboring Aii ACs, and itself. They extend laterally at the base of the IPL, collecting ribbon input from rod BCs. In contrast, mouse arboreal dendrites branch from a single primary dendrite, travel vertically through the IPL with limited self-interaction, and terminate at variable depths that align with the more broadly ramified axon terminals of mouse rod BCs. Synaptology reveals greater output in the OFF vs ON layer in mouse versus rabbit. Uniquely, descending axons of mouse ON cone BCs form gap junctions with Aii AC somas.
Conclusions: Lateral expansion of rabbit Aii ACs may be attributable to eccentricity. However, morphological differences correlate with connectivity differences. Comparative anatomy connectomics is essential for understanding implications of retinal structure on neuronal morphology and connectivity underlying network differences between the mouse and rabbit.
Cancer Data Exploration for the Public
Payne, Samuel; Paquette, Teancum; Lindgren, Caleb (Brigham Young University)
Faculty Advisor: Payne, Samuel (Brigham Young University, Life Sciences)
The National Cancer Institute’s Clinical Proteomic Tumor Analysis Consortium (CPTAC) generates comprehensive proteogenomic data for cancer cohorts. Our goal is to bring CPTAC data to researchers and the general public. A major difficulty in accomplishing this is the large amount of variability in the programming capabilities in the public. As a solution, we created a set of interactive tutorials that instructs users on exploring CPTAC data in a way that even novice programmers can understand. However, these tutorials still require software installation, which can be complicated. In order to empower more people to confidently use, access and analyze cancer data, we are making our tutorials accessible without any installation. We plan to do this by hosting the tutorials directly using a tool called Binder. In the end this project will not only improve the quality of user experience with CPTAC, but also improve the quality of their experience accessing a vast amount of cancer data.
Faculty Advisor: Payne, Samuel (Brigham Young University, Life Sciences)
The National Cancer Institute’s Clinical Proteomic Tumor Analysis Consortium (CPTAC) generates comprehensive proteogenomic data for cancer cohorts. Our goal is to bring CPTAC data to researchers and the general public. A major difficulty in accomplishing this is the large amount of variability in the programming capabilities in the public. As a solution, we created a set of interactive tutorials that instructs users on exploring CPTAC data in a way that even novice programmers can understand. However, these tutorials still require software installation, which can be complicated. In order to empower more people to confidently use, access and analyze cancer data, we are making our tutorials accessible without any installation. We plan to do this by hosting the tutorials directly using a tool called Binder. In the end this project will not only improve the quality of user experience with CPTAC, but also improve the quality of their experience accessing a vast amount of cancer data.
CD5 knockout mice display reduced ethanol consumption and resistance to ethanol induced sedation
Baptista, Gabriela; Payne, Andrew; Obray, J Daniel; Yorgason, Jordan; Weber, K Scott; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences, Psychology)
Cluster of differentiation 5 (CD5) is expressed in both T and B cells. CD5 has been found to display an altered expression profile following chronic ethanol use and during ethanol withdrawal. Specifically, the number of CD5+ B cells is reduced during withdrawal while the number of T cells is increased. Given the apparent sensitivity of these cells to ethanol and recent research suggesting that some ethanol effects are accounted for by neuroimmune interactions we assessed drinking behavior and ethanol induced sedation in CD5 knockout (KO) mice. We found that CD5 KO mice display decreased ethanol consumption as compared with wild-type controls and that ethanol consumption does not increase with repeated exposure in CD5 KO mice. Additionally, CD5 KO mice displayed considerable resistance to the sedating effects of ethanol. Further studies are underway to assess whether there are baseline differences in dopamine dynamics within the mesolimbic pathway between CD5 KO mice and wild-type controls as well as to whether neurons in the mesolimbic pathway differ in their response to ethanol in CD5 KO mice.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences, Psychology)
Cluster of differentiation 5 (CD5) is expressed in both T and B cells. CD5 has been found to display an altered expression profile following chronic ethanol use and during ethanol withdrawal. Specifically, the number of CD5+ B cells is reduced during withdrawal while the number of T cells is increased. Given the apparent sensitivity of these cells to ethanol and recent research suggesting that some ethanol effects are accounted for by neuroimmune interactions we assessed drinking behavior and ethanol induced sedation in CD5 knockout (KO) mice. We found that CD5 KO mice display decreased ethanol consumption as compared with wild-type controls and that ethanol consumption does not increase with repeated exposure in CD5 KO mice. Additionally, CD5 KO mice displayed considerable resistance to the sedating effects of ethanol. Further studies are underway to assess whether there are baseline differences in dopamine dynamics within the mesolimbic pathway between CD5 KO mice and wild-type controls as well as to whether neurons in the mesolimbic pathway differ in their response to ethanol in CD5 KO mice.
Cocoa Epicatechin Metabolites' affect on β Cell Proliferation and Cell Cycle
Ross, Mimi; Tessem, Jeffery; Orton, Emily; Ekpo, Idongesit; Beales, Joseph (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutrition, Dietetics, & Food Science)
In 2015 there were over 30 million Americans with diabetes and over 84 million Americans ages 18 and older had pre-diabetes. With diabetes being the seventh leading cause of death in the United States and becoming more prevalent the race is on to find a cure. One of the main problems with this disease is the decrease in functional β-cell mass. β-cells produce insulin to maintain blood glucose levels at healthy levels. Thus, if we can increase β-cell proliferation we are one step closer to curing diabetes. Cocoa epicatechins have been shown to be beneficial in blocking diabetes progression. Studies have shown that oligomeric and polymeric cocoa epicatechin extracts improve diabetes onset in a mouse model of Type 2 diabetes. We have demonstrated that the oligomeric fraction of cocoa epicatechins enhances β-cell proliferation in an in vitro model. Absorption studies have shown that while the oligomeric and polymeric forms are not readily absorbed in the gut, they are metabolized by gut bacteria and that these metabolites can be observed in circulation. Using flow cytometry we have studied how these phytochemicals: epicatechin, 5-phenylvaleric acid, Homovanilic acid, and Hippuric acid. Here we present the data regarding the effect of microbial cocoa flavanol metabolites on β-cell cell cycle during proliferation.
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutrition, Dietetics, & Food Science)
In 2015 there were over 30 million Americans with diabetes and over 84 million Americans ages 18 and older had pre-diabetes. With diabetes being the seventh leading cause of death in the United States and becoming more prevalent the race is on to find a cure. One of the main problems with this disease is the decrease in functional β-cell mass. β-cells produce insulin to maintain blood glucose levels at healthy levels. Thus, if we can increase β-cell proliferation we are one step closer to curing diabetes. Cocoa epicatechins have been shown to be beneficial in blocking diabetes progression. Studies have shown that oligomeric and polymeric cocoa epicatechin extracts improve diabetes onset in a mouse model of Type 2 diabetes. We have demonstrated that the oligomeric fraction of cocoa epicatechins enhances β-cell proliferation in an in vitro model. Absorption studies have shown that while the oligomeric and polymeric forms are not readily absorbed in the gut, they are metabolized by gut bacteria and that these metabolites can be observed in circulation. Using flow cytometry we have studied how these phytochemicals: epicatechin, 5-phenylvaleric acid, Homovanilic acid, and Hippuric acid. Here we present the data regarding the effect of microbial cocoa flavanol metabolites on β-cell cell cycle during proliferation.
Basal diet, green tea extract and gut microbiome interactions in a mouse multi-generation study.
Bartlett, Ashley; Phatak, Sumira; Hintze, Korry; Benninghoff, Abby (Utah State University)
Faculty Advisor: Benninghoff, Abby (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)
The gut microbiome modulates various physiological functions related to cancer development including inflammation, cell proliferation, apoptosis, and angiogenesis. Patients with inflammatory bowel disease have a microbiome distinct from healthy controls with consistent observations of reduced gut biomass, decreased diversity within the community, and altered relative abundance. Although a consensus cancer-related microbiome has not been identified, several pathogenic species play an instrumental role in the progression of colitis and tumorigenesis, including: Streptococcus bovis, Helicobacter pylori, Enterococcus faecalis, Clostridium septicum, and Escherichia coli. Gut microbial composition is highly responsive to diet and inadequate intake of micronutrients is a critical feature of the Western dietary pattern. Gut dysbiosis has been proposed to further limit mineral uptake and impair vitamin synthesis, predisposing the host to micronutrient deficiency. Dietary bioactives, such as those in green tea, may function as a mediator between the gut microbiome and basal diet to ultimately prevent colitis associated colorectal cancer (CAC). The overarching objective of our work is to determine the impact of ancestral or multi-generational consumption of the total Western diet (TWD) in a murine model of CAC. Our previous work is the first to investigate how diet induced transgenerational inheritance affects CAC outcome. Our data suggested that multigenerational patterns of exposure to the TWD altered both phenotype and gene expression in third generation offspring. Supplementation with green tea appeared to be most promising after consumption of TWD for multiple generations. Considering that gut microbes are inherited maternally after colonization during vaginal birth, the gut microbiome is a missing piece in this disease model puzzle. The hypothesis of our current project is to investigate whether intake of TWD influences the transmission of microbes and whether CAC outcome is reflected by altered gut microbial composition. Based on other work, we expect the healthy control to possess an abundance of varied bacterial taxa that maintain protective epithelial barrier function and overall homeostasis. On the other hand, a high fat diet would promote increased intestinal permeability, a substantial shift at the phyla level, and increased production of pro-inflammatory cytokines. After TWD consumption, we expect an overall negative phenotypic outcome within the gut microbiome, that includes a breakdown of the epithelial barrier and introduction of pathogenic bacteria. These harmful bacteria tend to thrive on simple sugars that are common in the Western dietary pattern and tend to produce metabolites known as endotoxins that promote dysbiosis.
Faculty Advisor: Benninghoff, Abby (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)
The gut microbiome modulates various physiological functions related to cancer development including inflammation, cell proliferation, apoptosis, and angiogenesis. Patients with inflammatory bowel disease have a microbiome distinct from healthy controls with consistent observations of reduced gut biomass, decreased diversity within the community, and altered relative abundance. Although a consensus cancer-related microbiome has not been identified, several pathogenic species play an instrumental role in the progression of colitis and tumorigenesis, including: Streptococcus bovis, Helicobacter pylori, Enterococcus faecalis, Clostridium septicum, and Escherichia coli. Gut microbial composition is highly responsive to diet and inadequate intake of micronutrients is a critical feature of the Western dietary pattern. Gut dysbiosis has been proposed to further limit mineral uptake and impair vitamin synthesis, predisposing the host to micronutrient deficiency. Dietary bioactives, such as those in green tea, may function as a mediator between the gut microbiome and basal diet to ultimately prevent colitis associated colorectal cancer (CAC). The overarching objective of our work is to determine the impact of ancestral or multi-generational consumption of the total Western diet (TWD) in a murine model of CAC. Our previous work is the first to investigate how diet induced transgenerational inheritance affects CAC outcome. Our data suggested that multigenerational patterns of exposure to the TWD altered both phenotype and gene expression in third generation offspring. Supplementation with green tea appeared to be most promising after consumption of TWD for multiple generations. Considering that gut microbes are inherited maternally after colonization during vaginal birth, the gut microbiome is a missing piece in this disease model puzzle. The hypothesis of our current project is to investigate whether intake of TWD influences the transmission of microbes and whether CAC outcome is reflected by altered gut microbial composition. Based on other work, we expect the healthy control to possess an abundance of varied bacterial taxa that maintain protective epithelial barrier function and overall homeostasis. On the other hand, a high fat diet would promote increased intestinal permeability, a substantial shift at the phyla level, and increased production of pro-inflammatory cytokines. After TWD consumption, we expect an overall negative phenotypic outcome within the gut microbiome, that includes a breakdown of the epithelial barrier and introduction of pathogenic bacteria. These harmful bacteria tend to thrive on simple sugars that are common in the Western dietary pattern and tend to produce metabolites known as endotoxins that promote dysbiosis.
Blue Streak on Uca Pugnax
Anderson, Lars; Baldwin, Haley; Christensen, Ben; Walker, Austen (Brigham Young University)
Faculty Advisor: Griffen, Blaine (Brigham Young University, Life Sciences)
This research looks at the blue coloration on uca pugnax crab carapace above the mouth and between the eyestalks and associates the coloration to the behavior, sexual maturity, and size of the crab, as well as the detection of metals in their environment. Up to ten crabs were photographed within twenty five isolated sites with the objective of gathering a high range of color difference among the uca pugnax. The photos of the crabs were set to match the same scale of light and RGB as to not have interference from external factors such as sunlight or overcast weather. The shade of blue on the carapace provides information about the surrounding environment where the uca pugnax are found.
Faculty Advisor: Griffen, Blaine (Brigham Young University, Life Sciences)
This research looks at the blue coloration on uca pugnax crab carapace above the mouth and between the eyestalks and associates the coloration to the behavior, sexual maturity, and size of the crab, as well as the detection of metals in their environment. Up to ten crabs were photographed within twenty five isolated sites with the objective of gathering a high range of color difference among the uca pugnax. The photos of the crabs were set to match the same scale of light and RGB as to not have interference from external factors such as sunlight or overcast weather. The shade of blue on the carapace provides information about the surrounding environment where the uca pugnax are found.
Beta Cell Heterogeneity: Nkx6.1 Binding Partners
Littlefield, Connor; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University, NDFS)
The transcription factor Nkx6.1 is essential for beta cell growth and function. Given that Nkx6.1 is expressed in beta cells undergoing high level expansion, our lab demonstrated that Nkx6.1 overexpression in primary rat islets was sufficient to induce beta cell proliferation and enhance glucose stimulated insulin secretion. However, while these phenotypes are evident in islets from young animals, islets from aged animals fail to induce proliferation or increased insulin secretion. One reason for why Nkx6.1 fails to drive proliferation or increase insulin secretion is due to lost binding partners that allow it to control gene transcription. We hypothesize that loss of Nkx6.1 binding partners curtails its ability to induce gene transcription that leads to proliferation and enhanced glucose stimulated insulin secretion. To test this hypothesis we have used Nkx6.1 BioID to define by mass spectrometry the proteins that interact with Nkx6.1 Here we define three novel interactors, Mef2D, Sirt7, PDX1. This finding will provide us with a greater understanding of Nkx6.1 function in the beta cell, provide us with new gene targets essential for Nkx6.1 function, and allow us to begin to apply these findings to aged beta cells.
Faculty Advisor: Tessem, Jeffery (Brigham Young University, NDFS)
The transcription factor Nkx6.1 is essential for beta cell growth and function. Given that Nkx6.1 is expressed in beta cells undergoing high level expansion, our lab demonstrated that Nkx6.1 overexpression in primary rat islets was sufficient to induce beta cell proliferation and enhance glucose stimulated insulin secretion. However, while these phenotypes are evident in islets from young animals, islets from aged animals fail to induce proliferation or increased insulin secretion. One reason for why Nkx6.1 fails to drive proliferation or increase insulin secretion is due to lost binding partners that allow it to control gene transcription. We hypothesize that loss of Nkx6.1 binding partners curtails its ability to induce gene transcription that leads to proliferation and enhanced glucose stimulated insulin secretion. To test this hypothesis we have used Nkx6.1 BioID to define by mass spectrometry the proteins that interact with Nkx6.1 Here we define three novel interactors, Mef2D, Sirt7, PDX1. This finding will provide us with a greater understanding of Nkx6.1 function in the beta cell, provide us with new gene targets essential for Nkx6.1 function, and allow us to begin to apply these findings to aged beta cells.
Bioinformatic comparison of peptidases in Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris
Wood, Branzen; Oberg, Taylor; Culumber, Michele; Oberg, Craig (Weber State University)
Faculty Advisor: Oberg, Taylor (Utah State University, Nutrition and Food Science); Culumber, Michele (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology)
The unique flavorings and textures of Cheddar cheese are produced by the degradation of the major milk proteins. One of those proteins, casein, is degraded by the enzyme chymosin and a series of peptidases produced by the starter Lactococcus added to the milk. As casein is degraded, several small peptides accumulate. One of these peptides, ß-casein, can have an adverse bitter taste that is non-desirable and considered a defect in Cheddar cheese. The two main starter cultures used industrially in Cheddar cheese making are Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris. L. lactis subsp. cremoris has been used traditionally in Cheddar cheese making, however, L. lactis subsp. lactis ferments more quickly and is becoming more popular in the cheese industry. With the transition creameries have seen a sharp rise in bitterness during production. Our hypothesis was that while closely related, cremoris synthesizes some peptidases that help with ß-casein degradation that lactis does not. Peptidases found in cremoris include PrtP I and II, Pep X, Pep C, Pep A, Pep T, Pep Q, Pep N, Pep V among others. We searched the genomes of both strains using RAST bioinformatic software, and the databases NCBI and UniProt. The peptidases common in cremoris were also found in lactis. We are now trying to determine if the location of the peptidases on the genomes change how they are regulated or produced. Further, we will begin looking into the genome for other, novel, enzymes that might have peptidase activity that influence bitterness.
Faculty Advisor: Oberg, Taylor (Utah State University, Nutrition and Food Science); Culumber, Michele (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology)
The unique flavorings and textures of Cheddar cheese are produced by the degradation of the major milk proteins. One of those proteins, casein, is degraded by the enzyme chymosin and a series of peptidases produced by the starter Lactococcus added to the milk. As casein is degraded, several small peptides accumulate. One of these peptides, ß-casein, can have an adverse bitter taste that is non-desirable and considered a defect in Cheddar cheese. The two main starter cultures used industrially in Cheddar cheese making are Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris. L. lactis subsp. cremoris has been used traditionally in Cheddar cheese making, however, L. lactis subsp. lactis ferments more quickly and is becoming more popular in the cheese industry. With the transition creameries have seen a sharp rise in bitterness during production. Our hypothesis was that while closely related, cremoris synthesizes some peptidases that help with ß-casein degradation that lactis does not. Peptidases found in cremoris include PrtP I and II, Pep X, Pep C, Pep A, Pep T, Pep Q, Pep N, Pep V among others. We searched the genomes of both strains using RAST bioinformatic software, and the databases NCBI and UniProt. The peptidases common in cremoris were also found in lactis. We are now trying to determine if the location of the peptidases on the genomes change how they are regulated or produced. Further, we will begin looking into the genome for other, novel, enzymes that might have peptidase activity that influence bitterness.
Collared Peccary (Pecari tajucu) Group Size at La Selva Biological Station, Costa Rica
Shin, Seungwon (Salt Lake Community College)
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
The abundance of collared peccaries (Pecari tajucu) is crucial to study because they are a keystone species that plays a large role in their ecosystems. They consume fallen fruits and nuts, disperse seeds, and provide food for predators. Additionally, they are ecosystem engineers altering the landscape for other species. Previous studies have shown that collared peccaries at La Selva Biological Station travel in smaller groups (averaging 10 individuals per group) compared to peccaries at other Neotropical sites. La Selva Biological Station is located in northeastern Costa Rica and it consists of both primary and secondary (i.e. degraded) forests surrounded on three sides by farmland. Due to the general decline of mammals in degraded habitats, I predicted that the average group size of collared peccaries at La Selva will be even smaller than previously reported. I collected data at La Selva Biological Station for two weeks in May 2019. I used three census methods: total count sampling (counting all the species in a certain area), line transect sampling (counting all the species I see when I walk through a trail), and point sampling (standing at selected viewpoints and recording the species visible from that location). I observed 39 peccaries in 17 separate sightings. Group size ranged from 1 to 7 peccaries with an average of 2.3 peccaries per group. Eight sightings (20%) were of single peccaries. My hypothesis that peccary group size would be smaller than 10 individuals was supported. Some limitations of the study were low visibility due to the dense forest and the dispersed social organization of peccary individuals within the group. Both of these factors would underestimate the actual group size of collared peccaries. Nevertheless, the results support previous findings that peccary group size at La Selva are smaller than at other Neotropical sites.
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
The abundance of collared peccaries (Pecari tajucu) is crucial to study because they are a keystone species that plays a large role in their ecosystems. They consume fallen fruits and nuts, disperse seeds, and provide food for predators. Additionally, they are ecosystem engineers altering the landscape for other species. Previous studies have shown that collared peccaries at La Selva Biological Station travel in smaller groups (averaging 10 individuals per group) compared to peccaries at other Neotropical sites. La Selva Biological Station is located in northeastern Costa Rica and it consists of both primary and secondary (i.e. degraded) forests surrounded on three sides by farmland. Due to the general decline of mammals in degraded habitats, I predicted that the average group size of collared peccaries at La Selva will be even smaller than previously reported. I collected data at La Selva Biological Station for two weeks in May 2019. I used three census methods: total count sampling (counting all the species in a certain area), line transect sampling (counting all the species I see when I walk through a trail), and point sampling (standing at selected viewpoints and recording the species visible from that location). I observed 39 peccaries in 17 separate sightings. Group size ranged from 1 to 7 peccaries with an average of 2.3 peccaries per group. Eight sightings (20%) were of single peccaries. My hypothesis that peccary group size would be smaller than 10 individuals was supported. Some limitations of the study were low visibility due to the dense forest and the dispersed social organization of peccary individuals within the group. Both of these factors would underestimate the actual group size of collared peccaries. Nevertheless, the results support previous findings that peccary group size at La Selva are smaller than at other Neotropical sites.
Determining the Function and Structure of Cms1, A Type V CRISPR Effector Endonuclease
Tonks, Adam; Domgaard, Hannah; Crowley, Valerie; Neumann, Gina; Keiser, Dylan; Metcalf, Josie; Guo, Hongjie; Zhou, Yi; Begemann, Mathew; Taylor, David; Jackson, Ryan (Utah State University)
Faculty Advisor: Jackson, Ryan (College of Science, Chemistry and Biochemistry)
Cms1 is a Type V endonuclease that contains a novel domain, shares little sequence homology with other Type V endonucleases, and in some organisms, is found near genes coding for other single-subunit nucleases. Studies in rice (Oryza sativa) have shown Cms1 capable of RNA-directed DNA editing. However, the mechanism of DNA cleavage remains unknown.
Here we present biochemical data that demonstrate Cms1 from Sulfuricurvum processes an RNA guide and binds/cleaves single- and double-stranded DNA through RuvC nuclease motifs. 2-D classification of structures obtained by negative staining electron microscopy show a major conformational change between SuCms1 bound and unbound to an RNA guide. The predicted global structure appears to be different than those reported for other Type V effectors. These data provide for a greater understanding of Type V endonucleases and may provide an alternative tool for genome editing applications.
Faculty Advisor: Jackson, Ryan (College of Science, Chemistry and Biochemistry)
Cms1 is a Type V endonuclease that contains a novel domain, shares little sequence homology with other Type V endonucleases, and in some organisms, is found near genes coding for other single-subunit nucleases. Studies in rice (Oryza sativa) have shown Cms1 capable of RNA-directed DNA editing. However, the mechanism of DNA cleavage remains unknown.
Here we present biochemical data that demonstrate Cms1 from Sulfuricurvum processes an RNA guide and binds/cleaves single- and double-stranded DNA through RuvC nuclease motifs. 2-D classification of structures obtained by negative staining electron microscopy show a major conformational change between SuCms1 bound and unbound to an RNA guide. The predicted global structure appears to be different than those reported for other Type V effectors. These data provide for a greater understanding of Type V endonucleases and may provide an alternative tool for genome editing applications.
CRISPR-based identification of Salmonella in local waterways
Hirschi-Forster, Jeanallie; Mendoza, Matthew; Van Oene; Nicholas ; Payton, Jullian (Weber State University)
Faculty Advisor: Clark, Daniel (Science, Microbiology)
The purpose of this research is to obtain quantitative data about possible sources for Salmonella contamination including tributaries to the Great Salt Lake, namely, the Jordan River, Weber River, and Bear River in Utah. We will also analyze specific water and soil sources near poultry farms for possible contamination. In recent studies, there is a greater number of produce items that have been found to contribute to Salmonella outbreaks. Contaminated water used for irrigation of these crops has been implicated as the causative agent for food contamination.
Bacteria found in these waterways are enriched using selective and differential media. This means, the media provides Salmonella species with required nutrients to grow effectively while differential media inhibits the growth of non-Salmonella species. The enrichment media that is used during this process is 3 X Tryptic Soy Broth and Gram-Negative broth. Gram negative broth is used as an enrichment step, but also selective in that it inhibits growth of other organisms. The two types of differential media would be XLT4 and MSRV. Salmonella is a motile bacterium and thus branches out from its original location of inoculation. This creates a halo-like growth pattern that makes it possible to differentiate Salmonella on MSRV plates. Once Salmonella is confirmed through the MSRV and XLT4 media, sequencing of its two CRISPR loci is completed. These two chromosomal regions have been shown to be distinct in different serovars, and as such, they can be used to distinct what subspecies is present in the sample.
Faculty Advisor: Clark, Daniel (Science, Microbiology)
The purpose of this research is to obtain quantitative data about possible sources for Salmonella contamination including tributaries to the Great Salt Lake, namely, the Jordan River, Weber River, and Bear River in Utah. We will also analyze specific water and soil sources near poultry farms for possible contamination. In recent studies, there is a greater number of produce items that have been found to contribute to Salmonella outbreaks. Contaminated water used for irrigation of these crops has been implicated as the causative agent for food contamination.
Bacteria found in these waterways are enriched using selective and differential media. This means, the media provides Salmonella species with required nutrients to grow effectively while differential media inhibits the growth of non-Salmonella species. The enrichment media that is used during this process is 3 X Tryptic Soy Broth and Gram-Negative broth. Gram negative broth is used as an enrichment step, but also selective in that it inhibits growth of other organisms. The two types of differential media would be XLT4 and MSRV. Salmonella is a motile bacterium and thus branches out from its original location of inoculation. This creates a halo-like growth pattern that makes it possible to differentiate Salmonella on MSRV plates. Once Salmonella is confirmed through the MSRV and XLT4 media, sequencing of its two CRISPR loci is completed. These two chromosomal regions have been shown to be distinct in different serovars, and as such, they can be used to distinct what subspecies is present in the sample.
Chemogenetic stimulation of connexin-36 expressing VTA GABA neurons enhances DA neuron firing rate
Tuttle, Jared; Payne, Andrew; Obray, J Daniel; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
A subpopulation of ventral tegmental area (VTA) GABA neurons express connexin-36 (Cx36) gap junctions (GJs). Activation of GJ-mediated electrical coupling between VTA GABA neurons supports brain stimulation reward and alcohol reward is lowered in Cx36 KO mice due to a hyper-dopamine (DA) state. The aim of this study was to further evaluate the role of a subpopulation of Cx36+ VTA GABA neurons in alcohol reward and dependence. To accomplish this study, we customized a Gq-coupled Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) viral vector to only express in Cx36+ neurons (AAV8.hCx36.hM3D(Gq)-mCherry.WPRE.rBG) in the VTA. The hM3Dq viral vector was infused into male CD-1 GAD GFP mice and male Wistar rats. The animals were then given 10-14 days to recover prior to experimentation. A control virus (AAV9.CB7.CI.mCherry.WPRE.rBG) was used for comparison. We implemented standard cell-attached mode electrophysiology to evaluate the effects of clozapine-n-oxide (CNO; the ligand for DREADDs) on VTA GABA and DA neuronal activity. We found a robust enhancement of VTA GABA neuron firing rate in hM3Dq+ neurons with 20 _M CNO ex vivo. Surprisingly, while investigating CNO effects on VTA DA neuron firing rate, we found that CNO activation of hM3Dq+ VTA GABA neurons increased DA neuron activity, suggesting that Cx36+ VTA GABA neurons indirectly modulate local VTA DA neurons. Intraperitoneal CNO (3 mg/kg) also enhanced the firing rate of VTA GABA neurons in vivo. Administration of CNO reduced ethanol consumption (drink-in-the-dark paradigm) in both ethanol naïve and ethanol dependent hM3Dq-injected mice as compared to controls, suggesting that activation of Cx36+ neurons in the VTA is enough to block ethanol consumption in both naïve and dependent animals. Taken together, these findings support previous studies indicating that enhanced electrical coupling between VTA GABA neurons is rewarding and promotes reward and lowers the hedonic value of ethanol.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
A subpopulation of ventral tegmental area (VTA) GABA neurons express connexin-36 (Cx36) gap junctions (GJs). Activation of GJ-mediated electrical coupling between VTA GABA neurons supports brain stimulation reward and alcohol reward is lowered in Cx36 KO mice due to a hyper-dopamine (DA) state. The aim of this study was to further evaluate the role of a subpopulation of Cx36+ VTA GABA neurons in alcohol reward and dependence. To accomplish this study, we customized a Gq-coupled Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) viral vector to only express in Cx36+ neurons (AAV8.hCx36.hM3D(Gq)-mCherry.WPRE.rBG) in the VTA. The hM3Dq viral vector was infused into male CD-1 GAD GFP mice and male Wistar rats. The animals were then given 10-14 days to recover prior to experimentation. A control virus (AAV9.CB7.CI.mCherry.WPRE.rBG) was used for comparison. We implemented standard cell-attached mode electrophysiology to evaluate the effects of clozapine-n-oxide (CNO; the ligand for DREADDs) on VTA GABA and DA neuronal activity. We found a robust enhancement of VTA GABA neuron firing rate in hM3Dq+ neurons with 20 _M CNO ex vivo. Surprisingly, while investigating CNO effects on VTA DA neuron firing rate, we found that CNO activation of hM3Dq+ VTA GABA neurons increased DA neuron activity, suggesting that Cx36+ VTA GABA neurons indirectly modulate local VTA DA neurons. Intraperitoneal CNO (3 mg/kg) also enhanced the firing rate of VTA GABA neurons in vivo. Administration of CNO reduced ethanol consumption (drink-in-the-dark paradigm) in both ethanol naïve and ethanol dependent hM3Dq-injected mice as compared to controls, suggesting that activation of Cx36+ neurons in the VTA is enough to block ethanol consumption in both naïve and dependent animals. Taken together, these findings support previous studies indicating that enhanced electrical coupling between VTA GABA neurons is rewarding and promotes reward and lowers the hedonic value of ethanol.
Changes in Islet Morphology Over the Axis of Age
Aitken, Talon; Jensen, Daelin; Baxter, Melanie (Brigham Young University)
Faculty Advisor: Tessem, Jeffrey (Brigham Young University, NDFS)
Diabetes Mellitus, a condition characterized by hyperglycemia resulting from defects in insulin secretion or effectiveness, affects over 8.5% of the adult US population. Both type one and type two diabetes have the common characteristic of a decrease of functional beta-cell mass from the islets of Langerhans, located within the pancreas. The upregulation of genes known to induce beta-cell growth and proliferation results in an increase of functional beta-cell mass in young cells but not in their aged counterparts. This age-related occurrence - under nonpathologic conditions — is poorly understood. For this study, the morphological differences between young islets and aged islets are studied to provide insight as to the reason behind this refractory behavior. Immunostaining methods show significant contrast been percentages of insulin-positive beta-cell area in the pancreata of young vs. old-aged rats.
Faculty Advisor: Tessem, Jeffrey (Brigham Young University, NDFS)
Diabetes Mellitus, a condition characterized by hyperglycemia resulting from defects in insulin secretion or effectiveness, affects over 8.5% of the adult US population. Both type one and type two diabetes have the common characteristic of a decrease of functional beta-cell mass from the islets of Langerhans, located within the pancreas. The upregulation of genes known to induce beta-cell growth and proliferation results in an increase of functional beta-cell mass in young cells but not in their aged counterparts. This age-related occurrence - under nonpathologic conditions — is poorly understood. For this study, the morphological differences between young islets and aged islets are studied to provide insight as to the reason behind this refractory behavior. Immunostaining methods show significant contrast been percentages of insulin-positive beta-cell area in the pancreata of young vs. old-aged rats.
Childhood experiences and adult health: The moderating effects of temperament
Miller, Jacob; Cheung, Aaron; Novilla, Kirsten; Crandall, Aliceann (Brigham Young University)
Faculty Advisor: Crandall, Aliceann (Life Sciences, Public Health)
Existing literature demonstrates a strong relationship between childhood experiences and adult health outcomes. The Differential Susceptibility to Environment Theory suggests that there are several factors, including personality and physiology, that effect a child's sensitivity to adverse and advantageous experiences. A sample of 246 adults (ages 19-57) were asked questions about extroverted personality characteristics, adverse and advantageous childhood experiences, and several measures of adult health, including executive functioning, perceived stress levels, depression, and past smoking habits. The sample was then stratified based on level of extroversion scores with the top quartile being labeled as "extroverts", the bottom quartile as "introverts", and those in between as "ambiverts". Regression analyses were then used to assess the relationship between childhood experiences and each adult health outcome. The results of the study showed that the extroverted individuals experienced more positive health outcomes after more advantageous childhood experiences, as well as decreases in adult health outcomes after more adverse childhood experiences. These results suggest that extroverts more than introverts are more sensitivity to environmental influences in childhood. More research is needed to understand the neurobiological mechanisms that increase environmental sensitivity among extroverts.
Faculty Advisor: Crandall, Aliceann (Life Sciences, Public Health)
Existing literature demonstrates a strong relationship between childhood experiences and adult health outcomes. The Differential Susceptibility to Environment Theory suggests that there are several factors, including personality and physiology, that effect a child's sensitivity to adverse and advantageous experiences. A sample of 246 adults (ages 19-57) were asked questions about extroverted personality characteristics, adverse and advantageous childhood experiences, and several measures of adult health, including executive functioning, perceived stress levels, depression, and past smoking habits. The sample was then stratified based on level of extroversion scores with the top quartile being labeled as "extroverts", the bottom quartile as "introverts", and those in between as "ambiverts". Regression analyses were then used to assess the relationship between childhood experiences and each adult health outcome. The results of the study showed that the extroverted individuals experienced more positive health outcomes after more advantageous childhood experiences, as well as decreases in adult health outcomes after more adverse childhood experiences. These results suggest that extroverts more than introverts are more sensitivity to environmental influences in childhood. More research is needed to understand the neurobiological mechanisms that increase environmental sensitivity among extroverts.
Characterizing Lampenflora Diversity in Great Basin National Park to Monitor Disturbances in Fragile Cave Ecosystems
Burgoyne, Jake; Leavitt, Steve (Brigham Young University)
Faculty Advisor: Leavitt, Steve (Life Sciences, Biology)
In show caves, artificially lighting is intended to highlight intricate cave formations for visitors. However, as an unintended consequence, artificial lighting promotes the growth of diverse biofilm communities termed Lampenflora that gain their energy from these novel light sources. Lampenflora, which generally consist of algae and cyanobacteria, discolor formations and introduce novel ecological interactions in simple cave ecosystems. Lampenflora communities have been understudied mainly due to technological limitations and difficult accessibility. However, by characterizing these communities, we can better monitor their impact and develop effective strategies for their removal. Using metagenomic high-throughput sequencing, this research provides the first molecular-based perspective into lampenflora diversity in cave systems in the Great Basin. The data collected, generated, and analyzed is vital in understanding Lampenflora biodiversity and how these communities develop. Furthermore, it offers ecologists a novel perspective on the use molecular detection to understand biodiversity within cave systems.
Faculty Advisor: Leavitt, Steve (Life Sciences, Biology)
In show caves, artificially lighting is intended to highlight intricate cave formations for visitors. However, as an unintended consequence, artificial lighting promotes the growth of diverse biofilm communities termed Lampenflora that gain their energy from these novel light sources. Lampenflora, which generally consist of algae and cyanobacteria, discolor formations and introduce novel ecological interactions in simple cave ecosystems. Lampenflora communities have been understudied mainly due to technological limitations and difficult accessibility. However, by characterizing these communities, we can better monitor their impact and develop effective strategies for their removal. Using metagenomic high-throughput sequencing, this research provides the first molecular-based perspective into lampenflora diversity in cave systems in the Great Basin. The data collected, generated, and analyzed is vital in understanding Lampenflora biodiversity and how these communities develop. Furthermore, it offers ecologists a novel perspective on the use molecular detection to understand biodiversity within cave systems.
Native seed density and priority effects drive invasion resistance against Phragmites in wetland restoration
Holdaway, Bailey; Emily, Martin; Kettenring, Karin (Utah State University)
Faculty Advisor: Kettenring, Karin (S.J. & Jessie E. Quinney College of Natural Resources, Watershed Sciences Department);
Seeds are the primary revegetation method for Great Salt Lake wetlands, however, the density and the priority timing to sow seeds are not clear to wetland managers due to a lack of Great Salt Lake specific revegetation research. Having too low a native seed sowing density could allow unwanted species like the non-native invasive plant Phragmites to reinvade. Too high of a density and density-dependent mortality of sown native seeds could occur, resulting in wasted seeds and unneeded costs for resource-limited managers. In addition, the priority timing (i.e., the relative time and order that seeds are sown) of sowing is also vital for revegetation to favor natives over unwanted invasives. Therefore, our research goal was to determine the optimal seed sowing density and timing priority for reestablishing Great Salt Lake native wetland plant communities. We conducted an outdoor mesocosm experiment with two native sowing densities (3 and 5x the standard sowing density in the region) and three native seed mix sowing timings (4, 2, or 0 weeks prior to sowing Phragmites seeds). We determined the cover of the native plant community and Phragmites at the end of the growing season across the 6 treatment combinations. We found the greatest reduction in Phragmites cover when the native seed mix was sown 4 weeks prior to Phragmites, particularly at the higher native sowing density. A 2-week priority effect did not significantly benefit native species over Phragmites. These results suggest that native seed mixes in Great Salt Lake wetland restorations need to sown much earlier in the summer growing season than when Phragmites seeds germinate and at a very high density to reduce Phragmites cover overall. Though, managers may need to greatly reduce Phragmites seed densities in the seed bank and in the vicinity of restoration sites before revegetation efforts begin.
Faculty Advisor: Kettenring, Karin (S.J. & Jessie E. Quinney College of Natural Resources, Watershed Sciences Department);
Seeds are the primary revegetation method for Great Salt Lake wetlands, however, the density and the priority timing to sow seeds are not clear to wetland managers due to a lack of Great Salt Lake specific revegetation research. Having too low a native seed sowing density could allow unwanted species like the non-native invasive plant Phragmites to reinvade. Too high of a density and density-dependent mortality of sown native seeds could occur, resulting in wasted seeds and unneeded costs for resource-limited managers. In addition, the priority timing (i.e., the relative time and order that seeds are sown) of sowing is also vital for revegetation to favor natives over unwanted invasives. Therefore, our research goal was to determine the optimal seed sowing density and timing priority for reestablishing Great Salt Lake native wetland plant communities. We conducted an outdoor mesocosm experiment with two native sowing densities (3 and 5x the standard sowing density in the region) and three native seed mix sowing timings (4, 2, or 0 weeks prior to sowing Phragmites seeds). We determined the cover of the native plant community and Phragmites at the end of the growing season across the 6 treatment combinations. We found the greatest reduction in Phragmites cover when the native seed mix was sown 4 weeks prior to Phragmites, particularly at the higher native sowing density. A 2-week priority effect did not significantly benefit native species over Phragmites. These results suggest that native seed mixes in Great Salt Lake wetland restorations need to sown much earlier in the summer growing season than when Phragmites seeds germinate and at a very high density to reduce Phragmites cover overall. Though, managers may need to greatly reduce Phragmites seed densities in the seed bank and in the vicinity of restoration sites before revegetation efforts begin.
Parks and Recreation Administrators' Role in the Food Environment: An Exploratory Qualitative Study
Spruance, Lori; Augustine, Madi (Brigham Young University)
Faculty Advisor: Spruance, Lori (Life Sciences, Public Health)
Youth sport programs are an opportunity to increase physical activity, but the food environment may be detrimental to improving and maintaining health. From a previous study, parents indicated that they would like guidance and direction in a top-down approach from coaches and administrators; yet, understanding the administrator experience relative to the youth sports food environment remains unclear. The purpose of this study is to understand that experience. Semi-structured qualitative interviews will take place with administrators across the state of Utah. Interviews will be recorded and transcribed. Thematic analysis will be conducted to identify salient themes. A peer-reviewed publication and multiple presentations will result from the study conducted.
Faculty Advisor: Spruance, Lori (Life Sciences, Public Health)
Youth sport programs are an opportunity to increase physical activity, but the food environment may be detrimental to improving and maintaining health. From a previous study, parents indicated that they would like guidance and direction in a top-down approach from coaches and administrators; yet, understanding the administrator experience relative to the youth sports food environment remains unclear. The purpose of this study is to understand that experience. Semi-structured qualitative interviews will take place with administrators across the state of Utah. Interviews will be recorded and transcribed. Thematic analysis will be conducted to identify salient themes. A peer-reviewed publication and multiple presentations will result from the study conducted.
Low dose alcohol enhances dopamine release in the nucleus accumbens via alpha6-containing nicotinic receptors on GABAergic inputs from the ventral tegmental area
Hansen, Wade; Stockard, Alyssa; Anderson, Elizabeth; Yorgason, Jordan; Sudweeks, Sterling; Wu, Jie; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology); Yorgason, Jordan (Life Sciences, Physiology & Developmental Biology); Sudweeks, Sterling (Life Sciences, Physiology & Developmental Biology)
The prevailing view is that enhancement of dopamine (DA) transmission in the mesolimbic underlies the rewarding properties of ethanol (EtOH) and nicotine (NIC). Although the dogma is that EtOH enhancement of DA neural activity contributes to enhancement of DA transmission, DA neurons are not sensitive to rewarding levels of EtOH. However, VTA GABA neurons are sensitive to low-dose EtOH. We have shown previously that EtOH modulation of DA release in the NAc is mediated by α6-containing nicotinic receptors (α6*-nAChRs), that α6*-nAChRs mediate low-dose EtOH effects on VTA GABA neurons and EtOH preference, and α6*-nAChRs may be a molecular target for low-dose EtOH. The aim of this study was to evaluate EtOH effects on VTA GABAergic input to CINs and DA release in the NAc. Using DIO channel rhodopsin-2 (ChR2) viral injections into the VTA of VGAT Cre mice, we found that VTA GABA neurons send an inhibitory projection to CINs, replicating what has been demonstrated by others. Low-dose EtOH (IC50 = 10 mM) decreased optically-evoked IPSCs (oIPSCs) on CINs and enhanced (EC50 = 10 mM) CIN-mediated spontaneous DA release. Surprisingly, oIPSCs on CINs were not blocked by typical GABAA receptor (GABAAR) antagonists, but by GABAR rho-1 antagonists, suggesting involvement of atypical GABARs on CINs that are postsynaptic to VTA GABAergic input. The α6-conotoxin MII blocked the effects of EtOH on spontaneous DA release and optically-evoked DA release in choline acetyltransferase (ChAT) ChR2 mice. Chronic administration of NIC enhanced EtOH consumption in the drink-in-the-dark procedure and EtOH preference in the CPP procedure and concomitantly enhanced expression of α6*-nAChRs in VTA GABA neurons, without affecting other nAChR subunits. Taken together, these findings suggest that VTA GABA neuron inhibitory input to CINs is modulated by α6*-nAChRs and sensitive to low-dose EtOH, which may underlie the rewarding properties of EtOH.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology); Yorgason, Jordan (Life Sciences, Physiology & Developmental Biology); Sudweeks, Sterling (Life Sciences, Physiology & Developmental Biology)
The prevailing view is that enhancement of dopamine (DA) transmission in the mesolimbic underlies the rewarding properties of ethanol (EtOH) and nicotine (NIC). Although the dogma is that EtOH enhancement of DA neural activity contributes to enhancement of DA transmission, DA neurons are not sensitive to rewarding levels of EtOH. However, VTA GABA neurons are sensitive to low-dose EtOH. We have shown previously that EtOH modulation of DA release in the NAc is mediated by α6-containing nicotinic receptors (α6*-nAChRs), that α6*-nAChRs mediate low-dose EtOH effects on VTA GABA neurons and EtOH preference, and α6*-nAChRs may be a molecular target for low-dose EtOH. The aim of this study was to evaluate EtOH effects on VTA GABAergic input to CINs and DA release in the NAc. Using DIO channel rhodopsin-2 (ChR2) viral injections into the VTA of VGAT Cre mice, we found that VTA GABA neurons send an inhibitory projection to CINs, replicating what has been demonstrated by others. Low-dose EtOH (IC50 = 10 mM) decreased optically-evoked IPSCs (oIPSCs) on CINs and enhanced (EC50 = 10 mM) CIN-mediated spontaneous DA release. Surprisingly, oIPSCs on CINs were not blocked by typical GABAA receptor (GABAAR) antagonists, but by GABAR rho-1 antagonists, suggesting involvement of atypical GABARs on CINs that are postsynaptic to VTA GABAergic input. The α6-conotoxin MII blocked the effects of EtOH on spontaneous DA release and optically-evoked DA release in choline acetyltransferase (ChAT) ChR2 mice. Chronic administration of NIC enhanced EtOH consumption in the drink-in-the-dark procedure and EtOH preference in the CPP procedure and concomitantly enhanced expression of α6*-nAChRs in VTA GABA neurons, without affecting other nAChR subunits. Taken together, these findings suggest that VTA GABA neuron inhibitory input to CINs is modulated by α6*-nAChRs and sensitive to low-dose EtOH, which may underlie the rewarding properties of EtOH.
Interleukin 10 increases dopamine neuron activity in the ventral tegmental area and increases dopamine release in the nucleus accumbens via reduction of GABA inhibition
Clarke, Eliza; Williams, Stephanie; Payne, Andrew; Obray, J Daniel; Yorgason, Jordan; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
Dopamine (DA) transmission is a key player in the rewarding aspects of ethanol as well as ethanol dependence. The current dogma is that DA transmission is increased during ethanol exposure via the inhibition of ventral tegmental area (VTA) GABA neurons and that excitation of VTA GABA neurons during withdrawal results in decreased DA transmission. Microglia, the major neuroimmune effector in the brain, may be a key mediator in this process by releasing cytokines following activation. It is also thought that BDNF may mediate this effect. We evaluated the effect of ethanol on cytokine concentrations in the VTA and nucleus accumbens (NAc), and found that low dose ethanol (1.0 g/kg) decreased interleukin (IL)-10 levels, but high dose ethanol (4.0 g/kg) increased IL-10 levels. We also used standard cell-attached mode electrophysiological techniques to evaluate the effects of select cytokines and BDNF on VTA neuron firing rate in vitro. We found no change in firing rate in response to IL-6 and BDNF, but an increase in firing rate in VTA DA neurons in response to IL-10. Consistent with the changes in firing rate, optically-evoked IPSCs were also found to be decreased in response to IL-10. Ex vivo voltammetry and in vivo microdialysis were done to determine whether IL-10 can directly result in an increase in DA release. Although ex vivo voltammetry showed no change in DA release, IL-10 increased DA release in vivo. These findings suggest that the rewarding and/or addictive effects of ethanol may be mediated by cytokines, specifically the anti-inflammatory cytokine IL-10.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
Dopamine (DA) transmission is a key player in the rewarding aspects of ethanol as well as ethanol dependence. The current dogma is that DA transmission is increased during ethanol exposure via the inhibition of ventral tegmental area (VTA) GABA neurons and that excitation of VTA GABA neurons during withdrawal results in decreased DA transmission. Microglia, the major neuroimmune effector in the brain, may be a key mediator in this process by releasing cytokines following activation. It is also thought that BDNF may mediate this effect. We evaluated the effect of ethanol on cytokine concentrations in the VTA and nucleus accumbens (NAc), and found that low dose ethanol (1.0 g/kg) decreased interleukin (IL)-10 levels, but high dose ethanol (4.0 g/kg) increased IL-10 levels. We also used standard cell-attached mode electrophysiological techniques to evaluate the effects of select cytokines and BDNF on VTA neuron firing rate in vitro. We found no change in firing rate in response to IL-6 and BDNF, but an increase in firing rate in VTA DA neurons in response to IL-10. Consistent with the changes in firing rate, optically-evoked IPSCs were also found to be decreased in response to IL-10. Ex vivo voltammetry and in vivo microdialysis were done to determine whether IL-10 can directly result in an increase in DA release. Although ex vivo voltammetry showed no change in DA release, IL-10 increased DA release in vivo. These findings suggest that the rewarding and/or addictive effects of ethanol may be mediated by cytokines, specifically the anti-inflammatory cytokine IL-10.
Patients' Perceptions of Stress During Hospitalization
Larson, Rebecca; Jimenez, Misty (Utah Valley University)
Faculty Advisor: Jensen, Francine (Utah Valley University, Nursing)
Stress is a known barrier to patient recovery. Patients experience increased emotions, such as stress, while hospitalized due to high stakes from risks to life, health and well-being. Patients' emotions can affect their perceptions, future intentions, and behaviors. In pediatrics, the way parents react to their child's illness may affect the children's compliance, emotional response to medical treatment, and even some development processes, demonstrating the premise that there are many possible stressors that can have significant impacts on patients. Hospitals have taken several measures to evaluate patient stress, such as encouraging hospital staff to discuss patient satisfaction surveys with their patient. However, not all patients recognize their own stressors, and some patients may not initially feel comfortable sharing them. For example, a study showed specific stressors that may experienced by patients of different demographics. These stressors may not always be apparent to nurses. Patients' stress can be reduced if the hospital environment fosters perceptions of control, social support and positive distraction. A change in patient environment can promote healing, as evidenced by a hospital, Navicent Health, that demonstrated in their neonatal intensive care unit that reducing stress and anxiety for both newborns and their parents facilitated healing growth and bonding. Nurses can improve the care they provide to patients by learning how to recognize and reduce stressors during the hospital stay.
Faculty Advisor: Jensen, Francine (Utah Valley University, Nursing)
Stress is a known barrier to patient recovery. Patients experience increased emotions, such as stress, while hospitalized due to high stakes from risks to life, health and well-being. Patients' emotions can affect their perceptions, future intentions, and behaviors. In pediatrics, the way parents react to their child's illness may affect the children's compliance, emotional response to medical treatment, and even some development processes, demonstrating the premise that there are many possible stressors that can have significant impacts on patients. Hospitals have taken several measures to evaluate patient stress, such as encouraging hospital staff to discuss patient satisfaction surveys with their patient. However, not all patients recognize their own stressors, and some patients may not initially feel comfortable sharing them. For example, a study showed specific stressors that may experienced by patients of different demographics. These stressors may not always be apparent to nurses. Patients' stress can be reduced if the hospital environment fosters perceptions of control, social support and positive distraction. A change in patient environment can promote healing, as evidenced by a hospital, Navicent Health, that demonstrated in their neonatal intensive care unit that reducing stress and anxiety for both newborns and their parents facilitated healing growth and bonding. Nurses can improve the care they provide to patients by learning how to recognize and reduce stressors during the hospital stay.
Influenza and Cancer: Shared Pathways and the Potential for New/Repurposed Therapeutics.
Edvalson, Logan; Davis, Morgan; Busath, David (Brigham Young University)
Faculty Advisor: Busath, David (Life Sciences, Physiology & Developmental Biology)
A significant research focus in influenza pathogenesis has been directed towards growth factor receptor tyrosine kinases (RTKs) and their respective phosphorylation cascades. Several recent studies have implicated RTK signaling cascades, that are classically associated with cancer, with increased viral titer. A portion of these studies have focused on early segments of the signaling cascade while others' efforts focus in the late segments. Experiments performed in our lab have identified two receptor pathways—PDGF and VEGF—that, when the receptor inhibited, reduces the efficiency of the influenza virus. These data were achieved using compounds, and variants of compounds, already approved for human use in cancer. Although the drug oseltamivir is already approved for influenza treatment, there is concern for the development of viral drug resistance. The introduction of several types of infection blockers similar to the ones identified by our, and others, laboratories can mitigate viral resistance; like the introduction of several types of antibiotics has reduced bacterial resistance. We hypothesize that these pathways work in multiple parts of the infection cycle ranging from viral endocytosis to the budding off of new virions. Experiments are now under way to determine the specific interactions in these pathways that are important in the viral life cycle.
Faculty Advisor: Busath, David (Life Sciences, Physiology & Developmental Biology)
A significant research focus in influenza pathogenesis has been directed towards growth factor receptor tyrosine kinases (RTKs) and their respective phosphorylation cascades. Several recent studies have implicated RTK signaling cascades, that are classically associated with cancer, with increased viral titer. A portion of these studies have focused on early segments of the signaling cascade while others' efforts focus in the late segments. Experiments performed in our lab have identified two receptor pathways—PDGF and VEGF—that, when the receptor inhibited, reduces the efficiency of the influenza virus. These data were achieved using compounds, and variants of compounds, already approved for human use in cancer. Although the drug oseltamivir is already approved for influenza treatment, there is concern for the development of viral drug resistance. The introduction of several types of infection blockers similar to the ones identified by our, and others, laboratories can mitigate viral resistance; like the introduction of several types of antibiotics has reduced bacterial resistance. We hypothesize that these pathways work in multiple parts of the infection cycle ranging from viral endocytosis to the budding off of new virions. Experiments are now under way to determine the specific interactions in these pathways that are important in the viral life cycle.
Mapping the Potential Distribution of an Invasive Plant, Lythrum salicaria, using Crowd-Sourced Survey Data.
Wertz, Parker (Weber State University)
Faculty Advisor: Dorsey, Bryan (Weber State University, Geography)
Prevention and predicting spread is the best method of control against invasive species. Land managers require accurate and reliable methods for containment and eradication to prevent land cover change and loss of biodiversity. Ecological niche models exist and are used by ecologists to map habitat suitability, but many rely on presence-absence samples which are difficult to obtain. Maximum entropy species distribution modeling (Maxent) is a popular model that has been increasingly used since it can make valid predictions using presence-only data. Many studies have used Maxent to model species distributions, but few have done so with crowdsourced data since it is more likely to be bias and unreliable. The purpose of this study is to test the robustness of Maxent using crowdsourced presence-only data on Lyrthum salicaria, a perennial herb that invades wetlands and pushes out native flora. The study is set in northern and central Utah, and uses environmental variables in climate, landcover, and topography, with landcover being the most contributive factor to the model. Model performance was very good, even with species data being bias towards areas of higher population, proving Maxent as a worthy method to use in species distribution modeling with crowdsourced species presence data. This results of this study show promise for use in modeling other invasive plants in the future.
Faculty Advisor: Dorsey, Bryan (Weber State University, Geography)
Prevention and predicting spread is the best method of control against invasive species. Land managers require accurate and reliable methods for containment and eradication to prevent land cover change and loss of biodiversity. Ecological niche models exist and are used by ecologists to map habitat suitability, but many rely on presence-absence samples which are difficult to obtain. Maximum entropy species distribution modeling (Maxent) is a popular model that has been increasingly used since it can make valid predictions using presence-only data. Many studies have used Maxent to model species distributions, but few have done so with crowdsourced data since it is more likely to be bias and unreliable. The purpose of this study is to test the robustness of Maxent using crowdsourced presence-only data on Lyrthum salicaria, a perennial herb that invades wetlands and pushes out native flora. The study is set in northern and central Utah, and uses environmental variables in climate, landcover, and topography, with landcover being the most contributive factor to the model. Model performance was very good, even with species data being bias towards areas of higher population, proving Maxent as a worthy method to use in species distribution modeling with crowdsourced species presence data. This results of this study show promise for use in modeling other invasive plants in the future.
Non-occupational Crystalline Silica Exposure from Sand and Gravel Pits in Utah
Greenhalgh, Mitchell; Merrill, Alex; Lopez, David; Lefevre, Sam; Williams, Greg (Brigham Young University)
Faculty Advisor: Abbott, Ben (Brigham Young University, Plant and Wildlife Sciences)
The presence of sand and gravel pits around Utah are usually accompanied by public complaints of increased negative health outcomes. The primary risk from these areas is crystalline silica (CS)—the molecule released into the air as a result of crushing rocks and sand. Literature has given mixed results of the potentially harmful effects of crystalline silica. To address the potential health risk of Utah residents from living near sand pits, we performed a meta-analysis on CS-related literature to estimate the true effects of CS on human health. We then used GIS software to estimate the total population in Utah that lives within a 5000-meter buffer of the sand pits in Utah. Using Utah cancer data, birth data, and hospital emergency department data, we created risk ratios for residents within the buffer. The meta-analysis concluded that CS is a weak lung carcinogen. Other research suggests that air pollution leads to low birth weight and preterm births. In our study, lung cancer rates were significantly lower in populations within the 5000-meter buffer. We found no evidence of significant adverse birth outcomes as a result of living in close proximity to sand and gravel pits. Non-malignant respiratory disease also had significantly lower rates within the buffer. These findings are important in determining the role of sand/gravel pit operations in disease incidence in surrounding communities. More research is needed to evaluate confounding factors such as smoking prevalence and socioeconomic status and to investigate crystalline silica in non-occupational settings.
Faculty Advisor: Abbott, Ben (Brigham Young University, Plant and Wildlife Sciences)
The presence of sand and gravel pits around Utah are usually accompanied by public complaints of increased negative health outcomes. The primary risk from these areas is crystalline silica (CS)—the molecule released into the air as a result of crushing rocks and sand. Literature has given mixed results of the potentially harmful effects of crystalline silica. To address the potential health risk of Utah residents from living near sand pits, we performed a meta-analysis on CS-related literature to estimate the true effects of CS on human health. We then used GIS software to estimate the total population in Utah that lives within a 5000-meter buffer of the sand pits in Utah. Using Utah cancer data, birth data, and hospital emergency department data, we created risk ratios for residents within the buffer. The meta-analysis concluded that CS is a weak lung carcinogen. Other research suggests that air pollution leads to low birth weight and preterm births. In our study, lung cancer rates were significantly lower in populations within the 5000-meter buffer. We found no evidence of significant adverse birth outcomes as a result of living in close proximity to sand and gravel pits. Non-malignant respiratory disease also had significantly lower rates within the buffer. These findings are important in determining the role of sand/gravel pit operations in disease incidence in surrounding communities. More research is needed to evaluate confounding factors such as smoking prevalence and socioeconomic status and to investigate crystalline silica in non-occupational settings.
Platelet-Derived Growth Factor Receptor (PDGFR) and Vascular Endothelial Growth Factor Receptor (VEGFR) Antagonists Impair Influenza Infection
Davis, Morgan; Edvalson, Logan; Busath, David (Brigham Young University)
Faculty Advisor: Busath, David (Life Science, Physiology and Developmental Biology)
Influenza infection, and subsequent pneumonias, are the cause of over fifty thousand deaths in the United States per year, and, according to the CDC, influenza is the 8th leading cause of death in this country. Research into the pathogenesis of influenza elucidates critical interactions that take place during different phases of infection which can be targeted by novel drug therapies. Our lab has focused on discovering the role of of PDGFR and VEGFR and other Receptor Tyrosine Kinases (RTKs) in aiding viral infection. RTK activation is reported to be important for successful viral infection, and our project has focused on three different RTKs: VEGFR, PDGFR, and endothelial growth factor receptor (EGFR). In these experiments, Madin Darby Canine Kidney (MDCK) cells were bathed in growth medium containing a specific RTK inhibitor, and then infected with the influenza virus. The vitality of the cells was measured using crystal violet staining and spectrophotometer results. The data showed that using a drug called imatinib—a potent PDGFR inhibitor—resulted in the highest cellular vitality while VEGFR inhibitors developed here at BYU also showed anti-influenza activity. This suggests that the influenza virus is at least partially dependent on PDGFR and VEGFR activation to enhance its life cycle. Future experimentation will study which of the many branches of these receptor's phosphorylation cascades are being utilized by the virus.
Faculty Advisor: Busath, David (Life Science, Physiology and Developmental Biology)
Influenza infection, and subsequent pneumonias, are the cause of over fifty thousand deaths in the United States per year, and, according to the CDC, influenza is the 8th leading cause of death in this country. Research into the pathogenesis of influenza elucidates critical interactions that take place during different phases of infection which can be targeted by novel drug therapies. Our lab has focused on discovering the role of of PDGFR and VEGFR and other Receptor Tyrosine Kinases (RTKs) in aiding viral infection. RTK activation is reported to be important for successful viral infection, and our project has focused on three different RTKs: VEGFR, PDGFR, and endothelial growth factor receptor (EGFR). In these experiments, Madin Darby Canine Kidney (MDCK) cells were bathed in growth medium containing a specific RTK inhibitor, and then infected with the influenza virus. The vitality of the cells was measured using crystal violet staining and spectrophotometer results. The data showed that using a drug called imatinib—a potent PDGFR inhibitor—resulted in the highest cellular vitality while VEGFR inhibitors developed here at BYU also showed anti-influenza activity. This suggests that the influenza virus is at least partially dependent on PDGFR and VEGFR activation to enhance its life cycle. Future experimentation will study which of the many branches of these receptor's phosphorylation cascades are being utilized by the virus.
Is behavioral lateralization in the tropical fish Xenophallus umbratilis related to morphological asymmetry?
Johnson, Erik; Johnson, Ellie; Johnson; Jerald (Brigham Young University)
Faculty Advisor: Johnson, Jerald (Brigham Young University, Biology)
It seems counterintuitive that organisms should evolve handedness (or what we might more broadly refer to as "lateralization"). Individuals who can forage equally well with both hands, who can kick equally well with both feet, who can detect stimuli and orient equally well in both directions, and so on—these individuals should be favored relative to those who are either right handed or left handed. Yet in humans, and in several other species, handedness is common, but we still no very little about why. Here we explore this question using a tropical freshwater fish species with an unusual anatomy. Males have a modified fin—the gonopodium—that they use to internally inseminate females. Interestingly, males are either right or left handed for this structure, which terminates with either a dextral or sinistral twist. In this study, we ask a simple question: is there a link between male gonopodium morphology and male behavioral lateralization. We use a detour test approach to determine how males approach different stimuli, turning either to the left or right to more clearly see each type of stimulus. We focus on how males approach potential mates, predators, and novel items. We predict that males with a dextral gonopodium will orient differently than those with a sinistral gonopodium, consistent with the idea that there is link between behavioral lateralization and morphological handedness. If true, it would suggest that reproductive morphology could be linked to brain and behavioral lateralization in vertebrates.
Faculty Advisor: Johnson, Jerald (Brigham Young University, Biology)
It seems counterintuitive that organisms should evolve handedness (or what we might more broadly refer to as "lateralization"). Individuals who can forage equally well with both hands, who can kick equally well with both feet, who can detect stimuli and orient equally well in both directions, and so on—these individuals should be favored relative to those who are either right handed or left handed. Yet in humans, and in several other species, handedness is common, but we still no very little about why. Here we explore this question using a tropical freshwater fish species with an unusual anatomy. Males have a modified fin—the gonopodium—that they use to internally inseminate females. Interestingly, males are either right or left handed for this structure, which terminates with either a dextral or sinistral twist. In this study, we ask a simple question: is there a link between male gonopodium morphology and male behavioral lateralization. We use a detour test approach to determine how males approach different stimuli, turning either to the left or right to more clearly see each type of stimulus. We focus on how males approach potential mates, predators, and novel items. We predict that males with a dextral gonopodium will orient differently than those with a sinistral gonopodium, consistent with the idea that there is link between behavioral lateralization and morphological handedness. If true, it would suggest that reproductive morphology could be linked to brain and behavioral lateralization in vertebrates.
Is Siphlonuridae Monophyletic: Phylogenetic Relationships of Minnow Mayflies (Ephemeroptera)
Backman, Natalia; Ogden, Heath (Utah Valley University)
Faculty Advisor: Ogden, Heath (Utah Valley University, Biology)
Siphlonuridae is a family of mayflies (order Ephemeroptera) that are nicknamed the "Primitive" Minnow Mayfly family. The family traditionally has consisted of four genera; Edmundsius, Parameletus, Siphlonisca, and Siphlonurus, representing twenty six described species. The family Dipteromimidae was described as a sister group to the family Siphlonuridae (Tojo & Matsukawa, 2003), however, past molecular evidence suggests that Dipteromimidae might nest within Siphlonuridae(T. H. Ogden et al., 2009; T. Heath Ogden & Whiting, 2005). This study aims to use more taxa and more molecular data in order to generate more robust phylogeny for these mayflies. Specifically our goals are to (i) test the monophyly of the family Siphlonuridae; and (ii) compare traditional Sanger sequencing loci to newly generated phylogenomic data from a targeted capture sequencing approach.
Faculty Advisor: Ogden, Heath (Utah Valley University, Biology)
Siphlonuridae is a family of mayflies (order Ephemeroptera) that are nicknamed the "Primitive" Minnow Mayfly family. The family traditionally has consisted of four genera; Edmundsius, Parameletus, Siphlonisca, and Siphlonurus, representing twenty six described species. The family Dipteromimidae was described as a sister group to the family Siphlonuridae (Tojo & Matsukawa, 2003), however, past molecular evidence suggests that Dipteromimidae might nest within Siphlonuridae(T. H. Ogden et al., 2009; T. Heath Ogden & Whiting, 2005). This study aims to use more taxa and more molecular data in order to generate more robust phylogeny for these mayflies. Specifically our goals are to (i) test the monophyly of the family Siphlonuridae; and (ii) compare traditional Sanger sequencing loci to newly generated phylogenomic data from a targeted capture sequencing approach.
Methylmercury exposure in orb weaver spiders (Neoscona oaxacensis) on Antelope Island State Park
Faulkner, Megan; Stoneham, Lisa; Brasso, Rebecka (Weber State University)
Faculty Advisor: Brasso, Rebecka (College of Science, Zoology)
Mercury is a toxic heavy metal that poses significant health risks to humans and wildlife. The organic form of mercury, methylmercury (MeHg), is converted from its inorganic form via microbial methylation primarily in aquatic systems. Methylmercury is dangerous because it attaches to proteins in blood and muscle and biomagnifies in food webs. The goal of this project, is to determine mercury concentrations in western spotted orb weaver spiders (Neoscona oaxacensis) collected from two sites on Antelope Island State Park. Previous studies have shown orb weavers associated with the Great Salt Lake ecosystem to accumulate significant methylmercury, connecting the aquatic and terrestrial ecosystems through a shared food web. The Great Salt Lake surrounding Antelope Island has historically shown some of the highest levels of Hg in surface waters in the United States and has both an abundance of orb weaver spiders and their preferred prey—brine flies. We tested the hypothesis that mercury concentrations in orb weaver spiders would differ between two sites on the island based on differences in environmental conditions (salinity) in the water where brine flies develop. All spiders were sexed, weighed, and individually analyzed for total mercury concentration using a Nippon MA-3000 Direct Mercury Analyzer.
Faculty Advisor: Brasso, Rebecka (College of Science, Zoology)
Mercury is a toxic heavy metal that poses significant health risks to humans and wildlife. The organic form of mercury, methylmercury (MeHg), is converted from its inorganic form via microbial methylation primarily in aquatic systems. Methylmercury is dangerous because it attaches to proteins in blood and muscle and biomagnifies in food webs. The goal of this project, is to determine mercury concentrations in western spotted orb weaver spiders (Neoscona oaxacensis) collected from two sites on Antelope Island State Park. Previous studies have shown orb weavers associated with the Great Salt Lake ecosystem to accumulate significant methylmercury, connecting the aquatic and terrestrial ecosystems through a shared food web. The Great Salt Lake surrounding Antelope Island has historically shown some of the highest levels of Hg in surface waters in the United States and has both an abundance of orb weaver spiders and their preferred prey—brine flies. We tested the hypothesis that mercury concentrations in orb weaver spiders would differ between two sites on the island based on differences in environmental conditions (salinity) in the water where brine flies develop. All spiders were sexed, weighed, and individually analyzed for total mercury concentration using a Nippon MA-3000 Direct Mercury Analyzer.
On the Relationship of Diabetes and Sleep Apnea: Evolution and Epigenetics
Wilson, Nancy; Johnson, Steven (Brigham Young University)
Faculty Advisor: Johnson, Steven (Life Sciences, Microbiology & Molecular Biology)
Diabetes is the seventh leading cause of death in the United States today. Between sixty and ninety percent of diabetics also have sleep apnea. Although both sleep apnea and diabetes engender weight gain, the comorbidity of the two conditions is higher than can be explained by obesity alone.
In this study we explore the advantages of and evidence for the coevolution of diabetes and sleep apnea.
There is a metabolic shift that takes place when the cells of the heart need repair. Normally, hypoxic events cause a shift in heart-cell metabolism toward a high-glucose energy use. This shift mechanism is still fully functional in a diabetic heart cell, but because the underlying diabetes shifts the cellular metabolism to a primarily fatty-acid-based energy use, even a normally functioning hypoxia-induced cascade does not lead to full glucose metabolism or normal cellular repair.
So sleep apnea might serve a useful function in instigating heart tissue repair in cells. This suggests that sleep apnea and diabetes are not just frequently found together, but one condition may be causing the other.
After discussing some of the possible evolutionary drivers for co-adaptation of sleep apnea and diabetes, we examine some of the epigenetic marks associated with the two conditions, laying the groundwork for a better understanding of the underlying etiology.
Faculty Advisor: Johnson, Steven (Life Sciences, Microbiology & Molecular Biology)
Diabetes is the seventh leading cause of death in the United States today. Between sixty and ninety percent of diabetics also have sleep apnea. Although both sleep apnea and diabetes engender weight gain, the comorbidity of the two conditions is higher than can be explained by obesity alone.
In this study we explore the advantages of and evidence for the coevolution of diabetes and sleep apnea.
There is a metabolic shift that takes place when the cells of the heart need repair. Normally, hypoxic events cause a shift in heart-cell metabolism toward a high-glucose energy use. This shift mechanism is still fully functional in a diabetic heart cell, but because the underlying diabetes shifts the cellular metabolism to a primarily fatty-acid-based energy use, even a normally functioning hypoxia-induced cascade does not lead to full glucose metabolism or normal cellular repair.
So sleep apnea might serve a useful function in instigating heart tissue repair in cells. This suggests that sleep apnea and diabetes are not just frequently found together, but one condition may be causing the other.
After discussing some of the possible evolutionary drivers for co-adaptation of sleep apnea and diabetes, we examine some of the epigenetic marks associated with the two conditions, laying the groundwork for a better understanding of the underlying etiology.
Lichens as bioindicators for air quality in the Intermountain West - creating a model for large-scale monitoring
Smith, Hayden; Leavitt, Steve (Brigham Young University)
Faculty Advisor: Leavitt, Steve (College of Life Sciences, Biology)
Brigham Young University's Lichen Air Quality Biomonitoring Program (Herbarium of Non-Vascular Cryptogams) represents one of the largest and longest-running lichen biomonitoring programs worldwide, with nearly 500 permanent reference plots distributed across the Intermountain West. At each reference site, sensitive indicator lichens are selected for elemental analyses of 25 potential pollutants, with the aim of subsequent resampling every five to 15 years for ongoing evaluations of ecological health. Using elemental analysis (EA) data from the past 30 years, a model for large-scale monitoring has been developed with the aim to (i) improve interpretation of air quality using lichen bioindicators, (ii) establish a framework to integrate future EA samples for comparison to historical data, and (iii) develop a platform in the future to more effectively share these data with land management agencies, research groups, and the broader public. The model will be field-tested with new EA samples collected along the Wasatch Front.
Faculty Advisor: Leavitt, Steve (College of Life Sciences, Biology)
Brigham Young University's Lichen Air Quality Biomonitoring Program (Herbarium of Non-Vascular Cryptogams) represents one of the largest and longest-running lichen biomonitoring programs worldwide, with nearly 500 permanent reference plots distributed across the Intermountain West. At each reference site, sensitive indicator lichens are selected for elemental analyses of 25 potential pollutants, with the aim of subsequent resampling every five to 15 years for ongoing evaluations of ecological health. Using elemental analysis (EA) data from the past 30 years, a model for large-scale monitoring has been developed with the aim to (i) improve interpretation of air quality using lichen bioindicators, (ii) establish a framework to integrate future EA samples for comparison to historical data, and (iii) develop a platform in the future to more effectively share these data with land management agencies, research groups, and the broader public. The model will be field-tested with new EA samples collected along the Wasatch Front.
Inhibitory Effect of Probiotics on Streptococcus Agalactiae Serotypes
L'Ecuyer, Katia (Utah Valley University)
Faculty Advisor: Gazdik Stofer, Michaela (Utah Valley University, Microbiology)
Streptococcus agalactiae most commonly known as Group B streptococcus (GBS), are encapsulated gram-positive bacteria encountered in approximately 15-40% of pregnant women's urogenital and gastrointestinal tracts. While most women are asymptomatic, GBS colonization of newborns as they pass through the birth canal can lead to sepsis. GBS bloodstream infections are the leading cause of mortality and morbidity amongst infants in the United States. In recent years, several studies have examined the benefits of oral probiotics to promote a healthy vaginal flora and assessed the inhibitory activity of lactobacilli against urogenital pathogens, with mixed results. The purpose of our research is to examine the effect of Lactobacilli on the growth of different GBS serotypes in the vaginal environment using in vitro culture competition experiments. Previously published microbiome studies were used to determine the dominant species found in the vaginal microbiota. We are examining the growth rate of GBS when co-cultured with vaginal microflora species, both individually and as a mixed community. This will provide a baseline regarding what strains of GBS could easily colonize the vagina in high levels when in competition with different normal flora communities. Different species of probiotic Lactobacilli will then be added to the vaginal culture collection to examine if there is an effect on GBS growth. Our goal is to identify probiotic species that prevent or slow the growth of GBS in a vaginal community.
Faculty Advisor: Gazdik Stofer, Michaela (Utah Valley University, Microbiology)
Streptococcus agalactiae most commonly known as Group B streptococcus (GBS), are encapsulated gram-positive bacteria encountered in approximately 15-40% of pregnant women's urogenital and gastrointestinal tracts. While most women are asymptomatic, GBS colonization of newborns as they pass through the birth canal can lead to sepsis. GBS bloodstream infections are the leading cause of mortality and morbidity amongst infants in the United States. In recent years, several studies have examined the benefits of oral probiotics to promote a healthy vaginal flora and assessed the inhibitory activity of lactobacilli against urogenital pathogens, with mixed results. The purpose of our research is to examine the effect of Lactobacilli on the growth of different GBS serotypes in the vaginal environment using in vitro culture competition experiments. Previously published microbiome studies were used to determine the dominant species found in the vaginal microbiota. We are examining the growth rate of GBS when co-cultured with vaginal microflora species, both individually and as a mixed community. This will provide a baseline regarding what strains of GBS could easily colonize the vagina in high levels when in competition with different normal flora communities. Different species of probiotic Lactobacilli will then be added to the vaginal culture collection to examine if there is an effect on GBS growth. Our goal is to identify probiotic species that prevent or slow the growth of GBS in a vaginal community.
Intravenous dopamine enhancement of dopamine release in the nucleus accumbens is peripheral dopamine 2 receptor dependent
Small, Christina; Obray, J Daniel; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
Recent studies have shown that administration of dopamine in the periphery (outside of the brain) produces a robust enhancement of dopamine release in the nucleus accumbens and alleviates cognitive deficits associated with schizophrenia-like and attention deficit hyperactivity disorder (ADHD)-like phenotypes in rodent models. Despite this, the mechanism whereby peripheral administration of dopamine produces these effects is unknown as dopamine does not cross the blood brain barrier. Activation of dopamine 2 receptors on circulating leukocytes encourages extravasation and can trigger production and release of cytokines such as TNF-_ and IL-10 as well as IL-1_. IL-1_ and IL-10 are both known to enhance dopamine release. In this study we demonstrate that the effects of intravenous dopamine on dopamine release in the NAc are mediated by peripheral dopamine 2 receptors. Additionally, we show that intravenous dopamine is rewarding and that these rewarding effects can be blocked by antagonism of peripheral dopamine 2 receptors. As many drugs of abuse enhance plasma dopamine levels this research elucidates a secondary pathway which may play a role in the development of substance use disorders.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)
Recent studies have shown that administration of dopamine in the periphery (outside of the brain) produces a robust enhancement of dopamine release in the nucleus accumbens and alleviates cognitive deficits associated with schizophrenia-like and attention deficit hyperactivity disorder (ADHD)-like phenotypes in rodent models. Despite this, the mechanism whereby peripheral administration of dopamine produces these effects is unknown as dopamine does not cross the blood brain barrier. Activation of dopamine 2 receptors on circulating leukocytes encourages extravasation and can trigger production and release of cytokines such as TNF-_ and IL-10 as well as IL-1_. IL-1_ and IL-10 are both known to enhance dopamine release. In this study we demonstrate that the effects of intravenous dopamine on dopamine release in the NAc are mediated by peripheral dopamine 2 receptors. Additionally, we show that intravenous dopamine is rewarding and that these rewarding effects can be blocked by antagonism of peripheral dopamine 2 receptors. As many drugs of abuse enhance plasma dopamine levels this research elucidates a secondary pathway which may play a role in the development of substance use disorders.
Loading a Novel Anti-biofilm Compound into Polyurethane Foam for Use in Negative Pressure Wound Therapy
Rawson, Kaden; Nueberger, Travis; Looper, Ryan; Sebahar, Paul; Williams, Dustin (University of Utah)
Faculty Advisor: Williams, Dustin (Engineering, Bioengineering)
Negative pressure wound therapy (NPWT) is commonly used to treat high energy, traumatic battlefield-related injuries, typically caused by an explosion. NPWT may be applied in the field at the time of injury or in the operating room as a therapeutic measure. Wounds are susceptible to contamination from the soil, which contains high amounts of bacteria (>10^9 colony forming units (CFU)/g of material). Greater than 99% of wild-type bacteria favor the biofilm phenotype in the natural world. Biofilms are aggregates of bacteria that are more resistant to traditional antibiotics due to their altered phenotypic and metabolic expressions. Thus, developed biofilms can potentially contaminate these wounds and lead to chronic infection. Furthermore, the lattice structure of polyurethane (PU) foam used in NPWT can potentially harbor and encourage increased biofilm growth. Since the introduction of NPWT as a standard of care for soldiers in 2004, "superficial and deep infections of soft tissue remain a clinical concern after sustaining combat-related trauma [while] using NPWT." To date, GRANUFOAM Silver by KCI is the only variation of PU foam for NPWT that possesses any degree of antimicrobial efficacy. However, silver nanoparticles are minimally effective against biofilms. Thus, the goal of this project is to develop a PU foam that is loaded with a biofilm-specific antimicrobial compound, CZ-01179 in order to decrease the rate of infection when NPWT is utilized in the field of battle.
To date, two prototypes have been developed: One prototype (V1) relies on THF and H2O to coat the Pu foam with CZ-01179 while the second prototype (V2) relies on a hydrogel scaffold to provide a sustained release of CZ-01179 over 24 hours. V1 has been shown to reduce MRSA AND A. baumanii by 7 Log10 CFU during in vitro dilution testing compared to a 1 Log10 reduction produced by GRANUFOAM Silver.
Faculty Advisor: Williams, Dustin (Engineering, Bioengineering)
Negative pressure wound therapy (NPWT) is commonly used to treat high energy, traumatic battlefield-related injuries, typically caused by an explosion. NPWT may be applied in the field at the time of injury or in the operating room as a therapeutic measure. Wounds are susceptible to contamination from the soil, which contains high amounts of bacteria (>10^9 colony forming units (CFU)/g of material). Greater than 99% of wild-type bacteria favor the biofilm phenotype in the natural world. Biofilms are aggregates of bacteria that are more resistant to traditional antibiotics due to their altered phenotypic and metabolic expressions. Thus, developed biofilms can potentially contaminate these wounds and lead to chronic infection. Furthermore, the lattice structure of polyurethane (PU) foam used in NPWT can potentially harbor and encourage increased biofilm growth. Since the introduction of NPWT as a standard of care for soldiers in 2004, "superficial and deep infections of soft tissue remain a clinical concern after sustaining combat-related trauma [while] using NPWT." To date, GRANUFOAM Silver by KCI is the only variation of PU foam for NPWT that possesses any degree of antimicrobial efficacy. However, silver nanoparticles are minimally effective against biofilms. Thus, the goal of this project is to develop a PU foam that is loaded with a biofilm-specific antimicrobial compound, CZ-01179 in order to decrease the rate of infection when NPWT is utilized in the field of battle.
To date, two prototypes have been developed: One prototype (V1) relies on THF and H2O to coat the Pu foam with CZ-01179 while the second prototype (V2) relies on a hydrogel scaffold to provide a sustained release of CZ-01179 over 24 hours. V1 has been shown to reduce MRSA AND A. baumanii by 7 Log10 CFU during in vitro dilution testing compared to a 1 Log10 reduction produced by GRANUFOAM Silver.
Micropropagation of Lepidium ostleri, an edaphic endemic plant species
DeNittis, Alyson; Larson, Joseph; Perez, June; Kopp, Olga R. (Utah Valley University)
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology)
Lepidium ostleri (Ostler's peppergrass) is an edaphic endemic plant species restricted to Ordovician limestone outcrops of the San Francisco Mountains in western Utah. L. ostleri is a species of conservation concern due to its restricted range and proximity to modern mining operations. The purpose of this research is to develop a micropropagation protocol to produce mature plants for population augmentation and introduction to support conservation efforts. De novo shoot organogenic response in tissue explants was highest with various concentrations and combinations of 6-Benzylaminopurine (BAP) and indole-3-acetic acid (IAA). In vitro and ex vitro rooting experiments were conducted on micropropagated plantlets supporting adequate number of shoots, with highest success in pulse treatments of indole-3 butyric acid (IBA). Plantlets were then acclimated to external environments for further propagation. Additional effects of different plant growth regulators, media, and growth conditions will be described. Methods for organogenesis for L. ostleri has not been published and this represents the first known instance of successful micropropagation of this rare plant species. Establishing a micropropagation protocol for L. ostleri provides valuable information for potential restoration or relocation efforts.
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology)
Lepidium ostleri (Ostler's peppergrass) is an edaphic endemic plant species restricted to Ordovician limestone outcrops of the San Francisco Mountains in western Utah. L. ostleri is a species of conservation concern due to its restricted range and proximity to modern mining operations. The purpose of this research is to develop a micropropagation protocol to produce mature plants for population augmentation and introduction to support conservation efforts. De novo shoot organogenic response in tissue explants was highest with various concentrations and combinations of 6-Benzylaminopurine (BAP) and indole-3-acetic acid (IAA). In vitro and ex vitro rooting experiments were conducted on micropropagated plantlets supporting adequate number of shoots, with highest success in pulse treatments of indole-3 butyric acid (IBA). Plantlets were then acclimated to external environments for further propagation. Additional effects of different plant growth regulators, media, and growth conditions will be described. Methods for organogenesis for L. ostleri has not been published and this represents the first known instance of successful micropropagation of this rare plant species. Establishing a micropropagation protocol for L. ostleri provides valuable information for potential restoration or relocation efforts.
Optimal chemotherapeutic combination of 9 putative natural compounds
Berlin, Ian; Kenealey, Jason. (Brigham Young University)
Faculty Advisor: Kenealey, Jason (Life Science; Nutrition, Dietetics, and Food Science)
Prostate cancer accounts for 9.9% of all new cancer cases in the United States annually, and thought it has high 5-year survival rate of 98%, but its prognosis changes if the cancer becomes drug resistant or metastases. Natural compounds are often used and studied for their potential chemotherapeutic effects or their sensitizing effects which increases the cancer cells susceptibility to treatment. Traditional Chinese medicine is a common source for finding bioactive small molecules which may have chemotherapeutic effects. This study focused on 9 putative natural compounds and their effectiveness of treating PC-3 prostate cancer cells. First their IC50s were calculated and then used in Mixture Design Response Surface Methodology (MDRSM) to determine the optimal mixture ratio and used in Chou Talalay statistical analysis to determine if combination effects were synergistic, antagonistic or additive. The compounds used in ascending order starting at the most potent or lowest IC50 to highest; Triptolide, .01819uM (Ttd), Shikonin, .6002uM (Shk), Curcumin 20.83uM (Cur), Emodin, 57.38uM (Em), Wogonin, 97.87uM (Wo) Berberine, 101.4uM (BB), Silibinin, 106.2uM (or Silybin) (Sy), Epigallocatechin gallate, 272.6uM (EGCG), and beta Elemene, 304.3uM (beta-E). Emodin, Silibinin and EGCG all appeared to act primarily via cell cycle inhibition and their effectiveness was found to increase in combination with other small molecules. The ideal combination was provided a multi-faced approach reduce cell viability which suggests it may help treat prostate cancer cells in vivo either in tandem or alone.
Faculty Advisor: Kenealey, Jason (Life Science; Nutrition, Dietetics, and Food Science)
Prostate cancer accounts for 9.9% of all new cancer cases in the United States annually, and thought it has high 5-year survival rate of 98%, but its prognosis changes if the cancer becomes drug resistant or metastases. Natural compounds are often used and studied for their potential chemotherapeutic effects or their sensitizing effects which increases the cancer cells susceptibility to treatment. Traditional Chinese medicine is a common source for finding bioactive small molecules which may have chemotherapeutic effects. This study focused on 9 putative natural compounds and their effectiveness of treating PC-3 prostate cancer cells. First their IC50s were calculated and then used in Mixture Design Response Surface Methodology (MDRSM) to determine the optimal mixture ratio and used in Chou Talalay statistical analysis to determine if combination effects were synergistic, antagonistic or additive. The compounds used in ascending order starting at the most potent or lowest IC50 to highest; Triptolide, .01819uM (Ttd), Shikonin, .6002uM (Shk), Curcumin 20.83uM (Cur), Emodin, 57.38uM (Em), Wogonin, 97.87uM (Wo) Berberine, 101.4uM (BB), Silibinin, 106.2uM (or Silybin) (Sy), Epigallocatechin gallate, 272.6uM (EGCG), and beta Elemene, 304.3uM (beta-E). Emodin, Silibinin and EGCG all appeared to act primarily via cell cycle inhibition and their effectiveness was found to increase in combination with other small molecules. The ideal combination was provided a multi-faced approach reduce cell viability which suggests it may help treat prostate cancer cells in vivo either in tandem or alone.
Understanding Drug Addiction Pathways Through Optogenetics
Bird, Devin; Nufer, Teresa; Wu, Bridget; Edwards, Jeffrey (Brigham Young University)
Faculty Advisor: Edwards, Jeffrey (Brigham Young University, Physiology and Developmental Biology)
Drug addiction is a consequence of neural plasticity in the ventral tegmental area (VTA), an area of the brain's reward system, in which higher levels of dopamine are expressed. Research suggests that decreased activity of inhibitory _-aminobutyric acid (GABA) neurons in the VTA could be the cause of increased activity of dopaminergic cells in the VTA, and thus mediate opiate addiction (Tan). However, not much additional research has been performed to evaluate the plasticity of VTA GABA neurons and the role they play in addiction. Why are VTA GABAergic cells being inhibited and how? We hypothesize that inhibitory inputs onto GABA neurons in the VTA directly affect the degree of inhibition of VTA dopaminergic cells. Additionally, we hypothesize that GABAergic neurons of the lateral hypothalamus (LH) is a source input that extends into the VTA and inhibits VTA GABAergic neurons. We believe that inhibition from these LH neurons induces plasticity of VTA GABAergic neurons.
Through the use of optogenetics we have been able to isolate precise GABAergic pathways that lead into the VTA. Specifically, we have isolated input sources from the LH. These optogenetic experiments, in combination with electrophysiology, have allowed us to measure the specific effects that LH GABA neurons have on VTA GABA neurons. Currently, our data suggests that LH GABAergic cells do induce long-term depression (LTD) in VTA GABAergic cells, however, it is too soon to make any conclusions. Although experiments are still underway, we believe that LH GABAergic neurons play an important role in the drug addiction pathway by inhibiting VTA GABAergic neurons and inducing plasticity.
Faculty Advisor: Edwards, Jeffrey (Brigham Young University, Physiology and Developmental Biology)
Drug addiction is a consequence of neural plasticity in the ventral tegmental area (VTA), an area of the brain's reward system, in which higher levels of dopamine are expressed. Research suggests that decreased activity of inhibitory _-aminobutyric acid (GABA) neurons in the VTA could be the cause of increased activity of dopaminergic cells in the VTA, and thus mediate opiate addiction (Tan). However, not much additional research has been performed to evaluate the plasticity of VTA GABA neurons and the role they play in addiction. Why are VTA GABAergic cells being inhibited and how? We hypothesize that inhibitory inputs onto GABA neurons in the VTA directly affect the degree of inhibition of VTA dopaminergic cells. Additionally, we hypothesize that GABAergic neurons of the lateral hypothalamus (LH) is a source input that extends into the VTA and inhibits VTA GABAergic neurons. We believe that inhibition from these LH neurons induces plasticity of VTA GABAergic neurons.
Through the use of optogenetics we have been able to isolate precise GABAergic pathways that lead into the VTA. Specifically, we have isolated input sources from the LH. These optogenetic experiments, in combination with electrophysiology, have allowed us to measure the specific effects that LH GABA neurons have on VTA GABA neurons. Currently, our data suggests that LH GABAergic cells do induce long-term depression (LTD) in VTA GABAergic cells, however, it is too soon to make any conclusions. Although experiments are still underway, we believe that LH GABAergic neurons play an important role in the drug addiction pathway by inhibiting VTA GABAergic neurons and inducing plasticity.
Using CRISPR and gRNA to Alter the HIV Genome
McRae, Elisa; Solis Leal, Antonio; Giler, Noemi; Karlinsey, Dalton; Quaye, Abraham; Berges, Bradford (Brigham Young University)
Faculty Advisor: Berges, Bradford (Brigham Young University, Microbiology and Molecular Biology)
HIV-1 infects CD4 T-cells by inserting its genome into a cell's genetic sequence. CRISPR technology allows for gene editing within the cell, causing a break in DNA sequences targeted by specific guide RNAs. Plasmids encoding CRISPR and guide RNA (gRNA) genes, in the context of lentiviral delivery vectors, will be transfected to produce two lentiviral vectors. In vitro experiments include human T cells that will be transduced with the lentiviral vectors and analyzed with flow cytometry to determine cells that express CRISPR and gRNAs. These cells will then be sorted to create a population of cells that express both the CRISPR and gRNA genes and will then be infected with the NL4-3 strain of HIV. For in vivo experiments, human hematopoietic stem cells will be transduced with the lentivirus vectors, after which they will be transplanted into humanized mice, thus producing a human-like immune system for testing the efficacy of our anti-HIV approach. After the human immune system has sufficiently developed in the mice, HIV-1 will be introduced. We expect that human immune cells with CRISPRs will be protected against HIV infection and death due to the use of gRNAs. These cells are postulated to no longer be susceptible to HIV-1 infection, thus preventing further cell lineages from becoming infected. We will analyze data for three main endpoints: 1. Cell killing of HIV, 2. HIV rebound due to the high mutation rate of the virus, 3. Amount of HIV replication, examined by assessing the viral RNA outside of cells using Q-RT-PCR. Data from this project will support whether cells transfected with CRISPR and guide RNAs offer cell lineages that adequately disrupt the HIV-1 genome. Efforts of this study hope to address HIV infection in humans following trials with humanized mice.
Faculty Advisor: Berges, Bradford (Brigham Young University, Microbiology and Molecular Biology)
HIV-1 infects CD4 T-cells by inserting its genome into a cell's genetic sequence. CRISPR technology allows for gene editing within the cell, causing a break in DNA sequences targeted by specific guide RNAs. Plasmids encoding CRISPR and guide RNA (gRNA) genes, in the context of lentiviral delivery vectors, will be transfected to produce two lentiviral vectors. In vitro experiments include human T cells that will be transduced with the lentiviral vectors and analyzed with flow cytometry to determine cells that express CRISPR and gRNAs. These cells will then be sorted to create a population of cells that express both the CRISPR and gRNA genes and will then be infected with the NL4-3 strain of HIV. For in vivo experiments, human hematopoietic stem cells will be transduced with the lentivirus vectors, after which they will be transplanted into humanized mice, thus producing a human-like immune system for testing the efficacy of our anti-HIV approach. After the human immune system has sufficiently developed in the mice, HIV-1 will be introduced. We expect that human immune cells with CRISPRs will be protected against HIV infection and death due to the use of gRNAs. These cells are postulated to no longer be susceptible to HIV-1 infection, thus preventing further cell lineages from becoming infected. We will analyze data for three main endpoints: 1. Cell killing of HIV, 2. HIV rebound due to the high mutation rate of the virus, 3. Amount of HIV replication, examined by assessing the viral RNA outside of cells using Q-RT-PCR. Data from this project will support whether cells transfected with CRISPR and guide RNAs offer cell lineages that adequately disrupt the HIV-1 genome. Efforts of this study hope to address HIV infection in humans following trials with humanized mice.
What in Tarnation? The Rozel Tar Seeps Impacts to Avian Fauna at the Great Salt Lake
Sanchez, Mary; Martin, Cayla; Butler, Jaimi; Parrott, David (Westminster College)
Faculty Advisor: Butler, Jaimi (Westminster College, Great Salt Lake Institute); Parrott, David (Westminster College, Biology)
The Great Salt Lake is one of the largest migratory stops for many species of birds in North America. Along the banks of the Great Salt Lake, at Rozel point, there are tar seeps, where some species of birds have gotten entrapped and died. These petroleum seeps are both naturally occurring and human created at Rozel Point. The temperature that the seeps become sticky, the possibility of prey animals drawing predators in, and the appearance of the tar seeps are all important aspects of why birds are drawn to these seeps. Using motion sensor cameras and temperature monitoring devices, the animals that are visiting the tar seeps and the temperature variation of the seeps were monitored. One of the largest human created oil wells at Rozel Point was recapped in January of 2019. The impacts of this recapping was monitored throughout the summer of 2019 to determine if there are fewer birds entrapped due to the reduction of oil escaping from the ground at this well.
Faculty Advisor: Butler, Jaimi (Westminster College, Great Salt Lake Institute); Parrott, David (Westminster College, Biology)
The Great Salt Lake is one of the largest migratory stops for many species of birds in North America. Along the banks of the Great Salt Lake, at Rozel point, there are tar seeps, where some species of birds have gotten entrapped and died. These petroleum seeps are both naturally occurring and human created at Rozel Point. The temperature that the seeps become sticky, the possibility of prey animals drawing predators in, and the appearance of the tar seeps are all important aspects of why birds are drawn to these seeps. Using motion sensor cameras and temperature monitoring devices, the animals that are visiting the tar seeps and the temperature variation of the seeps were monitored. One of the largest human created oil wells at Rozel Point was recapped in January of 2019. The impacts of this recapping was monitored throughout the summer of 2019 to determine if there are fewer birds entrapped due to the reduction of oil escaping from the ground at this well.
Understanding The Role Of Small Non-coding RNA In Bumble Bee Social Behavior
Figgins, Anna C.; Hunter, F. Kate; Kapheim, Karen M. (Utah State University)
Faculty Advisor: Kapheim, Karen (College of Science, Biology Department)
Certain species of ants, bees, and wasps have some of the most sophisticated forms of cooperative behavior known throughout the animal kingdom. These eusocial insects live in large family groups made up of castes (e.g., queens and workers) that specialize on different tasks within a colony. In many species, division of labor between queens and workers is associated with behavioral and physiological traits such as dominance interactions, ovary maturation, and lipid stores. Remarkably, these large phenotypic differences between castes emerge from a shared genome. This suggests caste differences stem from changes in how shared genes are regulated. We have been investigating the role of a small regulatory molecule (microRNA miR-13b) as a potential regulator of division of labor in bumble bees (Bombus impatiens). We tested the hypothesis that miR-13b regulates division of labor by inhibiting its function using small-interfering RNA (siRNA). We inhibited miR-13b function by injecting a synthetic antagonist of miR-13b (antagomir) into the abdomens of live bees. The average expression of miR-13b in the fat body of bees that received the antagomir injection was 0.52 relative to those that received the control injection (n = 7). This inhibition of miR-13b expression was accompanied by a significant decrease in fat body size. However, the influence of the antagomir only lasted 1 day. Future experiments will determine if the antagomir can influence gene expression longer than 1 day and assess physiological and behavioral changes in B. impatiens after miR-13b is inhibited. Knowledge gained from this study allows us to understand more about the mechanisms underlying social behavior in bees and helps us investigate how behavior is regulated by gene expression.
Faculty Advisor: Kapheim, Karen (College of Science, Biology Department)
Certain species of ants, bees, and wasps have some of the most sophisticated forms of cooperative behavior known throughout the animal kingdom. These eusocial insects live in large family groups made up of castes (e.g., queens and workers) that specialize on different tasks within a colony. In many species, division of labor between queens and workers is associated with behavioral and physiological traits such as dominance interactions, ovary maturation, and lipid stores. Remarkably, these large phenotypic differences between castes emerge from a shared genome. This suggests caste differences stem from changes in how shared genes are regulated. We have been investigating the role of a small regulatory molecule (microRNA miR-13b) as a potential regulator of division of labor in bumble bees (Bombus impatiens). We tested the hypothesis that miR-13b regulates division of labor by inhibiting its function using small-interfering RNA (siRNA). We inhibited miR-13b function by injecting a synthetic antagonist of miR-13b (antagomir) into the abdomens of live bees. The average expression of miR-13b in the fat body of bees that received the antagomir injection was 0.52 relative to those that received the control injection (n = 7). This inhibition of miR-13b expression was accompanied by a significant decrease in fat body size. However, the influence of the antagomir only lasted 1 day. Future experiments will determine if the antagomir can influence gene expression longer than 1 day and assess physiological and behavioral changes in B. impatiens after miR-13b is inhibited. Knowledge gained from this study allows us to understand more about the mechanisms underlying social behavior in bees and helps us investigate how behavior is regulated by gene expression.
Using Dendroclimatology To Study A Disjunct Population Of Pinus Ponderosa In Northern Utah
Stapleton, Michael; DeRose, Justin. (Utah State University)
Faculty Advisor: DeRose, Justin (S.J. & Jessie E. Quinney College of Natural Resources, Wildland Resources Department)
Ponderosa pine (Pinus ponderosa) is the most widespread coniferous tree in North America, occurring from Mexico to British Columbia and from California to Nebraska. Surprisingly, however, P. ponderosa is largely absent within the center of this range. Previous studies suggest that this absence may be linked to a range of climatic variables, but collectively fail to identify specific climate-growth responses. Using dendroclimatology, we will analyze how a disjunct population of P. ponderosa in northern Utah responds to local climate conditions. We seek to identify which of these variables the species is most sensitive to by correlating the population's average annual growth to a variety of climate composites. Similar tests will be conducted across three treatment blocks throughout the stand in order to distinguish if previous management altered the population's resilience to climate. Our results will help explain the current distribution of P. ponderosa and suggest how that distribution may respond to changing climate conditions.
Faculty Advisor: DeRose, Justin (S.J. & Jessie E. Quinney College of Natural Resources, Wildland Resources Department)
Ponderosa pine (Pinus ponderosa) is the most widespread coniferous tree in North America, occurring from Mexico to British Columbia and from California to Nebraska. Surprisingly, however, P. ponderosa is largely absent within the center of this range. Previous studies suggest that this absence may be linked to a range of climatic variables, but collectively fail to identify specific climate-growth responses. Using dendroclimatology, we will analyze how a disjunct population of P. ponderosa in northern Utah responds to local climate conditions. We seek to identify which of these variables the species is most sensitive to by correlating the population's average annual growth to a variety of climate composites. Similar tests will be conducted across three treatment blocks throughout the stand in order to distinguish if previous management altered the population's resilience to climate. Our results will help explain the current distribution of P. ponderosa and suggest how that distribution may respond to changing climate conditions.
YjbB encodes a phosphate exporter in E. coli
Funk, Stephen; Wood, Jacob; Catmull, Ashley; Martin, Brett (Brigham Young University)
Faculty Advisor: McCleary, Bill (Brigham Young University College of Life Sciences, Microbiology & Molecular Biology)
The survival of the model microorganism E. coli depends largely on its ability to regulate the concentration of nutrients in its cell. This regulation often relies on complex systems of both cooperative and competitive enzymes. One of these enzymes, encoded by the YjbB gene, was known to play some role in phosphate regulation. However, its exact function had not yet been characterized. We attempted to deduce the function of the YjbB-encoded protein in the context of two other phosphate regulatory systems: the PitA/B phosphate transport system and the PpK/X polyphosphate storage system. Using comparative growth curves in both phosphate-rich and phosphate-deprived media, we found that the YjbB-encoded protein protects against phosphate poisoning in the absence of phosphate exporters, suggesting that the protein in question functions primarily as a phosphate exporter as well.
Faculty Advisor: McCleary, Bill (Brigham Young University College of Life Sciences, Microbiology & Molecular Biology)
The survival of the model microorganism E. coli depends largely on its ability to regulate the concentration of nutrients in its cell. This regulation often relies on complex systems of both cooperative and competitive enzymes. One of these enzymes, encoded by the YjbB gene, was known to play some role in phosphate regulation. However, its exact function had not yet been characterized. We attempted to deduce the function of the YjbB-encoded protein in the context of two other phosphate regulatory systems: the PitA/B phosphate transport system and the PpK/X polyphosphate storage system. Using comparative growth curves in both phosphate-rich and phosphate-deprived media, we found that the YjbB-encoded protein protects against phosphate poisoning in the absence of phosphate exporters, suggesting that the protein in question functions primarily as a phosphate exporter as well.
Verification of microbial genes that affect host dietary preference in Drosophila melanogaster
Call, Tanner; Bean, Joseph; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)
The gut microbiome, or the microorganisms that colonize the GI tract of all macro-organisms, plays a significant role in host health and physiology. In a study last year, I found that the microbiome of D. melanogaster has a direct influence on dietary preference using a well-established, automated feeding assay. In this study, I extend these findings by performing a metagenome-wide association (MGWA) screen to predict bacterial genes responsible for the effect. Specifically, I measured dietary preferences in flies mono-associated with each of 40 different bacterial species. My mentor compared the dietary preference of these flies with the genomes of their associated bacteria using a MGWA. This analysis predicted 1932 bacterial genes that could be responsible for the feeding preference phenotype. I selected the top 22 genes, including all uncharacterized genes, for which we have knock-out mutants in a laboratory stock of bacterial mutants. I will test if these genes are necessary for inducing specific host feeding preferences by comparing feeding preferences of flies mono-associated with a bacterial mutant with controls, using a generalized mixed linear model. These results will help us understand how different members of the microbiota can influence animal feeding behaviors.
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)
The gut microbiome, or the microorganisms that colonize the GI tract of all macro-organisms, plays a significant role in host health and physiology. In a study last year, I found that the microbiome of D. melanogaster has a direct influence on dietary preference using a well-established, automated feeding assay. In this study, I extend these findings by performing a metagenome-wide association (MGWA) screen to predict bacterial genes responsible for the effect. Specifically, I measured dietary preferences in flies mono-associated with each of 40 different bacterial species. My mentor compared the dietary preference of these flies with the genomes of their associated bacteria using a MGWA. This analysis predicted 1932 bacterial genes that could be responsible for the feeding preference phenotype. I selected the top 22 genes, including all uncharacterized genes, for which we have knock-out mutants in a laboratory stock of bacterial mutants. I will test if these genes are necessary for inducing specific host feeding preferences by comparing feeding preferences of flies mono-associated with a bacterial mutant with controls, using a generalized mixed linear model. These results will help us understand how different members of the microbiota can influence animal feeding behaviors.
Quantification of Staphylococcus Biofilm Clearance
Kaneshiro, Alma; Jordan, Adam; Crompton, Rhees; Brailsford, Samantha; Spencer, Jonathan (Weber State University)
Faculty Advisor: Clark, Daniel (Science, Microbiology Department and Neuroscience Center); Chaston, John (Life Sciences, Plant & Wildlife Sciences)
Antibiotic resistance is of great concern in the medical community, with bacterial resistance increasing proportional to their use. Staphylococcus aureus, such as methicillin resistant S. aureus (MRSA), can cause fatal infections. Problems due to this resistance are compounded when the infecting bacteria form a biofilm, thick sticky layers of bacterial secretions, which are difficult for antibiotics to penetrate. Biofilm formation is common in hospital settings on stents, catheters, and IV lines. Biofilms make antibiotic treatment risky due to incomplete killing—the most resistant survive exposure. There is evidence that bacteriophage can break up biofilms, possibly making them more susceptible to antibiotics. We induced a S. aureus biofilm formation using chemicals that mimic a skin wound. Using bacteriophage K, we inoculated the biofilm and observed clearance. Samples of cell pellets and liquid supernatant were collected, and DNA was extracted. Real-time PCR was used to quantify the levels of bacteriophage K replication, representing clearance of the bacteria. This research can be used to find efficient ways to treat an infection caused by a S. aureus biofilm. Bacteriophage used in combination with antibiotics may be able to better clear a biofilm infection and reduce antibiotic resistance risk due to more complete infection clearance.
Faculty Advisor: Clark, Daniel (Science, Microbiology Department and Neuroscience Center); Chaston, John (Life Sciences, Plant & Wildlife Sciences)
Antibiotic resistance is of great concern in the medical community, with bacterial resistance increasing proportional to their use. Staphylococcus aureus, such as methicillin resistant S. aureus (MRSA), can cause fatal infections. Problems due to this resistance are compounded when the infecting bacteria form a biofilm, thick sticky layers of bacterial secretions, which are difficult for antibiotics to penetrate. Biofilm formation is common in hospital settings on stents, catheters, and IV lines. Biofilms make antibiotic treatment risky due to incomplete killing—the most resistant survive exposure. There is evidence that bacteriophage can break up biofilms, possibly making them more susceptible to antibiotics. We induced a S. aureus biofilm formation using chemicals that mimic a skin wound. Using bacteriophage K, we inoculated the biofilm and observed clearance. Samples of cell pellets and liquid supernatant were collected, and DNA was extracted. Real-time PCR was used to quantify the levels of bacteriophage K replication, representing clearance of the bacteria. This research can be used to find efficient ways to treat an infection caused by a S. aureus biofilm. Bacteriophage used in combination with antibiotics may be able to better clear a biofilm infection and reduce antibiotic resistance risk due to more complete infection clearance.
Role of CD5 in oral inflammation and periodontal disease
Townsend, Jessica; Freitas, Claudia; Weber, Scott; Cardon, Dallin (Brigham Young University)
Faculty Advisor: Weber, Scott (Brigham Young University / Life Sciences, Microbiology and Molecular Biology)
The World Health Organization reported in 2016 that oral diseases affected half of the world's population. Oral diseases are due to poor oral hygiene and tobacco use which can develop into periodontal disease. Periodontal disease is caused by an immune response to microbial challenge, which initiates an invasion of lymphocytes and other single-nucleated cells to the site of inflammation in the mouth that can cause tooth loss and is a risk factor for heart and lung disease. Patients with severe periodontitis have increased auto-reactive B lymphocytes that express the CD5 co-receptor and these cells are influenced by T cells. We propose to investigate the relationship between oral inflammation, CD5, and the T helper immune response. This will be done by comparing oral inflammation in mice with and without CD5. CD5 is a T cell co-receptor that regulates T cell development and function and we hypothesize CD5 plays an important role in periodontal disease. We will test this hypothesis by co-culturing T cells expressing or lacking CD5 with oral mucosal or gingival epithelial cells that have been exposed to LPS (lipopolysaccharide, a major component of gram-negative bacteria's wall) and will exam differences in cell number, T cell subtype, and cell function.
Faculty Advisor: Weber, Scott (Brigham Young University / Life Sciences, Microbiology and Molecular Biology)
The World Health Organization reported in 2016 that oral diseases affected half of the world's population. Oral diseases are due to poor oral hygiene and tobacco use which can develop into periodontal disease. Periodontal disease is caused by an immune response to microbial challenge, which initiates an invasion of lymphocytes and other single-nucleated cells to the site of inflammation in the mouth that can cause tooth loss and is a risk factor for heart and lung disease. Patients with severe periodontitis have increased auto-reactive B lymphocytes that express the CD5 co-receptor and these cells are influenced by T cells. We propose to investigate the relationship between oral inflammation, CD5, and the T helper immune response. This will be done by comparing oral inflammation in mice with and without CD5. CD5 is a T cell co-receptor that regulates T cell development and function and we hypothesize CD5 plays an important role in periodontal disease. We will test this hypothesis by co-culturing T cells expressing or lacking CD5 with oral mucosal or gingival epithelial cells that have been exposed to LPS (lipopolysaccharide, a major component of gram-negative bacteria's wall) and will exam differences in cell number, T cell subtype, and cell function.
Spatial variation in mercury concentrations of flying insects at Antelope Island
Stoneham, Lisa; Brasso, Dr. Rebecka (Weber State University)
Faculty Advisor: Brasso, Rebecka (Weber State University, Zoology)
Mercury is a toxic heavy metal that poses significant health threats to people and wildlife. The organic form of mercury, methylmercury, is converted from its inorganic form via microbial methylation. Methylmercury is dangerous because it attaches to proteins in the blood, muscle, and other tissues and can cross the blood-brain and placental barriers. Microbial methylation is enhanced in anoxic environments such as wetlands, which are increasingly being classified as mercury hotspots where animals accumulate elevated concentrations relative to those in terrestrial systems. This is concerning for the wetlands of the Great Salt Lake due to its history of anthropogenic inputs of pollutants and its importance as a breeding ground and rest stop for migrating avian species. Previous research has shown significant mercury methylation occurring within the Deep Brine Layer of the GSL. The purpose of this project was to investigate potential spatial variation in mercury concentration in different portions of the GSL. With a focus on invertebrates, we collected insects including brine flies, midges, damselflies, and crane flies from three sites of varying salinity around Antelope Island State Park: Farmington Bay, White Rock Bay, and the Antelope Island Marina. Mercury concentrations in insects were determined using a Nippon MA-3000 Direct Mercury Analyzer. Our results will provide a preliminary assessment of mercury concentrations in flying insects from different habitats around the island. This will help in determining differential risk to insectivorous songbirds, waterfowl, and shorebirds foraging on these common prey species in the GSL.
Faculty Advisor: Brasso, Rebecka (Weber State University, Zoology)
Mercury is a toxic heavy metal that poses significant health threats to people and wildlife. The organic form of mercury, methylmercury, is converted from its inorganic form via microbial methylation. Methylmercury is dangerous because it attaches to proteins in the blood, muscle, and other tissues and can cross the blood-brain and placental barriers. Microbial methylation is enhanced in anoxic environments such as wetlands, which are increasingly being classified as mercury hotspots where animals accumulate elevated concentrations relative to those in terrestrial systems. This is concerning for the wetlands of the Great Salt Lake due to its history of anthropogenic inputs of pollutants and its importance as a breeding ground and rest stop for migrating avian species. Previous research has shown significant mercury methylation occurring within the Deep Brine Layer of the GSL. The purpose of this project was to investigate potential spatial variation in mercury concentration in different portions of the GSL. With a focus on invertebrates, we collected insects including brine flies, midges, damselflies, and crane flies from three sites of varying salinity around Antelope Island State Park: Farmington Bay, White Rock Bay, and the Antelope Island Marina. Mercury concentrations in insects were determined using a Nippon MA-3000 Direct Mercury Analyzer. Our results will provide a preliminary assessment of mercury concentrations in flying insects from different habitats around the island. This will help in determining differential risk to insectivorous songbirds, waterfowl, and shorebirds foraging on these common prey species in the GSL.
Taller seedlings in about half the time: the effect of coyote ingestion on netleaf hackberry (Celtis reticulata) seeds
Hannah A. Veltkamp, Sydney Houghton, Michael T. Stevens (Utah Valley University)
Faculty Advisor: Stevens, Micheal (Utah Valley University, Biology)
Netleaf hackberry (Celtis reticulata) is a deciduous shrub native to the southwestern United States and northern Mexico. Individual shrubs can be long-lived, but newly established stands of hackberry are rare. The lack of juvenile hackberry in the wild could be due to low germination rates reported in both laboratory and field settings. The seeds of hackberry are embedded in drupes that are an important source of food for birds and small mammals. Animals likely play an important role in seed dispersal, and passing through a digestive tract could increase the germination rates of hackberry seeds. Passage through the digestive tract of a mammal can increase the germination rates for some plant species, but not for others. We hypothesized that passage through the digestive tract of a coyote would increase the germination rates of C. reticulata. To test this hypothesis, we collected 17 coyote scats containing visible hackberry fruits from along the Bonneville Shoreline Trail east of Provo, Utah, using latex gloves. Each scat location was recorded using a GPS unit. After collecting each scat, we found the closest hackberry shrub and picked a sample of fresh hackberry fruits from it. All samples were cleaned and cold stratified and then planted into cone-tainers containing a potting soil mix and placed in the Utah Valley University greenhouse. We sowed 20 seeds from each of the 17 coyote scats and
20 seeds from each of the neighboring hackberry bushes for a total of 680 seeds. The 680 cone-tainers were labeled with plastic stakes and randomly positioned into trays that were randomly distributed on a bench in the greenhouse. The seeds, and later seedlings, were watered as needed (typically three days/week). On watering days, we checked for newly-germinated hackberry seedlings and recorded their date of emergence. Near the end of the experiment, we measured the height of each seedling. The germination rate of hackberry seeds that had passed through the
digestive tract of a coyote did not differ from the germination rate of seeds from fresh-picked fruit (42.7% vs. 46.5%, respectively; _ 2 = 0.558, df = 1, p = 0.455). However, on average, the coyote-treatment seeds took just over half as many days to germinate as did the seeds from fresh-picked (undigested) fruit (35 days vs. 69 days, respectively; p < 0.001). The seedlings from coyote-treatment seeds were 9.5% taller than were the seedlings derived from seeds from undigested fruit (6.4 cm vs. 5.8 cm, respectively; p < 0.001). Our results show that consumption by coyotes can benefit hackberries by enabling their seeds to germinate earlier in the year when
conditions for establishment are good. The earlier start on germination that coyote-ingested hackberries get translates to increased height and likely a higher rate of survival in the field.
Faculty Advisor: Stevens, Micheal (Utah Valley University, Biology)
Netleaf hackberry (Celtis reticulata) is a deciduous shrub native to the southwestern United States and northern Mexico. Individual shrubs can be long-lived, but newly established stands of hackberry are rare. The lack of juvenile hackberry in the wild could be due to low germination rates reported in both laboratory and field settings. The seeds of hackberry are embedded in drupes that are an important source of food for birds and small mammals. Animals likely play an important role in seed dispersal, and passing through a digestive tract could increase the germination rates of hackberry seeds. Passage through the digestive tract of a mammal can increase the germination rates for some plant species, but not for others. We hypothesized that passage through the digestive tract of a coyote would increase the germination rates of C. reticulata. To test this hypothesis, we collected 17 coyote scats containing visible hackberry fruits from along the Bonneville Shoreline Trail east of Provo, Utah, using latex gloves. Each scat location was recorded using a GPS unit. After collecting each scat, we found the closest hackberry shrub and picked a sample of fresh hackberry fruits from it. All samples were cleaned and cold stratified and then planted into cone-tainers containing a potting soil mix and placed in the Utah Valley University greenhouse. We sowed 20 seeds from each of the 17 coyote scats and
20 seeds from each of the neighboring hackberry bushes for a total of 680 seeds. The 680 cone-tainers were labeled with plastic stakes and randomly positioned into trays that were randomly distributed on a bench in the greenhouse. The seeds, and later seedlings, were watered as needed (typically three days/week). On watering days, we checked for newly-germinated hackberry seedlings and recorded their date of emergence. Near the end of the experiment, we measured the height of each seedling. The germination rate of hackberry seeds that had passed through the
digestive tract of a coyote did not differ from the germination rate of seeds from fresh-picked fruit (42.7% vs. 46.5%, respectively; _ 2 = 0.558, df = 1, p = 0.455). However, on average, the coyote-treatment seeds took just over half as many days to germinate as did the seeds from fresh-picked (undigested) fruit (35 days vs. 69 days, respectively; p < 0.001). The seedlings from coyote-treatment seeds were 9.5% taller than were the seedlings derived from seeds from undigested fruit (6.4 cm vs. 5.8 cm, respectively; p < 0.001). Our results show that consumption by coyotes can benefit hackberries by enabling their seeds to germinate earlier in the year when
conditions for establishment are good. The earlier start on germination that coyote-ingested hackberries get translates to increased height and likely a higher rate of survival in the field.
Role of the CD5 T cell co-receptor in T cell metabolism
Haynie, Christopher; Freitas, Claudia M. Tellez; Whitley, Kiara V.; Weber, K. Scott (Brigham Young University)
Faculty Advisor: Weber, K. Scott (Life Sciences, Microbiology and Molecular Biology)
T cells play a critical role in the adaptive immune response and undergo significant metabolic changes upon activation. T cell co-receptors influence T cell activation and function, yet their influence on T cell metabolism remains unclear. CD5, an inhibitory co-receptor expressed on the surface of T cells, is known to regulate thymocyte selection and T cell receptor (TCR) signaling. We previously observed that CD5 plays a critical role in calcium signaling in naïve helper T cells. As calcium signaling influences metabolic changes in cells, our current work focuses on understanding the role of CD5 in T cell metabolism. To understand how CD5 regulates metabolism in T cells, we used CD5 deficient T cells and compared them to wildtype CD5 sufficient T cells. We have characterized their metabolic activity using glycolytic and mitochondrial respiration assays. Interestingly, CD5 deficient naïve T cells have increased glycolysis, mitochondrial respiration, and spare respiratory capacity in comparison to wildtype T cells. We hypothesize that this is due to CD5 altering mitochondrial membrane potential and mass, gene regulation, and the influence of different cellular fuels. Understanding how CD5 regulates T cell metabolism will provide critical insights for improved immunotherapeutic strategies.
Faculty Advisor: Weber, K. Scott (Life Sciences, Microbiology and Molecular Biology)
T cells play a critical role in the adaptive immune response and undergo significant metabolic changes upon activation. T cell co-receptors influence T cell activation and function, yet their influence on T cell metabolism remains unclear. CD5, an inhibitory co-receptor expressed on the surface of T cells, is known to regulate thymocyte selection and T cell receptor (TCR) signaling. We previously observed that CD5 plays a critical role in calcium signaling in naïve helper T cells. As calcium signaling influences metabolic changes in cells, our current work focuses on understanding the role of CD5 in T cell metabolism. To understand how CD5 regulates metabolism in T cells, we used CD5 deficient T cells and compared them to wildtype CD5 sufficient T cells. We have characterized their metabolic activity using glycolytic and mitochondrial respiration assays. Interestingly, CD5 deficient naïve T cells have increased glycolysis, mitochondrial respiration, and spare respiratory capacity in comparison to wildtype T cells. We hypothesize that this is due to CD5 altering mitochondrial membrane potential and mass, gene regulation, and the influence of different cellular fuels. Understanding how CD5 regulates T cell metabolism will provide critical insights for improved immunotherapeutic strategies.
Stress Sensitivity to Temperature in Plateau Side-blotched Lizards (Uta stansburiana uniformis): Implications for Immune Function
Lidgard, Audrey; French, Susannah; Hudson, Spencer (Utah State University)
Faculty Advisor: Lidgard, Susannah (College of Science, Biology Department)
Ectothermic organisms, such as reptiles, rely on the external environment for regulating internal temperatures necessary for vital physiological processes. When faced with environmental challenges, temperature may differentially affect how allostatic mediators (e.g., glucocorticoid hormones) are released to mediate energy allocation for handling stressors. Subsequent differences in energy mobilization and circulating metabolites during a stress response may ultimately influence self-maintenance processes such as immunity. The aims of this research were to determine how stress sensitivity varies with diurnal temperatures in the Plateau Side-blotched Lizard (Uta stansburiana uniformis) and to assess the potential implications for immune function. Both baseline and stress-induced levels of glucocorticoids (corticosterone) and energy metabolites (glucose) were compared to body temperature and the thermal environment. Variation in innate immune function (bactericidal ability) was then compared to both temperature and physiological parameters at baseline and stress-induced levels. Stress reactivity via glucocorticoid release positively corresponded with body and environmental temperatures, although glucose release did not. Bactericidal ability subsequent to a stressor negatively corresponded with body temperature and glucocorticoid release. Such findings provide further insight on how stress sensitivity and self-maintenance can vary across the thermal environment, posing potential fitness consequences for an ectothermic organism.
Faculty Advisor: Lidgard, Susannah (College of Science, Biology Department)
Ectothermic organisms, such as reptiles, rely on the external environment for regulating internal temperatures necessary for vital physiological processes. When faced with environmental challenges, temperature may differentially affect how allostatic mediators (e.g., glucocorticoid hormones) are released to mediate energy allocation for handling stressors. Subsequent differences in energy mobilization and circulating metabolites during a stress response may ultimately influence self-maintenance processes such as immunity. The aims of this research were to determine how stress sensitivity varies with diurnal temperatures in the Plateau Side-blotched Lizard (Uta stansburiana uniformis) and to assess the potential implications for immune function. Both baseline and stress-induced levels of glucocorticoids (corticosterone) and energy metabolites (glucose) were compared to body temperature and the thermal environment. Variation in innate immune function (bactericidal ability) was then compared to both temperature and physiological parameters at baseline and stress-induced levels. Stress reactivity via glucocorticoid release positively corresponded with body and environmental temperatures, although glucose release did not. Bactericidal ability subsequent to a stressor negatively corresponded with body temperature and glucocorticoid release. Such findings provide further insight on how stress sensitivity and self-maintenance can vary across the thermal environment, posing potential fitness consequences for an ectothermic organism.
Quantification of GAD 65/67 Proteins in Learning and Addiction Pathways
Edwards, Jeffrey; Friend, Lindsey; Weed, Jared; Sandova, Philipl; Nufer, Teresa; Ostlund, Isaac Ostlund (Brigham Young University)
Faculty Advisor: Edwards, Jeffrey (Life Sciences, Physiology and Developmental Biology)
Substance abuse is a widespread problem in the United States. Although there are some existing treatments for addiction, the neural mechanisms of addiction are not deeply understood. This study quantifies the expression of GAD65 and GAD67 in GABAergic cells in the VTA of adolescent mice to shed light on the subtypes of cells involved in learning and addiction pathways.
The ventral tegmental area (VTA) of the brain, a critical part of the dopamine reward system, has many dopamine cells that are inhibited by nearby GABAergic neurons. Formation of memories and addiction involve long-term potentiation (LTP) and long-term depression (LTD) of these inhibitory GABA cells. We studied potential pathways of learning and addiction by measuring levels of expression of GAD 65/67 proteins and quantifying the cells that express one or both of these proteins.
Our results will provide insight about which GABAergic neurons are involved in the addiction pathway, furthering our understanding of the cellular mechanism of addiction. This will pave the way for more educated, effective treatment of drug addicts in clinical settings.
Faculty Advisor: Edwards, Jeffrey (Life Sciences, Physiology and Developmental Biology)
Substance abuse is a widespread problem in the United States. Although there are some existing treatments for addiction, the neural mechanisms of addiction are not deeply understood. This study quantifies the expression of GAD65 and GAD67 in GABAergic cells in the VTA of adolescent mice to shed light on the subtypes of cells involved in learning and addiction pathways.
The ventral tegmental area (VTA) of the brain, a critical part of the dopamine reward system, has many dopamine cells that are inhibited by nearby GABAergic neurons. Formation of memories and addiction involve long-term potentiation (LTP) and long-term depression (LTD) of these inhibitory GABA cells. We studied potential pathways of learning and addiction by measuring levels of expression of GAD 65/67 proteins and quantifying the cells that express one or both of these proteins.
Our results will provide insight about which GABAergic neurons are involved in the addiction pathway, furthering our understanding of the cellular mechanism of addiction. This will pave the way for more educated, effective treatment of drug addicts in clinical settings.
Precipitation and Thunder Associated Vocalizations in Mantled Howler Monkeys (Alouatta palliata)
Pehkonen, Eliza (Salt Lake Community College)
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
Precipitation-associated behaviors have been observed in several species of primate including bonobos (e.g., building leafy shelters), chimpanzees (e.g., drinking, rain dancing displays), and mantled howler monkeys (e.g., licking rain from the air, altering typical behavior based on weather and season). The purpose of this study is to determine if mantled howler monkeys (Alouatta palliata) exhibit precipitation-associated vocalizations. A. palliata is well known for its vocalizations, which are the loudest sound made by any terrestrial mammal and are used for a wide variety of communicative purposes, such as attracting mates, defending territory, and deterring predation. Given the purpose with which A. palliata vocalizes and the existence of precipitation-associated behaviors within primate species, including A. palliata, it was hypothesized that A. palliata would vocalize in association with climatic events (precipitation and thunder). To test this hypothesis, 41.75 hours of data were collected on A. palliata over a two-week time period during the rainy season at La Selva Biological Station in Costa Rica. All-occurrence sampling was used to record the timing and duration of all A. palliata vocalizations, precipitation, and thunder events. Events were considered accompanied if they occurred within five minutes of one another. Of the 59 observed vocalization events 53% were associated with climatic events. Of the 20 observed precipitation events 90% were accompanied by vocalizations and of the 37 observed thunder events 57% were accompanied by vocalization. Associated vocalizations occurred before, during and after climatic events, however, during or after were most common. The data indicate an association between A. palliata vocalization and precipitation, confirming the hypothesis. Further research is warranted to investigate a possible purpose of precipitation-associated vocalizations, an understanding of which could provide further insight into A. palliata's behavioral interaction with climatic events.
Faculty Advisor: Seaboch, Melissa (Salt Lake Community College, Anthropology)
Precipitation-associated behaviors have been observed in several species of primate including bonobos (e.g., building leafy shelters), chimpanzees (e.g., drinking, rain dancing displays), and mantled howler monkeys (e.g., licking rain from the air, altering typical behavior based on weather and season). The purpose of this study is to determine if mantled howler monkeys (Alouatta palliata) exhibit precipitation-associated vocalizations. A. palliata is well known for its vocalizations, which are the loudest sound made by any terrestrial mammal and are used for a wide variety of communicative purposes, such as attracting mates, defending territory, and deterring predation. Given the purpose with which A. palliata vocalizes and the existence of precipitation-associated behaviors within primate species, including A. palliata, it was hypothesized that A. palliata would vocalize in association with climatic events (precipitation and thunder). To test this hypothesis, 41.75 hours of data were collected on A. palliata over a two-week time period during the rainy season at La Selva Biological Station in Costa Rica. All-occurrence sampling was used to record the timing and duration of all A. palliata vocalizations, precipitation, and thunder events. Events were considered accompanied if they occurred within five minutes of one another. Of the 59 observed vocalization events 53% were associated with climatic events. Of the 20 observed precipitation events 90% were accompanied by vocalizations and of the 37 observed thunder events 57% were accompanied by vocalization. Associated vocalizations occurred before, during and after climatic events, however, during or after were most common. The data indicate an association between A. palliata vocalization and precipitation, confirming the hypothesis. Further research is warranted to investigate a possible purpose of precipitation-associated vocalizations, an understanding of which could provide further insight into A. palliata's behavioral interaction with climatic events.