Life Sciences
Electrohydraulic Shockwaves as a Possible Treatment for Bacterial Biofilms
Brunetti, Bryce; Escarate, Ashley; Conway, Matthew; Slezak, Cyrill; Kopp, Olga (Utah Valley University)
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology); Slezak, Cyrill (Utah Valley University, Physics)
Purpose:
This study evaluates the effect of electrohydraulic shockwaves on Staphylococcus aureus biofilms. This system could be a great alternative to the use of antibiotics, and potentially life-saving technology that could save billions of dollars.
Background:
The rise of antibiotic-resistant bacteria is a global threat. Staphylococcus aureus is typically harmless, but this gram-positive species has become highly resistant and extremely pathogenic. Strains like MRSA and VRSA have the highest rate of drug resistance and are the leading cause of chronic bacterial infections via bacterial biofilms on medical devices. Biofilms are an aggregation of microbes that excrete an extracellular matrix providing an ideal environment for gene exchange and quorum sensing. Their complexity hinders the diffusion of antimicrobials. A proposed method to prevent device-associated infection is shockwave sterilization and therapy. A shockwave is a high-energy wave causing a sudden change in temperature, pressure and density in the medium. This study investigates the potential disruption of bacterial biofilms by electrohydraulic shockwaves.
Methods:
E. coli and S. aureus biofilms were grown on polystyrene plates. Biofilms were treated with shockwaves (0.19mJ/mm2, 300 pulses, 3 Hz) in a water bath and compared with those treated with Vancomycin. Cell viability was determined through XTT/menadione absorbance and specific biofilm formation through crystal violet absorbance.
Results:
Current testing has shown that electrohydraulic shockwaves have a bacteriostatic effect on biofilms. Other finding show potential for shockwaves to increase bacterial susceptibility to lower levels of antibiotics.
Conclusions:
Device-associated infections are a serious threat to patients' health. The diminishing effectiveness of antibiotics in treating and preventing infections along with evolution of mass resistance in bacteria have given rise to the term "post-antibiotic era." The better understanding of electrohydraulic shockwaves bacteriostatic effect could lead to more effective treatments for antibiotic resistant bacteria such as S. aureus.
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology); Slezak, Cyrill (Utah Valley University, Physics)
Purpose:
This study evaluates the effect of electrohydraulic shockwaves on Staphylococcus aureus biofilms. This system could be a great alternative to the use of antibiotics, and potentially life-saving technology that could save billions of dollars.
Background:
The rise of antibiotic-resistant bacteria is a global threat. Staphylococcus aureus is typically harmless, but this gram-positive species has become highly resistant and extremely pathogenic. Strains like MRSA and VRSA have the highest rate of drug resistance and are the leading cause of chronic bacterial infections via bacterial biofilms on medical devices. Biofilms are an aggregation of microbes that excrete an extracellular matrix providing an ideal environment for gene exchange and quorum sensing. Their complexity hinders the diffusion of antimicrobials. A proposed method to prevent device-associated infection is shockwave sterilization and therapy. A shockwave is a high-energy wave causing a sudden change in temperature, pressure and density in the medium. This study investigates the potential disruption of bacterial biofilms by electrohydraulic shockwaves.
Methods:
E. coli and S. aureus biofilms were grown on polystyrene plates. Biofilms were treated with shockwaves (0.19mJ/mm2, 300 pulses, 3 Hz) in a water bath and compared with those treated with Vancomycin. Cell viability was determined through XTT/menadione absorbance and specific biofilm formation through crystal violet absorbance.
Results:
Current testing has shown that electrohydraulic shockwaves have a bacteriostatic effect on biofilms. Other finding show potential for shockwaves to increase bacterial susceptibility to lower levels of antibiotics.
Conclusions:
Device-associated infections are a serious threat to patients' health. The diminishing effectiveness of antibiotics in treating and preventing infections along with evolution of mass resistance in bacteria have given rise to the term "post-antibiotic era." The better understanding of electrohydraulic shockwaves bacteriostatic effect could lead to more effective treatments for antibiotic resistant bacteria such as S. aureus.
Effects of Exercise on Subjective Feelings of Fatigue: A Randomized Crossover Study
Savage, Ryland; Stevens, Andrew; Hicks, Joseph (Brigham Young University)
Faculty Advisor: Bailey, Bruce (Brigham Young University, Exercise Science)
Purpose: This study examined the effect of different intensities of exercise on subjective feelings of fatigue both immediately following exercise and after 60 minutes of intense cognitive testing in men and woman.
Methods: These results come from a secondary analysis of a larger randomized crossover study examining the different levels of exercise intensity on cognitive performance. We recruited 228 men and woman to participate in the study. Each participant was tested after three exercise conditions: moderate (35% VO2 max), vigorous (70% VO2 max), and sedentary (no exercise). Each exercise condition was randomized and each test was separated by one week. After each 40-minute exercise condition, we administered a visual analog scale to evaluate subjective feelings of fatigue. This assessment of fatigue was delivered before treatment, immediately after treatment and then again after a variety of cognitive tasks were performed for the duration of one hour.
Results: For the sedentary condition, feelings of fatigue increased after the 40-minute intervention (p<0.001) and decreased after 60 minutes of cognitive work but remaining elevated compared to baseline (p<0.01). For the moderate condition, feelings of fatigue did not change immediately after exercise (p=0.063) but then increased after 60 minutes of cognitive work (p<0.001). For vigorous condition, feelings of fatigue increased after exercise(p<0.001) and remained elevated after performing the cognitive tasks (p=0.163). Following the 40-minute intervention, fatigue was highest after the sedentary condition, followed by the vigorous condition and did not change after completion of the moderate condition. Following 60 minutes of cognitive work, fatigue was highest in the vigorous condition (p<0.001) and did not differ between the sedentary and moderate conditions (p=0.063).
Conclusion: The results of this study show that moderate exercise has no significant impact on feelings of fatigue but exercise at a higher intensity and being sedentary increase fatigue. Doing cognitive work increases fatigue if exercise precedes the tasks but being sedentary beforehand decreases feelings of fatigue during cognitive tasks. This suggests cognitive work is usually draining, but if performed after sedentary or low activity states such as watching TV, cognitive work will be invigorating and help feel more alert.
Faculty Advisor: Bailey, Bruce (Brigham Young University, Exercise Science)
Purpose: This study examined the effect of different intensities of exercise on subjective feelings of fatigue both immediately following exercise and after 60 minutes of intense cognitive testing in men and woman.
Methods: These results come from a secondary analysis of a larger randomized crossover study examining the different levels of exercise intensity on cognitive performance. We recruited 228 men and woman to participate in the study. Each participant was tested after three exercise conditions: moderate (35% VO2 max), vigorous (70% VO2 max), and sedentary (no exercise). Each exercise condition was randomized and each test was separated by one week. After each 40-minute exercise condition, we administered a visual analog scale to evaluate subjective feelings of fatigue. This assessment of fatigue was delivered before treatment, immediately after treatment and then again after a variety of cognitive tasks were performed for the duration of one hour.
Results: For the sedentary condition, feelings of fatigue increased after the 40-minute intervention (p<0.001) and decreased after 60 minutes of cognitive work but remaining elevated compared to baseline (p<0.01). For the moderate condition, feelings of fatigue did not change immediately after exercise (p=0.063) but then increased after 60 minutes of cognitive work (p<0.001). For vigorous condition, feelings of fatigue increased after exercise(p<0.001) and remained elevated after performing the cognitive tasks (p=0.163). Following the 40-minute intervention, fatigue was highest after the sedentary condition, followed by the vigorous condition and did not change after completion of the moderate condition. Following 60 minutes of cognitive work, fatigue was highest in the vigorous condition (p<0.001) and did not differ between the sedentary and moderate conditions (p=0.063).
Conclusion: The results of this study show that moderate exercise has no significant impact on feelings of fatigue but exercise at a higher intensity and being sedentary increase fatigue. Doing cognitive work increases fatigue if exercise precedes the tasks but being sedentary beforehand decreases feelings of fatigue during cognitive tasks. This suggests cognitive work is usually draining, but if performed after sedentary or low activity states such as watching TV, cognitive work will be invigorating and help feel more alert.
Effects of Parenting Styles on Child Delinquency and Bullying
Leavitt, Amanda; Harris, Heather; Szanter, Kathryn; Stokes, Alexis (Weber State University)
Faculty Advisor: Dunn, Charles (Weber State University, Child and Family Studies); Hubler, Daniel (Weber State University, Child and Family Studies); Osai, Keith (Weber State University, Child and Family Studies)
How a child is parented can influence their success socially, mentally, physically, and emotionally. Agarwal (2017) states that "Parenting plays the most important part in any child's life, it can help a child by teaching them to deal with people, situations and adapt to a better living standard" (p.1335). Parents help form worldviews, shape a child's attitude towards personal achievement, teach how to approach adversity in life, and satisfy their needs whether it be psychological and/or physiological. Much is known about how the base of one's childhood is built upon one's primary caregivers' parenting style, however, less is known about the intergenerational transmission of parenting philosophies.
The current study was designed to assess how parenting styles affect child delinquency and bullying. Through social media invitations and snowball sampling, 200 people accepted invitations to participate in an online survey incorporating both open-ended and quantitative items. Participants were asked to reflect on how they were parented and then state what they have maintained or changed in their own parenting styles. Considering the rich nature of the responses to the qualitative items, a corroborative narrative came together through personal stories.
Through thematic analysis, several themes emerged. When participants reported being parented with: open communication, flexibility, love, trust, and higher expectations they wanted to keep those traits. When asked what participants carried on from their parents one participant stated, "Cherish the family and the moments we have together." Additionally, 28% (n = 113) of our participants shared the importance of spending quality time together. However, when participants' parents did not display those traits, most participants indicated that similar traits, namely communication, flexibility, love, quality time, trust, and higher expectations were desired in their homes. The findings from the current study can be used to inform future parenting research assessing influence on subsequent generations.
Faculty Advisor: Dunn, Charles (Weber State University, Child and Family Studies); Hubler, Daniel (Weber State University, Child and Family Studies); Osai, Keith (Weber State University, Child and Family Studies)
How a child is parented can influence their success socially, mentally, physically, and emotionally. Agarwal (2017) states that "Parenting plays the most important part in any child's life, it can help a child by teaching them to deal with people, situations and adapt to a better living standard" (p.1335). Parents help form worldviews, shape a child's attitude towards personal achievement, teach how to approach adversity in life, and satisfy their needs whether it be psychological and/or physiological. Much is known about how the base of one's childhood is built upon one's primary caregivers' parenting style, however, less is known about the intergenerational transmission of parenting philosophies.
The current study was designed to assess how parenting styles affect child delinquency and bullying. Through social media invitations and snowball sampling, 200 people accepted invitations to participate in an online survey incorporating both open-ended and quantitative items. Participants were asked to reflect on how they were parented and then state what they have maintained or changed in their own parenting styles. Considering the rich nature of the responses to the qualitative items, a corroborative narrative came together through personal stories.
Through thematic analysis, several themes emerged. When participants reported being parented with: open communication, flexibility, love, trust, and higher expectations they wanted to keep those traits. When asked what participants carried on from their parents one participant stated, "Cherish the family and the moments we have together." Additionally, 28% (n = 113) of our participants shared the importance of spending quality time together. However, when participants' parents did not display those traits, most participants indicated that similar traits, namely communication, flexibility, love, quality time, trust, and higher expectations were desired in their homes. The findings from the current study can be used to inform future parenting research assessing influence on subsequent generations.
Differences in anthropometric characteristics between intermediate and high level climbers
Miriam Reber; Ethan Del Toro; Holden Lyman; Weston Hargis; Travis Ficklin (Dixie State University)
Faculty Advisor: Ficklin, Travis (Dixie State University, Health and Human Performance)
In recent years sport climbing has increased in popularity, and with that have come questions regarding what factors improve performance. While multiple studies have examined the effect of anthropometric factors on injury in climbing, few have examined their relationship with climbing performance. Therefore, the purpose of this study is to compare the anthropometric measures of high-level to intermediate level climbers to determine if certain body types, or dimensions, lend themselves to better performance in climbing.
Twenty-one subjects were divided into an intermediate (INT) or high-level (HL) group based on highest graded climb achieved. Various measures including height, limb lengths, and experience. These were compared between the two groups using t-tests.
Significant differences were found in the number of days climbed per week (HL 2.9±1.1, INT 2.17±0.6, p = 0.03), and the ratios of upper arm to full arm length (HL 0.44±0.05, INT 0.41±0.02, p=0.03), and trunk to height (HL 0.26±0.03, INT 0.23±0.01, p=0.01). There were also statistical trends towards years of experience (HL 5.46±3.8, INT 3.39±1.8, p = 0.06), and trunk length (HL 44.96±7.1cm, INT 41.28±2.7cm, p=0.06).
The current data suggest that for the levels of climbing examined (5.9-5.13b on the Yosemite Decimal Scale) a greater trunk to height ratio, greater upper arm to arm length ratio, and increased frequency of practice (a variable within the climber's control) may positively impact climbing ability.
Faculty Advisor: Ficklin, Travis (Dixie State University, Health and Human Performance)
In recent years sport climbing has increased in popularity, and with that have come questions regarding what factors improve performance. While multiple studies have examined the effect of anthropometric factors on injury in climbing, few have examined their relationship with climbing performance. Therefore, the purpose of this study is to compare the anthropometric measures of high-level to intermediate level climbers to determine if certain body types, or dimensions, lend themselves to better performance in climbing.
Twenty-one subjects were divided into an intermediate (INT) or high-level (HL) group based on highest graded climb achieved. Various measures including height, limb lengths, and experience. These were compared between the two groups using t-tests.
Significant differences were found in the number of days climbed per week (HL 2.9±1.1, INT 2.17±0.6, p = 0.03), and the ratios of upper arm to full arm length (HL 0.44±0.05, INT 0.41±0.02, p=0.03), and trunk to height (HL 0.26±0.03, INT 0.23±0.01, p=0.01). There were also statistical trends towards years of experience (HL 5.46±3.8, INT 3.39±1.8, p = 0.06), and trunk length (HL 44.96±7.1cm, INT 41.28±2.7cm, p=0.06).
The current data suggest that for the levels of climbing examined (5.9-5.13b on the Yosemite Decimal Scale) a greater trunk to height ratio, greater upper arm to arm length ratio, and increased frequency of practice (a variable within the climber's control) may positively impact climbing ability.
Dopamine 2 receptors display rapid adaptation in response to acute ethanol administration
LeBaron, Josh; Obray, J Daniel; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences, Psychology)
Dopamine neurons in the substancia nigra (SN) and ventral tegmental area (VTA) are inhibited by dopamine (DA) via dopamine 2 receptor (D2R) activation. D2R expression in the striatum is a well-known biomarker for brain DA levels, drug abuse, and dependence. Markers of D2R expression are not only detectable in the brain but are also expressed in peripheral tissues, including the blood, where DA appears to play a pivotal role in mediating communication between the nervous and immune systems. Alteration in lympocytic D2Rs are seen in chronic psychostimulant use (Ersche et al., 2011). For the last two decades it has been generally accepted that D2R expression in the striatum is reduced by chronic ethanol use. Additionally, research has suggested that these changes mirror changes in DA levels in the striatum and predict risk of relapse. Despite this, the timecourse over which these changes occur has not been demonstrated. Further, recent research has challenged both the reduction in D2R expression produced by chronic ethanol and the mechanism whereby it was believed to be produced (reductions in striatal DA levels). This research has suggested that alterations in D2R levels may be due to disruption of sleep in individuals with substance use disorders. Here we demonstrate that dopamine 2 receptor expression in the brain and the blood follows brain and blood dopamine levels on a timescale of minutes to hours following an acute dose of ethanol. This research provides evidence for transient changes in D2R expression following a single dose of ethanol.
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences, Psychology)
Dopamine neurons in the substancia nigra (SN) and ventral tegmental area (VTA) are inhibited by dopamine (DA) via dopamine 2 receptor (D2R) activation. D2R expression in the striatum is a well-known biomarker for brain DA levels, drug abuse, and dependence. Markers of D2R expression are not only detectable in the brain but are also expressed in peripheral tissues, including the blood, where DA appears to play a pivotal role in mediating communication between the nervous and immune systems. Alteration in lympocytic D2Rs are seen in chronic psychostimulant use (Ersche et al., 2011). For the last two decades it has been generally accepted that D2R expression in the striatum is reduced by chronic ethanol use. Additionally, research has suggested that these changes mirror changes in DA levels in the striatum and predict risk of relapse. Despite this, the timecourse over which these changes occur has not been demonstrated. Further, recent research has challenged both the reduction in D2R expression produced by chronic ethanol and the mechanism whereby it was believed to be produced (reductions in striatal DA levels). This research has suggested that alterations in D2R levels may be due to disruption of sleep in individuals with substance use disorders. Here we demonstrate that dopamine 2 receptor expression in the brain and the blood follows brain and blood dopamine levels on a timescale of minutes to hours following an acute dose of ethanol. This research provides evidence for transient changes in D2R expression following a single dose of ethanol.
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.
Effect of Gender on Shear Wave Elastography and Cross Sectional Area of the Gastrocnemius in Senior Athletes.
Hutchison, Cortland; Preece, J. Caleb; Seibold, Tanner; Feland, J. Brent (Brigham Young University)
Faculty Advisor: Feland, Brent (Life Sciences, Exercise Science)
BACKGROUND: Muscle morphology changes with age and the fibrous/fatty infiltration should affect the overall stiffness of aging muscle. However, little info intrinsic stiffness as measured by SWE is available on aging muscle and the effect of gender using this measurement is not clear.
PURPOSE: This study aimed to determine how gender affects cross sectional area (CSA) and SWE of both the medial and lateral heads of the gastrocnemius muscle of the dominant leg in active older athletes participating in the HuntsmanWorld Senior Games.
METHODS: Data was collected from 116 volunteers (participants in the Huntsman World Senior Games) in St. George, Utah, 2019. Subjects (62 males: mean age = 68.9 ± 7.8 yrs, Ht = 177.4 ± 8.1 cm, Wt= 85.4±20 kg; 54 females: mean age = 66.9 ± 8.5 yrs, Ht =164.4 ± 7.2 cm, Wt= 69.8±18 kg) signed an approved consent form and then lay prone on a treatment table for ultrasonic measurement of both CSA and SWE of both heads of the gastrocnemius.
ANALYSIS: All data were analyzed a generalized linear model analysis using SPSS ver25 comparing CSA and SWE of both medial and lateral gastrocnemius heads by gender with age, height and weight as covariates.
RESULTS & CONCLUSION: We hypothesized that gender would affect both CSA and SWE with males exhibiting increases in both CSA and SWE values. We found a significant difference in CSA of both the medial and lateral gastroc (p=.000 for both) between genders, but no significant difference in lateral gastroc SWE (p=.337) or medial gastroc SWE (p=.320). Both age (p=.004) and weight (p=.000) were also found to significantly affect CSA and SWE between genders. The larger CSA values for men in this study may be more of a function of overall body size and weight since both populations are active athletes.
Faculty Advisor: Feland, Brent (Life Sciences, Exercise Science)
BACKGROUND: Muscle morphology changes with age and the fibrous/fatty infiltration should affect the overall stiffness of aging muscle. However, little info intrinsic stiffness as measured by SWE is available on aging muscle and the effect of gender using this measurement is not clear.
PURPOSE: This study aimed to determine how gender affects cross sectional area (CSA) and SWE of both the medial and lateral heads of the gastrocnemius muscle of the dominant leg in active older athletes participating in the HuntsmanWorld Senior Games.
METHODS: Data was collected from 116 volunteers (participants in the Huntsman World Senior Games) in St. George, Utah, 2019. Subjects (62 males: mean age = 68.9 ± 7.8 yrs, Ht = 177.4 ± 8.1 cm, Wt= 85.4±20 kg; 54 females: mean age = 66.9 ± 8.5 yrs, Ht =164.4 ± 7.2 cm, Wt= 69.8±18 kg) signed an approved consent form and then lay prone on a treatment table for ultrasonic measurement of both CSA and SWE of both heads of the gastrocnemius.
ANALYSIS: All data were analyzed a generalized linear model analysis using SPSS ver25 comparing CSA and SWE of both medial and lateral gastrocnemius heads by gender with age, height and weight as covariates.
RESULTS & CONCLUSION: We hypothesized that gender would affect both CSA and SWE with males exhibiting increases in both CSA and SWE values. We found a significant difference in CSA of both the medial and lateral gastroc (p=.000 for both) between genders, but no significant difference in lateral gastroc SWE (p=.337) or medial gastroc SWE (p=.320). Both age (p=.004) and weight (p=.000) were also found to significantly affect CSA and SWE between genders. The larger CSA values for men in this study may be more of a function of overall body size and weight since both populations are active athletes.
Effects of Flavanols on β-cell proliferation.
Tessem, Jeffery; Lloyd, Trevor; Brown, Nathan (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics and Food Science)
Diabetes is a global epidemic affecting millions of people. The total estimated cost of diabetes in the U.S. during 2017 was 327 billion dollars [1]. Diabetes is characterized by the loss of pancreatic β-cell function which is caused by an autoimmune disorder in Type 1 diabetes or insulin resistance and β-cell exhaustion in Type 2 (T2D) diabetes. Lifestyle changes in diet are beneficial in treating T2D. Phytochemicals are commonly utilized in these diets, and recent studies show diets high in flavanols exert beneficial bioactivity for β-cells. While flavanols demonstrate beneficial effects on β-cells, these flavanols are rarely observed in circulation, suggesting a necessary intermediate step. Flavanols are metabolized by gut bacteria to smaller metabolites that are absorbable. We hypothesize that these gut bacteria derived flavanol metabolites cross the gut and affect β-cell function. We have fed rats catechin supplemented or unsupplemented diets and collected urine as a means to isolate all absorbable gut flavanol metabolites. Here we present the effects of these absorbed metabolites on β-cell proliferation. This study begins to explain the mechanism by which flavanols exert their beneficial effect on glucose metabolism through the β-cell.
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics and Food Science)
Diabetes is a global epidemic affecting millions of people. The total estimated cost of diabetes in the U.S. during 2017 was 327 billion dollars [1]. Diabetes is characterized by the loss of pancreatic β-cell function which is caused by an autoimmune disorder in Type 1 diabetes or insulin resistance and β-cell exhaustion in Type 2 (T2D) diabetes. Lifestyle changes in diet are beneficial in treating T2D. Phytochemicals are commonly utilized in these diets, and recent studies show diets high in flavanols exert beneficial bioactivity for β-cells. While flavanols demonstrate beneficial effects on β-cells, these flavanols are rarely observed in circulation, suggesting a necessary intermediate step. Flavanols are metabolized by gut bacteria to smaller metabolites that are absorbable. We hypothesize that these gut bacteria derived flavanol metabolites cross the gut and affect β-cell function. We have fed rats catechin supplemented or unsupplemented diets and collected urine as a means to isolate all absorbable gut flavanol metabolites. Here we present the effects of these absorbed metabolites on β-cell proliferation. This study begins to explain the mechanism by which flavanols exert their beneficial effect on glucose metabolism through the β-cell.
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.
Effects of Grape Seed Extract Metabolites on ß-cell Proliferation and Function
Beales, Joseph; Lloyd, Trevor; Krueger, Emily; Barlow, Andrew (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutritional, Dietetics, and Food Science)
Worldwide, an estimated 415 million people suffer from diabetes.1 Diabetes is characterized by chronic dysfunction of the pancreatic ß-cell, which leads to unregulated insulin secretion and abnormal blood glucose levels. Therefore, methods which increase the number of ß-cells or improve their function have potential for complementary treatment of type 2 diabetes. Compounds such as antioxidants and their gut metabolites have received attention in literature as having potential ß-cell-regulating properties.2,3 Therefore, we hypothesize that supplementation of grape seed extract (GSE), which is rich in antioxidants, will enhance ß-cell proliferation and insulin secretion. Accordingly, we obtained metabolites, derived from rats on either a control or grape seed extract diet, to measure the metabolites' impact on ß-cell function through in vitro assays such as glucose stimulated insulin secretion (GSIS) and 3H-thymidine incorporation. Discoveries regarding GSE metabolites' effects on ß-cell function could be fundamental to understanding ß-cell regulation and potential pharmaceutical or dietary treatments for diabetes.
1 Ogurtsova, K., et al. "IDF Diabetes Atlas: Global Estimates for the Prevalence of Diabetes for 2015 and 2040." Diabetes Research and Clinical Practice, Elsevier, 31 Mar. 2017, www.sciencedirect.com/science/article/pii/S0168822717303753?via%3Dihub.
2 Bajaj, Sarita, and Afreen Khan. "Antioxidants and diabetes." Indian journal of endocrinology and metabolism vol. 16,Suppl 2 (2012): S267-71. Doi:10.4103/2230-8210.104057
3 Tsuda, Takanori. "Recent Progress in Anti-Obesity and Anti-Diabetes Effect of Berries." MDPI, Multidisciplinary Digital Publishing Institute, 6 Apr. 2016, www.mdpi.com/2076-3921/5/2/13.
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutritional, Dietetics, and Food Science)
Worldwide, an estimated 415 million people suffer from diabetes.1 Diabetes is characterized by chronic dysfunction of the pancreatic ß-cell, which leads to unregulated insulin secretion and abnormal blood glucose levels. Therefore, methods which increase the number of ß-cells or improve their function have potential for complementary treatment of type 2 diabetes. Compounds such as antioxidants and their gut metabolites have received attention in literature as having potential ß-cell-regulating properties.2,3 Therefore, we hypothesize that supplementation of grape seed extract (GSE), which is rich in antioxidants, will enhance ß-cell proliferation and insulin secretion. Accordingly, we obtained metabolites, derived from rats on either a control or grape seed extract diet, to measure the metabolites' impact on ß-cell function through in vitro assays such as glucose stimulated insulin secretion (GSIS) and 3H-thymidine incorporation. Discoveries regarding GSE metabolites' effects on ß-cell function could be fundamental to understanding ß-cell regulation and potential pharmaceutical or dietary treatments for diabetes.
1 Ogurtsova, K., et al. "IDF Diabetes Atlas: Global Estimates for the Prevalence of Diabetes for 2015 and 2040." Diabetes Research and Clinical Practice, Elsevier, 31 Mar. 2017, www.sciencedirect.com/science/article/pii/S0168822717303753?via%3Dihub.
2 Bajaj, Sarita, and Afreen Khan. "Antioxidants and diabetes." Indian journal of endocrinology and metabolism vol. 16,Suppl 2 (2012): S267-71. Doi:10.4103/2230-8210.104057
3 Tsuda, Takanori. "Recent Progress in Anti-Obesity and Anti-Diabetes Effect of Berries." MDPI, Multidisciplinary Digital Publishing Institute, 6 Apr. 2016, www.mdpi.com/2076-3921/5/2/13.
Drosophila melanogaster Determines Dietary Preference Through Volatile Detection
Walker, Carson; Burke, Tyler; Tanner, Call; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Brigham Young University, Plant and Wildlife Sciences)
Host-microbe interactions can dramatically influence Drosophila melanogaster phenotypes, but few studies have explained how these microbes are recognized by the host. For example, fruit flies from one area, Maine, prefer to consume diets inoculated with Lactic Acid Bacteria (LABs) over diets containing Acetic acid bacteria (AABs); whereas flies from another area, Florida, show no preference for either LAB or AAB. However, the bacterial mechanisms responsible for this preference are unknown. My follow-up analyses further suggest the hypothesis that Maine flies avoid AAB, rather than are attracted to LAB. Therefore, I propose a forward genetic approach to define the AAB processes that shape this fly preference. I will do this by comparing the feeding preferences of Maine and Florida flies to diets incorporated with LAB versus specific bacterial molecules, such as acetic acid, peptidoglycan, and lipopolysaccharides. Alternatively, if these molecules do not mediate the effects I will perform a metagenome wide association assay (MGWA) to identify genes linked to this preference. Together, these approaches will help to reveal the bacterial factors that influence fly feeding preferences.
Faculty Advisor: Chaston, John (Brigham Young University, Plant and Wildlife Sciences)
Host-microbe interactions can dramatically influence Drosophila melanogaster phenotypes, but few studies have explained how these microbes are recognized by the host. For example, fruit flies from one area, Maine, prefer to consume diets inoculated with Lactic Acid Bacteria (LABs) over diets containing Acetic acid bacteria (AABs); whereas flies from another area, Florida, show no preference for either LAB or AAB. However, the bacterial mechanisms responsible for this preference are unknown. My follow-up analyses further suggest the hypothesis that Maine flies avoid AAB, rather than are attracted to LAB. Therefore, I propose a forward genetic approach to define the AAB processes that shape this fly preference. I will do this by comparing the feeding preferences of Maine and Florida flies to diets incorporated with LAB versus specific bacterial molecules, such as acetic acid, peptidoglycan, and lipopolysaccharides. Alternatively, if these molecules do not mediate the effects I will perform a metagenome wide association assay (MGWA) to identify genes linked to this preference. Together, these approaches will help to reveal the bacterial factors that influence fly feeding preferences.
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.
The Beta Cell Struggle: How CDKIs and Age Affect Cell Proliferation in Type 1 Diabetes
Jensen, Daelin; Baxter, Melanie (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Nutrition, Dietetics, and Food Science; Life Sciences)
Approximately 1.25 million people are currently living with type 1 diabetes. By 2050, 5 million people are expected to be diagnosed with the disease1. The insulin secreting pancreatic beta cells are essential to control proper glucose absorption and storage in insulin sensitive peripheral tissue. Both type 1 and type 2 diabetes are characterized by decreased functional beta cell mass and, consequently, decreased insulin production. One potential intervention is the use of beta cell transplantation from cadaveric donors. A major impediment to greater application of this treatment is the scarcity of transplant ready beta cells. Increasing the quantity of functional beta cells for transplantation will lead to increased insulin production and better management of the disease. Various genes have been defined that can induce beta cell replication. A major caveat of these findings, however, is that these factors induce replication in young beta cells but not in aged beta cells. Age-dependent morphological changes in the beta cell are poorly understood, despite its relevance to type 1 diabetes: here, we show that insulin-positive tissue area changes with age. Given that the majority of beta cells that will be used for transplant will come from aged donors, it is imperative to understand why aged beta cells are refractory to the aforementioned proliferative mechanisms. The cell cycle is tightly regulated by cyclin-dependent kinases. Cyclin-dependent kinase inhibitors (CDKI's) bind to cyclin dependent kinases, inhibiting cell proliferation. We hypothesized that these CDKIs are responsible for the observed lack of proliferation in aged animals. We demonstrate the expression of the Ink4 and Cip/Kip family of CDKI's by mRNA, protein and histological expression in 5 week and 5 month old primary rat beta cells. In addition, we show how size-related expression differences of CDKIs relate to beta cell proliferation.
Faculty Advisor: Tessem, Jeffery (Nutrition, Dietetics, and Food Science; Life Sciences)
Approximately 1.25 million people are currently living with type 1 diabetes. By 2050, 5 million people are expected to be diagnosed with the disease1. The insulin secreting pancreatic beta cells are essential to control proper glucose absorption and storage in insulin sensitive peripheral tissue. Both type 1 and type 2 diabetes are characterized by decreased functional beta cell mass and, consequently, decreased insulin production. One potential intervention is the use of beta cell transplantation from cadaveric donors. A major impediment to greater application of this treatment is the scarcity of transplant ready beta cells. Increasing the quantity of functional beta cells for transplantation will lead to increased insulin production and better management of the disease. Various genes have been defined that can induce beta cell replication. A major caveat of these findings, however, is that these factors induce replication in young beta cells but not in aged beta cells. Age-dependent morphological changes in the beta cell are poorly understood, despite its relevance to type 1 diabetes: here, we show that insulin-positive tissue area changes with age. Given that the majority of beta cells that will be used for transplant will come from aged donors, it is imperative to understand why aged beta cells are refractory to the aforementioned proliferative mechanisms. The cell cycle is tightly regulated by cyclin-dependent kinases. Cyclin-dependent kinase inhibitors (CDKI's) bind to cyclin dependent kinases, inhibiting cell proliferation. We hypothesized that these CDKIs are responsible for the observed lack of proliferation in aged animals. We demonstrate the expression of the Ink4 and Cip/Kip family of CDKI's by mRNA, protein and histological expression in 5 week and 5 month old primary rat beta cells. In addition, we show how size-related expression differences of CDKIs relate to beta cell proliferation.
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.
The Effects of Glucolipotoxicity on Nkx6.1 Expression and Insulin Secretion in the Beta Cell
Elison, Weston; Bauchle, Casey; Bunker, Libby; Stephens, Samuel; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics and Food Science)
Type 2 Diabetes (T2D) effects hundreds of millions of people worldwide, with that number increasing rapidly. It is characterized by increased insulin resistance and dysfunctional insulin secretion. The beta cell of the pancreas is the primary insulin secreting tissue, found in the endocrine tissue of the pancreas called islets of Langerhans. In T2D beta cells become glucose intolerant and disease progression is characterized by loss of functional beta cell mass. Previous studies have shown that the transcription factor Nkx6.1 is vital for beta cell differentiation, identity, and insulin secretion. Research has indicated that Nkx6.1 expression and protein levels decrease in pancreatic islets from human donors with T2D. Our data indicates that glucolipotoxicity, a common model for obesity and diabetes in cell culture, leads to decreased Nkx6.1 mRNA expression, protein levels and nuclear localization in Ins-1 832/13 cells. Nkx6.1 regulates genes in the nucleus , and its loss inhibits proper insulin secretion. We propose that reactive oxygen species created by metabolism of excess fuel decreases Nkx6.1 expression and Nkx6.1 target gene expression, as measured by quantitative polymerase chain reaction (qPCR). Also, increased glucose concentrations causes increased Nkx6.1 protein degradation and translocation out of the nucleus. Protein levels will be measured by western blot and localization by confocal microscopy. In order to understand how these changes effect beta cell function, we will measure glucose stimulated insulin secretion by sandwich Enzyme Linked Immunosorbent Assay (ELISA). We further propose that Nkx6.1 overexpression will restore beta cell function. These results will assist in unraveling the cause of beta cell dysfunction in T2D.
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics and Food Science)
Type 2 Diabetes (T2D) effects hundreds of millions of people worldwide, with that number increasing rapidly. It is characterized by increased insulin resistance and dysfunctional insulin secretion. The beta cell of the pancreas is the primary insulin secreting tissue, found in the endocrine tissue of the pancreas called islets of Langerhans. In T2D beta cells become glucose intolerant and disease progression is characterized by loss of functional beta cell mass. Previous studies have shown that the transcription factor Nkx6.1 is vital for beta cell differentiation, identity, and insulin secretion. Research has indicated that Nkx6.1 expression and protein levels decrease in pancreatic islets from human donors with T2D. Our data indicates that glucolipotoxicity, a common model for obesity and diabetes in cell culture, leads to decreased Nkx6.1 mRNA expression, protein levels and nuclear localization in Ins-1 832/13 cells. Nkx6.1 regulates genes in the nucleus , and its loss inhibits proper insulin secretion. We propose that reactive oxygen species created by metabolism of excess fuel decreases Nkx6.1 expression and Nkx6.1 target gene expression, as measured by quantitative polymerase chain reaction (qPCR). Also, increased glucose concentrations causes increased Nkx6.1 protein degradation and translocation out of the nucleus. Protein levels will be measured by western blot and localization by confocal microscopy. In order to understand how these changes effect beta cell function, we will measure glucose stimulated insulin secretion by sandwich Enzyme Linked Immunosorbent Assay (ELISA). We further propose that Nkx6.1 overexpression will restore beta cell function. These results will assist in unraveling the cause of beta cell dysfunction in T2D.
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.
The Effects Of Invasive Common Carp On Invertebrate Food Sources For Diving Ducks In Great Salt Lake Wetlands
Karin, Kettenring; Robison, Talin; Leonard, Emily (Utah State University)
Faculty Advisor: Kettering, Karin (S.J. & Jessie E. Quinney College of Natural Resource, Watershed Sciences Department)
The Great Salt Lake (GSL) and its wetlands are important habitat for migrating birds. The GSL wetlands provide crucial habitat for nesting, food, and areas to recover from migration. Common carp are a threat to GSL wetlands. Carp disturb sediments in the water, blocking some of the sunlight from entering the water, which is utilized by aquatic macrophytes and algae. Carp also may be affecting invertebrate populations, which are critical food resources for migrating birds, but these effects have not been well-documented. My research addressed the question: what are the effects of invasive common carp on invertebrate food sources for diving ducks in the Great Salt Lake wetlands? I answered my research question by addressing the following objectives: (1) to identify the benthic, epiphytic, and water-column dwelling invertebrates in Farmington Bay Waterfowl Management Area (WMA), and (2) to determine if common carp are having an impact on the overall density, diversity, and abundance of the invertebrate communities fed on by diving ducks. I compared invertebrate communities (diversity and abundance) between carp-excluded boxes and control boxes. I constructed my carp exclosures of wire mesh and t-posts to prohibit carp from entering while still allowing invertebrates and water to freely move in and out of the exclosure. The control boxes were constructed of t-posts and allowed carp to freely enter and exit the box. I used dipnet and substrate core samples to determine what invertebrates are living in the water column and substrates at Farmington Bay wetlands. Although sample processing is on-going, early results indicate that carp reduce water column invertebrate abundance while effects on invertebrate diversity are thus far inconclusive. Given the importance of GSL wetlands and their invertebrate food sources to migrating diving, my research findings underscore the importance of aggressive carp management.
Faculty Advisor: Kettering, Karin (S.J. & Jessie E. Quinney College of Natural Resource, Watershed Sciences Department)
The Great Salt Lake (GSL) and its wetlands are important habitat for migrating birds. The GSL wetlands provide crucial habitat for nesting, food, and areas to recover from migration. Common carp are a threat to GSL wetlands. Carp disturb sediments in the water, blocking some of the sunlight from entering the water, which is utilized by aquatic macrophytes and algae. Carp also may be affecting invertebrate populations, which are critical food resources for migrating birds, but these effects have not been well-documented. My research addressed the question: what are the effects of invasive common carp on invertebrate food sources for diving ducks in the Great Salt Lake wetlands? I answered my research question by addressing the following objectives: (1) to identify the benthic, epiphytic, and water-column dwelling invertebrates in Farmington Bay Waterfowl Management Area (WMA), and (2) to determine if common carp are having an impact on the overall density, diversity, and abundance of the invertebrate communities fed on by diving ducks. I compared invertebrate communities (diversity and abundance) between carp-excluded boxes and control boxes. I constructed my carp exclosures of wire mesh and t-posts to prohibit carp from entering while still allowing invertebrates and water to freely move in and out of the exclosure. The control boxes were constructed of t-posts and allowed carp to freely enter and exit the box. I used dipnet and substrate core samples to determine what invertebrates are living in the water column and substrates at Farmington Bay wetlands. Although sample processing is on-going, early results indicate that carp reduce water column invertebrate abundance while effects on invertebrate diversity are thus far inconclusive. Given the importance of GSL wetlands and their invertebrate food sources to migrating diving, my research findings underscore the importance of aggressive carp management.
The Role of Bacterial Genotype in Persistence of the Microbiota of Drosophila melanogaster
Gottfredson, Sarah; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)
The microbiome of Drosophila melanogaster can have significant effects on the host, and many of these have been studied. However, the reason why the bacterial species associate with and persist in D. melanogaster has not been studied in depth. Here we define persistence as how long a microbe associates with a host. The early assumption has been that the D. melanogaster gut microbiome is established solely through diet, but recent work suggests that other factors may be at play in the microbiome establishment. This experiment aims to study the correlation between bacterial genotype and persistence in the D. melanogaster microbiome. In this study, a metagenome wide association (MGWAS) was done using 40 different strains of bacteria to find distinct bacterial genes that are significantly correlated with persistence. To do this, each strain was mono-associated with twenty-four individual flies. The flies were reared for fourteen days, transferred onto new food three times a day for two days, homogenized, and plated. Using the significant genes found through the MGWAS, the same experiment protocol will be used to test mutants of these genes for their effect on persistence. These data will provide us with distinct genes that are necessary for effective bacterial persistence.
Faculty Advisor: Chaston, John (Life Sciences, Plant and Wildlife Sciences)
The microbiome of Drosophila melanogaster can have significant effects on the host, and many of these have been studied. However, the reason why the bacterial species associate with and persist in D. melanogaster has not been studied in depth. Here we define persistence as how long a microbe associates with a host. The early assumption has been that the D. melanogaster gut microbiome is established solely through diet, but recent work suggests that other factors may be at play in the microbiome establishment. This experiment aims to study the correlation between bacterial genotype and persistence in the D. melanogaster microbiome. In this study, a metagenome wide association (MGWAS) was done using 40 different strains of bacteria to find distinct bacterial genes that are significantly correlated with persistence. To do this, each strain was mono-associated with twenty-four individual flies. The flies were reared for fourteen days, transferred onto new food three times a day for two days, homogenized, and plated. Using the significant genes found through the MGWAS, the same experiment protocol will be used to test mutants of these genes for their effect on persistence. These data will provide us with distinct genes that are necessary for effective bacterial persistence.
Temperature Effects on D. melanogaster Microbiota Content
Not yet published (Brigham Young University)
Faculty Advisor: Chaston, John (Brigham young University, Life Sciences)
Within an organism's gut are many strains of bacteria that are constantly interacting with their host. Microbiota composition has been shown to impact many aspects of host health such as metabolism, fat-storage, starvation resistance, and reproduction. Certain behaviors and outcomes have been correlated with certain microbial taxa present in the host gut.
D. melanogaster serves as a useful tool for studying this relationship because its microbiota contains relatively few bacterial strains and is both widely studied and largely understood. Previous research within our lab involving D. melanogaster has found trends in many life-history strategies (ie. reproduction, fecundity, lifespan) that correlate with the presence of certain gut bacteria. While there are many aspects of health that microbiota composition affects, there are also a variety of factors that impact microbiota composition thus leading to these end results.
This experiment seeks to further understand the role that environment has in determining microbiota composition. By rearing gnotobiotic flies in environments that differ in temperature, we can then analyze microbiota content to see if any fluctuations occur due to environmental temperature. If temperature is found to have an effect on the taxa present in fully developed D. melanogaster, we can then seek to determine whether or not there are evolutions taking place in host genotype that yield differing microbiota phenotypically.
Faculty Advisor: Chaston, John (Brigham young University, Life Sciences)
Within an organism's gut are many strains of bacteria that are constantly interacting with their host. Microbiota composition has been shown to impact many aspects of host health such as metabolism, fat-storage, starvation resistance, and reproduction. Certain behaviors and outcomes have been correlated with certain microbial taxa present in the host gut.
D. melanogaster serves as a useful tool for studying this relationship because its microbiota contains relatively few bacterial strains and is both widely studied and largely understood. Previous research within our lab involving D. melanogaster has found trends in many life-history strategies (ie. reproduction, fecundity, lifespan) that correlate with the presence of certain gut bacteria. While there are many aspects of health that microbiota composition affects, there are also a variety of factors that impact microbiota composition thus leading to these end results.
This experiment seeks to further understand the role that environment has in determining microbiota composition. By rearing gnotobiotic flies in environments that differ in temperature, we can then analyze microbiota content to see if any fluctuations occur due to environmental temperature. If temperature is found to have an effect on the taxa present in fully developed D. melanogaster, we can then seek to determine whether or not there are evolutions taking place in host genotype that yield differing microbiota phenotypically.
The search for Lactobacillus wasatchensis.
Thornton, Sherie; Cardona,Rebecca (Weber State University)
Faculty Advisor: Culumber, Michele (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology)
Lactobacillus wasatchensis was initially isolated from cheese produced at Utah State University and was found to be a Non-Starter Lactic Acid Bacteria (NSLAB) that causes late-gas production in cheese that can damage packaging and produce defects in the cheese. The goal of this project was to locate an environmental reservoir for Lactobacillus wasatchensis. Five samples of silage that were in different stages of fermentation and content and raw milk samples were obtained at the Utah State University dairy. Samples were serially diluted, plated on de Man, Rogosa and Sharpe agar supplemented with 1% D-Ribose (NRS-R) and incubated anaerobically for 5 days. Colonies that looked like potential Lb. wasatchensis were selected and regrown for isolation. All isolates were gram-positive rods. The isolates were further grown in broth for DNA extraction, sequencing, and analysis with API 50 carbohydrate panel (API 50CH). The API 50CH results were significantly different from Lb. wasatchensis, which only demonstrates use of ribose in this assay. Sequencing of the 16S rRNA gene, however, produced a match to three isolates from two different silage samples that had 99% sequence identity to Lb. wasatchensis. Further analysis of the isolates is being done to confirm this finding and describe the organism isolated from the soil. We hypothesize that these organisms are very closely related to Lb. wasatchensis and that silage could be an environmental source of contamination.
Faculty Advisor: Culumber, Michele (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology)
Lactobacillus wasatchensis was initially isolated from cheese produced at Utah State University and was found to be a Non-Starter Lactic Acid Bacteria (NSLAB) that causes late-gas production in cheese that can damage packaging and produce defects in the cheese. The goal of this project was to locate an environmental reservoir for Lactobacillus wasatchensis. Five samples of silage that were in different stages of fermentation and content and raw milk samples were obtained at the Utah State University dairy. Samples were serially diluted, plated on de Man, Rogosa and Sharpe agar supplemented with 1% D-Ribose (NRS-R) and incubated anaerobically for 5 days. Colonies that looked like potential Lb. wasatchensis were selected and regrown for isolation. All isolates were gram-positive rods. The isolates were further grown in broth for DNA extraction, sequencing, and analysis with API 50 carbohydrate panel (API 50CH). The API 50CH results were significantly different from Lb. wasatchensis, which only demonstrates use of ribose in this assay. Sequencing of the 16S rRNA gene, however, produced a match to three isolates from two different silage samples that had 99% sequence identity to Lb. wasatchensis. Further analysis of the isolates is being done to confirm this finding and describe the organism isolated from the soil. We hypothesize that these organisms are very closely related to Lb. wasatchensis and that silage could be an environmental source of contamination.
The effect of IL-1β on Nf-_B and ICAM-1 mechanism
Hendricks, Kyle; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
Over 30 million Americans suffer from type 1 (T1D) or type 2 diabetes (T2D), the seventh leading cause of death in the US. T1D and T2D is caused by a significant decrease in pancreatic β-cell mass, resulting in the body's inability to regulate blood glucose. Specifically, T1D is classified as an autoimmune disease due to pancreatic β-cell death by the body's T cells. Nf-κB is required for T cell mediated β-cell destruction. Nf-κB interacts with ICAM-1 on the T cell and acts in conjunction with IL-1β which acts as a T cell activator. This pathway is part of the mechanism that contributes to T cell mediated cell destruction. Here we hypothesize that IL-1β is involved in the mechanism that contributes to Nf-κB and ICAM-1 binding. We will begin with an electrophoretic mobility shift assay to identify the interactions between the ICAM-1 site on IL-1β treated cells and the Nf-κB binding complex. A better understanding of this pathology can, in the future, lead to a treatment that could regulate T cell mediated death of β-cells.
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
Over 30 million Americans suffer from type 1 (T1D) or type 2 diabetes (T2D), the seventh leading cause of death in the US. T1D and T2D is caused by a significant decrease in pancreatic β-cell mass, resulting in the body's inability to regulate blood glucose. Specifically, T1D is classified as an autoimmune disease due to pancreatic β-cell death by the body's T cells. Nf-κB is required for T cell mediated β-cell destruction. Nf-κB interacts with ICAM-1 on the T cell and acts in conjunction with IL-1β which acts as a T cell activator. This pathway is part of the mechanism that contributes to T cell mediated cell destruction. Here we hypothesize that IL-1β is involved in the mechanism that contributes to Nf-κB and ICAM-1 binding. We will begin with an electrophoretic mobility shift assay to identify the interactions between the ICAM-1 site on IL-1β treated cells and the Nf-κB binding complex. A better understanding of this pathology can, in the future, lead to a treatment that could regulate T cell mediated death of β-cells.
The role of Fibroblast Growth Factor 21 (FGF21) in Mitochondrial Disorders (MDs)
Almaw, Naredos; Chaudhuri, Dipayan (University of Utah)
Faculty Advisor: Chaudhuri, Dipayan (School of Medicine, Internal Medicine)
Fibroblast Growth Factor 21 (FGF21), a regulator of metabolism that is typically expressed in the liver, has recently been shown to be induced by other tissues in the body as a response to mitochondrial stress. Elevated levels of serum FGF21 was exhibited in children with mitochondrial mutation-induced mitochondrial dysfunctions. Similarly, in dilated cardiomyopathy, a common type of heart failure (HF) mitochondrial dysfunction is associated with mitochondrial DNA damage. This study aims to determine the signaling pathway that leads to the production and effects of FGF21 during mitochondrial dysfunction associated HF. We hypothesize that in left ventricular failure, cardiomyocytes experience oxidative stress, which initiates signaling pathways that leads to the production of FGF21 by other organs.
To test this hypothesis, HF was induced in four mice models via Transverse Aortic Constriction (TAC), and tissue samples were collected. Messenger RNA (mRNA) was extracted, and quantitative Polymerase Chain Reaction (qPCR) was performed to examine the FGF21 gene expression in control and experiment mice models. The qPCR data showed an upregulation of FGF21 in the heart, liver, and pancreas of experiment mice. qPCR results were confirmed through FGF21 protein expression via western blot. Our preliminary results appear to support our hypothesis that during heart failure, the heart sends stress signals to other organs to produce FGF21. Understanding the origin of FGF21 production could help better understand the critical role it plays in preventing disease progression in HF patients.
Faculty Advisor: Chaudhuri, Dipayan (School of Medicine, Internal Medicine)
Fibroblast Growth Factor 21 (FGF21), a regulator of metabolism that is typically expressed in the liver, has recently been shown to be induced by other tissues in the body as a response to mitochondrial stress. Elevated levels of serum FGF21 was exhibited in children with mitochondrial mutation-induced mitochondrial dysfunctions. Similarly, in dilated cardiomyopathy, a common type of heart failure (HF) mitochondrial dysfunction is associated with mitochondrial DNA damage. This study aims to determine the signaling pathway that leads to the production and effects of FGF21 during mitochondrial dysfunction associated HF. We hypothesize that in left ventricular failure, cardiomyocytes experience oxidative stress, which initiates signaling pathways that leads to the production of FGF21 by other organs.
To test this hypothesis, HF was induced in four mice models via Transverse Aortic Constriction (TAC), and tissue samples were collected. Messenger RNA (mRNA) was extracted, and quantitative Polymerase Chain Reaction (qPCR) was performed to examine the FGF21 gene expression in control and experiment mice models. The qPCR data showed an upregulation of FGF21 in the heart, liver, and pancreas of experiment mice. qPCR results were confirmed through FGF21 protein expression via western blot. Our preliminary results appear to support our hypothesis that during heart failure, the heart sends stress signals to other organs to produce FGF21. Understanding the origin of FGF21 production could help better understand the critical role it plays in preventing disease progression in HF patients.
The Effects of the Ketogenic Diet on Learning and Memory
Edwards, Jeffery; Saito, Erin; Blaylock, Tanner; Brantley, Adam; Winzenried, Eric (Brigham Young University)
Faculty Advisor: Edwards, Jeffrey (Life Sciences, Physiology and Developmental Biology)
The ketogenic diet initially began as a significant treatment to prevent epilepsy. More recently it has seen a rise in popularity again, with many attributing positive physiological and cognitive benefits. The purpose of this study is to assess the validity of those claims in an animal model in order to examine this at the cellular level as well as identify possible molecular mechanisms for the changes observed. To quantify this, mice will be fed a diet high in fats and low in carbohydrates. A Morris water maze, radial arm maze, and novel object recognition will then be used to assess the diets effect on behavioral memory. Field electrophysiology will then be performed in the CA1 region of the hippocampus, the region of the brain responsible for mediating memory, to measure two types of synaptic plasticity: long-term potentiation and long-term depression. It has been previously hypothesized that changes in BDNF concentration are a possible explanation for physiological changes caused by the keto diet. To assess this, ANA-12, a TrkB antagonist, will be used to block the effects caused by BDNF. Preliminary data gathered from bathed brain slices of both male and female animals have shown an enhancement of LTP, the cellular equivalent of learning and memory. These data lead us to our hypothesis that the ketogenic diet will cause significant changes in behavioral memory and CA1 synaptic plasticity through altered BDNF levels.
Faculty Advisor: Edwards, Jeffrey (Life Sciences, Physiology and Developmental Biology)
The ketogenic diet initially began as a significant treatment to prevent epilepsy. More recently it has seen a rise in popularity again, with many attributing positive physiological and cognitive benefits. The purpose of this study is to assess the validity of those claims in an animal model in order to examine this at the cellular level as well as identify possible molecular mechanisms for the changes observed. To quantify this, mice will be fed a diet high in fats and low in carbohydrates. A Morris water maze, radial arm maze, and novel object recognition will then be used to assess the diets effect on behavioral memory. Field electrophysiology will then be performed in the CA1 region of the hippocampus, the region of the brain responsible for mediating memory, to measure two types of synaptic plasticity: long-term potentiation and long-term depression. It has been previously hypothesized that changes in BDNF concentration are a possible explanation for physiological changes caused by the keto diet. To assess this, ANA-12, a TrkB antagonist, will be used to block the effects caused by BDNF. Preliminary data gathered from bathed brain slices of both male and female animals have shown an enhancement of LTP, the cellular equivalent of learning and memory. These data lead us to our hypothesis that the ketogenic diet will cause significant changes in behavioral memory and CA1 synaptic plasticity through altered BDNF levels.
Substrate specificity in variants of an aldehyde oxidoreductase
Carter, Riley; Hertig, Jess; Durrant, Doran (Southern Utah University)
Faculty Advisor: Pierce, Elizabeth (Science and Engineering, Physical Science)
Aldehyde oxidoreductases (AOR) are enzymes used to catalyze the conversion between aldehydes and carboxylic acids. Certain bacteria use these enzymes as a source of metabolism or to detoxify aldehydes to less toxic carboxylic acids: Desulfovibrio gigas uses a highly efficient enzyme (DgAOR) to oxidize benzaldehyde in metabolism while E. coli uses a periplasmic AOR (PaoABC) to detoxify aldehydes. These AORs are members of the xanthine oxidase family, but they don't metabolize many of the normal substrates characteristic of this enzyme family, namely purines. Moreover, the active sites of these enzymes have very different environments. Correia, et al (2014) characterized the kinetics and structure of DgAOR with several substrates and found that the Phe425 and Tyr535 residues at the active site likely stabilize aromatic aldehydes by pi stacking. This active site was also buried away from solvent. The active site of PaoABC lacked any significant aromatic residues and was positioned at the surface of the protein. The substrate stabilizing elements at this active site are Leu246 and Pro352. We are interested in why these active sites both are unreactive towards purines given their different chemical and location compared to the solvent. We propose that by mutating PaoABC to have smaller, nonpolar residues at the 246 and 352 position, we may be able to change the specificity of PaoABC to include purines. We also will mutate these residues to aromatic groups to probe at the chemical environment of the active site and its similarities to DgAOR.
Faculty Advisor: Pierce, Elizabeth (Science and Engineering, Physical Science)
Aldehyde oxidoreductases (AOR) are enzymes used to catalyze the conversion between aldehydes and carboxylic acids. Certain bacteria use these enzymes as a source of metabolism or to detoxify aldehydes to less toxic carboxylic acids: Desulfovibrio gigas uses a highly efficient enzyme (DgAOR) to oxidize benzaldehyde in metabolism while E. coli uses a periplasmic AOR (PaoABC) to detoxify aldehydes. These AORs are members of the xanthine oxidase family, but they don't metabolize many of the normal substrates characteristic of this enzyme family, namely purines. Moreover, the active sites of these enzymes have very different environments. Correia, et al (2014) characterized the kinetics and structure of DgAOR with several substrates and found that the Phe425 and Tyr535 residues at the active site likely stabilize aromatic aldehydes by pi stacking. This active site was also buried away from solvent. The active site of PaoABC lacked any significant aromatic residues and was positioned at the surface of the protein. The substrate stabilizing elements at this active site are Leu246 and Pro352. We are interested in why these active sites both are unreactive towards purines given their different chemical and location compared to the solvent. We propose that by mutating PaoABC to have smaller, nonpolar residues at the 246 and 352 position, we may be able to change the specificity of PaoABC to include purines. We also will mutate these residues to aromatic groups to probe at the chemical environment of the active site and its similarities to DgAOR.
Tbx2 Pigment Study by CRISPR/CAS9 Mutation
Bell, McKenzie; Porter, Tyrel; Naylor, Emily; Domyan, Eric (Utah Valley University)
Faculty Advisor: Domyan, Eric (Utah Valley University, Biology)
The domestic rock pigeon has been the subject of selective breeding for over a hundred years and so displays an immense variety of phenotypes. This variety provides opportunities to further understand the genetic basis of phenotypic evolution. Pigmentation of pigeon feathers is controlled by multiple alleles at different loci, which influences the type and amount of melanin deposited in the feathers. A specific phenotype, known as "recessive red", consists of distinctly red plumage and is caused by a mutation that greatly reduces the expression of the gene SOX10. This gene encodes a transcription factor, known to play a key role in melanocyte maturation and proliferation. SOX10 likely regulates the transcription of multiple downstream genes but the identities of these genes are largely unknown. To identify downstream targets of SOX10, we compared the transcriptomes of regenerating feathers from wild-type and recessive red birds to identify genes that had different expression levels between the two groups. We identified 46 genes that are expressed at different levels between wild-type and recessive red birds, and thus potential targets of SOX101. Of the 46 genes, Tbx2 was selected as a starter because it is one of the only transcription factors regulated by Sox10 in melanocytes. This mechanism makes it a plausible candidate given the critical role proteins play in phenotypic expression ("TBX2 T-box transcription factor 2—Gene—NCBI," n.d.).
Faculty Advisor: Domyan, Eric (Utah Valley University, Biology)
The domestic rock pigeon has been the subject of selective breeding for over a hundred years and so displays an immense variety of phenotypes. This variety provides opportunities to further understand the genetic basis of phenotypic evolution. Pigmentation of pigeon feathers is controlled by multiple alleles at different loci, which influences the type and amount of melanin deposited in the feathers. A specific phenotype, known as "recessive red", consists of distinctly red plumage and is caused by a mutation that greatly reduces the expression of the gene SOX10. This gene encodes a transcription factor, known to play a key role in melanocyte maturation and proliferation. SOX10 likely regulates the transcription of multiple downstream genes but the identities of these genes are largely unknown. To identify downstream targets of SOX10, we compared the transcriptomes of regenerating feathers from wild-type and recessive red birds to identify genes that had different expression levels between the two groups. We identified 46 genes that are expressed at different levels between wild-type and recessive red birds, and thus potential targets of SOX101. Of the 46 genes, Tbx2 was selected as a starter because it is one of the only transcription factors regulated by Sox10 in melanocytes. This mechanism makes it a plausible candidate given the critical role proteins play in phenotypic expression ("TBX2 T-box transcription factor 2—Gene—NCBI," n.d.).
The effect of Nr4a3 gene deletion on body weight, blood glucose levels, and glucose tolerance in mice
Yang, Haokun; Herring, Jacob; Elison, Weston; Wynn, Adam; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
Nearly 1 in 10 Americans have type 2 diabetes (T2D), a disease that is characterized by a loss of functional β-cell mass, resulting in decreased insulin secretion and glucose utilization. The pancreatic β-cell is responsible for producing and secreting insulin and monitoring blood glucose levels, and it is crucial to the understanding of T2D. The orphan nuclear receptor Nr4a3 (Nor1) has well-defined roles throughout the body, specifically with fuel utilization in the liver, muscle, and adipose tissues. Here we present data demonstrating that Nr4a3 KO mice have increased body weight, blood glucose levels (fasting and non-fasting), and impaired glucose tolerance when fed a standard diet. Respiration from adipose tissue is significantly impaired in male and female Nr4a3 KO animals. These data demonstrate that Nr4a3 is necessary for whole-body homeostasis. We believe that these data serve as a step toward understanding the pathway of T2D progression and finding a cure.
Faculty Advisor: Tessem, Jeffery (Brigham Young University; Nutrition, Dietetics, and Food Science)
Nearly 1 in 10 Americans have type 2 diabetes (T2D), a disease that is characterized by a loss of functional β-cell mass, resulting in decreased insulin secretion and glucose utilization. The pancreatic β-cell is responsible for producing and secreting insulin and monitoring blood glucose levels, and it is crucial to the understanding of T2D. The orphan nuclear receptor Nr4a3 (Nor1) has well-defined roles throughout the body, specifically with fuel utilization in the liver, muscle, and adipose tissues. Here we present data demonstrating that Nr4a3 KO mice have increased body weight, blood glucose levels (fasting and non-fasting), and impaired glucose tolerance when fed a standard diet. Respiration from adipose tissue is significantly impaired in male and female Nr4a3 KO animals. These data demonstrate that Nr4a3 is necessary for whole-body homeostasis. We believe that these data serve as a step toward understanding the pathway of T2D progression and finding a cure.
The Effects of High Glucose and Pterostilbene on the Oxidant Status of the Red Blood Cell
Garcia, Jessica; Hanks, Hana; Kist, Taylor; Suman, Tanner (Dixie State University)
Faculty Advisor: Meyer, Jennifer (Dixie State University, Physical Sciences)
Antioxidants in the human body regulate reactive oxygen species (ROS). If ROS are increased within the body it can potentially lead to oxidative stress and cell injury. Glucose-6-phosphate dehydrogenase (G6PD) is an enzyme found in the pentose phosphate pathway that assists in reducing the amount of ROS in the bloodstream. Increased concentrations of glucose, commonly found in patients that suffer from type 1 and type 2 diabetes mellitus have been shown to impair G6PD activity, thereby causing damage to erythrocytes (RBCs)2. It has been shown that hyperglycemic conditions decrease the activity of G6PD in RBCs, which is improved by the addition of pterostilbene, a potent antioxidant1. Glutathione, another antioxidant found in the pentose phosphate pathway, is decreased in RBCs within hyperglycemic conditions. The addition of pterostilbene is predicted to increase levels of glutathione under high glucose conditions.
References:
1. Richins, M., & Meyer, J. (2018). Pterostilbene Ameliorates Lipid Peroxidation and Increases Glucose-6-Phosphate Dehydrogenase Activity in Erythrocytes Subjected to High Glucose Conditions. American Heart Association Journals, 138.
2. Zang, Z., Apse, K., Pang, J., & Stanton, R. C. (2000). High glucose inhibits glucose-6-phosphate dehydrogenase via cAMP in aortic endothelial cells. The Journal of Biological Chemistry, 275(51), 40042-40047. Doi: 10.1074/jbc.M007505200
Faculty Advisor: Meyer, Jennifer (Dixie State University, Physical Sciences)
Antioxidants in the human body regulate reactive oxygen species (ROS). If ROS are increased within the body it can potentially lead to oxidative stress and cell injury. Glucose-6-phosphate dehydrogenase (G6PD) is an enzyme found in the pentose phosphate pathway that assists in reducing the amount of ROS in the bloodstream. Increased concentrations of glucose, commonly found in patients that suffer from type 1 and type 2 diabetes mellitus have been shown to impair G6PD activity, thereby causing damage to erythrocytes (RBCs)2. It has been shown that hyperglycemic conditions decrease the activity of G6PD in RBCs, which is improved by the addition of pterostilbene, a potent antioxidant1. Glutathione, another antioxidant found in the pentose phosphate pathway, is decreased in RBCs within hyperglycemic conditions. The addition of pterostilbene is predicted to increase levels of glutathione under high glucose conditions.
References:
1. Richins, M., & Meyer, J. (2018). Pterostilbene Ameliorates Lipid Peroxidation and Increases Glucose-6-Phosphate Dehydrogenase Activity in Erythrocytes Subjected to High Glucose Conditions. American Heart Association Journals, 138.
2. Zang, Z., Apse, K., Pang, J., & Stanton, R. C. (2000). High glucose inhibits glucose-6-phosphate dehydrogenase via cAMP in aortic endothelial cells. The Journal of Biological Chemistry, 275(51), 40042-40047. Doi: 10.1074/jbc.M007505200
The Synthesis and Characterization of Liposomes for Future Delivery of Gentamicin to Biofilms
Buehler, Nate; Hoehn, Nick; Stokes, Britt; Tyler, Areiann; Dr. Kopp, Olga (Utah Valley University)
Faculty Advisor: Kopp, Dr.Olga (Utah Valley University, Biology)
Bacterial infections are difficult to treat with antibiotics because of the protective nature of the biofilms produced by bacteria. Biofilms are a common cause of nosocomial and medical devices-related infections. The current treatments for biofilms include mechanically removing the biofilm itself or by treatments with antibiotics. Biofilms usually become resistant to drugs because of the higher frequency of mutation and horizontal gene transfer compared to planktonic cells. Liposomes are promising delivery systems because of their small size, surface characteristics and ability to encapsulate drugs and other molecules. Liposomal particles can slowly release the encapsulated drugs, increasing their distribution in targeted areas. Studies have shown that the fusion between liposomes and bacterial cells enhances the penetration of antibiotics. The purpose of this study is to form liposomes to encapsulate Gentamicin and characterize the formation and characteristics of these liposomes. Liposomes will be formed using the thin film hydration method and characterized using a scanning electron microscope. This project will present an analysis of the use of different ratios of phospholipids and cholesterol to evaluate the stability and ability to carry Gentamicin.
Faculty Advisor: Kopp, Dr.Olga (Utah Valley University, Biology)
Bacterial infections are difficult to treat with antibiotics because of the protective nature of the biofilms produced by bacteria. Biofilms are a common cause of nosocomial and medical devices-related infections. The current treatments for biofilms include mechanically removing the biofilm itself or by treatments with antibiotics. Biofilms usually become resistant to drugs because of the higher frequency of mutation and horizontal gene transfer compared to planktonic cells. Liposomes are promising delivery systems because of their small size, surface characteristics and ability to encapsulate drugs and other molecules. Liposomal particles can slowly release the encapsulated drugs, increasing their distribution in targeted areas. Studies have shown that the fusion between liposomes and bacterial cells enhances the penetration of antibiotics. The purpose of this study is to form liposomes to encapsulate Gentamicin and characterize the formation and characteristics of these liposomes. Liposomes will be formed using the thin film hydration method and characterized using a scanning electron microscope. This project will present an analysis of the use of different ratios of phospholipids and cholesterol to evaluate the stability and ability to carry Gentamicin.
Supplemental treatment options for diabetes: how DHE induces Nr4a1 expression and subsequent β-cell function
Brown, Nathan; Herring, Jacob; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham young University; Nutrition, Dietetics, and Food Science)
Diabetes is a global epidemic affecting millions of people. The total estimated cost of diabetes in the U.S. during 2017 was 327 billion dollars. Diabetes is characterized by the loss of pancreatic β-cell function which is caused by an autoimmune disorder in Type 1 diabetes or insulin resistance and β-cell exhaustion in Type 2 (T2D) diabetes.
It is shown that β-cell mitochondrial respiration is dependent on the nuclear receptor Nr4a1. Respiration rates of cells lacking Nr4a1 in the presence of 16 mM glucose resulted in a significant decrease in glucose-stimulated insulin secretion by impeding the production of ATP. It was also found that knockdown of Nr4a1 results in decreased expression of mitochondrial dehydrogenase subunits Idh3g and Sdhb. Thus, the orphan nuclear receptor Nr4a1 is critical for β-cell mitochondrial function and insulin secretion.
In subsequent studies it was shown that dihydroergotamine (DHE) induces Nr4a1 expression via recruitment of the super elongation complex to enable elongation of Nr4a1 promoter paused RNA polymerase II. While these experiments have been shown in cancer cells, I hypothesize that DHE will up-regulate Nr4a1 and other downstream targets. To test this I will use an in-vitro model to culture INS-1 832/3 rat insulinoma cell lines as a useful model for insulin secretion regulation and pancreatic islet beta-cell function studies. This study will shed further light on the regulation of the Nr4a1 nuclear receptor in pancreatic β-cells.
Faculty Advisor: Tessem, Jeffery (Brigham young University; Nutrition, Dietetics, and Food Science)
Diabetes is a global epidemic affecting millions of people. The total estimated cost of diabetes in the U.S. during 2017 was 327 billion dollars. Diabetes is characterized by the loss of pancreatic β-cell function which is caused by an autoimmune disorder in Type 1 diabetes or insulin resistance and β-cell exhaustion in Type 2 (T2D) diabetes.
It is shown that β-cell mitochondrial respiration is dependent on the nuclear receptor Nr4a1. Respiration rates of cells lacking Nr4a1 in the presence of 16 mM glucose resulted in a significant decrease in glucose-stimulated insulin secretion by impeding the production of ATP. It was also found that knockdown of Nr4a1 results in decreased expression of mitochondrial dehydrogenase subunits Idh3g and Sdhb. Thus, the orphan nuclear receptor Nr4a1 is critical for β-cell mitochondrial function and insulin secretion.
In subsequent studies it was shown that dihydroergotamine (DHE) induces Nr4a1 expression via recruitment of the super elongation complex to enable elongation of Nr4a1 promoter paused RNA polymerase II. While these experiments have been shown in cancer cells, I hypothesize that DHE will up-regulate Nr4a1 and other downstream targets. To test this I will use an in-vitro model to culture INS-1 832/3 rat insulinoma cell lines as a useful model for insulin secretion regulation and pancreatic islet beta-cell function studies. This study will shed further light on the regulation of the Nr4a1 nuclear receptor in pancreatic β-cells.
The Influence of Staphylococcus Aureus Biofilm-associated Gene Mutations on Biofilm Composition
Berges, Bradford; Wienclaw, Trevor; Ball, Ashley; Richmond, Bradley (Brigham Young University)
Faculty Advisor: Berges, Bradford (Life Sciences, Microbiology and Molecular Biology)
Staphylococcus Aureus (SA) biofilms are serious impediments to immune defenses and antibiotics, making them a major factor in SA infection. Such infections can be highly lethal even using current treatments, representing a major challenge to the healthcare industry. Previous genetic screenings of SA have revealed several genes that may be associated with biofilm formation. While the roles of many of these genes have been studied, little research has been done on how mutations of these genes impact biofilm composition. As several therapeutic options for treating mature SA biofilms require understanding of biofilm composition, a better understanding of how genes influence that composition is critical to improving current treatments and developing new ones.
In this project, we will study the biofilm phenotypes of SA with mutations in common biofilm-associated genes. By comparing the biofilm mass and composition of the wild-type (wt) Je2 strain to strains containing mutated biofilm-associated genes, we hope to uncover the impact that each mutation has on the composition of the biofilm matrix. We will utilize crystal violet assays as well as extracellular DNA and protein quantifying procedures to determine biofilm composition, after which meaningful comparisons can be made between mutant biofilms and wt biofilms.
Faculty Advisor: Berges, Bradford (Life Sciences, Microbiology and Molecular Biology)
Staphylococcus Aureus (SA) biofilms are serious impediments to immune defenses and antibiotics, making them a major factor in SA infection. Such infections can be highly lethal even using current treatments, representing a major challenge to the healthcare industry. Previous genetic screenings of SA have revealed several genes that may be associated with biofilm formation. While the roles of many of these genes have been studied, little research has been done on how mutations of these genes impact biofilm composition. As several therapeutic options for treating mature SA biofilms require understanding of biofilm composition, a better understanding of how genes influence that composition is critical to improving current treatments and developing new ones.
In this project, we will study the biofilm phenotypes of SA with mutations in common biofilm-associated genes. By comparing the biofilm mass and composition of the wild-type (wt) Je2 strain to strains containing mutated biofilm-associated genes, we hope to uncover the impact that each mutation has on the composition of the biofilm matrix. We will utilize crystal violet assays as well as extracellular DNA and protein quantifying procedures to determine biofilm composition, after which meaningful comparisons can be made between mutant biofilms and wt biofilms.
Census and Distribution Analysis of Alouatta palliata (mantled howler monkey) in La Selva Biological Station
Smith, Mick; Desdames, Chloe (University of Utah)
Faculty Advisor: Seaboch, Melissa (University of Utah, Anthropology)
Census and sampling work are important because they provide critical information on population size, distribution, and habitat preference — all important factors in conservation. La Selva Biological station is located on the north eastern side of Costa Rica and is a protected biological reserve. It is home to three species of primates including Alouatta palliata, commonly known as the mantled howler monkey. It is comprised of primary forest that has been undisturbed by human forces and secondary forest that is regenerating from past disturbances, such as deforestation. Past research conducted on howler monkeys shows they prefer primary forest over secondary because primary forests have higher species richness and a wider range of trees which provides food for howler monkey's selective diet. I predict that the majority of A. palliata in Costa Rica will be located in primary forest because it offers more resources. I censused 11 established trails at La Selva (two trails in primary forest, seven trails in secondary forest, and two trails crossing both forest types). For each howler monkey encountered, I recorded the location, forest type, and group size. I identified four different groups of A. palliata with group size ranging between 6-10 individuals. One group was located in primary forest and three groups in secondary forest; thus, my hypothesis that A. palliata would prefer primary forests was not supported. Anecdotally, A. palliata were observed more frequently around rivers or streams. This could indicate that rather than being concerned with primary versus secondary forest, they prefer riparian habitats found along the banks of rivers or other actively moving sources of water. A study conducted by Stoner found howler monkeys in riparian habits supporting this hypothesis. This type of research continues to provide critical information for understanding primate's habitats which helps with conservation of species.
Faculty Advisor: Seaboch, Melissa (University of Utah, Anthropology)
Census and sampling work are important because they provide critical information on population size, distribution, and habitat preference — all important factors in conservation. La Selva Biological station is located on the north eastern side of Costa Rica and is a protected biological reserve. It is home to three species of primates including Alouatta palliata, commonly known as the mantled howler monkey. It is comprised of primary forest that has been undisturbed by human forces and secondary forest that is regenerating from past disturbances, such as deforestation. Past research conducted on howler monkeys shows they prefer primary forest over secondary because primary forests have higher species richness and a wider range of trees which provides food for howler monkey's selective diet. I predict that the majority of A. palliata in Costa Rica will be located in primary forest because it offers more resources. I censused 11 established trails at La Selva (two trails in primary forest, seven trails in secondary forest, and two trails crossing both forest types). For each howler monkey encountered, I recorded the location, forest type, and group size. I identified four different groups of A. palliata with group size ranging between 6-10 individuals. One group was located in primary forest and three groups in secondary forest; thus, my hypothesis that A. palliata would prefer primary forests was not supported. Anecdotally, A. palliata were observed more frequently around rivers or streams. This could indicate that rather than being concerned with primary versus secondary forest, they prefer riparian habitats found along the banks of rivers or other actively moving sources of water. A study conducted by Stoner found howler monkeys in riparian habits supporting this hypothesis. This type of research continues to provide critical information for understanding primate's habitats which helps with conservation of species.
Cell Adhesion and Morphology in Relation to Neural Tube Defects
Lin, Jade; Park, Yeram; Ross, Micah; Stark, Michael; Hansen, Marc (Brigham Young University)
Faculty Advisor: Stark, Michael (Brigham Young University, Physiology and Developmental Biology); Hansen, Marc (Brigham Young University, Physiology and Developmental Biology)
Neural Tube Defects (NTDs) such as spina bifida and anencephaly are due to incomplete closure of neural tubes in developing embryos. While the etiology is still unknown, environmental and genetic factors, toxicants, and maternal health are implicated as potential causes. Previous research shows that NTDs are associated with increased levels of ceramide (C2) and possible exposure to fumonisin (FB1), and valproic acid (VPA). We hypothesize that these molecules may interfere with cell-to-cell interactions which are important for neural tube formation. To investigate the potential mechanisms by which these toxicants can induce NTDs, I will use a well-characterized MDCK cell model treated with C2, FB1, and VPA to assess their impact on cell adhesion.
Faculty Advisor: Stark, Michael (Brigham Young University, Physiology and Developmental Biology); Hansen, Marc (Brigham Young University, Physiology and Developmental Biology)
Neural Tube Defects (NTDs) such as spina bifida and anencephaly are due to incomplete closure of neural tubes in developing embryos. While the etiology is still unknown, environmental and genetic factors, toxicants, and maternal health are implicated as potential causes. Previous research shows that NTDs are associated with increased levels of ceramide (C2) and possible exposure to fumonisin (FB1), and valproic acid (VPA). We hypothesize that these molecules may interfere with cell-to-cell interactions which are important for neural tube formation. To investigate the potential mechanisms by which these toxicants can induce NTDs, I will use a well-characterized MDCK cell model treated with C2, FB1, and VPA to assess their impact on cell adhesion.
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.
Anti-Tumor Activity of Chalcone Derivatives
Allen, Brian; Covey, Tracy; Davies, Don; Eccles, Nick; Farnsworth, Brian; Ferguson, Parker; Hart, Sierra; Lowder, Jordan (Weber State University)
Faculty Advisor: Davies, Don (Weber State University, Chemistry and Biochemistry); Covey, Tracy (Weber State University, Chemistry and Biochemsitry)
Chalcones refer to biological molecules with the structure trans 1,3-diphenylprop-2-en-1-one. Biological chalcones and chalcone derivatives display anti-tumor, anti-fungal, anti-inflammatory and antibiotic properties. To understand the role of the chalcone structure in tumor cessation, derivatives to the original chalcone were synthesized using aldol condensation reactions. HeLa and HEK-293 cells were treated with the synthesized chalcone and an LD50, or the concentration of chalcone required to kill half of the cells, was calculated. The LD50 was then used to determine the efficiency of the chalcone derivative. Correlations between the structure and activity suggest that a Michael reaction occurs at the cell and indicate that that an aromatic ring at C3 is likely necessary. Further research will help determine the structures of more cytotoxic compounds.
Faculty Advisor: Davies, Don (Weber State University, Chemistry and Biochemistry); Covey, Tracy (Weber State University, Chemistry and Biochemsitry)
Chalcones refer to biological molecules with the structure trans 1,3-diphenylprop-2-en-1-one. Biological chalcones and chalcone derivatives display anti-tumor, anti-fungal, anti-inflammatory and antibiotic properties. To understand the role of the chalcone structure in tumor cessation, derivatives to the original chalcone were synthesized using aldol condensation reactions. HeLa and HEK-293 cells were treated with the synthesized chalcone and an LD50, or the concentration of chalcone required to kill half of the cells, was calculated. The LD50 was then used to determine the efficiency of the chalcone derivative. Correlations between the structure and activity suggest that a Michael reaction occurs at the cell and indicate that that an aromatic ring at C3 is likely necessary. Further research will help determine the structures of more cytotoxic compounds.
Antimicrobial Activity of Artemisia tridentata
Wasden, Kayla; Suisse, David; Kaundal, Amita (faculty mentor) (Utah State University)
Faculty Advisor: Kaundal, Amita (College of Agriculture and Applied Sciences; Plants, Soils, and Climate Department)
Many plants secrete substances to create a more favorable environment, including chemicals that kill pathogenic microbes or competing plants. Artemisia tridentata, also known as "Big Sagebrush," is prevalent in the Rocky Mountain region of the United States and is known to have antimicrobial capabilities. We will study the potential antimicrobial activity of Artemisia tridentata.
Studies report that chemicals released by the leaves and branches of A. tridentata affect bacteria native to deer rumen. Another study showed that 27 actinomycetes (anaerobic bacteria that form colonies) strains found in the rhizosphere of A. tridentata demonstrated antibacterial activities when tested on E. coli, Bacillus subtilis and Staphylococcus aureus. Native Americans traditionally used A. tridentata to relieve stomach pain, colds, coughs, sore eyes, snake bites and as an insect repellent. Researchers found several compounds, including flavonoids, that can affect antimicrobial activity. Articles regarding antimicrobial activities in A. tridentata were published between 1967 and 2004. With the chronological gaps and considering the progress that biological and molecular technology has made in recent years, knowledge of the chemicals released by A. tridentata lies largely untapped. In this study, we will investigate the antimicrobial activities of the leaves, stem, roots, and flowers of A. tridentata initially by the agar well diffusion method and followed by validating with the agar disk diffusion method. We will check the antimicrobial activity of the extract from different plant parts of A. tridentata on common bacteria such as E. coli, Bacillus subtilis, and some Pseudomonas spp. of plant pathogens.
The knowledge obtained from this research will further help in the identification and characterization of the secondary metabolites or chemicals involved in antimicrobial activity of sagebrush. Medicinal plants provide a healthy, natural alternative to conventional medication, and may lead to new insights on antibiotics and pharmaceuticals. Besides, Artemisia tridentata is a plant native to Utah and Idaho. It grows everywhere in the surrounding area, making it inexpensive (free) to produce.
Faculty Advisor: Kaundal, Amita (College of Agriculture and Applied Sciences; Plants, Soils, and Climate Department)
Many plants secrete substances to create a more favorable environment, including chemicals that kill pathogenic microbes or competing plants. Artemisia tridentata, also known as "Big Sagebrush," is prevalent in the Rocky Mountain region of the United States and is known to have antimicrobial capabilities. We will study the potential antimicrobial activity of Artemisia tridentata.
Studies report that chemicals released by the leaves and branches of A. tridentata affect bacteria native to deer rumen. Another study showed that 27 actinomycetes (anaerobic bacteria that form colonies) strains found in the rhizosphere of A. tridentata demonstrated antibacterial activities when tested on E. coli, Bacillus subtilis and Staphylococcus aureus. Native Americans traditionally used A. tridentata to relieve stomach pain, colds, coughs, sore eyes, snake bites and as an insect repellent. Researchers found several compounds, including flavonoids, that can affect antimicrobial activity. Articles regarding antimicrobial activities in A. tridentata were published between 1967 and 2004. With the chronological gaps and considering the progress that biological and molecular technology has made in recent years, knowledge of the chemicals released by A. tridentata lies largely untapped. In this study, we will investigate the antimicrobial activities of the leaves, stem, roots, and flowers of A. tridentata initially by the agar well diffusion method and followed by validating with the agar disk diffusion method. We will check the antimicrobial activity of the extract from different plant parts of A. tridentata on common bacteria such as E. coli, Bacillus subtilis, and some Pseudomonas spp. of plant pathogens.
The knowledge obtained from this research will further help in the identification and characterization of the secondary metabolites or chemicals involved in antimicrobial activity of sagebrush. Medicinal plants provide a healthy, natural alternative to conventional medication, and may lead to new insights on antibiotics and pharmaceuticals. Besides, Artemisia tridentata is a plant native to Utah and Idaho. It grows everywhere in the surrounding area, making it inexpensive (free) to produce.
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.
Analysis of binding affinity and molecular cloning of two PRMT1 substrates
Jones, Abigail; Hevel, Joan (Utah State University)
Faculty Advisor: Hevel, Joan (College of Science, Chemistry and Biochemistry Department)
PRMT1 is one of nine known mammalian Protein Arginine Methyltransferases (PRMTs) whose function are to transfer methyl groups from S-adenosyl methionine (SAM) to arginine residues of specific proteins. PRMT1 is known to methylate many different proteins in cells, but the mechanism of target recognition and binding is still unknown. Correct regulation of PRMT1 is critical to proper cellular function; thus, the action of PRMT1 is important to understand. In this study, we seek to elucidate how PRMT1 recognizes and binds its targets by identifying protein substrates of PRMT1 that form a stable complex with the enzyme. Such a protein would allow for additional studies (e.g. crystallographic or cryo-EM studies) to help visualize PRMT1-substrate interactions. Two substrates of PRMT1, TWIST1 and Smad6, have been purified, and the binding affinity of each to PRMT1 has been qualitatively assessed via pull-down assay and Western blot. Ligation-independent-cloning has been used to clone each substrate gene out of a GST-tagged vector and into a His-tagged vector, which will allow for further experiments assessing the stoichiometry of PRMT1-substrate binding.
Faculty Advisor: Hevel, Joan (College of Science, Chemistry and Biochemistry Department)
PRMT1 is one of nine known mammalian Protein Arginine Methyltransferases (PRMTs) whose function are to transfer methyl groups from S-adenosyl methionine (SAM) to arginine residues of specific proteins. PRMT1 is known to methylate many different proteins in cells, but the mechanism of target recognition and binding is still unknown. Correct regulation of PRMT1 is critical to proper cellular function; thus, the action of PRMT1 is important to understand. In this study, we seek to elucidate how PRMT1 recognizes and binds its targets by identifying protein substrates of PRMT1 that form a stable complex with the enzyme. Such a protein would allow for additional studies (e.g. crystallographic or cryo-EM studies) to help visualize PRMT1-substrate interactions. Two substrates of PRMT1, TWIST1 and Smad6, have been purified, and the binding affinity of each to PRMT1 has been qualitatively assessed via pull-down assay and Western blot. Ligation-independent-cloning has been used to clone each substrate gene out of a GST-tagged vector and into a His-tagged vector, which will allow for further experiments assessing the stoichiometry of PRMT1-substrate binding.
Antibiotic Resistance in Staphylococcus aureus: Effects of Biofilm Synthesis in Gene Transfer
Hirschi, Blake; Pickett, Brad; Thompson, Jared; Telford, Mady; Berges, Bradford (Brigham Young University)
Faculty Advisor: Berges, Brad (Life Sciences, Microbiology and Molecular Biology)
Staphylococcus aureus (S. aureus) is a commensal bacterium commonly found amongst livestock and near 30% of humans' nostrils. However, through acquisition of certain genes S. aureus may develop antibiotic resistance such as in methicillin-resistant Staphylococcus aureus (MRSA). One hypothesized component lending to acquisition of genetic resistance in S. aureus is the synthesis of colony biofilms. Biofilms are comprised of a variety of substances including secreted polysaccharides, protein and even extracellular DNA. Our work postulates that extracellular DNA-based biofilms will transfer genes for antibiotic resistance at a higher rate than in polysaccharide/protein biofilms. Through employment of polymerase chain reaction (PCR), we aim to characterize a wide sample of methicillin-susceptible S. aureus (MSSA) human associated strains and MRSA livestock associated strains for multiple antibiotic resistances. Co-inoculating pairs of human associated and livestock associated strains, each lacking the other's resistance genes, will provide an environment wherein biofilm-mediated gene transfer may occur. Further pairing based on biofilm composition (DNA or polysaccharide/protein) will yield data concerning which biofilm facilitates gene transfer more efficiently. Subsequent genotyping will confirm whether resulting isolates acquired new antibiotic resistance through biofilm-mediated transfer, thus increasing pathogenicity.
Faculty Advisor: Berges, Brad (Life Sciences, Microbiology and Molecular Biology)
Staphylococcus aureus (S. aureus) is a commensal bacterium commonly found amongst livestock and near 30% of humans' nostrils. However, through acquisition of certain genes S. aureus may develop antibiotic resistance such as in methicillin-resistant Staphylococcus aureus (MRSA). One hypothesized component lending to acquisition of genetic resistance in S. aureus is the synthesis of colony biofilms. Biofilms are comprised of a variety of substances including secreted polysaccharides, protein and even extracellular DNA. Our work postulates that extracellular DNA-based biofilms will transfer genes for antibiotic resistance at a higher rate than in polysaccharide/protein biofilms. Through employment of polymerase chain reaction (PCR), we aim to characterize a wide sample of methicillin-susceptible S. aureus (MSSA) human associated strains and MRSA livestock associated strains for multiple antibiotic resistances. Co-inoculating pairs of human associated and livestock associated strains, each lacking the other's resistance genes, will provide an environment wherein biofilm-mediated gene transfer may occur. Further pairing based on biofilm composition (DNA or polysaccharide/protein) will yield data concerning which biofilm facilitates gene transfer more efficiently. Subsequent genotyping will confirm whether resulting isolates acquired new antibiotic resistance through biofilm-mediated transfer, thus increasing pathogenicity.
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.
Analysis of the Gut Microbiome of Drosophila melanogaster Models of Parkinson's Disease
Marshman, Evan; Peterson, Samara; Call, Gerald; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Science, Plant and Wildlife Science)
In recent years the association between the human gut microbiome and the brain has become a promising field of study. Often referred to as the "gut-brain axis", this connection has greatly enriched our scientific understanding of many disorders that affect the brain and nervous system. A recent study showed the differing richness of bacteria in the microbiota of Parkinson's patients and healthy control subjects. Because recent research shows this connection, we predicted that we would detect variation in the microbiota of D. melanogaster (fruit flies) models of Parkinson's disease, relative to wild type flies. To test this hypothesis, I analyzed 16s rRNA sequence data, reporting the microbiota composition in flies that are a model of Parkinson's Disease, as well as wild type flies. I found one strain of the genus Acetobacter that was differentially abundant between the two fly types. Therefore, for my CURA I will extend my analysis by performing similar analyses by taking a larger set of Parkinson's fly models. Once they are sequenced, I will use QIIME, the same software I used in my preliminary analyses to further our understanding of the taxonomic differences between the gut bacteria of Parkinson's models and wild type flies.
Faculty Advisor: Chaston, John (Life Science, Plant and Wildlife Science)
In recent years the association between the human gut microbiome and the brain has become a promising field of study. Often referred to as the "gut-brain axis", this connection has greatly enriched our scientific understanding of many disorders that affect the brain and nervous system. A recent study showed the differing richness of bacteria in the microbiota of Parkinson's patients and healthy control subjects. Because recent research shows this connection, we predicted that we would detect variation in the microbiota of D. melanogaster (fruit flies) models of Parkinson's disease, relative to wild type flies. To test this hypothesis, I analyzed 16s rRNA sequence data, reporting the microbiota composition in flies that are a model of Parkinson's Disease, as well as wild type flies. I found one strain of the genus Acetobacter that was differentially abundant between the two fly types. Therefore, for my CURA I will extend my analysis by performing similar analyses by taking a larger set of Parkinson's fly models. Once they are sequenced, I will use QIIME, the same software I used in my preliminary analyses to further our understanding of the taxonomic differences between the gut bacteria of Parkinson's models and wild type flies.
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.