Microbiology Commons^™

Ebi2, Rodney Till, Dr. Brian Poole

Journal of Undergraduate Research

My project this year was a great learning experience and helped me to better understand the complexity of lab work, as well as, the tools used in troubleshooting problems; including collaboration with lab researchers and professors. I learned about flow cytometry and how to analyze migration data. I also learned how to do transfections with HEK cells and how to separate B cells from a fresh sample of blood. I was thankful that I could spend time doing lab work because it’s the best teaching environment I know of in academia. I came across genuine problems and studied topics with …

Examination Of The Role Of The Lana Protein In Persistence Of Kaposi’S Sarcoma-Associated Herpesvirus Infections, Marshall Sheide, Dr. Bradford Berges

Journal of Undergraduate Research

Kaposi’s sarcoma herpes virus (KSHV) is one of seven known cancer causing viruses which have the capacity to persist in the host for many years. This particular virus can undergo a long latent infection which ensures that the virus can hide by not expressing its genes. Last year, we proposed an experiment that would show the importance of a particular gene in the viral genome, LANA, in establishing a latent infection. The experiment required us to create stocks of a mutant virus that did not have the LANA gene inside (bac36deltaLANA) and then infect humanized mice with it. After infection, …

Search For Mitochondrial Dna Mutation In A Family With Myopathies And Lactic Acidosis, Mary Rennick, Dr. Brent Nielsen

Journal of Undergraduate Research

This research focused on the search for the probable mitochondrial DNA (mtDNA) mutation in one family affected with fibromyalgia and lactic acidosis. The hypothesis was that there is mixture of normal and mutated mtDNA, called heteroplasmy. I first obtained the four patients’ consent to participate in the study. The four patients were between the age of 19 and 25 years-­‐two males and two females. They donated urine samples to be used for DNA analysis. I isolated DNA using a Qiagen DNA extraction kit and obtained concentration measurements of 8-­‐ 10 nanograms per microliter using the nanodrop spectrophotometer in the RIC …

Antibody Production And Diversification In Humanized Mice, Stanton Nielsen, Dr. Brad Berges

Journal of Undergraduate Research

Many scientists are skeptical of using humanized mice to study the human adaptive immune response. The reasoning behind this skepticism stems from discrepancies in various studies seen previously. A study conducted by Traggiai et al. showed that humanized mice elicited a human adaptive immune response when challenged with the tetanus toxin (Traggiai). In addition, good antibody responses have been shown in humanized mice when challenged with Dengue virus (Kuruvilla). However, others have found only rare antibody responses to HIV when humanized mice were challenged (Baenziger). As mentioned earlier, a study conducted by Becker et al. showed the production of human …

Vitamin D3 As An Adjuvant To Produce Localized Immunity, Kaitlyn Vance, Dr. Eric Wilson

Journal of Undergraduate Research

The aim of our research was to help elucidate the degree of specificity employed by antibody secreting cells (lymphocytes) through a study of the role of CCR10 in the differential homing of these cells to mucosal issues. We used vitamin D3, known to upregulate expression of CCR10 on activated lymphocytes, as an adjuvant in our vaccines and tracked cell homing to mucosal tissues that express the CCR10 ligand CCL28 by detecting antigen specific antibody accumulation. Since mucosal tissues are ideal locations for vaccinations, the long-term aim was to develop a method for targeting antigen specific lymphocytes, and thus robust immune …

Characterizing Proteins That Interact With Pas Kinase, Jordan Mackay, Dr. Julianne Grose

Journal of Undergraduate Research

Characterizing the interacting protein partners of Per-Arnt-Sim (PAS) kinase has proven to be a formidable task. Since the beginning of 2012, I have been working hard with graduate student Desi DeMille on confirming the putative interactors uncovered by genetic screening done the previous year. This endeavor has yielded both successes and failures.

How ‘Bout Them Apples?: A Study On Antioxidants, Kylie Measom, Dr. Kim O'Neill

Journal of Undergraduate Research

They say “an apple a day keeps the doctor away.” Apples are laden with antioxidants which have been shown to prevent against cancer and other illnesses.¹ Antioxidants are molecules that prevent cancer by collecting excess free radicals in the body. Free radicals are highly reactive molecules. They may interfere with DNA and other biomolecules leading to DNA damage and mutations. Antioxidants prevent this by scavenging for free radicals before they can interact with other important biomolecules.² Apples, particularly the skins, have high phenolic content. Phenols are antioxidants that protect against oxidative damage caused by free radicals.³ Different …

Ebi2 Knockdown And Effects On B-Cell Migration, Jordan Mabey, Dr. Brian Poole

Journal of Undergraduate Research

B cells are specialized cells which participate in the normal immune response to an infection. As B cells encounter antigens in the body, they become activated and travel to what is known as a B cell follicle within a lymph node. At the outer follicle B cells combine with T cells and move to the inner follicle to become specialized immunity participating cells. Normally upon reaching the outer follicle, the B cell up-regulates Epstein-Barr virus induced molecule-2 (EBI2) which causes it to pause until the cell can complex with the proper T cell. Once this connection between the two cells …

Assessing The Effects Of The Irf5 Exon 1b On Translation, Jared Lambert, Dr. Brian Poole

Journal of Undergraduate Research

With our lab’s previous finding revealing a hairpin folding structure in exon 1B (IRF5 risk allele), not found in the other first exons, this project was focused towards more fully understanding the role of IRF5 exon 1B’s hairpin structure and its effect on the translation of downstream proteins. The data acquired through this project will provide much needed information regarding the mechanism through which the risk allele of IRF5 affects the development of Systemic Lupus Erythmatosus (SLE).

Determining The Role Of Kaposi’S Sarcoma-Associated Herpesvirus Lana Protein In The Development Of Persistent Infection, Jamie Gardiner, Dr. Brad Berges

Journal of Undergraduate Research

Kaposi’s Sarcoma-associated Herpesvirus (KSHV) is a human cancer virus that causes Kaposis’ sarcoma, the most common cancer found in AIDS patients. Our lab is working to develop humanized mice as a novel animal model to study KSHV infection and related diseases, since there has previously been no animal model established. The purpose of my project was to determine the role of the viral protein LANA (latencyassociated nuclear antigen) in establishing a latent KSHV infection in humanized mice. My project included two major aims:

Understanding The Molecular Targets Of Antimicrobial Chemokines, Brittany Kartchner, Dr. Eric Wilson

Journal of Undergraduate Research

For this project, I hypothesized that the amino acid composition of the C terminus of chemokine proteins allows some chemokines to bind to different sites on the bacterial cell wall and that the chemokines with a strong positive charge (CCL25 and CCL28) will bind to the same target, while CCL27, which has no positively charged C terminus region, will have a different binding site. In order to study the binding properties of each chemokine, I began by performing binding assays. In these assays, approximately 105 colony-forming units of exponentially growing bacteria were incubated on ice with the appropriate chemokine. This …

Identifying Proteins That Interact With Human Pas Kinase, Tacie Hall, Dr. Julianne Grose

Journal of Undergraduate Research

When PAS kinase is knocked out in mice placed on high-fat diets, these mice show such symptoms as decreased weight gain, hypermetabolic phenotype, decreased liver triglyceride accumulation, and retained insulin sensitivity when compared with their wild type littermates.¹ These symptoms are highly associated with chronic diseases commonly seen in humans today, such as diabetes, obesity, and heart disease. PAS kinase is a highly conserved protein from yeast to man which allows for the use of yeast as model organism to help discover the molecular mechanisms behind PAS kinase function. Since little is known about how PAS kinase functions, it …

Accumulation Of Attached Chemokines On Gram-Negative Bacterial Membranes, Peter Janzen, Dr. Eric Wilson

Journal of Undergraduate Research

The purpose of this project was to see if a genetic alteration to gram-negative bacteria would affect the phenotypic ability to resist the attachment of chemokines. In other words, we were trying to see if we could change the structure of a bacterium by altering the DNA. As a result of that alteration, would our immune systems fight the infection more easily with a chemokines response.

Casting For A Putative Dna Binding Sequence For Nbmp2, Brian Earley, Dr. Laura Bridgewater

Journal of Undergraduate Research

Bmp2 is a secretory mammalian protein that plays important roles in bone development and adult pathology. Recently, a nuclear variant (nBmp2) was discovered with an unknown function for this typically secretory protein in the nucleus (Felin et al., 2010). The purpose of this project is to find an in vitro DNA binding sequence for nBmp2. This discovery would lead to further research for how nBmp2 could affect transcription in vivo. However, since the conception of this CASTing project 16 months ago, I have faced challenges in plasmid construction, cell culture, and concessions with a fused nBmp2:GFP plasmid. While the plasmid …

Ex Vivo Infection And Re‐Isolation Of Human Herpesvirus 6, Stephanie Carlson, Dr. Bradford Berges

Journal of Undergraduate Research

Science rarely cooperates as anticipated, but that can often lead to unexpected insight and solution. I ran into that this semester with a certain aspect of our project. We are continuing to collect data for the American Society for Virology in late July and a future publication. We will need more replicates for a publication, but we have optimized our approach and protocol.

Genomic And Proteomic Analysis Of An Erwinia Amylovora Phage, Alicia Brighton, Dr. Julianne Grose

Journal of Undergraduate Research

Fireblight is a disease, caused by the bacteria Erwinia amylovora, that can infect some fruit trees, including apple and pear trees. When an infection occurs, the effects can be devastating—any infected part of the tree must be removed and destroyed, sometimes resulting in loss of the entire tree. Furthermore, in the warm and moist conditions typical of springtime, the spread of fireblight can be nearly impossible to control with current management methods.¹

Preservation Of Oncorhynchus Clarkii By Comparison Of Biomarkers, Chase Paulson, Dr. R. Paul Evans

Journal of Undergraduate Research

The purpose of this project is to analyze biomarkers present in cutthroat and cutbow (a mixture of cutthroat and rainbow trout) trout of the Payson Hatchery compared to typical cutthroat and rainbow markers to better understand if cutthroat trout are being preserved locally and in the Western United States.

The Effect Of The Overexpression Of Irf% In B-Cells On Inflammatory And Co-Stimulatory Activity, Reika Takita, Brian Poole

Journal of Undergraduate Research

Systemic Lupus Erythematosus is a chronic autoimmune disease characterized by inflammatory tissue damage inflicted by the body's rogue immune system (1). Although treatment is available for the disease, the pathogenesis of SLE remains unclear (2). However, researchers suspect the disease to be associated with environmental, genetic, and hormonal factors (1).

Antibiotic Resistance In Staphylococcus Aureus From Commercially Available Meat, Samuel Schriever, Bradford Berges

Journal of Undergraduate Research

Bacteria can be found everywhere. While some bacteria can help humans to make medicine and clean up environmental disasters, other bacteria can cause horrible disease. After the discovery of antibiotics in the early twentieth century humans have been using them to treat human and animal disease. At first antibiotics very effective at clearing bacterial infections. Bacteria that were able to survive the antibiotics were able to spread and become more common. Bacteria have the ability to share genes and those genes are very favorable for the bacteria that have them. Today, animals are given more antibiotics than humans are. This …

Engineering A Cancer Specific 3rd Generation Car Immunotherapy, Josie Tueller, K. Scott Weber

Journal of Undergraduate Research

This project aimed to develop a cancer-specific immunotherapy that will target and destroy cancer cells without killing healthy cells. Current cancer treatments struggle to successfully target cancer cells and commonly target all rapidly dividing cells (both healthy and cancerous). Chimeric antigen receptor (CAR) immunotherapy aims to harness T cells, the body’s natural defense system, to fight cancer by giving T cells via molecular engineering a non-native protein which targets cancer. We successfully produced a T cell that can effectively kill cancer cells while leaving normal cells untouched while minimizing the toxic side effects normally experienced by patients. Future steps involve …

Association Of Essential Tremor And Parkinson’S Disease Development, Alexander Gosch, Mary David

Journal of Undergraduate Research

Essential tremor (ET) is a neurological disorder characterized by symmetrical tremor of the upper limbs. Parkinson’s disease (PD) is a similar disorder, also characterized by tremor, with a few differences: ET tremor is mostly seen during action, while PD tremor is more prevalent while at rest. PD is characterized by problems with walking and balance, while ET is characterized primarily by tremor without balance problems. PD symptoms generally improve with the use of the drug levodopa while ET responds better to the drugs propranolol and primidone.

Examining How The Cd5 Co-Receptor Alters T Helper Cell Activation In Response To Bacterial Infection, Garrett Hamblin, Scott Weber

Journal of Undergraduate Research

Properly functioning helper T cells are crucial in a response to an infection. The adaptive immune response is orchestrated by T helper cells and their function is dependent upon interactions between the T cell receptor (TCR), peptide MHC (pMHC) and co-receptors. Upon TCR interaction with a foreign antigen, a calcium signaling cascade is initiated, which determines T cell activation, survival, proliferation and differentiation. CD5 is a T cell co-receptor that is a negative regulator of T cell activation. T cells with higher CD5 expression respond better to foreign antigen than those with lower CD5 expression. Cell-surface expression of CD5 is …

Development Of A Model For Adult T Cell Leukemia/Lymphoma In Htlv-1 Infected Humanized Mice, Kayleigh Ingersoll, Dr. Brad Berges

Journal of Undergraduate Research

Human T-lymphotropic virus type-1 (HTLV-1) is a virus that is linked to Adult T-cell leukemia/lymphoma (ATLL), causing cancer in T cells. There are an estimated 10-20 million people worldwide infected with HTLV-1. HTLV-1 is transmitted through contact of bodily fluid and usually occurs through breastfeeding, sexual transmission, and sharing needles. ATLL develops in only about 5% of infected persons after decades of infection. An ATLL diagnosis is grim as there is no cure, and most patients do not respond or only have a limited response to treatments. Current treatments include antiretrovirals or chemotherapy, but there has been little to no …

Identifying Novel Regulators Of Cellular Respiration, Sam Nicholes, Julianne Grose

Journal of Undergraduate Research

The impetus behind my project was to contribute in a small but meaningful way to the growing body of research for cures and treatments for cancer. Cancer is fundamentally unwanted cell growth and proliferation. Metabolic processes that take place on the cellular level are the key to finding solutions to the many worldwide problems of cancer. My project looked at some of these metabolic processes in order to help identify the control mechanisms that cause these processes to function normally or to behave aberrantly. The goal was to identify a cellular pathway that could be used as a potential target …

Borrelia Burgdorferi Invasion Of B Lymphocytes, Jacob Bailey, Richard Robinson

Journal of Undergraduate Research

The goal of this project was to determine if Borrelia Burgdorferi’s linear plasmid 36 is required for intracellular localization in B lymphocytes. A more complete understanding of the intracellular localization of B. burgdorferi may provide the means to effective Lyme treatment. It has been proposed that intracellular localization of Lyme spirochetes may be, at least in part, responsible for Post Treatment Lyme Disease. As the spirochetes are safe within human cells, Borrelia are able to evade host immune system responses and be protected from antibiotic treatment. A more complete understanding of the intracellular localization of B. burgdorferi may provide the …

Final Report For The 2015 Meg Entitled: Development Of Real-Time Polymerase Chain Reaction (Q-Pcr) Assays For The Detection And Identification Of Human Tick-Borne Pathogens, Richard Robison

Journal of Undergraduate Research

1. Develop singleplex q-PCR assays to identify the tick-borne pathogens Borrelia burgdorferi, Borrelia hermsii, Bartonella henselae, and Babesia microti. Primer generation software will be used to design primer and probe sequences that will theoretically perform optimally in PCR reactions. All sequences will be subjected to BLAST searches to confirm their specificity for a particular organism. Once primer sequences have been obtained, they will be synthesized by Integrated DNA Technologies, Inc. (IDT). These primers will be evaluated in q-PCR reactions using target DNA from multiple isolates and SyberGreen to detect nucleic acid amplification. Parameters for each reaction will be optimized using these …

Autoimmunity-Promoting B Cell Effects Of The Risk Factor Irf5, Brian Poole

Journal of Undergraduate Research

The academic objectives of this proposal were met. Using the funding provided by the MEG, we generated data that were used to publish two peer-reviewed journal articles:

The Chemotherapeutic Potential Of Resveratrol-Based Treatments, Connor Peck, Kim O'Neill

Journal of Undergraduate Research

Cancer is one of the most prevalent and deadly diseases in the world. In 2012 alone, there were approximately 14 million new cases of cancer and over 8 million cancer-related deaths worldwide. Despite extensive research in past decades, the burden of cancer continues to increase.

One of the most common treatments for cancer is chemotherapy, the use of various chemical substances that exhibit anti-cancer activity. Unfortunately, chemotherapy can be quite toxic and often produces inadequate results. There remains a great need for improved chemotherapeutic treatment strategies in order to improve outcomes for cancer patients.

The purpose of my project was …

Effect Of High Expression Of Irf5 On B Cell Activation After Antigen Stimulation, Kalarelyn Eberting, Brian Poole

Journal of Undergraduate Research

While autoimmune diseases are very prevalent in the United States, little is understood about the cause of autoimmunity, making it difficult to take preventative measures and develop treatments. Previous studies have determined that a common genetic risk factor across many autoimmune diseases leads to B cell overexpression of the transcription factor IRF5 (1,2,4). Researchers have also discovered that high IRF5 levels are strongly associated with autoantibody production, suggesting that IRF5 may play a role in actually causing autoimmunity. At this point, however, it is unclear how IRF5 promotes the production of autoantibodies.

Cellular Mechanism Of Thymidine Kinase 1 Secretion, Wei Meng, Kim O'Neill

Journal of Undergraduate Research

Cancer remains the second most common cause of death in the US, accounting for nearly 1 of every 4 deaths. Studies have shown that the early detection of cancer leads to better patient prognosis and a greater five-year survival rate. [1] Diagnostic and prognostic markers play a key role in classifying tumors and determining the best treatment plan for a patient. Among these biomarkers, Thymidine kinase 1 (TK1) has been studied extensively, primarily as a diagnostic biomarker for a variety of cancer types. TK1 plays a role in regulating the intracellular thymidine pools throughout the cell cycle, and thus regulates …