The central nervous system demonstrates its functional reliance on glial cells in support of neurological function primarily through its use of oligodendrocytes and the myelin they produce. Damage to myelin is caused by different pathways, one exemplified in Multiple Sclerosis (MS), where the immune system incorrectly attacks healthy tissue of the CNS. Most research focusing on these biological processes of demyelination has relied on rodent models. However, zebrafish, Danio rerio, are an ideal model for these goals because they are transparent and therefore are easy to monitor for demyelination and remyelination progression. In this study, the viability of zebrafish as a model for demyelination and remyelination research studies was evaluated. The initial steps were the development of cuprizone-infused food and the design of a cuprizone drug feeding protocol. Following treatment, myelin staining techniques were used to assess the level of myelin damage. The goal of this study is to develop a reliable new demyelination research model that will improve imaging of myelin damage and repair during demyelinating diseases.

Uninfected human blood free of anti-malarial medications is essential for the laboratory culture of Plasmodium falciparum, the causative agent of severe malaria, but is of limited availability in sub-Saharan Africa where research is conducted by the Macha Research Trust.  Cryopreservation of erythrocytes obtained from uninfected donors in the US would provide an adequate supply.  Our goal has been to identify methods of freezing that will preserve the capacity of RBCs to support asexual propagation and gametocyte formation by P. falciparum strain NF54 in serial asynchronous culture, as well as the shelf life of RBCs stored at -80ºC.  We have investigated RBCs cryopreserved using two types of freezing solutions: 1) a mixture of hydroxy ethyl starch (HES) and polyvinyl alcohol (PVA); 2) glycerol supplemented with human serum.  Our results reveal that blood frozen with hydroxy ethyl starch and polyvinyl alcohol failed to support asexual propagation of P. falciparum beyond three serial passages. Blood frozen using the glycerol & serum solution was able to support asexual propagation through at least nine serial passages, and later passages were capable of gametocyte generation.  NF54 parasites maintained in serial culture using glycerol/serum-frozen remained unable to proliferate when serially passaged using HES/PVA-frozen blood, confirming the absence of mutations that may have adapted the parasite to long-term culture in frozen blood.  These results will be discussed with respect to essential functions of the apicoplast, a parasite organelle required for erythrocyte invasion.

Pancreatic cancer has a 5-year survival rate of 8%, and the average prognosis has not improved in recent history. Chemotherapy is the primary clinical approach but carries poor efficacy and offers little improvement in patient survival due to the highly fibrotic collagen-rich tumor microenvironment. Our lab has previously confirmed that the gastrin receptor antagonist, proglumide, significantly decreases pancreatic tumor fibrosis in vivo.  In vitro efforts to elucidate the cellular target of proglumide have been unable to demonstrate a decrease in the mRNA expression of collagen (COL1A) or its post-translational processing enzymes, in either myofibroblast-like pancreatic stellate cells or cancer cells.  We hypothesize that quantification of deposited collagen matrix in vitro will better reflect the decreased fibrosis apparent in vivo. Utilizing the Sirius Red/Fast Green protein staining assay, collagen deposition was measured in cultured cells.  To normalize collagen to potentially disparate cell proliferation rates, we confirmed that the alamarBlue cell quantification assay does not interfere with subsequent Sirius Red/Fast Green protein staining.  Human pancreatic stellate cells (RLT-PSC), mouse pancreatic cancer cells (Panc02), and human pancreatic cancer cells (PANC-1) were stained 4 days after seeding.  Significantly higher collagen was evident in Panc02 and PANC-1 wells compared to stellate cells (p<0.05).  To test the impact of proglumide treatment, cells were treated with proglumide (200 μg/ml).  Lines showed no change in normalized collagen deposition after 4 days compared to untreated controls.  These preliminary data reflect the unchanged COL1A mRNA levels previously apparent in the stellate and cancer monocultures.  Further inhibition studies will utilize co-cultures to address potential heterotypic intercellular communication between these cell types, characteristic of the tumor microenvironment.

While heavy metals such as zinc and nickel are necessary for several life-sustaining processes, elevated levels of these metals pose a threat to humans, plants, and other forms of life. Researchers have found that certain plants known as hyperaccumulators tolerate higher levels of heavy metals than non-hyperaccumulating plants. They can be planted in heavy metal contaminated soil to take up excess heavy metals, thus decontaminating the soil in a process known as phytoremediation. In this research project, we will examine the altered heavy metal stress response of two Columbia-line Arabidopsis thaliana mutants that we hypothesize are hyperaccumulators of heavy metal divalent cations. This hypothesis will be tested using nickel tolerance and accumulation assays. The tolerance assays will compare the root length, a key indicator of plant health, of the two mutant A. thaliana plants against the Columbia-line wildtype plant in varying concentrations of nickel. The accumulation assay will be used to quantify the concentration of heavy metals present in the plant tissue. These analyses will allow us to determine the impact of the mutations on the nickel heavy metal stress response of the plant.

The cryopreservation of fresh, uninfected human red blood cells (RBCs) is fundamental in culturing Plasmodium falciparum, to create better preventive measures in combatting malaria. We hypothesize that human blood cryopreserved with hydroxyethyl starch and polyvinyl alcohol (HES/PVA) or glycerol and serum will support asexual parasite growth and gametocyte formation in culture following prolonged freezing at -80°C of A+ or O+ blood. These freezing techniques utilized multiple conditions to evaluate each of the ability for the culture to propagate sexually and asexually with the NF54 parasite strain. The conditions tested through serial propagation: fresh, never frozen blood, which acted as the control; short-term cryopreserved blood, which was stored at -80ºC for less than two weeks; and long-term frozen blood, which was stored at -80ºC from six months to one year. This shelf-life study found that HES/PVA cryopreservation technique supported the asexual propagation of P. falciparum through only two serial passages; parasites could not be rescued by subsequent addition of non-frozen blood. Blood frozen using the glycerol & serum solution was able to support NF54 propagation beyond a ninth dilution and re-plating (passage). Blood cryopreserved by either method supported the generation of mature gametocytes in the first passage.  Additionally, our results suggest that the NF54 strain did not accumulate mutations that abnormally enhanced or sustained growth on cyropreserved blood.

Vaccines are an integral part of modern-day disease prevention. Healthcare professionals should be aware of the basic immunological foundations of how vaccines work. Students in past offerings of Messiah University's Microbiology course were able to carry out an exercise in which commercially obtained polyclonal neutralizing antisera (samples from simulated patients) were compared for ability to inhibit infection of E. coli B cells by bacteriophage T4.  Unfortunately, the antiserum is no longer available for purchase.  Therefore, we seek to produce similar antibodies using monoclonal technologies.  Hybridoma clones capable of generating T4 neutralizing monoclonal antibodies will be produced from T4-immunized Balb/c mice. T4 virus stocks of known titer with reduced concentrations of endotoxin must be produced to vaccinate the mice.  Our approach has been to reduce endotoxin concentrations in commercially obtained T4 phage stocks by mild organic extraction, followed by dialysis and concentration using Centricon centrifugal filtration units.  titers of the concentrated stocks were determined by infection of E. coli B cells.  The endotoxin levels in the virus concentrates were measured to confirm reduction relative to viral titer.

That pancreatic cancer is unusually aggressive and deadly has prompted extensive research into its mechanisms and options for therapy. Our research seeks to develop a vaccine that will activate the tumor-bearing host immune system against a novel protein expressed by aggressive pancreatic tumors. Our approach is to engineer immortalized syngeneic cells to express a derivative of the Simian virus 40 Large Tumor antigen protein (SV40 T ag) that contains a 20 amino acid insertion corresponding to a unique sequence found in the CCK2i4svR pancreatic-cancer associated growth factor receptor. These cells will be used as a vaccine to cause the host to develop cellular or humoral immune responses against the cancer-associated CCK2i4svR epitope. Immortalized cell lines were produced by transfection of C57Bl/6 primary kidney cells with four plasmids, each encoding a different derivative of the SV40 T antigen with respect to presence and insertion site of the CCK2i4svR 20 codon sequence.  Transfected cultures were monitored for growth of dense foci which result from immortalization as a result of expression of the SV40 T ag. Foci were harvested, expanded and evaluated by indirect immunofluorescence using two SV40 T ag-specific monoclonal antibodies to confirm that they had been successfully transformed and expressed the SV40 T antigen protein derivative. SV40 T ag-positive lines were cryopreserved for characterization in subsequent immunization and tumor immunotherapy studies.

Oligodendrocytes are the myelinating cells of the central nervous system, and these cells and their myelin sheaths are the autoimmune targets in multiple sclerosis (MS). Increasing the differentiation of OPCs to myelinating OLs is a promising method for treating MS. Recent clinical trials have revealed a positive effect of clemastine, an antihistamine/muscarinic antagonist, in stimulating myelin repair in patients with MS. This study is investigating the combinatorial effects of muscarinic receptor antagonists darifenacin (M1) and pirenzepine (M3) on the rate of oligodendrocyte (OL) maturation.  Cultured oligodendrocyte progenitors were used to assess receptor subtype effects on myelin-specific gene expression and the rate of OL maturation during treatment for 4 days.  Combination treatments were also studied in the larval zebrafish model.  RNA was isolated and analyzed for changes in expression of myelin-specific genes MPZ and MAG. Several dosages of combinatorial treatment and time points were tested, but no significant changes in larval zebrafish gene expression were identified.  However, muscarinic agonist, cevimeline, was shown to reduce the expression of myelin-specific genes MPZ and MAG, which supports the hypothesis of the involvement of the muscarinic pathway in myelination. Muscarinic antagonists do not appear to enhance the vigorous myelination program of zebrafish larvae.

Post-Traumatic Stress Disorder (PTSD) can develop after experiencing an intensely distressing event. The condition exhibits as a maladaptation of the fear response, causing frequent activation of the sympathetic nervous system (fight and flight behavior) in response to benign stimuli. Persistent sympathetic activity results in chronic stress and precipitates other physical and psychological conditions—severely impacting the individual’s quality of life. Current treatments yield low recovery rates and poor prognosis, particularly when initiated long after the original trauma. New research explores opioids as a prospective preventative medication, as morphine seems to diminish PTSD acquisition if received during the critical period of memory consolidation. However, PTSD is often comorbid with increased rates of substance abuse, including opioid addiction.  Our research examined if PTSD treatment with morphine increases drug seeking behavior using a rodent model. Rats underwent Stress-Enhanced Fear Learning (SEFL) to induce PTSD like symptoms. After SEFL was completed, the rats were given two injections of morphine in a unique “Context A” and two injections of saline in “Context B”.  The rats were then allowed to pass between the chambers, and the relative amounts of time spent in either Context A or Context B was measured to determine if the rodents exhibited a Conditioned Place Preference (CPP). If SEFL subjected rats exhibit CPP for Context A, this indicates a correlation between receiving morphine treatment for PTSD and increased drug seeking behavior.

Malaria is caused by multiple species of the parasite Plasmodium and disproportionately affects people living in the developing world where effective control of or protection from the parasite is lacking. This study seeks to support ongoing research at the Macha Research Trust (MRT) which is located in the Southern Province of Zambia where the virulent species, Plasmodia falciparum is prevalent.  Our goal is to support the capacity of the laboratory at MRT to culture (propagate and preserve) locally-isolated or laboratory strains of Plasmodium.  Laboratory cultivation of P. falciparum requires fresh human blood.  However, it is difficult to assure the steady supply of fresh, uninfected human blood needed to sustain culture experiments at MRT because blood from local residents cannot be used, and because many visiting scientists and physicians routinely take prophylactic anti-malarial drugs which can make their erythrocytes unable to support asexual propagation of or gametocyte generation by P. falciparum in culture.  We are investigating methods that should allow for the cryopreservation of erythrocytes obtained in the US from uninfected individuals, with subsequent shipment to Zambia.  We have cryopreserved preparations of fresh, leukocyte-depleted erythrocyte suspensions using minimal aqueous volumes of solutions containing the hydroxyethyl starch (HES) and polyvinyl alcohol (PVA), or with glycerol-based solutions. This presentation will describe ongoing efforts to determine if cryopreserved RBCs can effectively support asexual propagation or gametocyte formation for the P. falciparum laboratory strain NF54, and how well this capacity is maintained following long-term storage at -80°C.

Post-traumatic stress disorder (PTSD) is a stress-induced disorder that may be established after a traumatic experience. Individuals with PTSD may develop extensive fear and anxiety in non-threatening environments. The stress-enhanced fear learning animal model (SEFL) mimics the symptoms of PTSD in humans by observing the effects of an animal exposed to a prolonged stressor. SEFL is measuring the increase of subsequent learning about fearful stimuli that occurs after a severe stressor. Using the SEFL animal model, rats were exposed to two different environments, Context A and Context B. In Context A, the stressor was 15-foot shocks administered over 90 minutes induces PTSD-like symptoms. Context B, which had a different scent and noise than Context A, would be exposed to a single shock in an unfamiliar environment. The two reminder shocks were delivered in Context B, one followed the injection of morphine or saline, while the other shock was administered three weeks later. Morphine is a promising treatment for PTSD as a method to prevent the formation of the fear learning response in the animal model. The rats were recorded for seven weeks in Context B and the rats’ degree of freezing was assessed. Freezing behavior in rats is indicative of a fear response. We hypothesize that morphine-treated rats would exhibit less freezing behavior than saline-treated rates following both reminder shocks.

Pancreatic cancer is currently the fourth most deadly form of cancer in the United States. One factor implicated in pancreatic cancer growth is the hormone signaling pathway between gastrin and its receptor, CCK2R. In the early 2000’s, it was also discovered that pancreatic cancer cells can contain a variant (CCK2_i4svR) of the normal receptor protein. Crucially, this longer variant has been shown to be hyper-stimulated and to drive increased cancer growth. Measuring the relative abundance of these two receptors at the protein level can help us understand their role in pancreatic cancer and may represent prognostic value as a biomarker. However, while RNA detection and measurement have been reproducible, protein detection has been problematic. Using western blot analysis, we have been able to detect the receptors in cells expressing them at high levels; however, detection in wild type and stably-transfected lines more representative of physiological expression has been unclear. Thus, we hypothesize that low, natural abundance of the receptors requires enrichment for reliable quantification. Herein, we report our initial attempt to enrich for green fluorescent protein-tagged variants, CCK2R-GFP and CCK2_i4svR-GFP, utilizing a membrane extraction protocol based on the non-ionic detergent, Triton X-114. Following enrichment, western analysis demonstrated a significant decrease in cytosolic protein in control cells transfected with untagged GFP. Efforts to verify retention of membrane proteins in the hydrophobic fraction, and subsequent specific detection of the CCK2R variants, are ongoing.

Post-traumatic stress disorder (PTSD) is a chronic, debilitating disorder in which the heightened fear and arousal experienced during a traumatic event generalizes to nonthreatening stimuli. Previous research has suggested that administering morphine within 24-48 hours of a traumatic event inhibits memory consolidation and prevents future development of PTSD. The goal of this study was to block the reconsolidation of previously stored stress memories by administering morphine within 24-48 hours of conscious recall of the original stressor. A rat model of PTSD called stress-enhanced fear learning (SEFL) was used, which employed two distinct contexts, Context A and Context B. The severe stressor, which mimics trauma experienced by humans, occurred in Context A, during which 15 foot shocks were administered within a 90-minute period. Two reminder shocks were delivered in Context B. The first was followed by an injection of morphine or saline, and the second was delivered three weeks later without a subsequent pharmacological injection. The animals were video recorded for seven weeks in Context B to assess for freezing behavior, a classic fear response in rats. We hypothesized that the morphine-treated rats would exhibit less freezing behavior following both reminder shocks, whereas the saline-treated rats would exhibit a heightened fear response.

Pancreatic cancer is projected to become the third leading cause of cancer-related deaths in the United States and has a low 5-year relative survival of 10%. The poor response to treatment is attributed to the immunosuppressive and highly fibrotic nature of the pancreatic tumor microenvironment. Previous studies have demonstrated that proglumide treatment results in decreased fibrosis and may increase T-lymphocyte infiltration. Additionally, our lab has demonstrated that peptide immunization targeting the human CCK2_i4sv receptor, a cancer-associated splice variant of CCK2R, evokes a potent T-cell response in mice. We hypothesized that combination therapy utilizing concomitant immunization and anti-fibrotic treatment will further enhance T-cell infiltration and decrease tumor burden. To address this, we first analyzed T-cell infiltration in a pilot group of orthotopic murine pancreatic tumors (Panc02) obtained from mice treated +/- proglumide. Tumors were analyzed by immunohistochemistry (n=3 per grp), and infiltrating CD-3+ T-cells were numerated in five random photomicrographs for each tumor. Tumors treated with proglumide exhibited increased T-cell infiltrate and significantly less fibrosis. To assess the efficacy of combination therapy, mice (C57Bl/6) were immunized weekly with CCK2_i4sv and control peptides starting three weeks prior to orthotopic injection of syngeneic cancer cells (Panc02) engineered to express human CCK2_i4svR. Both groups received oral proglumide following tumor establishment. Upon necropsy (≤6 wks), mice exhibited no significant difference in tumor burden. Immunohistochemical analysis showed the number of infiltrating CD-3+ T-cells per tissue area trended higher in these proglumide-treated mice compared to the previous untreated pilot controls. However, no difference in infiltration was observed in tumors from CCK2_i4svR-immunized mice compared to control immunization. Our data confirm proglumide treatment may enhance immune access to the tumor; however, simultaneous analysis of the circulating lymphocyte response is necessary to thoroughly determine the efficacy of combined immunotherapy on T-cell infiltration.

Plant cells uptake metals from the soil by operating specific cellular pumps to transport metal nutrients. These pumps are crucial for maintaining the healthy concentration of metal elements. Metal Micronutrients, like zinc or nickel, are essential to plant growth and their uptake is controlled by the MTP1 transporter on the vacuoles in cells. Using Arabidopsis Thaliana as a model organism, the effect of mutations on the MTP1 gene can provide a window into MTP1 transporter function and metal nutrient uptake pathways. Two preliminary metal tolerance experiments were conducted on Columbia line wild-type seeds with zinc and nickel contamination to learn what effects the metal ions were having on the plants’ growth. Unfortunately, a problem found during a preliminary wild-type experiment highlighted problems somewhere in the experiment. After that result, the primary purpose became to troubleshooting the experimental method. Both experiments with zinc and nickel contamination yielded results that unexpectedly appeared to be hyper-concentrations of metal ions. Hyper-concentration pointed to a problem with the media, the metal ion solutions, or the seeds. The media was examined by chemical digestion and atomic absorption spectroscopy to quantify metal ion concentration. The analysis demonstrated the concentrations were not hyper-concentrated in the media meaning the problem was with the metal ion solutions. This semester began with a complete remake of metal ion stock solutions and growing a new batch of seeds that would hopefully fix the problems. Early experiments showed the same apparent hyper-concentration problem but with a noticeable consistency. The plants were reacting consistently to the presence of metal ions in a range between 20-60 uM with nickel contamination. This consistency is shown over multiple experiments and means an adjusted experiment range to can be used to test MTP1 mutant tolerance.

Carolina Laboratory Supplies was the sole commercial supplier of polyclonal antisera specific to bacteriophage T4, an essential item used in the Phage Neutralization Lab in Microbiology courses at Messiah University. Carolina ceased production of this antiserum, necessitating discontinuation the Phage Neutralization Lab. the goal of this study is to produce monoclonal T4-neutralizing antiserum so that the Phage Neutralization Lab can be restored. The T4 suspensions used need to have confirmed titers and be depleted of lipopolysaccharide (endotoxin) which are lethal to mammals. Determining the titer of the bacteriophage suspension, purifying it, and verifying that the endotoxin levels are below lethal concentrations must be done before immunization of the mice. The successful generation of antibodies against bacteriophage T4 would enable the project to continue to the next stage, where monoclonal antiserum can be produced using hybridoma technology. 

Oligodendrocytes are the myelinating cell of the central nervous system. Myelination of axons aids in the propagation of neuronal signals and are essential to the function of the nervous system. In this study, oligodendrocyte cultures from neonatal rat pups were used to analyze the signaling protein Lck and its role in oligodendrocyte differentiation. We hypothesized that Lck signaling promotes the normal development and differentiation of oligodendrocytes, thus the inhibition of Lck will result in decreased ability to express markers associated with myelinating oligodendrocytes. Differentiation of oligodendrocytes was assessed by immunocytochemistry to identify maturation-specific markers and expression of myelin-specific genes following treatment with an Lck-specific inhibitor. Untreated oligodendrocytes expressed more O1, a cell-surface marker expressed by mature oligodendrocytes, than oligodendrocytes treated with a Lck inhibitor at four and five days after treatment. No significant difference was observed in A2B5, a cell-surface marker of immature oligodendrocytes, four days after treatment. Oligodendrocyte precursor cells (OPCs) were transfected with a mammalian green fluorescent protein (GFP) plasmid and grown under conditions that either encouraged or discouraged OPC maturation to investigate the efficacy and longevity of the transfected GFP protein expression. Future studies will investigate the signaling pathways impacted by loss of Lck activity and their role in oligodendrocyte differentiation.

The Atelopus zeteki, or Panamanian Golden Frog, is extinct in wild populations and only survives in captivity. Reintroduction is the future of Atelopus, but due to their vulnerability, a low impact method of observation and population monitoring are required. With recent advances in Unmanned Aircraft Systems (UAS: drones), a much-needed low impact solution for golden frog population survey for continued ensure post-reintroduction success. This semester we have dedicated time and planning to certify, train, and practice on drone flight. We are planning to set up experiments with frog models close to that of the Atelopus in Panama, their country of origin. These experiments will allow us to utilize an ecological drone to test its effectiveness for golden frog population monitoring in the future.

The purpose of this study was to determine the viability of growing potatoes hydroponically with chicken waste as an alternative, more sustainable nutrient source in substitute for fish waste in an aquaponics system. Hydroponics is a growing form of sustainable agriculture which utilizes a water-based method to deliver nutrients to plants. Hydroponics is a preferable alternative as it uses 90% less water than traditional agriculture and does not contribute to common agricultural issues such as land degradation. A popular form of hydroponics is aquaponics, which combines hydroponic technology with aquaculture. The fish waste in this system is the sole source of nutrients for the plants. The cost and difficulty of maintaining the fish in this system can be an obstacle to farming communities, however. A proposed alternative to the use of aquaculture is the use of chicken waste as a nutrient source. Chicken waste is a much cheaper and more accessible resource for less developed farming communities. It is also not as high maintenance as aquaculture and contains a high content of essential nutrients such as nitrogen, potassium, and phosphorus. Potatoes are the crop of choice for this study due to their relatively high nutrient content, making them a popular staple crop in less developed countries. If chicken waste proves to be an effective nutrient source for staple crops such as potatoes in a hydroponic system, it may be a viable option for less developed farming communities who are in need of sustainable alternatives to traditional farming practices.

Though antibiotics have served as a powerful tool in humanity’s fight against disease, the overuse of antibiotics has caused some bacteria to develop resistances to antibiotics, making these drugs less effective. One means by which bacteria may be exposed to antibiotics and develop resistance is through wastewater treatment plant effluent, which carries antibiotics from human wastes into local streams. We hypothesize that increased levels of antibiotics downstream of treatment plants have selected for increased antibiotic resistance in bacteria living in these downstream regions. As such, when both upstream and downstream samples of bacteria are exposed to the same antibiotic treatments, downstream samples should exhibit higher survival. To test our hypothesis, we sampled bacteria from Dogwood Run and Stony Run in Dillsburg, PA, selecting collection sites from both upstream and downstream of the local wastewater treatment plants. We cultured these bacteria in the lab, classified them based on colony morphology and Gram-stain results, and exposed them to antibiotics through the Kirby-Bauer and replica plating methods. When comparing all bacteria, there did not appear to be a clear trend of increased resistance in either the upstream or downstream samples. However, when comparing bacteria only within our classified groups, initial results showed 26 instances of increased downstream resistance and only 13 instances of increased upstream resistance. These results are promising for our future work in this area, and they may indicate that increased regulations on wastewater treatment plants are needed to combat further antibiotic resistance development.

Goldenseal (Hydrastis canadensis) is a plant native to woodlands of east-central North America. Goldenseal roots contain antimicrobial compounds which promote respiratory and digestive health and the plant has been wild-harvested almost to the point of extinction. Plants are products of their environments and the concentration of medicinal compounds within a plant are affected by the conditions in which the plant was grown. Wild harvested plants are subject to the variations in their natural environment and it is almost impossible to accurately know the concentrations of desired medicinal compounds within plants that are wild-harvested.  With all medicinal compounds there exists a therapeutic dosage in which the compounds are beneficial for the body. Outside of this window, the compounds are either ineffective or toxic. It is crucial for supplements made from medicinal plants to contain known concentrations of the desired medicinal compounds in order to ensure the effectiveness and safety of the supplement. Due to these reasons it is crucial to develop a sustainable and controlled method of growing goldenseal, and aquaponic technology is a form of agriculture that serves as a hopeful solution to these concerns. Previously, goldenseal was aquaponically grown at Messiah University somewhat successfully, but many of the plants died prematurely of root rot. Other than the work completed at Messiah, there are no published studies that include growing goldenseal aquaponically. It is proposed that goldenseal be grown in an aquaponic system at various water levels to find a growing method that maximizes the plant's survival and productivity.

The world is rapidly approaching a point where they need to find a source of sustainable clean energy.  Something that could aid this problem is new efficient catalysts, one potential catalyst is Cobalt substituted Holocytochrome C.  Holocytochrome c is a protein that is attached to a heme group at a CXXCH amino acid motif.  These are found naturally occurring in the mitochondria and are part of the electron transport chain. These enzymes have the capability to catalyze the reduction of water into a diatomic hydrogen which provides a fuel source for energy production.  Cytochrome c naturally has an iron heme cofactor, but this wild type does not have the same enzymatic capabilities as a cobalt substituted heme to catalyze the reduction of water.  In order to synthesize a large quantity of the desired coenzyme, they must be created in vitro using E. coli expression vectors.  The primary objective of my project was to create these expression vectors that would be capable of expressing large amounts of desired product by having the cytochrome c be expressed by cytochrome c heme lyase, the naturally existing enzyme responsible for holocytochrome c synthesis.  These expression vectors can utilize mutations of both the cytochrome c and cytochrome heme lyase genes to express different mutants.  These mutant products can then be quantitatively analyzed in different combinations to find what combination gives the highest enzymatic capability to combine cytochrome c and a cobalt substituted heme.  Once the ideal combination is found, we will attempt to make a microbe and utilize the mutated proteins to effectively perform the process in vivo. Utilizing this could provide a catalyst that leads to efficient production of fuels from sustainable energy sources. 

Type 2 diabetes is a serious metabolic disease with a growing need for improved therapeutics. Traditional antidiabetic drugs can assist with the maintenance of blood glucose levels in diabetics but are not optimal treatments due to side effects. PTP1B is a validated therapeutic target for type 2 diabetes, as it acts as a negative regulator in the insulin signaling pathway. Inhibiting PTP1B may allow for increased insulin sensitivity, leading to glucose homeostasis and improved metabolism. In the past three allosteric sites were identified as targets for PTP1B inhibition. This study analyzes the validity of these sites and the possibility of other allosteric sites through computational methods and enzymatic assays. A new allosteric site was identified based on the determination of the anti-diabetic drug Lobeglitazone’s binding site and used to improve the computational method for testing possible inhibitors of PTP1B. Correlation of the computational method with known natural product PTP1B inhibitors revealed that the computational method is independent of kinetic properties. This study provided a new allosteric site to target and a computational model to eliminate non-inhibiting compounds in the search for type 2 diabetes therapeutics.

Cytochrome c is a mitochondrial enzyme that is responsible for catalyzing many important chemical reactions in many different species of animals. My research was primarily concerned with the creation of a mutant variant to Horse Cytochrome c (HCc), with potentially different electrochemistry than the original. The target amino acid for the project was HCc’s axial histidine, which I attempted to replace with an Unnatural Amino Acid (UAA) that is simply a methylated variant of histidine. At this point, I have successfully transformed BL21 cells with engineered plasmids to create cells with the potential to express the mutant HCc protein. Once the new protein is confirmed to have been created, its properties will be compared to the original to gauge the impact of the methylated, axial histidine on the electrochemistry of HCc.

Molecular dynamics is a computational method that models a macromolecule in a solvent over time. Using the molecular dynamics software GROMACS to engineer metal-binding sites in proteins, precious time and energy can be saved by computational modeling of the metal-binding sites first. In this project, molecular dynamics was used to simulate metal binding sites with different combinations of ligands to obtain optimized geometries to ensure that the results of the simulations were realistic. Specifically, iron and zinc sites involving cysteine and histidine ligands were analyzed. Using GROMACS, these metal sites were subjected to simulated annealing, and the resulting bond angles and bond distances from the stable conformations were compared to the average values in the Protein Data Bank (PDB). By adjusting force constants, bond distances, and bond angles in the GROMACS force fields, two particular sets of parameters were identified for an iron site with four histidine ligands that produced realistic bond distances. However, when aligning these conformations to the metal binding site in Troponin C, the lowest RMSD value calculated was 0.72 which is not low enough to indicate good overlap, indicating that the specific combinations of amino acids in the model would not likely generate a good iron binding site in Troponin C. A lower RSMD value would indicate better alignment between the metal binding site backbone atoms and the protein backbone atoms. The bond angles will need to be optimized in addition to the bond distances to produce geometries that overlap favorably with proteins. This procedure can be applied to metal binding sites with different combinations of cysteine and histidine to determine what specific set of ligands will produce an iron binding site ligated by amino acids whose backbones overlay with the existing backbone structure of Troponin C.  To increase the efficiency of engineering metal binding sites in proteins, only binding sites with favorable backbone atom overlap would be produced and tested in the laboratory.

Protein phosphatases have been considered a potential target for drug-based therapy since their discovery. Mutations of phosphatases found in cell signaling pathways have been linked to type II diabetes, obesity, and certain types of cancer. One of these phosphatases, protein tyrosine phosphatase 1B (PTP1B), functions as a negative regulator of the insulin pathway. A gene knockout study in mice confirmed that mutations to this protein result in a dampened insulin sensitivity. Previous attempts at competitive inhibition through drug-based therapy have been unsuccessful due to the highly conserved active site across the phosphatase family. This study explores the plethora of natural products available as potential inhibitors for PTP1B in hopes of designing a molecular scaffold to generate site-specific inhibitors. Chalcones and stilbenes are two compound families that have previously exhibited inhibitory binding. Hydroxychalcone derivatives, specifically those in the para position, show promise as a drug scaffold for specific active site inhibitors. In addition, isoprenyl groups attached to branching aromatic rings have a significantly higher binding affinity to PTP1B and a lower IC50. Initial in silico results showed moderate to high binding affinities for a small hydroxychalcone library. Currently, enzyme activity levels in the presence of these hydroxychalcones are determined using absorbance assays. Duplicate assays are performed with detergent to account for non-specific inhibition caused by aggregation. Preliminary assays using commercially available hydroxychalcones display slight to moderate inhibition of PTP1B at the active site. In future studies, twelve hydroxychalcones will be synthesized and introduced into an absorbance assay to evaluate their potential as a scaffold for PTP1B specific inhibitors.

Heme is a molecule that plays a vital role in many biological processes. Because of its central iron atom, heme proteins are a class of metalloproteins, which are renowned for their ability to enable many of the daily processes such as oxygen transport to begin in an organism. Our research aims to substitute the central iron atom with a cobalt atom due to its unique chemistry that will allow us to expand the metalloprotein cofactors living systems employ to fine tune their catalytic properties. Modifying this molecule is best done using the metabolic pathways of E. coli which has an expression vector that we can manipulate to see if the mutants of the cytochrome c heme lyase enzyme, responsible for ligating heme to its attachment motif, would allow us to replace the central iron atom with a cobalt atom. Our experiments consist of raising E. coli colonies and extracting their DNA via MiniPrep so that we can induce mutations to the bacterial plasmid using several types of PCR based mutagenesis. We can then use MiniPrep to extract the newly transformed DNA and confirm that the bacterial cell successfully integrated the mutation into its plasmid with Sanger Sequencing. This part of the project was performed by Cornell Genomics Center. Out of the 4 variants created, only one, W97A, expressed the desired mutation that would allow for the possibility of heme substitution. However, further tests will be performed on the additional 3 variants after their PCR products have been placed through a Dpn1 digest. 

Metal ion imbalances in the brain can lead to the formation of amyloid plaques, groups of misfolded proteins, characteristic of neurodegenerative diseases like Alzheimer’s. The current method for detecting metal ions, the Patch-Clamp, requires excised tissue, expensive equipment, and skilled workers. This study seeks to develop a less labor-intensive technique by genetically encoding the binding site of a fluorescent calcium-sensing protein, troponin-C, to bind to iron and copper. The hypothesis of this research is that if enough mutations are introduced to the ten amino acids in the calcium-binding site of troponin-C, one mutant will be able to bind to iron and copper respectively. 1,024 possible mutant combinations were introduced to create a mutant library through a series of PCR reactions resulting in five mutated fragments. The five mutated fragments were then consecutively reconstructed through a series of COE-PCR reactions to rebuild the mutated troponin-C gene. Gel electrophoresis was used to verify the resulting fragment lengths after each reaction. Certain fragment combinations did not combine successfully despite altering COE-PCR parameters, while other combinations were successful. Gibbon Assembly was also used to reconstruct the troponin-C gene. In the future the mutant library will be verified through Sanger DNA sequencing and successfully mutated DNA will then be inserted into E.coli cells via transformation. The library will be screened based on fluorescence and successful sequences will be compared to fine-tune future mutation libraries.

Zinc selenide (ZnSe) has drawn particular interest as a possible supporting substrate for self-assembled monolayers (SAMs) due to its transparency in the infrared region and its functionality as a semiconductor. The naturally forming oxide layer on bulk ZnSe has been shown to support SAMs; however, the effect of modifications to this oxide layer through chemical means has not been well-established. In this study, the oxide layer and choice of adsorbate molecular head group are optimized towards the formation of high-quality SAMs. This is achieved by stripping the oxide later from optically polished ZnSe substrates using varying chemical surface treatments prior to SAM deposition. The resulting surfaces are characterized via contact angle analysis, Fourier transform infrared spectroscopy, and atomic force microscopy. While SAMs with thiolate head groups formed readily on both etched and un-etched substrates, we find that acid head groups show affinity only for certain etched surfaces. This can be reconciled with the surface compositions under various etching conditions, as determined via X-Ray photoelectron spectroscopy, to determine the surface compounds conducive to the formation of these SAMs.

One proposed solution to reduce global carbon dioxide emissions is to replace the traditional engine with one that does not produce harmful emissions. A hydrogen fuel cell meets this requirement and utilizes the oxygen reduction reaction (ORR) at the cathode of the cell. One major reason that they are not widely implemented is that the main catalyst used, platinum, is expensive and breaks down. Carbon nitride has been explored as a possible replacement for the catalyst of the ORR. Physical vapor deposition via DC magnetron sputtering was used to create thin films of CN­_x for analysis by cyclic voltammetry, serving as a model ORR in the lab. It is necessary to deposit a metallic layer between substrate and CN­_x for the thin films to be stable. However, upon heating the carbon nitride thin films become non-conductive and therefore cannot be measured by cyclic voltammetry. Future work will focus on solving the conductivity issues associated with heating in the deposition process as well as structural determinations of nitrogen content in CN_x to determine the optimal doping mode.

We helped build a website application that is meant to store and track information concerning users, sponsors, and children for ease of looking up information. The Burkina’s Promise organization started when several short-term mission trips to the country of Burkina Faso revealed a need for helping the children of pastors become educated. The need for the site itself arose when the Burkina’s Promise organization outgrew its use of Microsoft Excel to handle all of its data and information. During our time developing the site, we added features such as a User Messaging feature, more visitor pages, and other features. Lastly, we tested the entire site and fixed various bugs such as getting the French translator API to work throughout the user pages and being able to delete children in the database on the frontend via a button.

Our application was built to aid special education teachers in the monitoring and tracking of students’ behavior. Every day, these teachers are asked to record student’s performance of specific tasks to track their progression over time. Teachers will then use this data to generate reports to share with other teachers, parents, or counselors of that student. Currently, teachers are using complex and unmanageable spreadsheets to record student’s ratings on specific tasks within specific classes. Behavv serves to make reporting this data easier and more user-friendly by simplifying data entry and presenting clear visual representations of that information. Our application allows teachers to focus less on stressful spreadsheets and more on students’ success.

The iSeek team has built an application to aid people that are visually impaired.  If you are visually impaired and have misplaced something you own then you could spend an unwanted amount of time trying to find your object.  iSeek intends to solve this problem. Through the use of your phone’s camera, alongside a trained chatbot, helps users find misplaced or new objects in their surroundings.  iSeek is built with the user in mind, all actions inside the app are voice-activated alongside the classic touch controls.  We hope to have created an app that is unique to the market and that has real-world applications to help people live a more enjoyable, easy life.  

We created an application that helps provide coping mechanisms for individuals struggling with anxiety. Everyone experiences stress and anxiety in different ways, so we sought after a solution, tailor made to the user, to help them find better ways that work for them to manage their stressors. By taking a few short surveys, our algorithm learns about the user's likes, dislikes, and past experiences with stress to offer handpicked suggestions generated from that data. Each suggestion comes with an excerpt on why it's effective for anxiety, and a playlist from Spotify to help calm the mind during the activity. The more they interact with the app by reviewing those suggestions, the better our algorithm gets at personalizing them for each user. 

Effective degree planning, course scheduling, and advising are predicated upon an understandable and clear curricular hierarchy. Presently, there is no tool at Messiah University that allows students and advisors to simultaneously visualize course requirements, prerequisite structure, and course offerings in an intuitive format.

To this end, I built an application that lets the user—faculty or student—visualize these requirements for degrees in the Department of Computing, Mathematics and Physics. The output of this program is a tree diagram that provides “at a glance” mapping of course sequencing and availability.

We built a mobile app that quizzes you on peoples’ names and faces in order to help users remember them. Humans are bad at remembering people’s names. Despite really wanting to learn someone’s name, many people struggle to recall it, which can lead to feelings of awkwardness. According to Charan Ranganath, the director of the Dynamic Memory Lab at UC Davis, people underestimate the work that needs to be done in order to commit somebody’s name to memory: “People are often overconfident, and they underestimate how hard it will be later on.” It is easy to focus on making a good impression, rather than learning a new name and face. Ranganath asserts that one way to solve this problem is to test your name memory skills: “The act of actually testing yourself on the name will help you retain it better in the long term.” Our application implements this solution by providing a way to create and take quizzes on people’s names and faces. Users can group people into categories in order to associate people with certain characteristics, e.g., classmates or coworkers. Additionally, managers can create lists and share them with other users, making onboarding a new group of people a breeze.

Amputations, specifically lower limb amputations, are common in sub-Saharan Africa and across the broader global community largely due to infection and disease. All amputees struggle with the negative social stigma surrounding their missing limb and have difficulty getting from one place to another. Our project, The Prosthetic Knee Team, partners with the orthopedic workshop at the CURE International Hospital in Kijabe, Kenya to create a prosthetic knee design for a specific type of amputation known as a Knee Disarticulation (also called through-knee). Currently, the orthopedic workshop is only able to provide one very expensive prosthetic knee option for these patients. Often, these patients choose to undergo a second surgery, a trans-femoral amputation, because it is more affordable. The goal of our project is to provide the orthopedic workshop with a manufacturable prosthetic knee design that provides through-knee amputees with a cheaper prosthetic option and removes the need to have a second amputation above the knee. Throughout the past two semesters, our focus has been to modify and improve our current prosthetic design to best meet all of the specifications laid out by our partner. Those specifications include minimized thigh-lengthening, low weight, maximized stability and durability, and aesthetically pleasing. We also have performed simulated tests to determine the overall strength and lifespan of the design. Moving forward, our team is presently working on manufacturing the prosthetic knees at Messiah University’s machine shop.

Caffeine is the most commonly used psychoactive drug in the world and has wide implications in medicine, athletics, and public health. Studies have shown that metabolism, clearance, reception, and response to caffeine vary significantly among individuals. The pharmacokinetics are primarily dictated by the cytochrome p450 enzyme CYP1A2 while the adenosine neuroreceptor ADORA2A heavily influences the drug’s pharmacodynamics. Polymorphisms of the -163 A>C CYP1A2 and the 1976 T>C ADORA2A are thought to influence these interindividual responses; therefore, the purpose of this project is to determine the effect of caffeine consumption on anaerobic exercise. 12-15 female college athletes completed two maximal WAnT30 anaerobic bike tests on a Velotron cycle ergometer.  The participants ingested a capsule of caffeine (5mg·kg^-1 bodyweight) or a placebo capsule (maltodextrin) one hour prior to testing. The order of the bolus was randomized, counterbalanced, and administered in a double-blind manner. Peak power (W·kg^-1), anaerobic capacity (W·kg^-1), and total power output (W·kg^-1) were recorded during each test.  Buccal epithelial cells were collected using a 0.9% NaCl mouth rinse with DNA extraction conducted using proteinase k to lyse cells and collection using QiAmp Mini spin columns.  Allelic discrimination was obtained using TaqMan® SNP Assay for CYP1A2 (rs762551) and ADORA2A (rs5751876) and a One-Step qPCR. Each sample was run in duplicate positive and negative quality controls. Each variable was analyzed using a factorial ANOVA with repeated measures (p > 0.05).  

The effects of caffeine on task performance and attentional focus during movement have been suggested to be influenced by the interaction between the adenosine (ADORA2A) and dopamine (DRD2) receptors in the dopaminergic pathways and specific centers in the brain. Individual polymorphisms of 1976 T/C (ADORA2A) and C957T (DRD2) are thought to influence the varied response of caffeine during movement. PURPOSE: The purpose of this study is to determine the influence of caffeine, 1976 T>C ADORA2A polymorphism, and C957T (DRD2) polymorphism on measures of unilateral and bilateral motor tasks. METHODS: The Purdue Pegboard Test and the Minnesota Dexterity Test were used to assess fine motor abilities following both placebo or caffeine ingestion (5mg·kg-1 bodyweight). Buccal epithelial cells of each subject were retrieved by a 0.9% NaCl mouth rinse. To collect the DNA samples, the cells were lysed via proteinase k and isolated with QiAmp Mini spin columns. The polymorphisms were identified using a TaqMan® SNP Assay for ADORA2A (rs5751876) and DRD2 (rs1800497) and allelic discrimination via One-Step qPCR amplification. All samples were run in duplicate with negative and positive controls. RESULTS/DISCUSSION: The Results and Discussion will be presented at the Symposium.

Individual responses to caffeine are suggested to be genetically influenced by polymorphisms of the cytochrome P450 enzymes, specifically the -163 A>C CYP1A2, for metabolism in the liver and through the adenosine receptor, 1976 T>C ADORA2A, for sensitivity of specific target cells. Individuals with the AA variant are caffeine responders, while those with the AC/CC variants are caffeine non-responders. ADORA2A TT variants demonstrate an increased sensitivity to caffeine compared to TC/CC variants. Purpose To examine the effect of caffeine and CYP1A2 and ADORA2A polymorphisms on anaerobic power during exhaustive exercise. Methods Fifteen elite NCAA male athletes (age=20.1 yrs, weight=77.4 kg, height=176.7 cm) participated in a double-blind study. Subjects performed two separate 90-s Wingate Tests (WAnT90) separated by two to four days on a Velotron cycle ergometer, resistance=0.05 kg·BW(kg)^-1. Subjects ingested a bolus of caffeine, 5mg·kg^-1BW, or a placebo (maltodextrin) one hour prior to each trial that were administered in a randomized/counterbalanced design. Peak power (W·kg^-1), total power (W·kg^-1), and average power (W·kg^-1) were calculated for the 90-s and each 30-s interval. Buccal epithelial cells were collected using a mouth rinse, 0.9% NaCl, and DNA was extracted via spin columns and proteinase k. Allelic discriminations for CYP1A2 (rs762551) and ADORA2A (rs5751876) were procured via an assay and a One-Step qPCR amplification. Samples were run in duplicate, with positive and negative controls. The data was analyzed using a factorial ANOVA with repeated measures (p > 0.05) for each variable. Results/Discussion The results and the discussion will be presented at the Symposium.

Flexibility and power output are two critical components of dance performance; however, recent research suggests that acute bouts of stretching may decrease muscular power. PURPOSE: The purpose of this study was to compare the effects of dynamic stretching (DS) and static stretching (SS) on muscular power in dancers. METHODS: 12 female, collegiate dance majors volunteered for this study. The subjects attended a familiarization session, gave informed consent, and were oriented to all testing procedures. Three different experimental sessions were conducted in randomized order for each stretching condition: DS, SS, and no stretching (NS), with a minimum of 48 hours between testing days. The sessions began with a warm-up (5 min walk) followed by a guided stretching protocol for each condition. Muscular power (peak torque, Watts) was obtained for the plantar/dorsiflexors with an isokinetic dynamometer (60o/s and 180o/s) and vertical power (W/kg BW) was obtained using both a squat (SJ) and countermovement jump (CMJ). One-way ANOVA with repeated measures with Tukey HSD post-hoc tests was conducted to determine significance (p< 0.05) for each variable. RESULTS: The results indicated that there were no significant differences for the power output of dancers after incorporating DS, SS, or NS as part of a warm-up. Isokinetic muscular power isolating the plantar flexors demonstrated no significant difference at 60o/s (DS, 57.1 + 22.7 W; SS, 57.8 +28.4 W; NS, 62.7 + 19.3 W, p = 0.82) or at 180o/s (DS, 28.3 + 9.50 W; SS, 30.5 + 14.7 W; NS, 32.2 +16.2 W, p = 0.76). In addition, measurements of power incorporating whole body power demonstrated no significant difference in jump height for the SJ (DS, 21.05 + 3.36 cm; SS, 20.83 +3.55 cm; NS, 20.9 + 3.14 cm, p = 0.893) or the CMJ (DS, 23.8 + 3.9 cm; SS, 23.7 + 5.7 cm; NS, 24.7 +6.8 cm, p = 0.98). CONCLUSION: Although some research suggests that varying forms of stretching may decrease muscular power, this study suggests that neither acute static stretching nor dynamic stretching will elicit a significant change in muscular power in dancers.  The jump athlete data is still being collected at the time of abstract submission.

Options Greeks are embedded in the definition of the Black-Scholes equation and have direct applications to the financial markets, specifically through the pricing of options contracts. This presentation endeavors to provide an overview of the Greeks by contextualizing increased retail investor participation and its collective influence on short-term price volatility for select equities. In conjunction with a consideration of the underlying theoretical mathematics, a practical example is presented to demonstrate contemporary relevance and to evaluate the implications for established hedge funds and other financial firms.

In this presentation, we will discuss our research on staircase tableaux, which are mathematical objects in the field of combinatorics that have applications in physics and biochemistry. Staircase tableaux are constructed in a similar way to Sudoku puzzles. Boxes are aligned in a staircase shape and these boxes are filled with alphas and betas depending on a few simple rules. In our research, we determined the probability of staircase tableaux that have an alpha followed by a beta on the main diagonal as well as a beta followed by an alpha. These results are interesting because of the applications staircase tableaux have in other disciplines.

The conditions of a given hypothesis test should always be taken into account when deciding what test best fits the given data. But what happens when the conditions of normal, rank based, or sign based tests are not fully met? After an introduction to the nuances and power of the usual t, Wilcoxon sign rank, and Fisher sign test, this paper will analyze the efficiencies of these tests and analyze how each responds to violations of the conditions. Through the use of a significant number of iterations of SAS created data sets, it will be shown how frequently normal, symmetric, and non-symmetric data affect the theoretical outcomes of each test. These results will play an important role in understanding the impact conditional violations have on the conclusions and overall which test is best in each data setting.

In 1971, the Society for Baseball Statistical Research (SABR) was founded. This society was the first to coin the term “sabermetrics” which refers to the empirical analysis of baseball. Prior to sabermetrics, decisions about players, teams, and coaches in baseball have always been made based upon a subjective interpretation. However, this society started a massive movement of applying an objective-based science such as mathematics to the subjective world of baseball. This combination of an objective science and a subjective sport makes sabermetrics an interesting part of mathematics to explore.

Fisher’s and Neyman’s tests are both randomization-based tests that can be used to detect effects of experimental treatments; however, they differ significantly, particularly in the construction of the hypotheses. As a result, each test has specific limitations: in certain cases, Fisher’s test can result in either Type I or Type II errors, and Neyman’s test relies on approximations of variance and normality. This project offers a comparison of these two tests and an analysis of their limitations through simulations.

The purpose of this presentation is to give an example of applied mathematics being used to enhance mathematics instruction. We show several examples of game theory applications in mathematics where connections are made to the mathematical topics of saddle points on three-dimensional graphs and the use of matrices in representing payoff functions. The main examples explored are two player games where we explore crucial information about each scenario such as the Nash Equilibrium point. The discussion follows as to how these examples can be worked into a mathematics curriculum for enrichment. We conclude by evaluating how effective practices such as game theory applications encourage appreciation, engagement, and performance in instructional settings.

Since its conception in the early 18th Century, graph theory, or the mathematical study of graphs, has become a useful tool in solving and analyzing a variety of problems. One of these problems is the seemingly simple pen-and-paper game of Sprouts created by John Conway and Michael Paterson in 1967. Sprouts is an easy game to learn and play for the casual player, but it has proven to be a much more difficult game from a competitive point of view. In order to get a better understanding of how to win any game of Sprouts, the game's properties can be analyzed and studied mathematically with the help of graph theory. The results of such analyses can be helpful in determining each game's outcome as well as understanding new variants of the game when changing a few mechanics.

Deus Absconditus is a Latin phrase used by Martin Luther and John Calvin in explaining the hiddenness, or unknowability of our God. In Isaiah 45, the prophet Isaiah writes "Truly you are a God who has been hiding himself, the God and Savior of Israel." Why is it that Christian faith accepts that God cannot be known to us? The study of Game Theory might have an answer. Game Theory is a fascinating extension of Mathematics that seeks to understand risk, analyze social interaction, and provide researchers with an understanding of why "players" behave the way they do. By researching Game Theory, I hope to support my Christian faith by providing an understanding of why our creator has chosen to be unknowable, and require belief without seeing. 

Polynomial rings are a foundational concept in understanding the characteristics of number systems. Through the examination of specific polynomial rings and their properties, it is possible to develop methods of ordering the polynomials within these rings. In this presentation, a broad introduction to polynomial rings will be provided in addition to an in-depth exploration of ordered polynomial rings and their properties. An understanding of ordered polynomial rings is useful for further development in both number theory and calculus. 

The broad applications of the Central Limit Theorem cannot be understated. It allows one to approximate other distributions with that of a normal distribution, which is crucial in hypotheses testing and modern day statistical analysis. The properties of this theorem have been observed for hundreds of years but its actual discovery and proof did not happen until 1810. The elegance of this mathematical proof is shown in this paper, along with its applicability to modern day hypotheses testing using real world data.

The Tower of Hanoi is a famous puzzle invented by a French mathematician Edouard Lucas in 1883. The task of that is we are given a tower of eight disks, initially stacked in decreasing size on one of three pegs. The objective is to transfer the entire tower to one of the other pegs, moving only one disk at a time and never moving a large one onto a smaller. How do we solve this problem? This problem may not appear to be related to mathematics, but we can actually use a recursive sequence to solve it. Sequences are mathematical objects discussed in Calculus II and the Tower of Hanoi is just one example of how sequences can be applied in the real world. 

In this presentation, I will discuss the Black-Scholes Model which is an important mathematical model in financial theory for pricing options. I will discuss the background of this model and an example of how it is used. I also will discuss some contradictions that have been found when using the model and how it is still being used.

With the speculation that electoral maps are divided into boundaries that favor one election outcome over another, the Supreme Court has deemed political gerrymandering as unconstitutional. Various mathematical models have been proposed in an attempt to combat the lack of standardized parameters for detecting the extent to which this occurs. The efficiency gap metric is a simple formula introduced as an evaluation of how extensive districting has favored one party over another within a specific boundary. With the desire to produce a more statistical-heavy method, a hypothetical state is explored and various iterations of boundaries are illustrated in an attempt to find the most effective algorithm. It is concluded that Markov chains could prove useful in the area of redistricting.

Dark matter is hard to detect because it does not interact with normal matter with a few exceptions. A dark matter candidate of interest is the dark photon which could convert with some probability to a regular photon. The dark photon is not directly detectable due to small interaction. There is theoretical motivation however to increased conversion to regular photon when it is in the presence of a plasma. We have explored what dark photon masses and what plasma conditions create favorable conditions for its detection. Previous works look at the extreme cases of mass being much larger or smaller than the plasma frequency, while we look for plasma frequency at a similar magnitude to the dark photon mass. We use the earth’s ionosphere as the medium for this project. The areas of focus are the regions of plasma frequency matching the dark photon mass where high conversions might occur due to resonance.

This is a computational research project, which uses many concepts from computer science in addition to physics. The analysis for this project is done using the python programming language.

Plant root systems are inherently 3-dimensional structures. To effectively understand these root systems—such as the model organism Arabidopsis Thaliana— it is necessary to develop and employ imaging methods capable of generating 3-D data sets. One such method is laser sheet imaging of plant roots grown in transparent media, such as in hydroponic/aquaponic cultures or hydrogels. We have been working on a custom-built laser sheet system to achieve this end. This modality requires the coordination of smooth translation stage motion, camera exposure, and camera frame rate settings. Multiple graphical user interfaces (GUIs) written in custom Python software are necessary to guide the user through the imaging process. This software communicates with the stage, controls laser power, and triggers the camera in order to synchronize motion and imaging to get clear images with definite dimensions. This system can then be used to perform time-lapse imaging of plant root structures to analyze how environmental factors affect root structure and development.

Dark Matter is assumed to exist because of the gravitational effects on stars as they move around galaxies. Efforts have been made to discover its properties in ways such as studying possible inelastic collisions in particles, but they have not yielded conclusive results. Dark photons have been motivated from theory as dark matter candidates. They can convert to regular photons at a small rate. Therefore, my study will strive to discover the regular photons emitted by the dark photons. This can be done by setting up an experimental area shielded from all light sources where the existence of a dark photon would be manifested by the appearance of regular photons in a completely dark area. To increase the chance of detection, a spherical mirror will be used to reflect the photons towards our detector, which increases our effective surface area. So far, I have worked with the photodetector and the data acquisition system in order to understand detector performance in a vacuum, which allows for photons of a shorter wavelength to last longer before they are absorbed, and how to determine the background noise from actual signals.

Despite the rapid progression of knowledge present in the field of particle physics, many mysteries still abound that have yet to be fully solved and understood; one such example is that of dark matter as there is relatively little known about it and many experiments today endeavor to detect it. This project focuses on the use of a relatively new technology, Silicon Photomultipliers (SiPMs), to detect high energy particles incident from space. It is currently hypothesized that dark matter serves as the source of high energy particles that are particularly more energetic than those incident from the sun and supernovae. The Silicon Photomultipliers are coupled with deionized water and scintillating materials to generate and absorb Cherenkov Radiation. While classic photomultipliers (PMTs) can solely be used for this experiment, they cost more and require significantly more power than that of SiPMs. While SiPMs are significantly smaller than PMTs, use of a large volume of water can increase the effective detection range of the SiPM. We detail the design, construction and overall development of multiple Cherenkov Radiation detectors. Moreover, we report on the performance of each design and discuss comparisons among each combination of models. Finally, we present the new data programming tool that has been developed to aid in the analysis and visualization of all collected data.

One of the distinguishing features of hepadnaviruses is the presence of partially double-stranded DNA. Hepatitis B is one of the well-known examples of this virus family, yet there remains a paucity of effective methods for probing this nucleic acid structure. This study investigates the use of the dye Acridine Orange (AO) as a possible probe for understanding these genomes. AO is of interest for identifying partially double-stranded DNA because of its unique spectral properties. Specifically, AO fluoresces red (~650 nm) if bonded to single-stranded DNA (ssDNA), or green (~525 nm) if bonded to double-stranded DNA (dsDNA) by intercalation. To test this system, confocal optics and single-photon counting modules were used to probe dsDNA or ssDNA at the single-molecule level.  The ratios of green/red fluorescence by AO-stained DNA in varying concentrations of AO were assessed.