2020 Abstracts

Wasden, Kayla; Suisse, David; Kaundal, Amita (faculty mentor) (Utah State University)
Faculty Advisor: Kaundal, Amita (College of Agriculture and Applied Sciences; Plants, Soils, and Climate Department)

Many plants secrete substances to create a more favorable environment, including chemicals that kill pathogenic microbes or competing plants. Artemisia tridentata, also known as "Big Sagebrush," is prevalent in the Rocky Mountain region of the United States and is known to have antimicrobial capabilities. We will study the potential antimicrobial activity of Artemisia tridentata.

Studies report that chemicals released by the leaves and branches of A. tridentata affect bacteria native to deer rumen. Another study showed that 27 actinomycetes (anaerobic bacteria that form colonies) strains found in the rhizosphere of A. tridentata demonstrated antibacterial activities when tested on E. coli, Bacillus subtilis and Staphylococcus aureus. Native Americans traditionally used A. tridentata to relieve stomach pain, colds, coughs, sore eyes, snake bites and as an insect repellent. Researchers found several compounds, including flavonoids, that can affect antimicrobial activity. Articles regarding antimicrobial activities in A. tridentata were published between 1967 and 2004. With the chronological gaps and considering the progress that biological and molecular technology has made in recent years, knowledge of the chemicals released by A. tridentata lies largely untapped. In this study, we will investigate the antimicrobial activities of the leaves, stem, roots, and flowers of A. tridentata initially by the agar well diffusion method and followed by validating with the agar disk diffusion method. We will check the antimicrobial activity of the extract from different plant parts of A. tridentata on common bacteria such as E. coli, Bacillus subtilis, and some Pseudomonas spp. of plant pathogens.

The knowledge obtained from this research will further help in the identification and characterization of the secondary metabolites or chemicals involved in antimicrobial activity of sagebrush. Medicinal plants provide a healthy, natural alternative to conventional medication, and may lead to new insights on antibiotics and pharmaceuticals. Besides, Artemisia tridentata is a plant native to Utah and Idaho. It grows everywhere in the surrounding area, making it inexpensive (free) to produce.

Baptista, Gabriela; Payne, Andrew; Obray, J Daniel; Yorgason, Jordan; Weber, K Scott; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences, Psychology)

Cluster of differentiation 5 (CD5) is expressed in both T and B cells. CD5 has been found to display an altered expression profile following chronic ethanol use and during ethanol withdrawal. Specifically, the number of CD5+ B cells is reduced during withdrawal while the number of T cells is increased. Given the apparent sensitivity of these cells to ethanol and recent research suggesting that some ethanol effects are accounted for by neuroimmune interactions we assessed drinking behavior and ethanol induced sedation in CD5 knockout (KO) mice. We found that CD5 KO mice display decreased ethanol consumption as compared with wild-type controls and that ethanol consumption does not increase with repeated exposure in CD5 KO mice. Additionally, CD5 KO mice displayed considerable resistance to the sedating effects of ethanol. Further studies are underway to assess whether there are baseline differences in dopamine dynamics within the mesolimbic pathway between CD5 KO mice and wild-type controls as well as to whether neurons in the mesolimbic pathway differ in their response to ethanol in CD5 KO mice.

Ross, Mimi; Tessem, Jeffery; Orton, Emily; Ekpo, Idongesit; Beales, Joseph (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Life Sciences; Nutrition, Dietetics, & Food Science)

In 2015 there were over 30 million Americans with diabetes and over 84 million Americans ages 18 and older had pre-diabetes. With diabetes being the seventh leading cause of death in the United States and becoming more prevalent the race is on to find a cure. One of the main problems with this disease is the decrease in functional β-cell mass. β-cells produce insulin to maintain blood glucose levels at healthy levels. Thus, if we can increase β-cell proliferation we are one step closer to curing diabetes. Cocoa epicatechins have been shown to be beneficial in blocking diabetes progression. Studies have shown that oligomeric and polymeric cocoa epicatechin extracts improve diabetes onset in a mouse model of Type 2 diabetes. We have demonstrated that the oligomeric fraction of cocoa epicatechins enhances β-cell proliferation in an in vitro model. Absorption studies have shown that while the oligomeric and polymeric forms are not readily absorbed in the gut, they are metabolized by gut bacteria and that these metabolites can be observed in circulation. Using flow cytometry we have studied how these phytochemicals: epicatechin, 5-phenylvaleric acid, Homovanilic acid, and Hippuric acid. Here we present the data regarding the effect of microbial cocoa flavanol metabolites on β-cell cell cycle during proliferation.

Bartlett, Ashley; Phatak, Sumira; Hintze, Korry; Benninghoff, Abby (Utah State University)
Faculty Advisor: Benninghoff, Abby (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)

The gut microbiome modulates various physiological functions related to cancer development including inflammation, cell proliferation, apoptosis, and angiogenesis. Patients with inflammatory bowel disease have a microbiome distinct from healthy controls with consistent observations of reduced gut biomass, decreased diversity within the community, and altered relative abundance. Although a consensus cancer-related microbiome has not been identified, several pathogenic species play an instrumental role in the progression of colitis and tumorigenesis, including: Streptococcus bovis, Helicobacter pylori, Enterococcus faecalis, Clostridium septicum, and Escherichia coli. Gut microbial composition is highly responsive to diet and inadequate intake of micronutrients is a critical feature of the Western dietary pattern. Gut dysbiosis has been proposed to further limit mineral uptake and impair vitamin synthesis, predisposing the host to micronutrient deficiency. Dietary bioactives, such as those in green tea, may function as a mediator between the gut microbiome and basal diet to ultimately prevent colitis associated colorectal cancer (CAC). The overarching objective of our work is to determine the impact of ancestral or multi-generational consumption of the total Western diet (TWD) in a murine model of CAC. Our previous work is the first to investigate how diet induced transgenerational inheritance affects CAC outcome. Our data suggested that multigenerational patterns of exposure to the TWD altered both phenotype and gene expression in third generation offspring. Supplementation with green tea appeared to be most promising after consumption of TWD for multiple generations. Considering that gut microbes are inherited maternally after colonization during vaginal birth, the gut microbiome is a missing piece in this disease model puzzle. The hypothesis of our current project is to investigate whether intake of TWD influences the transmission of microbes and whether CAC outcome is reflected by altered gut microbial composition. Based on other work, we expect the healthy control to possess an abundance of varied bacterial taxa that maintain protective epithelial barrier function and overall homeostasis. On the other hand, a high fat diet would promote increased intestinal permeability, a substantial shift at the phyla level, and increased production of pro-inflammatory cytokines. After TWD consumption, we expect an overall negative phenotypic outcome within the gut microbiome, that includes a breakdown of the epithelial barrier and introduction of pathogenic bacteria. These harmful bacteria tend to thrive on simple sugars that are common in the Western dietary pattern and tend to produce metabolites known as endotoxins that promote dysbiosis.

Anderson, Lars; Baldwin, Haley; Christensen, Ben; Walker, Austen (Brigham Young University)
Faculty Advisor: Griffen, Blaine (Brigham Young University, Life Sciences)

This research looks at the blue coloration on uca pugnax crab carapace above the mouth and between the eyestalks and associates the coloration to the behavior, sexual maturity, and size of the crab, as well as the detection of metals in their environment. Up to ten crabs were photographed within twenty five isolated sites with the objective of gathering a high range of color difference among the uca pugnax. The photos of the crabs were set to match the same scale of light and RGB as to not have interference from external factors such as sunlight or overcast weather. The shade of blue on the carapace provides information about the surrounding environment where the uca pugnax are found.

Sagers, Rachel; Nguyen, Andy; Weston, Jake; Grooms, Nicholas; Eggleston, Morgan; Martin, Randy; Coulombe, Roger (Utah State University)
Faculty Advisor: Coulombe, Roger (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)

The scenic mountain views of Cache Valley in Northern Utah stand in stark contrast with the valley's high concentrations of fine particulate air pollution (PM2.5), some of the worst reported in the United States. The unique geography promotes formation of ammonium nitrate (NH4NO3) from nitrogen oxides produced by motor vehicles and ammonia from dairy cow excreta. Winter atmospheric inversions, exacerbated by the mountainous terrain, trap and concentrate air pollutants. Epidemiological studies have revealed an association between PM exposure and early all-cause mortality. Exposure to PM2.5 is also associated with a variety of cardiovascular, cardiopulmonary, and neurodegenerative diseases, including myocardial infarction, stroke, COPD, lung cancer, Alzheimer's disease, and Parkinson's disease. Previous studies have shown that Cache Valley PM (CVPM) has pro-inflammatory effects, which has been linked to enhanced activation of Akt in human pulmonary epithelial cells. This research examined the cellular responses of human lung (BEAS-2B) cells exposed to CVPM and diesel exhaust particles (DEP), at 1 and 12 µg/ml concentrations of each particle type for a 24 hour exposure period. The CVPM used was collected onto stainless steel plates by a Tisch impactor. Assessment by the comet assay reveal genetic damage to CVPM exposed cells with equal potency to DEP exposed cells. Flow cytometry (p < 0.05) showed CVPM exposed cells had a significant increase in the number of actively-dividing cells compared to control cells. Whole-genome microarray identified affected genes related to inflammatory pathways, as well as activated Akt-dependent pathways. Subsequent qRT-PCR showed that CVPM exposure significantly increased expression of inflammatory markers, including IL-6, CD40LG, PLAG27, and cytochrome P450 (CYP) 1A1 (p < 0.05). Immunoblotting confirmed activation of Akt by phosphorylation of Thr308 in both CVPM and DEP exposed cells. This data supports the hypothesis that CVPM may induce pro-carcinogenic pathways with potency similar to DEP.

Littlefield, Connor; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University, NDFS)

The transcription factor Nkx6.1 is essential for beta cell growth and function. Given that Nkx6.1 is expressed in beta cells undergoing high level expansion, our lab demonstrated that Nkx6.1 overexpression in primary rat islets was sufficient to induce beta cell proliferation and enhance glucose stimulated insulin secretion. However, while these phenotypes are evident in islets from young animals, islets from aged animals fail to induce proliferation or increased insulin secretion. One reason for why Nkx6.1 fails to drive proliferation or increase insulin secretion is due to lost binding partners that allow it to control gene transcription. We hypothesize that loss of Nkx6.1 binding partners curtails its ability to induce gene transcription that leads to proliferation and enhanced glucose stimulated insulin secretion. To test this hypothesis we have used Nkx6.1 BioID to define by mass spectrometry the proteins that interact with Nkx6.1 Here we define three novel interactors, Mef2D, Sirt7, PDX1. This finding will provide us with a greater understanding of Nkx6.1 function in the beta cell, provide us with new gene targets essential for Nkx6.1 function, and allow us to begin to apply these findings to aged beta cells.

Wood, Branzen; Oberg, Taylor; Culumber, Michele; Oberg, Craig (Weber State University)
Faculty Advisor: Oberg, Taylor (Utah State University, Nutrition and Food Science); Culumber, Michele (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology)

The unique flavorings and textures of Cheddar cheese are produced by the degradation of the major milk proteins. One of those proteins, casein, is degraded by the enzyme chymosin and a series of peptidases produced by the starter Lactococcus added to the milk. As casein is degraded, several small peptides accumulate. One of these peptides, ß-casein, can have an adverse bitter taste that is non-desirable and considered a defect in Cheddar cheese. The two main starter cultures used industrially in Cheddar cheese making are Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris. L. lactis subsp. cremoris has been used traditionally in Cheddar cheese making, however, L. lactis subsp. lactis ferments more quickly and is becoming more popular in the cheese industry. With the transition creameries have seen a sharp rise in bitterness during production. Our hypothesis was that while closely related, cremoris synthesizes some peptidases that help with ß-casein degradation that lactis does not. Peptidases found in cremoris include PrtP I and II, Pep X, Pep C, Pep A, Pep T, Pep Q, Pep N, Pep V among others. We searched the genomes of both strains using RAST bioinformatic software, and the databases NCBI and UniProt. The peptidases common in cremoris were also found in lactis. We are now trying to determine if the location of the peptidases on the genomes change how they are regulated or produced. Further, we will begin looking into the genome for other, novel, enzymes that might have peptidase activity that influence bitterness.

Jones, Abigail; Hevel, Joan (Utah State University)
Faculty Advisor: Hevel, Joan (College of Science, Chemistry and Biochemistry Department)

PRMT1 is one of nine known mammalian Protein Arginine Methyltransferases (PRMTs) whose function are to transfer methyl groups from S-adenosyl methionine (SAM) to arginine residues of specific proteins. PRMT1 is known to methylate many different proteins in cells, but the mechanism of target recognition and binding is still unknown. Correct regulation of PRMT1 is critical to proper cellular function; thus, the action of PRMT1 is important to understand. In this study, we seek to elucidate how PRMT1 recognizes and binds its targets by identifying protein substrates of PRMT1 that form a stable complex with the enzyme. Such a protein would allow for additional studies (e.g. crystallographic or cryo-EM studies) to help visualize PRMT1-substrate interactions. Two substrates of PRMT1, TWIST1 and Smad6, have been purified, and the binding affinity of each to PRMT1 has been qualitatively assessed via pull-down assay and Western blot. Ligation-independent-cloning has been used to clone each substrate gene out of a GST-tagged vector and into a His-tagged vector, which will allow for further experiments assessing the stoichiometry of PRMT1-substrate binding.

Hirschi, Blake; Pickett, Brad; Thompson, Jared; Telford, Mady; Berges, Bradford (Brigham Young University)
Faculty Advisor: Berges, Brad (Life Sciences, Microbiology and Molecular Biology)

Staphylococcus aureus (S. aureus) is a commensal bacterium commonly found amongst livestock and near 30% of humans' nostrils. However, through acquisition of certain genes S. aureus may develop antibiotic resistance such as in methicillin-resistant Staphylococcus aureus (MRSA). One hypothesized component lending to acquisition of genetic resistance in S. aureus is the synthesis of colony biofilms. Biofilms are comprised of a variety of substances including secreted polysaccharides, protein and even extracellular DNA. Our work postulates that extracellular DNA-based biofilms will transfer genes for antibiotic resistance at a higher rate than in polysaccharide/protein biofilms. Through employment of polymerase chain reaction (PCR), we aim to characterize a wide sample of methicillin-susceptible S. aureus (MSSA) human associated strains and MRSA livestock associated strains for multiple antibiotic resistances. Co-inoculating pairs of human associated and livestock associated strains, each lacking the other's resistance genes, will provide an environment wherein biofilm-mediated gene transfer may occur. Further pairing based on biofilm composition (DNA or polysaccharide/protein) will yield data concerning which biofilm facilitates gene transfer more efficiently. Subsequent genotyping will confirm whether resulting isolates acquired new antibiotic resistance through biofilm-mediated transfer, thus increasing pathogenicity.

Tuttle, Jared; Payne, Andrew; Obray, J Daniel; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)

A subpopulation of ventral tegmental area (VTA) GABA neurons express connexin-36 (Cx36) gap junctions (GJs). Activation of GJ-mediated electrical coupling between VTA GABA neurons supports brain stimulation reward and alcohol reward is lowered in Cx36 KO mice due to a hyper-dopamine (DA) state. The aim of this study was to further evaluate the role of a subpopulation of Cx36+ VTA GABA neurons in alcohol reward and dependence. To accomplish this study, we customized a Gq-coupled Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) viral vector to only express in Cx36+ neurons (AAV8.hCx36.hM3D(Gq)-mCherry.WPRE.rBG) in the VTA. The hM3Dq viral vector was infused into male CD-1 GAD GFP mice and male Wistar rats. The animals were then given 10-14 days to recover prior to experimentation. A control virus (AAV9.CB7.CI.mCherry.WPRE.rBG) was used for comparison. We implemented standard cell-attached mode electrophysiology to evaluate the effects of clozapine-n-oxide (CNO; the ligand for DREADDs) on VTA GABA and DA neuronal activity. We found a robust enhancement of VTA GABA neuron firing rate in hM3Dq+ neurons with 20 _M CNO ex vivo. Surprisingly, while investigating CNO effects on VTA DA neuron firing rate, we found that CNO activation of hM3Dq+ VTA GABA neurons increased DA neuron activity, suggesting that Cx36+ VTA GABA neurons indirectly modulate local VTA DA neurons. Intraperitoneal CNO (3 mg/kg) also enhanced the firing rate of VTA GABA neurons in vivo. Administration of CNO reduced ethanol consumption (drink-in-the-dark paradigm) in both ethanol naïve and ethanol dependent hM3Dq-injected mice as compared to controls, suggesting that activation of Cx36+ neurons in the VTA is enough to block ethanol consumption in both naïve and dependent animals. Taken together, these findings support previous studies indicating that enhanced electrical coupling between VTA GABA neurons is rewarding and promotes reward and lowers the hedonic value of ethanol.

Aitken, Talon; Jensen, Daelin; Baxter, Melanie (Brigham Young University)
Faculty Advisor: Tessem, Jeffrey (Brigham Young University, NDFS)

Diabetes Mellitus, a condition characterized by hyperglycemia resulting from defects in insulin secretion or effectiveness, affects over 8.5% of the adult US population. Both type one and type two diabetes have the common characteristic of a decrease of functional beta-cell mass from the islets of Langerhans, located within the pancreas. The upregulation of genes known to induce beta-cell growth and proliferation results in an increase of functional beta-cell mass in young cells but not in their aged counterparts. This age-related occurrence - under nonpathologic conditions — is poorly understood. For this study, the morphological differences between young islets and aged islets are studied to provide insight as to the reason behind this refractory behavior. Immunostaining methods show significant contrast been percentages of insulin-positive beta-cell area in the pancreata of young vs. old-aged rats.

Miller, Jacob; Cheung, Aaron; Novilla, Kirsten; Crandall, Aliceann (Brigham Young University)
Faculty Advisor: Crandall, Aliceann (Life Sciences, Public Health)

Existing literature demonstrates a strong relationship between childhood experiences and adult health outcomes. The Differential Susceptibility to Environment Theory suggests that there are several factors, including personality and physiology, that effect a child's sensitivity to adverse and advantageous experiences. A sample of 246 adults (ages 19-57) were asked questions about extroverted personality characteristics, adverse and advantageous childhood experiences, and several measures of adult health, including executive functioning, perceived stress levels, depression, and past smoking habits. The sample was then stratified based on level of extroversion scores with the top quartile being labeled as "extroverts", the bottom quartile as "introverts", and those in between as "ambiverts". Regression analyses were then used to assess the relationship between childhood experiences and each adult health outcome. The results of the study showed that the extroverted individuals experienced more positive health outcomes after more advantageous childhood experiences, as well as decreases in adult health outcomes after more adverse childhood experiences. These results suggest that extroverts more than introverts are more sensitivity to environmental influences in childhood. More research is needed to understand the neurobiological mechanisms that increase environmental sensitivity among extroverts.

Burgoyne, Jake; Leavitt, Steve (Brigham Young University)
Faculty Advisor: Leavitt, Steve (Life Sciences, Biology)

In show caves, artificially lighting is intended to highlight intricate cave formations for visitors. However, as an unintended consequence, artificial lighting promotes the growth of diverse biofilm communities termed Lampenflora that gain their energy from these novel light sources. Lampenflora, which generally consist of algae and cyanobacteria, discolor formations and introduce novel ecological interactions in simple cave ecosystems. Lampenflora communities have been understudied mainly due to technological limitations and difficult accessibility. However, by characterizing these communities, we can better monitor their impact and develop effective strategies for their removal. Using metagenomic high-throughput sequencing, this research provides the first molecular-based perspective into lampenflora diversity in cave systems in the Great Basin. The data collected, generated, and analyzed is vital in understanding Lampenflora biodiversity and how these communities develop. Furthermore, it offers ecologists a novel perspective on the use molecular detection to understand biodiversity within cave systems.

Marshman, Evan; Peterson, Samara; Call, Gerald; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Science, Plant and Wildlife Science)

In recent years the association between the human gut microbiome and the brain has become a promising field of study. Often referred to as the "gut-brain axis", this connection has greatly enriched our scientific understanding of many disorders that affect the brain and nervous system. A recent study showed the differing richness of bacteria in the microbiota of Parkinson's patients and healthy control subjects. Because recent research shows this connection, we predicted that we would detect variation in the microbiota of D. melanogaster (fruit flies) models of Parkinson's disease, relative to wild type flies. To test this hypothesis, I analyzed 16s rRNA sequence data, reporting the microbiota composition in flies that are a model of Parkinson's Disease, as well as wild type flies. I found one strain of the genus Acetobacter that was differentially abundant between the two fly types. Therefore, for my CURA I will extend my analysis by performing similar analyses by taking a larger set of Parkinson's fly models. Once they are sequenced, I will use QIIME, the same software I used in my preliminary analyses to further our understanding of the taxonomic differences between the gut bacteria of Parkinson's models and wild type flies.

Kaneshiro, Alma; Jordan, Adam; Crompton, Rhees; Brailsford, Samantha; Spencer, Jonathan (Weber State University)
Faculty Advisor: Clark, Daniel (Science, Microbiology Department and Neuroscience Center); Chaston, John (Life Sciences, Plant & Wildlife Sciences)

Antibiotic resistance is of great concern in the medical community, with bacterial resistance increasing proportional to their use. Staphylococcus aureus, such as methicillin resistant S. aureus (MRSA), can cause fatal infections. Problems due to this resistance are compounded when the infecting bacteria form a biofilm, thick sticky layers of bacterial secretions, which are difficult for antibiotics to penetrate. Biofilm formation is common in hospital settings on stents, catheters, and IV lines. Biofilms make antibiotic treatment risky due to incomplete killing—the most resistant survive exposure. There is evidence that bacteriophage can break up biofilms, possibly making them more susceptible to antibiotics. We induced a S. aureus biofilm formation using chemicals that mimic a skin wound. Using bacteriophage K, we inoculated the biofilm and observed clearance. Samples of cell pellets and liquid supernatant were collected, and DNA was extracted. Real-time PCR was used to quantify the levels of bacteriophage K replication, representing clearance of the bacteria. This research can be used to find efficient ways to treat an infection caused by a S. aureus biofilm. Bacteriophage used in combination with antibiotics may be able to better clear a biofilm infection and reduce antibiotic resistance risk due to more complete infection clearance.

Townsend, Jessica; Freitas, Claudia; Weber, Scott; Cardon, Dallin (Brigham Young University)
Faculty Advisor: Weber, Scott (Brigham Young University / Life Sciences, Microbiology and Molecular Biology)

The World Health Organization reported in 2016 that oral diseases affected half of the world's population. Oral diseases are due to poor oral hygiene and tobacco use which can develop into periodontal disease. Periodontal disease is caused by an immune response to microbial challenge, which initiates an invasion of lymphocytes and other single-nucleated cells to the site of inflammation in the mouth that can cause tooth loss and is a risk factor for heart and lung disease. Patients with severe periodontitis have increased auto-reactive B lymphocytes that express the CD5 co-receptor and these cells are influenced by T cells. We propose to investigate the relationship between oral inflammation, CD5, and the T helper immune response. This will be done by comparing oral inflammation in mice with and without CD5. CD5 is a T cell co-receptor that regulates T cell development and function and we hypothesize CD5 plays an important role in periodontal disease. We will test this hypothesis by co-culturing T cells expressing or lacking CD5 with oral mucosal or gingival epithelial cells that have been exposed to LPS (lipopolysaccharide, a major component of gram-negative bacteria's wall) and will exam differences in cell number, T cell subtype, and cell function.

Spruance, Lori; Augustine, Madi (Brigham Young University)
Faculty Advisor: Spruance, Lori (Life Sciences, Public Health)

Youth sport programs are an opportunity to increase physical activity, but the food environment may be detrimental to improving and maintaining health. From a previous study, parents indicated that they would like guidance and direction in a top-down approach from coaches and administrators; yet, understanding the administrator experience relative to the youth sports food environment remains unclear. The purpose of this study is to understand that experience. Semi-structured qualitative interviews will take place with administrators across the state of Utah. Interviews will be recorded and transcribed. Thematic analysis will be conducted to identify salient themes. A peer-reviewed publication and multiple presentations will result from the study conducted.