2020 Abstracts
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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.
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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.
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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.
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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.
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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.
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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.
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Tree of Trees
Lahy, Neve (Westminster College)
Faculty Advisor: Kruback, Matt (Westminster College, Art)
Tree of Trees
Neve Lahy:
The Uinta-Wasatch-Cache National Forest is a group of national forests within our Wasatch Mountains. The forest is a host to many species of trees, both coniferous and flowering. I have chosen seven of each of the most prominent local cone bearing and flowering trees to focus on.
The trees are a crucial part of the ecosystem and participate in many symbiotic relationships. Without the trees the ecosystem wouldn't survive. It is important to acknowledge the vitality of our local forests not only for us, but for all other organisms dependent on the trees.
A phylogenetic tree is a branching diagram showing the evolutionary relationships among various biological species. Phylogeny is based upon similarities and differences in their physical or genetic characteristics. The branches indicate closeness in relation to one another. The species nearest to each other on the phylogeny are the most closely related organisms based on special derived traits.
Learning how to identify the trees that I am so often surrounded by has brought me a deeper appreciation and love for the organisms around me. Now that I can spend time in trees and know exactly what I am looking at has given me a broader understanding of how the trees not only interact with me but the other ecological factors present in the area. I feel more keenly aware of what it takes to protect these trees and enjoy them more respectfully.
Faculty Advisor: Kruback, Matt (Westminster College, Art)
Tree of Trees
Neve Lahy:
The Uinta-Wasatch-Cache National Forest is a group of national forests within our Wasatch Mountains. The forest is a host to many species of trees, both coniferous and flowering. I have chosen seven of each of the most prominent local cone bearing and flowering trees to focus on.
The trees are a crucial part of the ecosystem and participate in many symbiotic relationships. Without the trees the ecosystem wouldn't survive. It is important to acknowledge the vitality of our local forests not only for us, but for all other organisms dependent on the trees.
A phylogenetic tree is a branching diagram showing the evolutionary relationships among various biological species. Phylogeny is based upon similarities and differences in their physical or genetic characteristics. The branches indicate closeness in relation to one another. The species nearest to each other on the phylogeny are the most closely related organisms based on special derived traits.
Learning how to identify the trees that I am so often surrounded by has brought me a deeper appreciation and love for the organisms around me. Now that I can spend time in trees and know exactly what I am looking at has given me a broader understanding of how the trees not only interact with me but the other ecological factors present in the area. I feel more keenly aware of what it takes to protect these trees and enjoy them more respectfully.
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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.).
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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.
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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
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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.
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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.
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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.
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Understand whether folic acid can rescue fumonisin, ceramide, and valproic acid induced NTDs
Park, Yeram; Lin, Jade; Ross, Micah; Stark, Michael; (Brigham Young University)
Faculty Advisor: Stark, Michael (Life Sciences, Physiological and Developmental Biology); Hansen, Marc (Life Sciences, Physiological and Developmental Biology)
Neural tube defects (NTDs), which result from failure to close the neural tube during embryonic development, are one of the most widespread and common congenital malformations. Variance in these malformations can range from anencephaly (failure of the neural tube to close on the cranial end) to spina bifida (failure of closure on the posterior/dorsal end). Over the years, scientists have explored this field and have found different environmental factors that may attribute to the likelihood of NTDs. Some of these include fumonisin, valproic acid and more recently discovered, ceramide. To help counter NTDs, studies have shown that folic acid supplementation given to pregnant women has reduced the risk of NTDs and this has become a recommended suggestion by doctors. With its known preventative effects, this study aims to determine whether the preventative effects of folic acid can counter the harmful effects of fumonisin, valproic acid, or ceramide.
Faculty Advisor: Stark, Michael (Life Sciences, Physiological and Developmental Biology); Hansen, Marc (Life Sciences, Physiological and Developmental Biology)
Neural tube defects (NTDs), which result from failure to close the neural tube during embryonic development, are one of the most widespread and common congenital malformations. Variance in these malformations can range from anencephaly (failure of the neural tube to close on the cranial end) to spina bifida (failure of closure on the posterior/dorsal end). Over the years, scientists have explored this field and have found different environmental factors that may attribute to the likelihood of NTDs. Some of these include fumonisin, valproic acid and more recently discovered, ceramide. To help counter NTDs, studies have shown that folic acid supplementation given to pregnant women has reduced the risk of NTDs and this has become a recommended suggestion by doctors. With its known preventative effects, this study aims to determine whether the preventative effects of folic acid can counter the harmful effects of fumonisin, valproic acid, or ceramide.
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Understanding Drug Addiction Pathways Through Optogenetics
Bird, Devin; Nufer, Teresa; Wu, Bridget; Edwards, Jeffrey (Brigham Young University)
Faculty Advisor: Edwards, Jeffrey (Brigham Young University, Physiology and Developmental Biology)
Drug addiction is a consequence of neural plasticity in the ventral tegmental area (VTA), an area of the brain's reward system, in which higher levels of dopamine are expressed. Research suggests that decreased activity of inhibitory _-aminobutyric acid (GABA) neurons in the VTA could be the cause of increased activity of dopaminergic cells in the VTA, and thus mediate opiate addiction (Tan). However, not much additional research has been performed to evaluate the plasticity of VTA GABA neurons and the role they play in addiction. Why are VTA GABAergic cells being inhibited and how? We hypothesize that inhibitory inputs onto GABA neurons in the VTA directly affect the degree of inhibition of VTA dopaminergic cells. Additionally, we hypothesize that GABAergic neurons of the lateral hypothalamus (LH) is a source input that extends into the VTA and inhibits VTA GABAergic neurons. We believe that inhibition from these LH neurons induces plasticity of VTA GABAergic neurons.
Through the use of optogenetics we have been able to isolate precise GABAergic pathways that lead into the VTA. Specifically, we have isolated input sources from the LH. These optogenetic experiments, in combination with electrophysiology, have allowed us to measure the specific effects that LH GABA neurons have on VTA GABA neurons. Currently, our data suggests that LH GABAergic cells do induce long-term depression (LTD) in VTA GABAergic cells, however, it is too soon to make any conclusions. Although experiments are still underway, we believe that LH GABAergic neurons play an important role in the drug addiction pathway by inhibiting VTA GABAergic neurons and inducing plasticity.
Faculty Advisor: Edwards, Jeffrey (Brigham Young University, Physiology and Developmental Biology)
Drug addiction is a consequence of neural plasticity in the ventral tegmental area (VTA), an area of the brain's reward system, in which higher levels of dopamine are expressed. Research suggests that decreased activity of inhibitory _-aminobutyric acid (GABA) neurons in the VTA could be the cause of increased activity of dopaminergic cells in the VTA, and thus mediate opiate addiction (Tan). However, not much additional research has been performed to evaluate the plasticity of VTA GABA neurons and the role they play in addiction. Why are VTA GABAergic cells being inhibited and how? We hypothesize that inhibitory inputs onto GABA neurons in the VTA directly affect the degree of inhibition of VTA dopaminergic cells. Additionally, we hypothesize that GABAergic neurons of the lateral hypothalamus (LH) is a source input that extends into the VTA and inhibits VTA GABAergic neurons. We believe that inhibition from these LH neurons induces plasticity of VTA GABAergic neurons.
Through the use of optogenetics we have been able to isolate precise GABAergic pathways that lead into the VTA. Specifically, we have isolated input sources from the LH. These optogenetic experiments, in combination with electrophysiology, have allowed us to measure the specific effects that LH GABA neurons have on VTA GABA neurons. Currently, our data suggests that LH GABAergic cells do induce long-term depression (LTD) in VTA GABAergic cells, however, it is too soon to make any conclusions. Although experiments are still underway, we believe that LH GABAergic neurons play an important role in the drug addiction pathway by inhibiting VTA GABAergic neurons and inducing plasticity.
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Uncovering Data for Susceptible Populations: STR DNA findings on Male Rape Victims
Pugh, Sam; Valentine, Julie; Miles, Leslie (Brigham Young University)
Faculty Advisor: Valentine, Julie (Brigham Young University, College Of Nursing); Miles, Leslie (Brigham Young University, College of Nursing)
Rape is generally recognized as a sexual assault by a male perpetrator to a female victim. However, sexual assault is a crime that affects all genders. Although the majority of rapes are male to female, current findings indicate that one in seventy-one men will be raped in their lifetime. Over time, research has recognized the psychological effects and underreporting that ail male rape victims. However, very little has been reported regarding short tandem repeat (STR) DNA findings from sexual assault kits of male victim rapes. These STR DNA profiles prove to be highly influential in the detainment and prosecution of perpetrators. After an extensive search for earlier publications concerning the topic, only three articles were found to have relative correlation to this topic. Current best practice is to obtain STR DNA profiles from sexual assault kit samples to enter into the FBI Combined DNA Index System (CODIS). The purpose of this study was to evaluate DNA analysis findings from 266 sexual assault kits collected from male sexual assault victims and compare predictors for the development of CODIS-eligible STR DNA profiles of male victims to female victims. Our study methodology is an exploratory, retrospective design to identify male rape victims from a sample size of 5,758 victims who received sexual assault forensic examinations with sexual assault kit evidence collection. Approximately 5% of the victims in our study were male (N=266). Male victims were found to have more physical or mental impairments. Male victim cases revealed significantly less development of STR DNA profiles and CODIS-eligible DNA profiles of the perpetrator (p=.007). Due to low STR DNA profile yields and increased targeting of mentally impaired or otherwise vulnerable male victims, we must improve our response to male victims to ensure justice to all victims of sexual assault.
Faculty Advisor: Valentine, Julie (Brigham Young University, College Of Nursing); Miles, Leslie (Brigham Young University, College of Nursing)
Rape is generally recognized as a sexual assault by a male perpetrator to a female victim. However, sexual assault is a crime that affects all genders. Although the majority of rapes are male to female, current findings indicate that one in seventy-one men will be raped in their lifetime. Over time, research has recognized the psychological effects and underreporting that ail male rape victims. However, very little has been reported regarding short tandem repeat (STR) DNA findings from sexual assault kits of male victim rapes. These STR DNA profiles prove to be highly influential in the detainment and prosecution of perpetrators. After an extensive search for earlier publications concerning the topic, only three articles were found to have relative correlation to this topic. Current best practice is to obtain STR DNA profiles from sexual assault kit samples to enter into the FBI Combined DNA Index System (CODIS). The purpose of this study was to evaluate DNA analysis findings from 266 sexual assault kits collected from male sexual assault victims and compare predictors for the development of CODIS-eligible STR DNA profiles of male victims to female victims. Our study methodology is an exploratory, retrospective design to identify male rape victims from a sample size of 5,758 victims who received sexual assault forensic examinations with sexual assault kit evidence collection. Approximately 5% of the victims in our study were male (N=266). Male victims were found to have more physical or mental impairments. Male victim cases revealed significantly less development of STR DNA profiles and CODIS-eligible DNA profiles of the perpetrator (p=.007). Due to low STR DNA profile yields and increased targeting of mentally impaired or otherwise vulnerable male victims, we must improve our response to male victims to ensure justice to all victims of sexual assault.
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Using CRISPR and gRNA to Alter the HIV Genome
McRae, Elisa; Solis Leal, Antonio; Giler, Noemi; Karlinsey, Dalton; Quaye, Abraham; Berges, Bradford (Brigham Young University)
Faculty Advisor: Berges, Bradford (Brigham Young University, Microbiology and Molecular Biology)
HIV-1 infects CD4 T-cells by inserting its genome into a cell's genetic sequence. CRISPR technology allows for gene editing within the cell, causing a break in DNA sequences targeted by specific guide RNAs. Plasmids encoding CRISPR and guide RNA (gRNA) genes, in the context of lentiviral delivery vectors, will be transfected to produce two lentiviral vectors. In vitro experiments include human T cells that will be transduced with the lentiviral vectors and analyzed with flow cytometry to determine cells that express CRISPR and gRNAs. These cells will then be sorted to create a population of cells that express both the CRISPR and gRNA genes and will then be infected with the NL4-3 strain of HIV. For in vivo experiments, human hematopoietic stem cells will be transduced with the lentivirus vectors, after which they will be transplanted into humanized mice, thus producing a human-like immune system for testing the efficacy of our anti-HIV approach. After the human immune system has sufficiently developed in the mice, HIV-1 will be introduced. We expect that human immune cells with CRISPRs will be protected against HIV infection and death due to the use of gRNAs. These cells are postulated to no longer be susceptible to HIV-1 infection, thus preventing further cell lineages from becoming infected. We will analyze data for three main endpoints: 1. Cell killing of HIV, 2. HIV rebound due to the high mutation rate of the virus, 3. Amount of HIV replication, examined by assessing the viral RNA outside of cells using Q-RT-PCR. Data from this project will support whether cells transfected with CRISPR and guide RNAs offer cell lineages that adequately disrupt the HIV-1 genome. Efforts of this study hope to address HIV infection in humans following trials with humanized mice.
Faculty Advisor: Berges, Bradford (Brigham Young University, Microbiology and Molecular Biology)
HIV-1 infects CD4 T-cells by inserting its genome into a cell's genetic sequence. CRISPR technology allows for gene editing within the cell, causing a break in DNA sequences targeted by specific guide RNAs. Plasmids encoding CRISPR and guide RNA (gRNA) genes, in the context of lentiviral delivery vectors, will be transfected to produce two lentiviral vectors. In vitro experiments include human T cells that will be transduced with the lentiviral vectors and analyzed with flow cytometry to determine cells that express CRISPR and gRNAs. These cells will then be sorted to create a population of cells that express both the CRISPR and gRNA genes and will then be infected with the NL4-3 strain of HIV. For in vivo experiments, human hematopoietic stem cells will be transduced with the lentivirus vectors, after which they will be transplanted into humanized mice, thus producing a human-like immune system for testing the efficacy of our anti-HIV approach. After the human immune system has sufficiently developed in the mice, HIV-1 will be introduced. We expect that human immune cells with CRISPRs will be protected against HIV infection and death due to the use of gRNAs. These cells are postulated to no longer be susceptible to HIV-1 infection, thus preventing further cell lineages from becoming infected. We will analyze data for three main endpoints: 1. Cell killing of HIV, 2. HIV rebound due to the high mutation rate of the virus, 3. Amount of HIV replication, examined by assessing the viral RNA outside of cells using Q-RT-PCR. Data from this project will support whether cells transfected with CRISPR and guide RNAs offer cell lineages that adequately disrupt the HIV-1 genome. Efforts of this study hope to address HIV infection in humans following trials with humanized mice.
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A New Beginning
Mitchell, Emily (Weber State University)
Faculty Advisor: Arnold, Kristen (Engineering, Applied Science & Technology, Interior Design)
The design in this space will be used to keep the building fresh and calm with all of the changing that will be happening. This non-profit is here to help out women who are running from abusive relationships. Union Station is large enough so that they can house many different families or individuals. Giving them a space where they can get checkup when they are too afraid. There will be security in this locations so women will feel safe. (Aolain) There will be checkup rooms as well as therapy rooms where they can talk with someone private. There will be plenty of space to sleep for everyone that comes into this facility. To create a sense of home is to give them a space where they can have their own time and own space. (Falk, Wijk and Persson) There will be a small cafe and small store where they can buy small items for themselves or for their children. There will be classrooms for both the women and the children where everyone can learn to better themselves. Using the calming color of soft blue and soft green the space will be there for the women. (Instablogs.com) It will help with their fears of the unknown. The space will reflect the way that the women want with their time at the location. To be happy and healthy with themselves or their families. The space will have energy efficiency for this historical building. (Martinez-Molina, Tort-Ausina and Cho).
Faculty Advisor: Arnold, Kristen (Engineering, Applied Science & Technology, Interior Design)
The design in this space will be used to keep the building fresh and calm with all of the changing that will be happening. This non-profit is here to help out women who are running from abusive relationships. Union Station is large enough so that they can house many different families or individuals. Giving them a space where they can get checkup when they are too afraid. There will be security in this locations so women will feel safe. (Aolain) There will be checkup rooms as well as therapy rooms where they can talk with someone private. There will be plenty of space to sleep for everyone that comes into this facility. To create a sense of home is to give them a space where they can have their own time and own space. (Falk, Wijk and Persson) There will be a small cafe and small store where they can buy small items for themselves or for their children. There will be classrooms for both the women and the children where everyone can learn to better themselves. Using the calming color of soft blue and soft green the space will be there for the women. (Instablogs.com) It will help with their fears of the unknown. The space will reflect the way that the women want with their time at the location. To be happy and healthy with themselves or their families. The space will have energy efficiency for this historical building. (Martinez-Molina, Tort-Ausina and Cho).
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