Health and Medicine
Aging-associated accumulation of myocardial protein aggregates in mice is attenuated by late-in-life exercise training.
Symons, J. David; Thompson, Lauren; Ramous, Caroline; Cho, JaeMin; Boudina, Sihem; Margetts, Alex; Buzianis, Skye; Park, Seul Ki; Luu, Kiana; Hansen, Michele; Pires, Karla; Whitehead, Kevin; Carter, Kandis; Buffolo, Marcio (University of Utah)
Faculty Advisor: Symones, J. David (University of Utah, Nutrition and Integrative Physiology)
Protein aggregates accumulate and organelles become damaged and / or dysfunctional during the process of healthy aging. A progressive loss of the cellular quality control mechanism autophagy (i.e., "self-eating") contributes to this age-associated decline in cellular function in many organs. Evidence for an age-associated repression in cardiac autophagy is not consistent. We hypothesized that 24-month old (old) male C57Bl6/J mice exhibit repressed autophagosome formation in the heart, an accumulation of cardiac protein aggregates, myocardial dysfunction, and reduced exercise capacity vs. 6-month old (adult) mice. First, cardiac lysates from old mice displayed reduced (p<0.05) accumulation of LC3II / GAPDH and degradation of p62 vs. adult animals (assessed via immunoblotting; n=12 per group). Second, the lysosomal acidification inhibitor chloroquine (CQ) induced accrual (p<0.05) of LC3II / GAPDH and p62 in hearts from adult but not old mice (quantified by immunoblotting; n=7 per group). Third, the number and size of protein aggregates was higher (p<0.05) in hearts from old vs. adult mice (measured via scanning electron microscopy; n=5 per group). Fourth, left ventricular mass / tibial length was greater (p<0.05), and indices of systolic, diastolic, and global left ventricular function (measured via transthoracic echocardiography) were impaired (p<0.05), in old vs. adult animals (n=12 per group). Finally, maximal workload performed during a treadmill-test, and soleus muscle oxidative enzyme capacity (citrate synthase activity assessed via ELISA), were less (p<0.05) in aged (n=11) vs. adult (n=12) mice. To determine whether late-in-life exercise training improves cardiac autophagy to an extent that demonstrates functional relevance, separate cohorts of older male mice completed a progressive-resistance treadmill-running program (old-ETR) or remained sedentary (old-SED) from 21-24 months. Body composition (estimated via nuclear magnetic resonance), exercise performance during a maximal workload test, soleus muscle citrate synthase activity, indices of cardiac antioxidant enzyme activity (quantified via immunoblotting), markers of cardiac autophagy, accumulation of cardiac protein aggregates, and indices of myocardial function, all improved (p<0.05) in old-ETR (n=11) vs. old-SED (n=12) mice. These data are the first to demonstrate that markers of cardiac autophagy are elevated, and indicators of protein aggregate removal and myocardial function are improved, in older mice that complete a treadmill-training regimen that is sufficient to increase skeletal muscle CS activity and maximal exercise capacity. Our results provide strong proof of concept to evaluate cause and effect relationships among exercise-training, myocardial autophagy, and cardiac function using genetic approaches in pre-clinical models and these studies are ongoing in our laboratory.
Faculty Advisor: Symones, J. David (University of Utah, Nutrition and Integrative Physiology)
Protein aggregates accumulate and organelles become damaged and / or dysfunctional during the process of healthy aging. A progressive loss of the cellular quality control mechanism autophagy (i.e., "self-eating") contributes to this age-associated decline in cellular function in many organs. Evidence for an age-associated repression in cardiac autophagy is not consistent. We hypothesized that 24-month old (old) male C57Bl6/J mice exhibit repressed autophagosome formation in the heart, an accumulation of cardiac protein aggregates, myocardial dysfunction, and reduced exercise capacity vs. 6-month old (adult) mice. First, cardiac lysates from old mice displayed reduced (p<0.05) accumulation of LC3II / GAPDH and degradation of p62 vs. adult animals (assessed via immunoblotting; n=12 per group). Second, the lysosomal acidification inhibitor chloroquine (CQ) induced accrual (p<0.05) of LC3II / GAPDH and p62 in hearts from adult but not old mice (quantified by immunoblotting; n=7 per group). Third, the number and size of protein aggregates was higher (p<0.05) in hearts from old vs. adult mice (measured via scanning electron microscopy; n=5 per group). Fourth, left ventricular mass / tibial length was greater (p<0.05), and indices of systolic, diastolic, and global left ventricular function (measured via transthoracic echocardiography) were impaired (p<0.05), in old vs. adult animals (n=12 per group). Finally, maximal workload performed during a treadmill-test, and soleus muscle oxidative enzyme capacity (citrate synthase activity assessed via ELISA), were less (p<0.05) in aged (n=11) vs. adult (n=12) mice. To determine whether late-in-life exercise training improves cardiac autophagy to an extent that demonstrates functional relevance, separate cohorts of older male mice completed a progressive-resistance treadmill-running program (old-ETR) or remained sedentary (old-SED) from 21-24 months. Body composition (estimated via nuclear magnetic resonance), exercise performance during a maximal workload test, soleus muscle citrate synthase activity, indices of cardiac antioxidant enzyme activity (quantified via immunoblotting), markers of cardiac autophagy, accumulation of cardiac protein aggregates, and indices of myocardial function, all improved (p<0.05) in old-ETR (n=11) vs. old-SED (n=12) mice. These data are the first to demonstrate that markers of cardiac autophagy are elevated, and indicators of protein aggregate removal and myocardial function are improved, in older mice that complete a treadmill-training regimen that is sufficient to increase skeletal muscle CS activity and maximal exercise capacity. Our results provide strong proof of concept to evaluate cause and effect relationships among exercise-training, myocardial autophagy, and cardiac function using genetic approaches in pre-clinical models and these studies are ongoing in our laboratory.
An Exploration of Nurse Perceptions of Workplace Violence and Factors that Make Them Feel Safer
Gardner, Emily; Floyd, Eden; Taylor, Shaylee (Utah Valley University)
Faculty Advisor: Jensen, Francine (Utah Valley University, Nursing)
Problem Statement: Nurses experience a high incidence of workplace violence.
Purpose: The purpose of this qualitative research study aims to examine nurses' perceptions of workplace violence, including nurse-to-nurse, patient-to-nurse, and coworker-to-nurse violence, and explore nurses' perceptions of safety measures in their work environment.
Research Question: What are nurses' perceptions and experiences of workplace violence and what factors make them feel safer?
Background: Safety in healthcare is a concern that needs active improvement; "67% of all nonfatal workplace violence injuries occur in healthcare, but healthcare represents only 11.5% of the U.S. workforce" (Locke, Bromley, & Derspiel, 2018). The Joint Commission (2018), an accrediting body for many healthcare organizations, reported that nurses are among the most victimized by violence in healthcare. Certain locations such as the emergency department, psychiatric units, geriatric units, and intensive care units may experience a greater number of violent incidents (Camerino, Estryn-Behar, Conway, van der Heijden, & Hasselhorn, 2008). One study showed that 64% of the nurse respondents felt like violence, both physical or verbal, was an expected part of their jobs (Copeland & Henry, 2017). In addition, it was demonstrated that many nurses accept a level of violence on the job and do not see the importance of reporting incidents and looking for solutions (Copeland & Henry, 2017).
Hope to accomplish: The course of this research will explore how nurses perceive their workplace safety, shed light on experiences of workplace violence they have witnessed, and their perceptions of on-the-job safety. This research will also examine what makes newer and older nurses feel a greater sense of security while performing their jobs. Since some of the factors that make nurses feel safer may involve unit or hospital policies, these findings may offer ideas for potential changes to policies to help improve workplace safety in healthcare.
Faculty Advisor: Jensen, Francine (Utah Valley University, Nursing)
Problem Statement: Nurses experience a high incidence of workplace violence.
Purpose: The purpose of this qualitative research study aims to examine nurses' perceptions of workplace violence, including nurse-to-nurse, patient-to-nurse, and coworker-to-nurse violence, and explore nurses' perceptions of safety measures in their work environment.
Research Question: What are nurses' perceptions and experiences of workplace violence and what factors make them feel safer?
Background: Safety in healthcare is a concern that needs active improvement; "67% of all nonfatal workplace violence injuries occur in healthcare, but healthcare represents only 11.5% of the U.S. workforce" (Locke, Bromley, & Derspiel, 2018). The Joint Commission (2018), an accrediting body for many healthcare organizations, reported that nurses are among the most victimized by violence in healthcare. Certain locations such as the emergency department, psychiatric units, geriatric units, and intensive care units may experience a greater number of violent incidents (Camerino, Estryn-Behar, Conway, van der Heijden, & Hasselhorn, 2008). One study showed that 64% of the nurse respondents felt like violence, both physical or verbal, was an expected part of their jobs (Copeland & Henry, 2017). In addition, it was demonstrated that many nurses accept a level of violence on the job and do not see the importance of reporting incidents and looking for solutions (Copeland & Henry, 2017).
Hope to accomplish: The course of this research will explore how nurses perceive their workplace safety, shed light on experiences of workplace violence they have witnessed, and their perceptions of on-the-job safety. This research will also examine what makes newer and older nurses feel a greater sense of security while performing their jobs. Since some of the factors that make nurses feel safer may involve unit or hospital policies, these findings may offer ideas for potential changes to policies to help improve workplace safety in healthcare.
A Chronic Inflammatory Model In Human Hepatocytes And Curcumin's Ability to Attenuate Its Effects
Chandler L Eyre. David A. Eastley (Brigham Young University)
Faculty Advisor: Hancock, Chad (Brigham Young University, NDFS)
Purpose: To determine the effects of H2O2 exposure to human hepatocytes on the iron regulatory proteins such as transferrin receptor (TfR), Ferritin Light-Chain (FLC). Methods: Cells were cultured at an initial density of 35,000 cells/cm2 in 6-well plates, grown until confluent, and then harvested. Iron treatments were done using FeCl3 solution with media to desired concentrations, and then incubating for 24 hours. Groups that received both iron and H2O2 treatments were exposed to 12 hours of iron, followed by incubation with media that included both iron and GOX at 4.8 mU/mL and 0.165 mg/mL of CAT (2-5 KU/mg). BCA protein assay was used to assess protein content, and then normalize each sample to each other for subsequent preparation for western blot. Western blots were carried out were analyzed by fluorescence following densitometry. MTT assays were carried out utilizing the mitochondrial reductase enzyme, and colorimetrically measured to assess cell viability. Results: Iron treatments of 10, 50, and 100 µM for 24-hrs did not result in any significant cell death. Treatment with the same concentrations resulted in a significant decrease (n=12) in TfR for all three groups when compared against control cells that were cultured and harvested simultaneously without any addition the growth media. (10 µM p<0.01), (50 µM p<0.01), and (100 µM p<0.01). FTL in the 10 µM group was significantly decreased (n=11, p<0.01), but not for the 50 µM, or 100 µM groups with p values of 0.9867, and 0.9612, respectively. H2O2 treatments produced concentrations of 5-10 µM, mimicking neutrophil release during inflammatory response. Conclusion: Our assay is sufficient to mimic neutrophil release of H2O2. Iron treatments are able to induce TfR decrease but seem to have a threshold at 10 µM for increasing FTL for storage. We hypothesize that this may be due to a dysregulation at higher concentrations.
Faculty Advisor: Hancock, Chad (Brigham Young University, NDFS)
Purpose: To determine the effects of H2O2 exposure to human hepatocytes on the iron regulatory proteins such as transferrin receptor (TfR), Ferritin Light-Chain (FLC). Methods: Cells were cultured at an initial density of 35,000 cells/cm2 in 6-well plates, grown until confluent, and then harvested. Iron treatments were done using FeCl3 solution with media to desired concentrations, and then incubating for 24 hours. Groups that received both iron and H2O2 treatments were exposed to 12 hours of iron, followed by incubation with media that included both iron and GOX at 4.8 mU/mL and 0.165 mg/mL of CAT (2-5 KU/mg). BCA protein assay was used to assess protein content, and then normalize each sample to each other for subsequent preparation for western blot. Western blots were carried out were analyzed by fluorescence following densitometry. MTT assays were carried out utilizing the mitochondrial reductase enzyme, and colorimetrically measured to assess cell viability. Results: Iron treatments of 10, 50, and 100 µM for 24-hrs did not result in any significant cell death. Treatment with the same concentrations resulted in a significant decrease (n=12) in TfR for all three groups when compared against control cells that were cultured and harvested simultaneously without any addition the growth media. (10 µM p<0.01), (50 µM p<0.01), and (100 µM p<0.01). FTL in the 10 µM group was significantly decreased (n=11, p<0.01), but not for the 50 µM, or 100 µM groups with p values of 0.9867, and 0.9612, respectively. H2O2 treatments produced concentrations of 5-10 µM, mimicking neutrophil release during inflammatory response. Conclusion: Our assay is sufficient to mimic neutrophil release of H2O2. Iron treatments are able to induce TfR decrease but seem to have a threshold at 10 µM for increasing FTL for storage. We hypothesize that this may be due to a dysregulation at higher concentrations.
Baseline effects of lysophosphatidylcholine and nerve growth factor in a rat model of sciatic nerve regeneration after crush injury
Boatright, Greggory; Medrano, Braxton; Goeckeritz, Joel (Brigham Young University)
Faculty Advisor: Roeder, Beverly (Brigham Young University, Life Sciences)
Schwann cells play a major role in helping heal injured nerves. They help clear debris, produce neurotrophins, upregulate neurotrophin receptors, and form bands of Büngner to guide the healing nerve. But nerves do not always produce enough neurotrophins and neurotrophin receptors to repair themselves. Nerve growth factor (NGF) is an important neurotrophin for promoting nerve healing and lysophosphatidylcholine (LPC) has been shown to stimulate NGF receptors (NGFR). This study tested the administration of a single intraneural injection of LPC (1 mg/mL for single LPC injection and 10 mg/mL for multiple LPC injections) at day 0 and one (day 7), two (days 5 and 7), or three (days 5, 7, and 9) injections of NGF (160 ng/mL for single injections and 80 ng/mL for multiple injections) to determine baseline effects on crushed sciatic nerves in rats. The rats were randomly divided into four groups: control, crush, crush-NGF, and crush-LPC-NGF. The healing of the nerves was measured weekly by monitoring gait; electrophysiological parameters: compound muscle action potential (CMAP) amplitudes; and morphological parameters: total fascicle areas, myelinated fiber counts, fiber densities, fiber packing, and mean g-ratio values at weeks 3 and 6. The crush, crush-NGF, and crush-LPC-NGF groups statistically differed from the control group for all six weeks for the electrophysiological parameters but only differed from the control group at week 3 for the morphological parameters. The crush, crush-NGF, and crush LPC-NGF groups did not differ from each other over the course of the study. Single injections of LPC and NGF one week apart or multiple treatments of NGF at 5, 7 and 9 days post-injury did not alter the healing rate of the sciatic nerves during weeks 1-6 of the study. These findings are important to define the baseline effects of NGF and LPC injections, as part of a larger effort to determine the minimal dose regimen of NGF to regenerate peripheral nerves.
Faculty Advisor: Roeder, Beverly (Brigham Young University, Life Sciences)
Schwann cells play a major role in helping heal injured nerves. They help clear debris, produce neurotrophins, upregulate neurotrophin receptors, and form bands of Büngner to guide the healing nerve. But nerves do not always produce enough neurotrophins and neurotrophin receptors to repair themselves. Nerve growth factor (NGF) is an important neurotrophin for promoting nerve healing and lysophosphatidylcholine (LPC) has been shown to stimulate NGF receptors (NGFR). This study tested the administration of a single intraneural injection of LPC (1 mg/mL for single LPC injection and 10 mg/mL for multiple LPC injections) at day 0 and one (day 7), two (days 5 and 7), or three (days 5, 7, and 9) injections of NGF (160 ng/mL for single injections and 80 ng/mL for multiple injections) to determine baseline effects on crushed sciatic nerves in rats. The rats were randomly divided into four groups: control, crush, crush-NGF, and crush-LPC-NGF. The healing of the nerves was measured weekly by monitoring gait; electrophysiological parameters: compound muscle action potential (CMAP) amplitudes; and morphological parameters: total fascicle areas, myelinated fiber counts, fiber densities, fiber packing, and mean g-ratio values at weeks 3 and 6. The crush, crush-NGF, and crush-LPC-NGF groups statistically differed from the control group for all six weeks for the electrophysiological parameters but only differed from the control group at week 3 for the morphological parameters. The crush, crush-NGF, and crush LPC-NGF groups did not differ from each other over the course of the study. Single injections of LPC and NGF one week apart or multiple treatments of NGF at 5, 7 and 9 days post-injury did not alter the healing rate of the sciatic nerves during weeks 1-6 of the study. These findings are important to define the baseline effects of NGF and LPC injections, as part of a larger effort to determine the minimal dose regimen of NGF to regenerate peripheral nerves.
Correlation between Chronic Pain Symptoms and Brain Structure
McQuinn, Sophie; Love, Tiffany (University of Utah)
Faculty Advisor: Love, Tiffany (University of Utah, Psychiatry)
Chronic pain is a major health crisis, and is considered the second major cause of disability in the world. People with chronic pain that lasts 6 months or longer often experience other symptoms as well, including depression. While the causes of chronic pain are often unknown, it has been shown that people with chronic pain exhibit brain structure differences compared to those who do not. It is important to know how chronic pain and brain structure are interconnected so that we can find a better way to treat patients. Gaining more knowledge of this connection can lead to a better understanding of the underlying causes. Brain structure goes hand in hand with neuroplasticity, which plays a key role in normal brain development. While it has been shown that chronic pain can have a significant effect on brain structure, it is unknown whether different symptoms affect different areas of the brain. The effects of chronic pain on the brain have only been looked at in a holistic sense and have not been quantified according to symptoms. Because chronic pain can have a variety of causes, this makes it difficult to determine a good method of treatment for individuals. Our aim was to determine how different symptoms of chronic pain affect the brain individually and evaluate possible overlap. The categories we tested were sensory, affective, unpleasantness, intensity, and depression. We used MRI scans from both healthy individuals and participants experiencing chronic lower back pain. The relative levels of each symptom that the participants were experiencing were determined via the McGill Pain Questionnaire and the Center for Epidemiologic Studies Depression Scale. Having a better understanding of how different symptoms of chronic pain affect the brain can aid in finding more personalized treatment for those experiencing it.
Faculty Advisor: Love, Tiffany (University of Utah, Psychiatry)
Chronic pain is a major health crisis, and is considered the second major cause of disability in the world. People with chronic pain that lasts 6 months or longer often experience other symptoms as well, including depression. While the causes of chronic pain are often unknown, it has been shown that people with chronic pain exhibit brain structure differences compared to those who do not. It is important to know how chronic pain and brain structure are interconnected so that we can find a better way to treat patients. Gaining more knowledge of this connection can lead to a better understanding of the underlying causes. Brain structure goes hand in hand with neuroplasticity, which plays a key role in normal brain development. While it has been shown that chronic pain can have a significant effect on brain structure, it is unknown whether different symptoms affect different areas of the brain. The effects of chronic pain on the brain have only been looked at in a holistic sense and have not been quantified according to symptoms. Because chronic pain can have a variety of causes, this makes it difficult to determine a good method of treatment for individuals. Our aim was to determine how different symptoms of chronic pain affect the brain individually and evaluate possible overlap. The categories we tested were sensory, affective, unpleasantness, intensity, and depression. We used MRI scans from both healthy individuals and participants experiencing chronic lower back pain. The relative levels of each symptom that the participants were experiencing were determined via the McGill Pain Questionnaire and the Center for Epidemiologic Studies Depression Scale. Having a better understanding of how different symptoms of chronic pain affect the brain can aid in finding more personalized treatment for those experiencing it.
Genetic engineering of cells for treatment of autoimmune disorders
Passey, Abigail; Domyan, Eric (Utah Valley University)
Faculty Advisor: Domyan, Eric (Utah Valley University, Department Of Biotechnology/Biology)
In the United States approximately 3 million people are living with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). Approximately 75,000 of those patients represent pediatric cases. We aim to create a new drug delivery system with the intention of establishing a more feasible, stable, and effective form of administering treatments to those with the aforementioned autoimmune diseases, specifically attempting to provide a more ideal treatment for juvenile patients. We are focusing on first providing a rudimentary proof of concept. For the project, we will attempt to engineer mammalian cells that will produce the fusion protein CTLA4-Ig, commonly known as abatacept, a current treatment for RA, and a potential treatment for SLE. Thus far, we have completed and verified success of the molecular cloning necessary to create the recombinant molecule. We have successfully induced expression of the fusion protein in mammalian cell lines COS-7 and B16F10 via lipofections. We are currently working to optimize lipofection conditions and test for successful cellular production of CTLA4-Ig. Ideally, we aim to engineer red blood cells (RBCs) to produce the molecule. If we can complete our proof of concept, we will then attempt to reprogram myeloid and lymphoid progenitors into induced hematopoietic stem cells (iHSCs), and culture the cells ex vivo to allow for massive expansion of these iHSCs, which can then be genetically engineered. Additionally, the iHSCs will be cultured in such a way that, once in vivo, will result in them committing explicitly to erythroid lineages, and secreting the target protein as they mature into fully functional, adult RBCs. Causing RBCs to secrete CTLA4-Ig throughout the body would eliminate the need for regular injections of the drug, and thus potentially improve the quality of pediatric patients' lives.
Faculty Advisor: Domyan, Eric (Utah Valley University, Department Of Biotechnology/Biology)
In the United States approximately 3 million people are living with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). Approximately 75,000 of those patients represent pediatric cases. We aim to create a new drug delivery system with the intention of establishing a more feasible, stable, and effective form of administering treatments to those with the aforementioned autoimmune diseases, specifically attempting to provide a more ideal treatment for juvenile patients. We are focusing on first providing a rudimentary proof of concept. For the project, we will attempt to engineer mammalian cells that will produce the fusion protein CTLA4-Ig, commonly known as abatacept, a current treatment for RA, and a potential treatment for SLE. Thus far, we have completed and verified success of the molecular cloning necessary to create the recombinant molecule. We have successfully induced expression of the fusion protein in mammalian cell lines COS-7 and B16F10 via lipofections. We are currently working to optimize lipofection conditions and test for successful cellular production of CTLA4-Ig. Ideally, we aim to engineer red blood cells (RBCs) to produce the molecule. If we can complete our proof of concept, we will then attempt to reprogram myeloid and lymphoid progenitors into induced hematopoietic stem cells (iHSCs), and culture the cells ex vivo to allow for massive expansion of these iHSCs, which can then be genetically engineered. Additionally, the iHSCs will be cultured in such a way that, once in vivo, will result in them committing explicitly to erythroid lineages, and secreting the target protein as they mature into fully functional, adult RBCs. Causing RBCs to secrete CTLA4-Ig throughout the body would eliminate the need for regular injections of the drug, and thus potentially improve the quality of pediatric patients' lives.
Effect of dietary blueberry on vascular function in aged C57BL/6J mice
Petersen, Chrissa; Velaytham, Anandh; Saldivar, Miguel (University of Utah)
Faculty Advisor: Velayutham, Anandh (Integrative Physiology and Nutrition)
Aging is a major risk factor for cardiovascular diseases such as atherosclerosis, which are major causes of disability and mortality in the elderly. Endothelial dysfunction plays a major role in aging-associated vascular complications. Dietary change may be one of the novel strategies to ameliorate endothelial dysfunction and aging-associated complications. Our lab recently showed that dietary supplementation of blueberries improves vascular inflammation and dysfunction in diabetic mice. In our present study, we investigated the effect of dietary blueberries on vascular function in aged mice. Adult male mice (two months old) and old male mice (17 months old) were fed a control rodent diet (Y and O respectively). The subgroups of Y and O mice were fed a diet supplemented with 3.8% freeze-dried blueberries (Y+BB, O+BB respectively) for 15 weeks. Based on normalization to body surface area, this dose in mice is equivalent to ~1.5 servings of blueberries (~240 g) in humans. Mesenteric arteries were collected and used to assess vascular function using a wire myograph system. After arteries were precontracted to ~65% of maximal phenylephrine-induced contraction and tension was stable, responses to acetylcholine (ACh, 10-8-10-6 M) were evaluated to determine endothelium-dependent vasorelaxation. In our study, there is no difference existed between Y vs. O and O vs. O+BB indicating the vascular function was similar among the groups. Our ongoing studies are focused on identifying the effect of dietary blueberries on vascular inflammation in aged mice and the possible molecular mechanisms involved.
Faculty Advisor: Velayutham, Anandh (Integrative Physiology and Nutrition)
Aging is a major risk factor for cardiovascular diseases such as atherosclerosis, which are major causes of disability and mortality in the elderly. Endothelial dysfunction plays a major role in aging-associated vascular complications. Dietary change may be one of the novel strategies to ameliorate endothelial dysfunction and aging-associated complications. Our lab recently showed that dietary supplementation of blueberries improves vascular inflammation and dysfunction in diabetic mice. In our present study, we investigated the effect of dietary blueberries on vascular function in aged mice. Adult male mice (two months old) and old male mice (17 months old) were fed a control rodent diet (Y and O respectively). The subgroups of Y and O mice were fed a diet supplemented with 3.8% freeze-dried blueberries (Y+BB, O+BB respectively) for 15 weeks. Based on normalization to body surface area, this dose in mice is equivalent to ~1.5 servings of blueberries (~240 g) in humans. Mesenteric arteries were collected and used to assess vascular function using a wire myograph system. After arteries were precontracted to ~65% of maximal phenylephrine-induced contraction and tension was stable, responses to acetylcholine (ACh, 10-8-10-6 M) were evaluated to determine endothelium-dependent vasorelaxation. In our study, there is no difference existed between Y vs. O and O vs. O+BB indicating the vascular function was similar among the groups. Our ongoing studies are focused on identifying the effect of dietary blueberries on vascular inflammation in aged mice and the possible molecular mechanisms involved.
Engineering Cell-free Protein Synthesis as a Biosensor for Endocrine Disrupting Chemicals
Nelson, Andrew; Foutz, Isaac; Hunt, Porter; Wood, David; Bundy, Bradley; (Brigham Young University)
Faculty Advisor: Bundy, Bradley (Brigham Young University, Chemical Engineering)
Cell-free Protein Synthesis (CFPS), an in vitro system for producing recombinant protein, is a rapidly expanding field. To date, applications of this technology, among others, include unnatural amino acid incorporation, protein microarray fabrication, genome engineering, and the production of therapeutics, vaccines, and biocatalysts. Here, we further engineer cell-free protein synthesis as a biosensor for endocrine-disrupting chemicals (EDCs), compounds that mimic hormones and thus disrupt endocrine system physiology in the body.
Faculty Advisor: Bundy, Bradley (Brigham Young University, Chemical Engineering)
Cell-free Protein Synthesis (CFPS), an in vitro system for producing recombinant protein, is a rapidly expanding field. To date, applications of this technology, among others, include unnatural amino acid incorporation, protein microarray fabrication, genome engineering, and the production of therapeutics, vaccines, and biocatalysts. Here, we further engineer cell-free protein synthesis as a biosensor for endocrine-disrupting chemicals (EDCs), compounds that mimic hormones and thus disrupt endocrine system physiology in the body.
Daily Experiences of Children with Cancer Reported Through a Game-Based App
Linder, Caitlin; Linder, Lauri (University of Utah)
Faculty Advisor: Linder, Lauri (College of Nursing, Nursing)
Childhood cancer disrupts children's day-to-day experiences. The purpose of this study was to analyze children's responses to two questions included in a daily symptom reporting app: "What is the best thing about today?" and "What is bothering you the most today?" Responses were part of a larger study evaluating the feasibility and acceptability of the app. Children used the app to record daily symptoms and answer short questions about their day.
Children completed a trial of the app between visits at the hospital for chemotherapy. Daily responses to each question were analyzed using descriptive qualitative content analysis with each response serving as a unit of analysis. Coding was completed by each author and reviewed together to reach agreement. Children's responses were organized into categories and subcategories.
Participants were 19 children 6-12 years of age (median 8 years) (12 boys) receiving chemotherapy who used the app for a total of 83 days (median 4.5 days/child). Children provided 72 responses about the best thing about their day that were organized into nine categories: Activities (n=22), People (n=14), Food (n=9), Well-Being (n=9), School (n=7), Nothing (n=5), Object (n=4), Going Home (n=3), and Don't Know (n=1). Children provided 60 responses about the most bothersome aspect of their day that were organized into six categories: Nothing (n=22), Symptoms (n=17), Port (n=7), Cancer Treatment (n=5), Day-to-Day Stuff (n=5), and People (n=4).
Children's responses provide perspective of the impact of cancer on their daily lives. Their responses indicate the importance of maintaining developmentally normal activities and family relationships. Children's responses further indicate the pervasiveness of the cancer experience, such as symptoms, even on days when children are away from the hospital. Mobile health apps can help children not only track symptoms but also reflect on their day. Clinicians can use children's information to better understand children's experiences.
Faculty Advisor: Linder, Lauri (College of Nursing, Nursing)
Childhood cancer disrupts children's day-to-day experiences. The purpose of this study was to analyze children's responses to two questions included in a daily symptom reporting app: "What is the best thing about today?" and "What is bothering you the most today?" Responses were part of a larger study evaluating the feasibility and acceptability of the app. Children used the app to record daily symptoms and answer short questions about their day.
Children completed a trial of the app between visits at the hospital for chemotherapy. Daily responses to each question were analyzed using descriptive qualitative content analysis with each response serving as a unit of analysis. Coding was completed by each author and reviewed together to reach agreement. Children's responses were organized into categories and subcategories.
Participants were 19 children 6-12 years of age (median 8 years) (12 boys) receiving chemotherapy who used the app for a total of 83 days (median 4.5 days/child). Children provided 72 responses about the best thing about their day that were organized into nine categories: Activities (n=22), People (n=14), Food (n=9), Well-Being (n=9), School (n=7), Nothing (n=5), Object (n=4), Going Home (n=3), and Don't Know (n=1). Children provided 60 responses about the most bothersome aspect of their day that were organized into six categories: Nothing (n=22), Symptoms (n=17), Port (n=7), Cancer Treatment (n=5), Day-to-Day Stuff (n=5), and People (n=4).
Children's responses provide perspective of the impact of cancer on their daily lives. Their responses indicate the importance of maintaining developmentally normal activities and family relationships. Children's responses further indicate the pervasiveness of the cancer experience, such as symptoms, even on days when children are away from the hospital. Mobile health apps can help children not only track symptoms but also reflect on their day. Clinicians can use children's information to better understand children's experiences.
Determining Potential Abuse Liability of a Novel Pain Therapy
Bobeck, Erin; McDermott, Max (Utah State University)
Faculty Advisor: Bobeck, Erin (College of Science, Biology Department)
TBD - Updated for publication
Faculty Advisor: Bobeck, Erin (College of Science, Biology Department)
TBD - Updated for publication
cAMP Secretion of mutated M.smegmatis
Oram, Kathryn; Griffiths, Alayna (Utah Valley University)
Faculty Advisor: Gazdik-Stofer, Michaela (Utah Valley University, Biology)
The World Health Organization currently estimates that 4,384 individuals die per day due to complications of Mycobacterium Tuberculosis and affects 1.8 million people worldwide as it infects individuals through air droplets from a cough or sneeze. Cyclic adenosine monophosphate (cAMP) is known to be a major component in TB because it acts as a macrophage inhibitor that is responsible for blocking the immune defense allowing the M. Tuberculosis to rapidly replicated within cells. The function of cAMP in TB patients is known but the focal point of our research is why and how the increased levels of cAMP effects patients that are infected with TB. Our team uses mutated M. Smegmatis bacteria due to the comparable levels of cAMP secretion and high pathogenicity of M. Tuberculosis. We are currently screening the cAMP secretion in 1,000 mutated M. Smegmatis colonies to identify secretion differences from the wild-type M. Smegmatis. The mutants samples with high variation from the wild-type will be sequenced to identify the genes and determine the proteins that are present. Finding the genes and proteins can help understand why and what causes the inflation on cAMP secretion in TB patients.
Faculty Advisor: Gazdik-Stofer, Michaela (Utah Valley University, Biology)
The World Health Organization currently estimates that 4,384 individuals die per day due to complications of Mycobacterium Tuberculosis and affects 1.8 million people worldwide as it infects individuals through air droplets from a cough or sneeze. Cyclic adenosine monophosphate (cAMP) is known to be a major component in TB because it acts as a macrophage inhibitor that is responsible for blocking the immune defense allowing the M. Tuberculosis to rapidly replicated within cells. The function of cAMP in TB patients is known but the focal point of our research is why and how the increased levels of cAMP effects patients that are infected with TB. Our team uses mutated M. Smegmatis bacteria due to the comparable levels of cAMP secretion and high pathogenicity of M. Tuberculosis. We are currently screening the cAMP secretion in 1,000 mutated M. Smegmatis colonies to identify secretion differences from the wild-type M. Smegmatis. The mutants samples with high variation from the wild-type will be sequenced to identify the genes and determine the proteins that are present. Finding the genes and proteins can help understand why and what causes the inflation on cAMP secretion in TB patients.
Clinical and molecular epidemiology of invasive Haemophilus influenzae disease in Utah, 2007-2017
Varghese, Alyssa; Crandall, Hillary; Blaschke, Anne (University of Utah)
Faculty Advisor: Crandall, Hillary (University of Utah, Department of Pediatrics); Blaschke, Anne (University of Utah, Department of pediatrics)
Haemophilus influenzae serotype b (Hib) causes serious bacterial infections in children associated with high morbidity and mortality. The incidence of Hib disease decreased with widespread vaccination in the mid-1980's. However, other H. influenzae serotypes, such as serotype a (Hia), have emerged in specific pediatric populations, including Utah and indigenous populations in Alaska and Canada. Hia appears to cause disease similar to Hib in both severity and disease presentations.
Cases of invasive H. influenzae disease in children <18 years were identified via a search of electronic medical records within the Intermountain Healthcare system. Phylogenetic division was determined using sodC PCR. Whole genome sequencing (WGS) was performed on available isolates. Gene presence and absence data from resulting assemblies were utilized to build a relationship tree from all available Hia isolates, as well as select Hib, H. influenzae serotype f (Hif), and non-typeable H. influenzae (NTHi) isolates. Multi-locus sequence type (ST) was extracted from WGS data for each isolate.
118 cases of invasive H. influenzae disease were identified from 2007 to 2017. Fifty-one (43.2%) cases were Hia and 11 (9.3%) were Hib. Twenty-eight of 51 (56%) Hia isolates were available for further molecular analysis. Three STs were identified: ST56, ST62, and ST576. Twenty-one isolates (75%) belonged to ST62, clonal division II. The relationship tree indicates that ST62 Hia isolates are most closely related to each other and more closely related to Hif than to other Hia and Hib.
The molecular epidemiology of invasive Hia disease in Utah is unique, with a predominance of ST62 strains, a ST that has been infrequently reported in other studies of invasive Hia. Further genomic analysis will help us understand genetic determinants of virulence. These analyses will be critical in characterizing the clinical and molecular features of invasive H. influenzae disease in Utah.
Faculty Advisor: Crandall, Hillary (University of Utah, Department of Pediatrics); Blaschke, Anne (University of Utah, Department of pediatrics)
Haemophilus influenzae serotype b (Hib) causes serious bacterial infections in children associated with high morbidity and mortality. The incidence of Hib disease decreased with widespread vaccination in the mid-1980's. However, other H. influenzae serotypes, such as serotype a (Hia), have emerged in specific pediatric populations, including Utah and indigenous populations in Alaska and Canada. Hia appears to cause disease similar to Hib in both severity and disease presentations.
Cases of invasive H. influenzae disease in children <18 years were identified via a search of electronic medical records within the Intermountain Healthcare system. Phylogenetic division was determined using sodC PCR. Whole genome sequencing (WGS) was performed on available isolates. Gene presence and absence data from resulting assemblies were utilized to build a relationship tree from all available Hia isolates, as well as select Hib, H. influenzae serotype f (Hif), and non-typeable H. influenzae (NTHi) isolates. Multi-locus sequence type (ST) was extracted from WGS data for each isolate.
118 cases of invasive H. influenzae disease were identified from 2007 to 2017. Fifty-one (43.2%) cases were Hia and 11 (9.3%) were Hib. Twenty-eight of 51 (56%) Hia isolates were available for further molecular analysis. Three STs were identified: ST56, ST62, and ST576. Twenty-one isolates (75%) belonged to ST62, clonal division II. The relationship tree indicates that ST62 Hia isolates are most closely related to each other and more closely related to Hif than to other Hia and Hib.
The molecular epidemiology of invasive Hia disease in Utah is unique, with a predominance of ST62 strains, a ST that has been infrequently reported in other studies of invasive Hia. Further genomic analysis will help us understand genetic determinants of virulence. These analyses will be critical in characterizing the clinical and molecular features of invasive H. influenzae disease in Utah.
Glutens Impact on the Microbiome of the Human G.I. Tract
Hansen, Riley; Hooper, Victoria; Rawson, Clayton (Utah Valley University)
Faculty Advisor: Gazdik-Stofer, Michaela (Utah Valley University, Biology)
The human microbiome is the community of microbes associated with the human body. Disruption and imbalances in these microbiomes can cause diverse symptoms from depression to eczema. Celiac Disease (CD) is characterized by an immune response in the small intestine that is triggered by the protein gluten, found in wheat. Previous studies have identified microbial dysbiosis in patients with CD. To better understand how these dysbiosis can affect the disease itself we will exam the microbiome diversity and composition in CD patients and healthy individuals on gluten-free diets. This will help identify what microbiome changes are associated with CD compared to those that are caused by the CD patients lack of gluten in their diet. The microbial populations of three different groups will be analyzed: 1. those diagnosed with CD and on a gluten-free diet, 2. those who do not have CD and are on a gluten-free diet, and 3. those who do not have CD and are not on a specific diet. The dietary and disease influences on the microbiome of the gastrointestinal tract will be analyzed via a single stool sample collected from each study subject. Microbial DNA was extracted using the QIAamp PowerFecal DNA kit following manufacturer's specifications (Qiagen) V3/V4 region of 16s rRNA will be sequenced using the 600 cycle v3 MiSeq kit (Illumina) The results of the analysis will be compared to existing microbial genome databases to determine the composition of microbes present in the study groups.
Faculty Advisor: Gazdik-Stofer, Michaela (Utah Valley University, Biology)
The human microbiome is the community of microbes associated with the human body. Disruption and imbalances in these microbiomes can cause diverse symptoms from depression to eczema. Celiac Disease (CD) is characterized by an immune response in the small intestine that is triggered by the protein gluten, found in wheat. Previous studies have identified microbial dysbiosis in patients with CD. To better understand how these dysbiosis can affect the disease itself we will exam the microbiome diversity and composition in CD patients and healthy individuals on gluten-free diets. This will help identify what microbiome changes are associated with CD compared to those that are caused by the CD patients lack of gluten in their diet. The microbial populations of three different groups will be analyzed: 1. those diagnosed with CD and on a gluten-free diet, 2. those who do not have CD and are on a gluten-free diet, and 3. those who do not have CD and are not on a specific diet. The dietary and disease influences on the microbiome of the gastrointestinal tract will be analyzed via a single stool sample collected from each study subject. Microbial DNA was extracted using the QIAamp PowerFecal DNA kit following manufacturer's specifications (Qiagen) V3/V4 region of 16s rRNA will be sequenced using the 600 cycle v3 MiSeq kit (Illumina) The results of the analysis will be compared to existing microbial genome databases to determine the composition of microbes present in the study groups.
Cell surface changes that influence size based dissemination of a fungal pathogen
Brammer, Brianna; Denham, Steven; Wambaugh, Morgan; Brown, Jessica. (University of Utah)
Faculty Advisor: Brown, Jessica (University of Utah, Pathology)
Cryptococcus neoformans is an opportunistic fungal pathogen, which primarily affects those with compromised immune systems, contributing to 15% of global AIDS-related deaths. Initial human exposure occurs after inhalation of desiccated cryptococcal cells, which undergo morphological changes in the lungs, including altering cell body and polysaccharide capsule size. Fatality occurs after C. neoformans disseminates to extrapulmonary organs, including the brain where it causes cryptococcal meningitis. Preliminary data show that fungal cells decrease in size throughout the course of infection; we hypothesize that this shift increases the ability of fungal cells to exit lung epithelium, either extracellularly or via macrophage, as small cryptococcal cells exhibit enhanced extrapulmonary dissemination. We later determined that this effect was not solely due to fungal cell size by inoculating mice with fluorescent beads corresponding to cryptococcal cell size groups. The beads showed similar dissemination trends, but were significantly less efficient at extrapulmonary dissemination, suggesting the necessity of cell surface factors. By measuring the exposure of various fungal factors relative to size, we have identified mannose as a potential key factor in dissemination, as small cells exhibit higher levels of exposed mannose relative to size. The hypothesized role of mannose in cryptococcal dissemination is investigated throughout this project utilizing a variety of techniques, including the addition of polysaccharides to macrophage-cryptococcal association assays to determine mannose-specific recognition, in vivo co-inoculation with mannose and cryptococcal cells, and identifying differential gene expression between cell sizes using RNA sequencing and gene ontology. These data have contributed to our working model of cryptococcal infection, where cryptococcal cells undergo morphological changes in the lungs, yielding a higher prevalence of small cells. Cell size is an important, but not determinant factor in dissemination, suggesting the role of cell surface factors, such as mannose, that increase virulence by promoting phagocytosis and intracellular survival.
Faculty Advisor: Brown, Jessica (University of Utah, Pathology)
Cryptococcus neoformans is an opportunistic fungal pathogen, which primarily affects those with compromised immune systems, contributing to 15% of global AIDS-related deaths. Initial human exposure occurs after inhalation of desiccated cryptococcal cells, which undergo morphological changes in the lungs, including altering cell body and polysaccharide capsule size. Fatality occurs after C. neoformans disseminates to extrapulmonary organs, including the brain where it causes cryptococcal meningitis. Preliminary data show that fungal cells decrease in size throughout the course of infection; we hypothesize that this shift increases the ability of fungal cells to exit lung epithelium, either extracellularly or via macrophage, as small cryptococcal cells exhibit enhanced extrapulmonary dissemination. We later determined that this effect was not solely due to fungal cell size by inoculating mice with fluorescent beads corresponding to cryptococcal cell size groups. The beads showed similar dissemination trends, but were significantly less efficient at extrapulmonary dissemination, suggesting the necessity of cell surface factors. By measuring the exposure of various fungal factors relative to size, we have identified mannose as a potential key factor in dissemination, as small cells exhibit higher levels of exposed mannose relative to size. The hypothesized role of mannose in cryptococcal dissemination is investigated throughout this project utilizing a variety of techniques, including the addition of polysaccharides to macrophage-cryptococcal association assays to determine mannose-specific recognition, in vivo co-inoculation with mannose and cryptococcal cells, and identifying differential gene expression between cell sizes using RNA sequencing and gene ontology. These data have contributed to our working model of cryptococcal infection, where cryptococcal cells undergo morphological changes in the lungs, yielding a higher prevalence of small cells. Cell size is an important, but not determinant factor in dissemination, suggesting the role of cell surface factors, such as mannose, that increase virulence by promoting phagocytosis and intracellular survival.
Does It Wash Away? The Impact of Bathing or Showering on DNA Analysis Findings from Sexual Assault Kits
Richardson, Deborah; Valentine, Julie; Miles, Leslie (Brigham Young University)
Faculty Advisor: Valentine, Julie (Brigham Young University, Nursing); Miles, Leslie (Brigham Young university, Nursing)
Purpose: Describe the minimal effect of post-assault bathing or showering on the development of FBI Combined DNA Index System (CODIS) eligible DNA profiles from sexual assault kit evidence.
Sexual assault kit (SAK) submission rates significantly decrease in cases wherein the victim has bathed or showered. The belief that these actions diminish the possibility of finding evidence heavily contributes to be a significant negative predictor of SAK submissions by law enforcement (LE). Now there are improved DNA analysis methods that can yield CODIS eligible DNA profiles from skin regardless of whether the victim bathed or showered.
SAK submission rates and DNA analysis findings from 5,423 cases were evaluated in this retrospective study. 36% of victims reported post-assault bathing or showering. These washing actions was found to be highly correlated with time between assault and sexual assault forensic examination. In a generalized estimating equation (GEE) logistic regression analysis on SAK submission rates, victim reports of post-assault bathing or showering was a consistent predictor of law enforcement not submitting kits. Yet, bivariable statistical analysis determined that victim bathing or showering post-assault was not associated with the lack of development of a DNA probative profile from SAK evidence.
The effect of post-assault bathing or showering must be reconsidered in the forensic science and criminal justice community as DNA analysis can yield probative DNA profiles, even after a victim has bathed or showered. The presentation of these research findings will encourage the submission of SAK by law enforcement and potentially increase SAK submission rates by eradicating any doubt regarding the minimal effect of bathing or showering on the development of CODIS eligible DNA profiles. New research data supports the collection, submission and testing of all SAK especially those with a victim history of post-assault bathing or showering.
Faculty Advisor: Valentine, Julie (Brigham Young University, Nursing); Miles, Leslie (Brigham Young university, Nursing)
Purpose: Describe the minimal effect of post-assault bathing or showering on the development of FBI Combined DNA Index System (CODIS) eligible DNA profiles from sexual assault kit evidence.
Sexual assault kit (SAK) submission rates significantly decrease in cases wherein the victim has bathed or showered. The belief that these actions diminish the possibility of finding evidence heavily contributes to be a significant negative predictor of SAK submissions by law enforcement (LE). Now there are improved DNA analysis methods that can yield CODIS eligible DNA profiles from skin regardless of whether the victim bathed or showered.
SAK submission rates and DNA analysis findings from 5,423 cases were evaluated in this retrospective study. 36% of victims reported post-assault bathing or showering. These washing actions was found to be highly correlated with time between assault and sexual assault forensic examination. In a generalized estimating equation (GEE) logistic regression analysis on SAK submission rates, victim reports of post-assault bathing or showering was a consistent predictor of law enforcement not submitting kits. Yet, bivariable statistical analysis determined that victim bathing or showering post-assault was not associated with the lack of development of a DNA probative profile from SAK evidence.
The effect of post-assault bathing or showering must be reconsidered in the forensic science and criminal justice community as DNA analysis can yield probative DNA profiles, even after a victim has bathed or showered. The presentation of these research findings will encourage the submission of SAK by law enforcement and potentially increase SAK submission rates by eradicating any doubt regarding the minimal effect of bathing or showering on the development of CODIS eligible DNA profiles. New research data supports the collection, submission and testing of all SAK especially those with a victim history of post-assault bathing or showering.
Is intact endothelial cell autophagy necessary to observe training-induced vascular adaptations?
Symons, John David; Cho, Jae Min; Ly, Kellsey; Thompson, Lauren; Lee, Sebastian; Hansen, Michele; Carter, Kandis (University of Utah)
Faculty Advisor: Symons, John David (University of Utah; Nutrition and Integrative Physiology)
The process of macroautophagy is operational during basal conditions to maintain organelle and protein quality control, but is upregulated during cellular stress to adapt to changing nutritional and energy demands. We tested the hypothesis that intact endothelial cell (EC) autophagy is required to observe exercise training-induced vascular improvements. Rationale for this hypothesis was provided by an earlier report that obese mice with germline, whole body mutation of a protein requisite for autophagy i.e., Bcl2-AAA mice were refractory to training-induced improvements concerning glucose homeostasis. First we demonstrated that : (i) workload achieved during a maximal treadmill test; (ii) soleus muscle citrate synthase activity; (iii) vascular indices of autophagy; and (iv) endothelium-dependent function were greater (p<0.05) in male C57Bl/6 mice that completed 10-weeks of treadmill-training vs. age-matched sedentary animals. These findings indicate that an efficacious training protocol improves vascular autophagy and arterial function. Next, age-matched male mice on a C75Bl/6 background with tamoxifen-inducible Cre/LoxP-based impairment of autophagy-related gene 3 (Atg3) specifically in ECs (iecAtg3KO mice) or wild-type (WT) littermates were trained (ETR) identically or remained sedentary (SED). Atg3 mRNA was minimal (p<0.05) in ECs obtained from iecAtg3KO vs. WT mice, but vascular smooth muscle cell Atg3 was similar between groups. These data verify that specific knockdown of Atg3 existed in ECs but not vascular smooth muscle of iecAtg3KO mice. As expected, intraluminal flow-mediated vasodilation (FMD) improved (p<0.05) in WT-ETR vs. WT-SED mice, while vascular smooth muscle responses to sodium nitroprusside were similar between groups. Further, as anticipated, intraluminal FMD was blunted (p<0.05) in iecAtg3KO-SED vs. WT-SED mice, indicating the importance of EC autophagy to FMD induced vasodilation. Contrary to our hypothesis, however, training-induced vascular adaptations were observed (p<0.05) in iecAtg3KO-ETR vs. iecAtg3KO-SED mice, while vascular smooth muscle responses were similar between groups. Indeed, training-induced vascular improvements concerning intraluminal FMD were not different between WT-ETR and iecAtg3KO-ETR mice. These findings are not congruent with our original hypothesis, and indicate that intact EC Atg3 is not required for training-induced vascular adaptations to occur.
Faculty Advisor: Symons, John David (University of Utah; Nutrition and Integrative Physiology)
The process of macroautophagy is operational during basal conditions to maintain organelle and protein quality control, but is upregulated during cellular stress to adapt to changing nutritional and energy demands. We tested the hypothesis that intact endothelial cell (EC) autophagy is required to observe exercise training-induced vascular improvements. Rationale for this hypothesis was provided by an earlier report that obese mice with germline, whole body mutation of a protein requisite for autophagy i.e., Bcl2-AAA mice were refractory to training-induced improvements concerning glucose homeostasis. First we demonstrated that : (i) workload achieved during a maximal treadmill test; (ii) soleus muscle citrate synthase activity; (iii) vascular indices of autophagy; and (iv) endothelium-dependent function were greater (p<0.05) in male C57Bl/6 mice that completed 10-weeks of treadmill-training vs. age-matched sedentary animals. These findings indicate that an efficacious training protocol improves vascular autophagy and arterial function. Next, age-matched male mice on a C75Bl/6 background with tamoxifen-inducible Cre/LoxP-based impairment of autophagy-related gene 3 (Atg3) specifically in ECs (iecAtg3KO mice) or wild-type (WT) littermates were trained (ETR) identically or remained sedentary (SED). Atg3 mRNA was minimal (p<0.05) in ECs obtained from iecAtg3KO vs. WT mice, but vascular smooth muscle cell Atg3 was similar between groups. These data verify that specific knockdown of Atg3 existed in ECs but not vascular smooth muscle of iecAtg3KO mice. As expected, intraluminal flow-mediated vasodilation (FMD) improved (p<0.05) in WT-ETR vs. WT-SED mice, while vascular smooth muscle responses to sodium nitroprusside were similar between groups. Further, as anticipated, intraluminal FMD was blunted (p<0.05) in iecAtg3KO-SED vs. WT-SED mice, indicating the importance of EC autophagy to FMD induced vasodilation. Contrary to our hypothesis, however, training-induced vascular adaptations were observed (p<0.05) in iecAtg3KO-ETR vs. iecAtg3KO-SED mice, while vascular smooth muscle responses were similar between groups. Indeed, training-induced vascular improvements concerning intraluminal FMD were not different between WT-ETR and iecAtg3KO-ETR mice. These findings are not congruent with our original hypothesis, and indicate that intact EC Atg3 is not required for training-induced vascular adaptations to occur.
Dissecting the tumor microenvironment in zebrafish models of melanoma
Carrington, James; Xue, Qian; Roh-Johnson, Minna (University of Utah)
Faculty Advisor: Roh-Johnson, Minna (University of Utah, Biochemistry)
Metastasis of melanoma to distant sites of the body result in poor patient prognosis with a high mortality rate (76%). Cell migration has been studied in vitro and focal adhesions, which allow cells to move forward by attaching to extracellular matrix (ECM) on the front of the cell and breaking down at the back of the cell, have been shown to play important roles in locomotion. However, it has been difficult to visualize these structures in vivo, especially during tumor cell dissemination. Understanding how cancer cells are utilizing focal adhesions could play an important role in developing therapeutics to counteract metastasis using focal adhesion inhibitors and lead to improved patient outcomes. This research focuses on identifying if a previously observed focal adhesion marker in melanoma is formed at surfaces where cells are in contact with ECM. Because zebrafish share 80% disease homology with humans and are transparent during early embryonic development, they provide an optimal model for visualizing cell migration while still maintaining physiological significance. To identify if migrating melanoma cells are in contact with ECM, we injected fluorescently labeled melanoma cells in zebrafish. We allowed the melanoma cells to migrate and labeled components of the ECM (laminin, collagen, and fibronectin). We then imaged the zebrafish and determined the proximity of melanoma cells to the ECM. We also used transmission electron microscopy to identify the location of melanoma cells in respect to the ECM. We found that melanoma cells are in contact with ECM in vivo. The in vivo contact of focal adhesion markers in melanoma with ECM suggests that focal adhesions can be visualized and studied in zebrafish. Future studies will examine how focal adhesion formation is regulated and how inhibiting their function will impact tumor cell dissemination.
Faculty Advisor: Roh-Johnson, Minna (University of Utah, Biochemistry)
Metastasis of melanoma to distant sites of the body result in poor patient prognosis with a high mortality rate (76%). Cell migration has been studied in vitro and focal adhesions, which allow cells to move forward by attaching to extracellular matrix (ECM) on the front of the cell and breaking down at the back of the cell, have been shown to play important roles in locomotion. However, it has been difficult to visualize these structures in vivo, especially during tumor cell dissemination. Understanding how cancer cells are utilizing focal adhesions could play an important role in developing therapeutics to counteract metastasis using focal adhesion inhibitors and lead to improved patient outcomes. This research focuses on identifying if a previously observed focal adhesion marker in melanoma is formed at surfaces where cells are in contact with ECM. Because zebrafish share 80% disease homology with humans and are transparent during early embryonic development, they provide an optimal model for visualizing cell migration while still maintaining physiological significance. To identify if migrating melanoma cells are in contact with ECM, we injected fluorescently labeled melanoma cells in zebrafish. We allowed the melanoma cells to migrate and labeled components of the ECM (laminin, collagen, and fibronectin). We then imaged the zebrafish and determined the proximity of melanoma cells to the ECM. We also used transmission electron microscopy to identify the location of melanoma cells in respect to the ECM. We found that melanoma cells are in contact with ECM in vivo. The in vivo contact of focal adhesion markers in melanoma with ECM suggests that focal adhesions can be visualized and studied in zebrafish. Future studies will examine how focal adhesion formation is regulated and how inhibiting their function will impact tumor cell dissemination.
Exploring relationships between practice history, performance, and injury risk in a sample of developmental alpine skiers
Taylor, Sarah; Fawver, Bradley; DeCouto, Brady; Lohse, Keith R.; Williams, A. Mark� (University of Utah)
Faculty Advisor: Lohse, Keith (University of Utah; Health, Kinesiology, and Recreation); Williams, Mark (University of Utah; Health, Kinesiology, and Recreation)
Achieving elite status in sport often requires athletes to overcome significant physical injuries. However, to date, there has been a paucity of studies exploring how hours engaged in practice and early developmental milestones influence injury rates in "high-risk" winter sports, such as alpine skiing. Moreover, despite numerous published reports on injury epidemiology, a lack of objective measures of performance has been a notable oversight. The purpose of this study was to assess how previous sport engagement and performance are related to injury in a sample of sub-elite youth alpine skiers. Adolescent skiers enrolled in U.S. academies (N = 169, males = 81) were given questionnaires assessing practice/injury history and sport-specific milestones, while performance in speed and technical disciplines were derived from participants' National points (i.e., ranking) for each year available. Simple correlations, MANOVAs, and linear mixed-effect regressions were used to assess relationships between predictors: age, gender, sport-specific milestones (e.g., age of first competition), practice hours, ranking; and the outcomes of interest: injury incidence (i.e., proportion of seasons an athlete sustained injuries causing them to miss > 4 weeks) and injury impact (i.e., average weeks missed due to injury each year). Results revealed that while older athletes had accumulated more injury weeks across their career (p = .020), female skiers reported greater injury incidence (p = .049). Neither injury incidence nor injury impact was associated with performance trends (all p's > .05), but they were negatively associated with time spent in group and individual practice (both p's < .05). Finally, the age of first competition was positively correlated with injury impact (p = .014). These and other findings are discussed in relation to previous studies of sport-injury, as well as applied implications for working with developmental athlete populations in high-risk domains.
Faculty Advisor: Lohse, Keith (University of Utah; Health, Kinesiology, and Recreation); Williams, Mark (University of Utah; Health, Kinesiology, and Recreation)
Achieving elite status in sport often requires athletes to overcome significant physical injuries. However, to date, there has been a paucity of studies exploring how hours engaged in practice and early developmental milestones influence injury rates in "high-risk" winter sports, such as alpine skiing. Moreover, despite numerous published reports on injury epidemiology, a lack of objective measures of performance has been a notable oversight. The purpose of this study was to assess how previous sport engagement and performance are related to injury in a sample of sub-elite youth alpine skiers. Adolescent skiers enrolled in U.S. academies (N = 169, males = 81) were given questionnaires assessing practice/injury history and sport-specific milestones, while performance in speed and technical disciplines were derived from participants' National points (i.e., ranking) for each year available. Simple correlations, MANOVAs, and linear mixed-effect regressions were used to assess relationships between predictors: age, gender, sport-specific milestones (e.g., age of first competition), practice hours, ranking; and the outcomes of interest: injury incidence (i.e., proportion of seasons an athlete sustained injuries causing them to miss > 4 weeks) and injury impact (i.e., average weeks missed due to injury each year). Results revealed that while older athletes had accumulated more injury weeks across their career (p = .020), female skiers reported greater injury incidence (p = .049). Neither injury incidence nor injury impact was associated with performance trends (all p's > .05), but they were negatively associated with time spent in group and individual practice (both p's < .05). Finally, the age of first competition was positively correlated with injury impact (p = .014). These and other findings are discussed in relation to previous studies of sport-injury, as well as applied implications for working with developmental athlete populations in high-risk domains.
Idle-Free Schools: Perceptions and Efficacy
Contreras, John; Forrest, Rachel T.; Bayles, Madelyn (Westminster College)
Faculty Advisor: Contreras, John (Westminster College, Public Health); Forrest; Rachel T. (Westminster College, Public Health)
Air pollution, much of which results from human activities like transportation, may significantly harm children's health, from worsening asthma to potentially even lowering I.Q. This makes idling near schools a serious concern. In response, the EPA has developed an idle-free schools campaign. This campaign will be implemented during the 2019-2020 school year at a Salt Lake City elementary school. Ground-level ozone and PM2.5 levels will be measured before and after the campaign in order to determine its efficacy. Additionally, during the spring and summer of 2019, parents' perceptions of idling, and, by extension, air pollution, were examined using a focus group and a survey. The resulting data, which was analyzed using R statistical software and Excel, found associations between gender and beliefs about air pollution, as well as between education level and beliefs about air pollution. A better understanding of the effectiveness of the EPA's campaign and of parents' perceptions may be helpful in combating air pollution, leading to improved community health.
Faculty Advisor: Contreras, John (Westminster College, Public Health); Forrest; Rachel T. (Westminster College, Public Health)
Air pollution, much of which results from human activities like transportation, may significantly harm children's health, from worsening asthma to potentially even lowering I.Q. This makes idling near schools a serious concern. In response, the EPA has developed an idle-free schools campaign. This campaign will be implemented during the 2019-2020 school year at a Salt Lake City elementary school. Ground-level ozone and PM2.5 levels will be measured before and after the campaign in order to determine its efficacy. Additionally, during the spring and summer of 2019, parents' perceptions of idling, and, by extension, air pollution, were examined using a focus group and a survey. The resulting data, which was analyzed using R statistical software and Excel, found associations between gender and beliefs about air pollution, as well as between education level and beliefs about air pollution. A better understanding of the effectiveness of the EPA's campaign and of parents' perceptions may be helpful in combating air pollution, leading to improved community health.
Effects of Indigenous Diet and Location on Hemoglobin Levels of Ghanaians
Gaona-Partida, Paul; Chan, Julian; Aguilar-Alvarez, David (Weber State University)
Faculty Advisor: Chan, Julian (Weber State University, Mathematics); Aguilar-Alvarez, David (Weber State University, Exercise and Nutrition Sciences)
PURPOSE/HYPOTHESIS:
Low hemoglobin (HB) levels are used to indicate anemia, which increases your risk for contracting infectious diseases and overall mortality. The rate of iron deficiency anemia in Ghana is more than triple the world average, making this condition a main health concern for the country. In this study, we sought to identify the influence of location and iron consumption on HB levels of Ghanaians.
METHODOLOGY:
Food frequency questionnaire was given to 140 participants in 5 different locations of Ghana (Jamestown, King Tower, Mole, Larabanga, University of Ghana). All food was classified as low, medium, or high iron content according to the USDA food database. Participants were categorized as adequate or inadequate iron consumers according to this classification. ANOVA and t-test analysis was conducted to determine the differences in hemoglobin level means. P-value was set at µ0.05 and HSD was used as the post-hoc test.
RESULTS/PREDICTED RESULTS:
Mole National Park population had higher HB (µ=13.3 ± 2.51) compared to Jamestown (µ=12.1 ± 1.97) (p=0.034) and Larabanga (µ=12.0 ± 1.97) (p =.032). Individuals classified as high iron consumers had significantly higher HB that their low iron consumers counterparts (µ=13.8 ± 2.42 vs µ=12.1 ± 1.97, p=0.001).
CONCLUSION:
We were able to identify foods associated with increased Ghanaians HB. Mole had higher access to iron-rich foods such as game meat, which was reflected in their HB levels. This study illustrates the importance of promoting consumption of indigenous iron-rich foods in the current Ghanaian diet to prevent iron deficiency anemia in this population.
Faculty Advisor: Chan, Julian (Weber State University, Mathematics); Aguilar-Alvarez, David (Weber State University, Exercise and Nutrition Sciences)
PURPOSE/HYPOTHESIS:
Low hemoglobin (HB) levels are used to indicate anemia, which increases your risk for contracting infectious diseases and overall mortality. The rate of iron deficiency anemia in Ghana is more than triple the world average, making this condition a main health concern for the country. In this study, we sought to identify the influence of location and iron consumption on HB levels of Ghanaians.
METHODOLOGY:
Food frequency questionnaire was given to 140 participants in 5 different locations of Ghana (Jamestown, King Tower, Mole, Larabanga, University of Ghana). All food was classified as low, medium, or high iron content according to the USDA food database. Participants were categorized as adequate or inadequate iron consumers according to this classification. ANOVA and t-test analysis was conducted to determine the differences in hemoglobin level means. P-value was set at µ0.05 and HSD was used as the post-hoc test.
RESULTS/PREDICTED RESULTS:
Mole National Park population had higher HB (µ=13.3 ± 2.51) compared to Jamestown (µ=12.1 ± 1.97) (p=0.034) and Larabanga (µ=12.0 ± 1.97) (p =.032). Individuals classified as high iron consumers had significantly higher HB that their low iron consumers counterparts (µ=13.8 ± 2.42 vs µ=12.1 ± 1.97, p=0.001).
CONCLUSION:
We were able to identify foods associated with increased Ghanaians HB. Mole had higher access to iron-rich foods such as game meat, which was reflected in their HB levels. This study illustrates the importance of promoting consumption of indigenous iron-rich foods in the current Ghanaian diet to prevent iron deficiency anemia in this population.
Inducible disruption of endothelial cell ceramide biosynthesis: Vascular implications
Thompson, Lauren; Ramous, Caroline; Ly, Kellsey; Luu, Kiana; Margetts, Alex; Warren, Tahno; Tippetts, Trevor; Choi, Ran Hee; Symons, JD (University of Utah)
Faculty Advisor: Symons, J. David (University of Utah, Nutrition and Integrative Physiology)
Type II diabetes mellitus (T2DM) is an epidemic worldwide. Cardiovascular complications (e.g. endothelial dysfunction and hypertension) are associated with T2DM. T2DM affects the quality of life for the patient and their caregivers, and the costs for treating cardiovascular complications are unsustainable. An urgent need exists to elucidate new therapeutic targets for intervention. Our laboratory is interested in defining the contribution from the sphingolipid ceramide. We reported earlier that arterial dysfunction and hypertension that otherwise develop in mice that consume an obesogenic diet is attenuated by pharmacological inhibition of ceramide using myriocin and by germline haploinsufficiency for dihydroceramide desaturase (DES1), an enzyme required for ceramide biosynthesis. However, each study had limitations. Myriocin improved systemic glucose homeostasis, and DES1 inhibition elevated dihydroceramides, both of which could impact arterial function. In the present study, we used a novel murine model to inhibit the rate-limiting enzyme responsible for ceramide biosynthesis (serine palmitoyl transferase light chain 2; Sptlc2) specifically in endothelial cells (ECs). We hypothesized that EC specific inhibition of ceramide biosynthesis would preserve arterial function in obese mice. Six-week-old male mice with intact Sptlc2 (wild-type; WT) and EC specific deletion of Sptlc2 (iecSptlc2KO mice) consumed either standard (CON) or high fat diet (HFD) for 14 weeks. qPCR results indicated Sptlc2 was knocked down > 80% in ECs but not media and adventitia from iecSptlc2KO vs. WT mice. In general, results were similar between WT and iecSptlc2KO mice concerning glucose, insulin, and pyruvate tolerance tests (indicating intact glucose homeostasis) and lean mass, fat mass, and fluid mass (indicating body composition was unaltered). Of note, intraluminal flow-mediated vasodilation was greater in femoral arteries from iecSptlc2KO vs. WT mice that consumed high-fat chow. Preventing ceramide biosynthesis specifically in ECs from mice that consume an obesogenic diet might be vasculoprotective.
Faculty Advisor: Symons, J. David (University of Utah, Nutrition and Integrative Physiology)
Type II diabetes mellitus (T2DM) is an epidemic worldwide. Cardiovascular complications (e.g. endothelial dysfunction and hypertension) are associated with T2DM. T2DM affects the quality of life for the patient and their caregivers, and the costs for treating cardiovascular complications are unsustainable. An urgent need exists to elucidate new therapeutic targets for intervention. Our laboratory is interested in defining the contribution from the sphingolipid ceramide. We reported earlier that arterial dysfunction and hypertension that otherwise develop in mice that consume an obesogenic diet is attenuated by pharmacological inhibition of ceramide using myriocin and by germline haploinsufficiency for dihydroceramide desaturase (DES1), an enzyme required for ceramide biosynthesis. However, each study had limitations. Myriocin improved systemic glucose homeostasis, and DES1 inhibition elevated dihydroceramides, both of which could impact arterial function. In the present study, we used a novel murine model to inhibit the rate-limiting enzyme responsible for ceramide biosynthesis (serine palmitoyl transferase light chain 2; Sptlc2) specifically in endothelial cells (ECs). We hypothesized that EC specific inhibition of ceramide biosynthesis would preserve arterial function in obese mice. Six-week-old male mice with intact Sptlc2 (wild-type; WT) and EC specific deletion of Sptlc2 (iecSptlc2KO mice) consumed either standard (CON) or high fat diet (HFD) for 14 weeks. qPCR results indicated Sptlc2 was knocked down > 80% in ECs but not media and adventitia from iecSptlc2KO vs. WT mice. In general, results were similar between WT and iecSptlc2KO mice concerning glucose, insulin, and pyruvate tolerance tests (indicating intact glucose homeostasis) and lean mass, fat mass, and fluid mass (indicating body composition was unaltered). Of note, intraluminal flow-mediated vasodilation was greater in femoral arteries from iecSptlc2KO vs. WT mice that consumed high-fat chow. Preventing ceramide biosynthesis specifically in ECs from mice that consume an obesogenic diet might be vasculoprotective.
Giving the VIP Treatment to Prevent Missed Injuries After a Fall
Winters, Blaine; Dustin, Tracy; Escobar, Jeana; Ostlund, Megan (Brigham Young University)
Faculty Advisor: Blaine, Winters (Brigham Young University, Biology)
Background:
Evaluation of patients who have fallen varies significantly between nurses as well as health care facilities. In many cases, injuries go unrecognized or their identification is delayed due to poor or incomplete assessments and follow-up.
Purpose:
The purpose of this project was to design a standardized post-fall assessment for geriatric patients residing in long-term care and community settings.
Methods:
A literature review revealed that in the past 10 years, very few studies were conducted on post-fall assessments, and the majority of these studies were conducted outside of the United States. Due to the lack of current literature on the subject, the Advanced Trauma Life Support algorithm was used as a framework for the development of the post-fall assessment.
Outcomes:
A post-fall assessment evaluation was developed which includes the production of a standardized post-fall assessment flowchart, accompanied by a documentation guide. A training video was also produced to illustrate the use of the assessment tools and to allow nurses to practice their new skills.
Implications for Practice:
The standardized post-fall assessment tools will lead to more thorough evaluation and follow-up of patients who have fallen in long-term care or the community setting. This will lead to more rapid treatment of identified injuries as well as fewer missed injuries.
Teaching Methods:
Participants will receive training on the post-fall evaluation method, how it was developed and how it will be implemented.
Faculty Advisor: Blaine, Winters (Brigham Young University, Biology)
Background:
Evaluation of patients who have fallen varies significantly between nurses as well as health care facilities. In many cases, injuries go unrecognized or their identification is delayed due to poor or incomplete assessments and follow-up.
Purpose:
The purpose of this project was to design a standardized post-fall assessment for geriatric patients residing in long-term care and community settings.
Methods:
A literature review revealed that in the past 10 years, very few studies were conducted on post-fall assessments, and the majority of these studies were conducted outside of the United States. Due to the lack of current literature on the subject, the Advanced Trauma Life Support algorithm was used as a framework for the development of the post-fall assessment.
Outcomes:
A post-fall assessment evaluation was developed which includes the production of a standardized post-fall assessment flowchart, accompanied by a documentation guide. A training video was also produced to illustrate the use of the assessment tools and to allow nurses to practice their new skills.
Implications for Practice:
The standardized post-fall assessment tools will lead to more thorough evaluation and follow-up of patients who have fallen in long-term care or the community setting. This will lead to more rapid treatment of identified injuries as well as fewer missed injuries.
Teaching Methods:
Participants will receive training on the post-fall evaluation method, how it was developed and how it will be implemented.
Developing Antibodies for Studying Structures and Functions of Polycystic Kidney Disease Proteins
DeNiro, Adara; Cao, Erhu; Wang, Qinzhe; Wang, Jinliang (University of Utah)
Faculty Advisor: Cao, Erhu (University of Utah, School of Medicine)
The kidneys are responsible for several essential bodily functions critical to our survival. These include filtering blood and excreting metabolic waste into urine. Two proteins, polycystin-1 and polycystin-2, interact in renal tubules and promote the normal development and function of the kidneys. However, the proteins' actions are not well understood. Polycystin-1 and Polycystin-2 are encoded by the PKD1 gene and the PKD2 gene respectively. A mutation in either of these genes can lead to Autosomal Dominant Polycystic Kidney Disease (ADPKD), a common inherited disease leading to progressive renal failure. Several extrarenal manifestations occur as a result of ADPKD including hepatic cysts and intracranial aneurysms. Ultimately, ADPKD often leads to end-stage renal disease that requires dialysis or transplantation. Currently, there is no cure for ADPKD due to setbacks regarding structural determination of the Polycystic Kidney Disease Proteins. Some setbacks are the inability to view the regulatory cytosolic domains of the PKD1/PKD2 channel in atomic detail and understand the function(s) of the polycystic kidney disease proteins. My project examines the ability of Fabs and nanobodies to bind to the PKD2 channel and the PKD1/2 complex as this potentially facilitates structural determination. This is done by stabilizing cytosolic domains and/or promoting a more uniform orientation distribution of these proteins essential for imaging by cryo-electron microscopy.
Faculty Advisor: Cao, Erhu (University of Utah, School of Medicine)
The kidneys are responsible for several essential bodily functions critical to our survival. These include filtering blood and excreting metabolic waste into urine. Two proteins, polycystin-1 and polycystin-2, interact in renal tubules and promote the normal development and function of the kidneys. However, the proteins' actions are not well understood. Polycystin-1 and Polycystin-2 are encoded by the PKD1 gene and the PKD2 gene respectively. A mutation in either of these genes can lead to Autosomal Dominant Polycystic Kidney Disease (ADPKD), a common inherited disease leading to progressive renal failure. Several extrarenal manifestations occur as a result of ADPKD including hepatic cysts and intracranial aneurysms. Ultimately, ADPKD often leads to end-stage renal disease that requires dialysis or transplantation. Currently, there is no cure for ADPKD due to setbacks regarding structural determination of the Polycystic Kidney Disease Proteins. Some setbacks are the inability to view the regulatory cytosolic domains of the PKD1/PKD2 channel in atomic detail and understand the function(s) of the polycystic kidney disease proteins. My project examines the ability of Fabs and nanobodies to bind to the PKD2 channel and the PKD1/2 complex as this potentially facilitates structural determination. This is done by stabilizing cytosolic domains and/or promoting a more uniform orientation distribution of these proteins essential for imaging by cryo-electron microscopy.