Minority Health Disparities (MHD) ProgramCoordinator: Holly Lopez Instructors: Sponsors: The University of Arizona, The University of Arizona Graduate College, Western Alliance to Expand Student Opportunities (WASEO), Building Undergraduate Infrastructure Leading to Diversity: Southwest Consortium of Health-Oriented Education Leaders and Research Scholars(BUILDing SCHOLARS)-University of Texas, El Paso

Minority Health Disparities (MHD) ProgramRICHARD BOTKINRun-On Oligomerization Related to Sgr AI EnzymeThe University of Arizona, ChemistryMentor: Dr. Nancy Horton, Biochemistry

AbstractSgrAI, is a “rare-cutting” sequence specific type II restriction endonuclease (REase), which can recognize three degenerate primary octanucleotide sequences in duplex DNA, and unlike other REases, in the presence of activating primary site DNA, can cleave any of 14 additional secondary site DNA sequences. The unexpected enzyme behavior is due to the allosteric activation of 200- to 1000- fold by DNA containing a primary site sequence, leading to the expansion of its DNA sequence specificity. Previously, it has been shown that, in the presence of activator DNA, SgrAI forms an oligomeric structure with DNA bound SgrAI complexes arranged in a regular, repeating left-handed helical arrangement known as a run-on oligomer, which possesses the ability to grow to different sizes. Using the three-dimensional cryo-electron microscopy structure, mutations were designed to interrupt run-on oligomer formation, allowing for the testing of its role in vivo. SgrAI has the ability to protect host bacteria DNA from phage invasion. The effects of different mutants were tested using a phage challenge assay.

KIANA BURNETTThe Relationship Between Lipedema and Venoarterial ReflexUniversity of Texas-El Paso, Cellular Molecular BiochemistryMentor: Dr. Karen Herbst, Medicine

AbstractLipedema is an adipose tissue disease in which fluid develops in fat tissue making it painful and persistent; body mass index increases by Stage 1->3. It is hypothesized that lipedema is caused by leaking blood vessels that pool fluid into the lymphatic system though the mechanism of this behavior is unknown. The purpose of this project was to investigate the Venoarterial Reflex (VAR), a signal veins use to communicate with arteries to control blood flow throughout the body and prevent fluid leakage from blood vessels. The VAR was measured the arm and leg using the finger and toe. Data was collected at the Fat Disorders Research Conference in Dallas Texas April 27-29, 2018 from 47 women with lipedema, and 7 controls, with additional participants enrolling. We used an iphone app called iPhysioMeter to measure pulse volume. The VAR was calculated as the means of pulse volume of the dependent limb (down) over the means of the pulse volume of the neutral limb (up). VAR was considered normal if it was less than one

because pulse volume of a limb when it is down should be lower than when it is up. AVAR>1 was determined to be abnormal. The average VAR was normal in the hand among all the stages and controls. Among the feet the VAR increased as the stages increased. Stage 3 lipedema displayed abnormal average VAR in the feet (43% subjects had VAR>1), indicating that treatment of VAR may help reduce progression within and amongst stages.

ANGELICA ESCOTONF-kB in HER-Driven Cancers Using a Stapled EGFR PeptideNorthern Arizona University, Biological and Ecological SciencesMentor: Dr. Joyce Schroeder, Molecular and Cellular Biology

AbstractEGF has been shown to induce NF-kB activation, however the mechanism remains to be well understood. The Epidermal Growth Factor Receptor (EGFR; ERBB1, HER-1) is a protein receptor for the extracellular protein ligands belonging to the epidermal growth factor (EGF) family. Dimerization of these receptors can drive transformation and metastasis. It has been demonstrated that the uxtamembrane domain of EGFR controls various aspects of ERBB-dependent biology, including the facilitation of receptor dimerization. A hydrocarbon-stapled peptide, SAH5-EJ1, mimicking the internal juxtamembrane domain of EGFR has been shown to inhibit dimerization of ERBB1. Treatment with SAH5-EJ1 significantly suppresses EGF-induced autophosphorylation of ERBB1, promotes non-functional ERBB1 multimers, and suppresses downstream signaling pathways. Therefore, the inhibition of ERBB1 with SAH5-EJI and the subsequent downregulation of EGF-induced NF-kB activation may affect its role in inflammation and cell proliferation in inflammatory breast cancer.

AIBHLIN ESPARZAElav-GAL4 Gene Switch Drives TDP-43 Protein Expression in RU486 Induced Drosophila Model of ALSUniversity of Texas-El Paso, Mechanical EngineeringMentor: Dr. Daniela Zarnescu, Molecular and Cellular Biology

AbstractAmyotrophic Lateral Sclerosis (ALS) is a fatal progressive neurodegenerative disease that affects motor neurons. Although this disease is poorly understood, an aggregated protein, TAR DNA Binding Protein 43 (TDP-43), has been identified in familial and sporadic ALS patients. Various research models, such as mouse, zebrafish, Drosophila, and worms, have been used to better understand TDP-43 and its role in ALS. Drosophila, known as a fruit fly, is an inexpensive model, easy to obtain and maintain in laboratories, and shares similar metabolic pathways with humans. For this reason, Drosophila has been

used for the last century as a model for genetics. In this project, we use Elav-GAL4 Gene Switch System. This system works by crossing driver lines that express Elav with UAS-reporter lines with inserts of a target gene. With an activator present, Elav then attaches to the GAL4 binding sites and the target gene is expressed. In this experiment, the activator is the drug food (RU486). We created three genetic crosses with a white control group (w1118), human TDP-wild type (hTDPwt), and disease causing variant of TDP-43 (G298S9). By placing these genetic crosses in standard fly food for 5 days and collecting 1st and 2nd instar larvae and transferring them into varying concentrations of RU486 food for 2 days. After this exposure time, we collected larvae samples for Western Blot Analysis. This analysis demonstrated that exposure to higher concentration of RU486 resulted in greater TDP-43 expression, further indicating that Elav-GAL4 Gene Switch System successful induced TDP-43 protein.

MIGUEL FERMAINTDecrease in Gamma Oscillation Precedes Transient Dopamine Release OnsetThe University of Arizona, Neuroscience and Cognitive ScienceMentor: Dr. Stephen Cowen, Psychology

AbstractDopamine plays an important role in reward valuation, reinforcement learning, memory, and movement. Dopamine is a neurotransmitter produced by dopaminergic (DA) cells within the VTA. These cells project to the nucleus accumbens (NAc), which is known for its role in processing reinforcing stimuli. VTA activity is known to mediate dopamine release downstream in the NAc, however, it remains unclear what specific patterns of local-field potential activity facilitate dopamine release. To investigate how local-field oscillations contribute to dopamine release, we used a novel technology developed in the Cowen laboratory capable of measuring dopamine release and electrophysiology simultaneously. Because dopamine neurons are under the control of local GABAergic interneurons, it was hypothesized that a decrease in gamma-frequency oscillations (i.e., a decrease in GABAergic activity) would result in the disinhibition of dopamine cell activity and an increase in dopamine release. It was found there was a broad-band decrease in LFP frequency around the time of dopamine release events.

ASHLEY FLORESEffects of Sleep Disruption by Environmental Noise on Food Preference, Weight Gain, and Estrous Cycle in Female and Male Sprague-Dawley RatsThe University of Arizona, Neuroscience and Cognitive ScienceMentor: Dr. Jen Teske, Nutritional Science

AbstractNoise exposure reduces sleep and increases obesity. Women are more obese, more sensitive to noise, and have worse sleep quality than men. Consumption of a variety of palatable unhealthy foods stimulates calorie intake and weight gain in humans and rodents. In a rodent model of sleep disruption-induced obesity where male rats had ad libitum access to hedonic foods, noise-induced sleep disruption increased calorie intake from non-preferred hedonic foods. Preference for hedonic foods among sleep disrupted females and the effect of hedonic foods on the estrous cycle is unclear. Objective: To determine whether binge eating affects estrous cycle and sleep disruption affects preferred calorie intake in a rodent binge eating model. Methods: 3-month old Sprague-Dawley rats (n = 12 (male) and 20 (female) completed three study periods (staging, acclimation, and treatment). Staging: Rats were fed a standard diet (chow) for 9d. Acclimation: Rats had3h access to hedonic foods (Cheetos, Vienna Sausage, Sugar Cookie) daily plus chow for 9d. Treatment: Rats were randomized to sleep undisturbed or sleep disruption (environmental noise exposure, 8h/d during light cycle hen rats normally sleep) for 9d. Estrous cycle phase, bodyweight, and calorie intake were measured daily. Results: Binge eating hadno effect an impact on estrous cycle length. Bodyweight gain was lowest in proestrus. Males gained significantly more weight throughout the study. In this binge eating model, sleep disruption did not stimulate weight gain. Based on the last 3d of treatment, sleep disruption reduced intake of non-preferred foods and increased their intake of preferred foods.

ISABEL FORLASTROThe Type Six Secretion System in the Entomopathogenic Bacterium Xenorhabdus Bovienii Plays a Key Role in Bacterial CompetitionThe University of Arizona, Veterinary ScienceMentor: Dr. Patricia Stock, Animal and Comparative Biomedical Science

AbstractThe Type Six Secretion System (T6SS) is a molecular mechanism that is employed by multifarious gram-negative bacteria to inject effector proteins into a target cell. T6SSs have been shown to play key roles in pathogenesis with eukaryotic cells, and in competition with other bacteria. Xenorhabdus bovienii (XB) is an entomopathogenic gram-negative bacteria that exhibits an unusual dual-lifestyle: living in a mutualistic relationship within the gut of the Steinernema nematode, while also being pathogenic to a wide variety of

insects. Much is still unknown about the T6SS in XB. Therefore, we employed a homologous recombination approach to generate a XB mutant lacking hcp, a crucial structural gene of the T6SS, and conducted bioassays to assess the role of this secretion system. Specifically, we evaluated the virulence in competing wild-type, the hcp mutant, as well as an hcp complement with a “prey” bacteria. To determine if the T6SS plays a role in bacterial competition, we GFP labeled an XB strain that naturally lacks a T6SS (prey), mixed it with the XB wild-type or T6SS mutant (predators), and observed GFP fluorescence. While wild-type XB decreases fluorescence of the prey strain, the hcp mutant does not seem to have an affect on fluorescence. The re-introduction of the hcp gene in the complement reflects the affect of wild-type in that fluorescence diminishes. Quantifiable assays were also conducted with wild-type and two hcp mutants against the prey. CFU counts were taken after 24 hours of plating and the results show a significant decrease in the number of prey colonies growing when competed with wild-type, while a significant number of prey colonies survive during competition with the hcp mutants.

VALERIA GARCIAGPR68 MRNA Expression in Normal Rat TissuesNorthern Arizona University, Biological and Ecological SciencesMentor: Dr. Jason Yuan, Medicine

AbstractIdiopathic pulmonary arterial hypertension (IPAH) is a progressive and multifactorial disease which if left untreated usually culminates in right heart failure, and early mortality. Regardless of the initial pathogenic factors that may include genetic and/or environmental stimuli, pulmonary vascular remodeling, sustained pulmonary vasoconstriction, in situ thrombosis and increased pulmonary vascular wall stiffness are the major contributors to the elevated pulmonary vascular resistance (PVR) in patients with IPAH. Although the pathology of IPAH in the lungs is well known, the underlying cellular and molecular processes remain unclear. GPR68 is expressed in a subset of vascular endothelial cells that have a significant response to shear stress. GPR68 gene is believed to play a role in flow-mediated vasodilation, and is a critical sensor for blood flow. This research was based on the hypothesis that GPR68 gene is expressed in the lungs. In order to determine GPR68 mRNA expression, several rat tissues from two normal different rats were tested: brain, heart, kidney, liver, and lung. Polymerase chain reaction (PCR) technique was used to test this hypothesis. By isolating total RNA and converting it to cDNA we will be able to run a PCR in which its products will be visualized using gel electrophoresis. Findings showed that GPR68 mRNA expression in rats is tissue specific, meaning GRP68 gene is expressed at different levels in the different normal rat tissues. The highest GPR68 mRNA expression was detected in the brain and lung while the lowest expression was in the heart, kidney, and liver. With the data collected future experiments can be ran in order to determine GPR68 mRNA expression in human cells, such as pulmonary arterial endothelial cells and pulmonary arterial smooth muscle cells.

NIKOLE GOMEZInvestigation of Potential Fasciated Inflorescence Deletion Mutants in TomatoesSouthern Oregon University, Chemistry, ACS Concentration in BiochemistryMentor: Dr. Frans Tax, Molecular and Cellular Biology

AbstractIn higher plants, including Arabidopsis thaliana, the floral meristem gives rise to the plant’s floral organs and the fruits. In Arabidopsis, the wild type fruit has two compartments separated by the septum, but mutations in the Clavata (CLV) pathway can led to the proliferation of stem cells and more than two compartments. Errors in this pathway can lead to larger meristems and larger fruits which can be seen in fasciated inflorescence (FIN) mutants in tomatoes. FIN encodes an enzyme that adds sugar groups to CLV3-type peptides in tomatoes. Using PCR and various primers, three contiguous sections of the gene itself were amplified and the DNA bands were visualized using gel electrophoresis. Since previous research led the laboratory to believe that there was a deletion in the FIN gene, the site of interest was the middle of the gene. Costoluto Florentino, did not contain a deletion, and neither did 17 other strains. Two strains, Big Rainbow and Ananas Noire, had potential deletions of protein-coding parts of the gene (F2/R2 and/or F3/R3). More re-testing and more primers will be needed to confirm and characterize these potential deletions.

KEVIN HURTADOInvestigation of the Possible Off-Targets Effects of Blocking CRMP2 SUMOylation to Reduce NaV1.7 Currents in PainNorthern Arizona University, Biological and Ecological SciencesMentor: Dr. Raj Khanna, Medical Pharmacology

AbstractAbstract submitted for publication

CIARA LUGOReverse Osmosis Byproduct Management Through Membrane DistillationThe University of Arizona, Chemical Engineering and MathematicsMentor: Dr. Kerri Hickenbottom, Chemical and Environmental Engineering

AbstractAs the population increases, the availability of potable water decreases. Arizona is dependent upon the Colorado river for its water supply; however, a reservoir on the Colorado River, Lake Mead, is less than 40% full due to severe drought conditions [1]. Despite our current water crisis, Arizona has been considered an acclaimed leader in water management for over a decade [1]. One reason for that acclamation is the following proposed water purification process; the process goes as

such: waste water effluent goes through ultrafiltration (UF), then reverse osmosis (RO), then UV/advanced oxidation (UV/AO), then granular activated carbon (GAC) and lastly chlorine disinfection in order to become pure water [3]. The second stage, RO is an advanced membrane process that isolates dissolved solids, such as ions, from the water [4]. Though RO produces high quality water, its recovery ranges from 50% to 85% with most of the systems’ recovery at 75% [4]. The balance water (25% of water entered in most cases) that is not recovered - RO byproduct, as referenced above - is highly salinized water that needs to be managed, because it is does not meet water quality standards. In this study, we aim to filter the RO byproduct through a hybrid bacterially activated filtration (BAF) and membrane distillation (MD). Though the BAF system is crucial to this process, we are in the beginning stages of characterizing the MD system and plan on combining them to use the BAF system as a pre-treatment to MD. MD is a somewhat new process that is similar to distillation and RO in the ways that it is economical and conserves energy when being compared to standard separation processes; this is why it is being explored by scientists all over the world [2]. The overall goal of this project is to manage the concentrated brine by converting it into water that can be safely disposed, consumed or both. One obstacle that we have faced is scaling and fouling of the membrane being used. This is one of the reasons we plan to use BAF as a pre-treatment.

CELISA MARTINEZAnalysis of Mechanisms in an Isoproterenol Study Involving a Hematology Analyzer and Flow CytometryThe University of Arizona, MicrobiologyMentor: Dr. Richard Simpson, Nutritional Sciences

Abstract

Complete blood count (CBC) is one of the most common tests in both medical and clinical settings. By referencing the results, doctors and scientists can gather a comprehensive picture of health and immune status of an individual. This information can then be used to help make diagnoses or gauge clinical progression. A hematology analyzer can be utilized to obtain CBC, which includes White Blood Cells (WBC), Red Blood Cells (RBC), platelets, and differential counts1. Flow cytometry can also be used to obtain similar data; although using a different principle. Within this poster, a comparison between data obtained from a hematology analyzer and flow cytometer is presented within an isoproterenol study.

KAREN MOLINAA Comparative Analysis of Kidney and Pancreas Morphology in Diabetic and Non-Diabetic MinipigsWestmont College, BiologyMentor: Dr. Klearchos Papas and Dr. Leah Penrod, Surgery

AbstractType I Diabetes (T1D) is a chronic autoimmune condition in which insulin-producing β-cells are attacked and destroyed. As A result, insulin production becomes impaired and blood glucose levels cannot be sufficiently regulated. Individuals diagnosed with diabetes may also experience diabetic nephropathy. Understanding the effects on insulin-production and the complications associated with T1D has implications for future studies and will provide better treatment options for patients. Animal models mimicking diabetes are widely used to evaluate different treatment options. These models must be artificially induced with the drug streptozotocin (STZ) in order to create a diabetic model. Streptozotocin works through the glucose transport protein, GLUT2, and causes damage to the cell’s DNA. Side effects of using STZ in rodents and pigs include potential kidney and liver damage. However, a large enough dose must be administered to ensure pancreatic β-cells are destroyed. A diabetic pig model can be used to test treatment options, but also determine damage caused to the kidneys compared to non-diabetic pigs. The objective of this study was to analyze the morphological differences between diabetic and non-diabetic pig pancreas and kidneys using immunohistochemistry methods.

WENDY NUNEZT Cell-Mediated Post Menopausal HypertensionThe University of Arizona, BiologyMentor: Dr. Heddwen Brooks, Physiology

AbstractPre-menopausal women are protected from hypertension in comparison to age matched males. Interestingly, this protection is lost following menopause. Our laboratory uses the novel drug 4-vinylcyclohexene diepoxide (VCD) to induce menopause in mice and study how susceptibility to hypertension changes throughout the transition. The VCD model is a transitional ovarian failure model, uniquely preserving the pre-menopause, perimenopause, and post-menopause stages seen in human females. Recently, we have used this model to investigate the role of the adaptive immune system in mediating pre- and post-menopausal susceptibility to hypertension. Our lab has previously shown that male and female Rag-1-/-mice, which lack an adaptive immune system, are unable to develop Ang II-induced hypertension. However, following T cell adoptive transfer, both male and post-menopausal female mice show a robust

hypertensive response while pre-menopausal females remain protected. The purpose of this study was to investigate changes in blood pressure and renal inflammatory markers in both pre-and post-menopausal mice in response to Ang II infusion following the adoptive transfer of T cells from hypertensive donor mice.

MAGGIE ROMEROIn Vitro Evaluation of Metformin Hydrochloride Dose-Response on Transdermal, Nasal, and Buccal Human Cell LinesUniversity of Texas-El Paso, Biology: Biomedical Sciences EmphasisMentor: Dr. Heidi Mansour, Pharmacy Practice and Science

AbstractCell viability plays a major role in pharmaceutical studies considering the most effective drug delivery system in efforts to achieve desired dose response. Metformin Hydrochloride (HCl) is an oral antidiabetic drug widely used in the United States to treat type 2 diabetes mellitus. This study evaluates cell viability and dose-response of Metformin HCl on transdermal, nasal and buccal mammalian cell lines. Metformin HCl is characterized as having a low systemic bioavailability due to being challenged by the drugs’ high water solubility and its variable Intestina labsorption. The aim of this study was to provide an invitro evaluation to ultimately determine if the given cell lines may be proven effective in the absorption of Metformin HCl. Cell images were taken to reveal the morphology of each cell line before conducting a cell viability assessment utilizing a resazurin assay. In addition, Air-Liquid Interface Culture Condition (ALI) and Liquid Covered Culture Condition (LCC) systems were performed to mimic human epithelial cellular absorption in RPMI2650 nasal epithelium cell line. ALI and LCC conditions were compared to determine if conditions resulted more significantly reliable in determining cell viability in RPMI 2650 cell line. Among the cell lines reviewed in this study, all resulted promising with viability values ranging from 87.76% to 144.36% within the drug concentrations delivered into each cell line. Consequently, with the transdermal, nasal, and buccal cell lines resulting in high cell viability and effective dose-response treated with Metformin HCl, may allow for future developmental formulation studies to treat hyperinsulinemia derived cancers.

CESAR SALAZARUsing Fruit Flies and Lymphoblastoid Cells to Study RNA Binding Protein Interactions in ALSThe University of Arizona, Neuroscience and Cognitive ScienceMentor: Dr. Daniela Zarnescu, Molecular and Cellular Biology

Abstract

AYESHA SWANNIGANEngineering Cre Recombinase Secreting Toxoplasma to Study Toxoplasma-Neuron InteractionsThe University of Arizona, Neuroscience and Cognitive ScienceMentor: Dr. Anita Koshy, Neurology

AbstractToxoplasma gondii is an intracellular parasite that most often infects, and persists, in the central nervous system (CNS).1,2 In the CNS, Toxoplasma almost exclusively persists in neurons. Dogma holds that parasites persist in neurons because neurons are not able to mount the innate immune responses that non-neuronal cells use to clear intracellular parasites. The Koshy lab’s recent work using a system that allows one to track host cells injected with parasite proteins in vivo questions this dogma. This system relies upon the fact that Toxoplasma injects proteins into host cells prior to invasion. Thus, by engineering a strain of Toxoplasma (Pru-TCre) to have Cre recombinase fused to toxofilin, one of these seCreted proteins, Cre is injected into host cells. Using these Pru-TCre parasites to infect transgenic mice that express green fluorescent protein (GFP) only after Cre mediated recombination revealed that the majority of GFP+ (injected) neurons did not harbor an active infection. This leads to the question of whether neurons are capable of clearing Toxoplasma. Here we have further refined the system to address this question.

ALEJANDRO VILLALOBOSExpression and Purification of TEV ProteaseThe University of Arizona, BiochemistryMentor: Dr. Wolfgang Peti, Chemistry & Biochemistry - Med

AbstractTobacco etch virus (TEV) protease is a member of the potyviridae virus family that infects a variety of plants, fruits, and vegetables. The major mediators of the virus are known as picornaviral 3C type proteases. In addition to them being virus mediators, these proteases can serve as enzymatic proteins that can cleave affinity tags from recombinant proteins so that the tags do not interfere with further protein studies.

TEV’s structure is composed of two β-sheets, β1 and β2. These β-sheets organize the active site (or catalytic triad) that allow for substrate recruitment and catalysis. The catalytic triad relies on an asparagine residue in β1, and a histidine and cysteine residue in β2. Cysteine is the amino acid responsible for the nucleophilic attack towards the peptide substrate. In addition, TEV is also highly specific. TEV most commonly identifies the sequence, ENLYFQS and cleaves between Q and S. This method is often used in the Peti Lab to cleave affinity tags, such as His6-tags, to study the protein without the interference of a purification tag. My responsibility was to learn how to express and purify recombinant protein, especially TEV protease. The production of the TEV protease will facilitate individuals in the lab to cleave affinity tags, fused proteins, and advance their research in protein study. Thus, activity tests had to be performed to ensure that only active protein was produced by me.