Pennsylvania Academy of Science

Ahmed, Biyar*, and Debra Wohl Elizabethtown College, Elizabethtown, PA 17022. Microbiome integrity of unprocessed Canis familiaris stool samples prior to storage for fecal matter transplants.- Fecal microbiota transplants (FMT) are used for patients with dysbiosis of their gut microbiome or are facing endogenous pathogens. While FMTs are promising, it has become apparent that the gut microbiome varies between individuals and in the same individuals with diet and age. Therefore, it is difficult to establish a microbiome baseline and assess the compatibility of donor stool. A recent approach to address this issue is the storage of stool samples. This storage would prove useful for patients prone to dysbiosis pathologies. The primary goal of this experiment was to analyze changes in the stool microbiome overtime prior to stool sample storage. The alternative hypothesis was that as time passes, endogenous bacteria that are poorly adapted for the external environment would decrease while opportunistic exogenous bacteria would increase. The study was performed with stool samples passively obtained from Canis familiaris. Samples were set out in room temperature for 0 hours, 1 hour, 2 hours, 3 hours, and 2 days. Once the allotted time passed, the samples were homogenized and underwent DNA extraction and PCR amplification of the V4 16S DNA region. Preliminary findings show that there was a substantial change in the microbiome for the samples left out for 2 days. These results suggest that the microbiome does change as time passes prior to processing. Finalized data will assist in understanding the time sensitivity of sample collection and processing along with microbiome integrity, which can guide future practices of stool collection and processing. (2)

Aidoo, Emmanuel*, and Nicholas Sizemore University of Scranton, Scranton, PA 18510. Transannular cyclizations of medium-ring amido alkenes – A computational study.- Transannular cyclization reactions are an important method for transforming macrocyclic molecules into fused bicyclic structures. This type of reaction using amido alkenes as substrates has been reported, though the current scope is limited. The products formed through this type of transannular cyclization (indolizidones and related fused heterocycles) are useful synthetic intermediates for many biologically active molecules. Therefore, a better understanding of the factors governing the stereochemical and regiochemical outcomes of this sub-class of transannular cyclizations would be beneficial. As such, a computational study was undertaken to investigate the cyclization of amido alkene-derived bromonium ions. Both E and Z alkene geometries were modeled in an effort to determine their effect on the stereochemistry of the fused bicyclic products. Additionally, the spacing between the alkene and amide functionalities were varied to investigate the regioselectivity of cyclization. Initial conformational analysis calculations of halonium intermediates, transition state estimates, and bicyclic ammonium products using Merck molecular force field (MMFF) were performed in Spartan’18. Subsequent optimization and frequency calculations were performed in Gaussian’16 using density functional theory (DFT) at the B3LYP 6-31G+(d) level of theory. The results of this computational study will be described herein.   (17)

Alanazi, Razan S.*, and Amy Faivre Cedar Crest College, Allentown, PA 18104. Potential bee pollinators on the Cedar Crest College campus in Allentown, PA.- Bees and flowering plants have a mutualistic relationship; bees collect nectar and pollen from flowers and contribute to the spread of pollen, which eventually helps the plants to reproduce. However, bees often collect pollen to feed their larvae.  Thus, depending upon where on the bee’s body the pollen is collected, it may or may not be distributed to another flower, it may, instead, be groomed off and fed to young. In the fall of 2019 and fall of 2020 we collected bees visiting flowering plants that were in a garden on the Cedar Crest College campus. Using fuchsin gel cubes we collected samples of pollen from three body parts (abdomen, leg, head) in 2019 and (head, abdomen and the entire body) in 2020 of each bee. The cubes were melted on microscope slides and observed using a compound microscope. The cubes were all the same size so that we were able to quantify how much pollen each bee carried on each part of its body. We also collected the pollen from flowering plants to identify which species of plants the bees had on their bodies.  Bees could be divided into four major groups: honey bees (Apis mellifera), bumble bees (Bombus spp.), carpenter bees (Xylocopa virginica) and native sweat bees (several different species of Halictidae).  Most of the pollen on their bodies was from flowers open in the garden at the time of the study.  Information from this study can be used to determine which bees may be the most effective pollinators for plant species in our campus gardens. (58)

Armstrong, Kathryn*, Scott Kieffer, Micheal Shin, and Micheal Sedhom Messiah University, Mechanicsburg, PA 17055. The role of CYP1A2 and ADORA2A in Individual Response to Caffeine Consumption under Anaerobic Conditions.- 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 varies 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 Results and Discussion will be presented at the conference. (35)

Ayub, Hassan* Germantown Academy, Fort Washington, PA 19034. The viability of potatoes and rice as biofuel alternatives.- Throughout the past decade, there has been a tremendous increase in the research and use of renewable energy. One common renewable energy source is ethanol, which is a biofuel that primarily consists of corn, a crop with a high starch content. In order for biofuels to become a stronger renewable energy source, more alternatives are needed. In this experiment, I tested potatoes and rice, two plants that also have a high starch content, to measure their viability as biofuels. I also tested corn as a control within the experiment. The experimental setup consisted of an aluminum tumbler and a Labquest with temperature probes for gathering the experimental data. Multiple trials should be conducted for each substance and then the average and individual temperatures for each substance should be compared with each other. My experimental results rejected my hypothesis because the biofuels were unable to burn in the form I wished to test them. I did not have sufficient resources for modifying the experimental setup to try and get the biofuels to burn. Although my experiment was not successful, further research can be done for the purpose of attaining viable data through using the resources that were unavailable to me to modify the experimental setup. (75)

Baker, Taylor*, Brad Engle, and M. Dana Harriger Wilson College, Chambersburg, PA 17201. The effects of environmental stress on the equine immune response to Sarcocystis neurona and the development of Equine Protozoal Myeloencephalitis.- Pennsylvania’s horse racing industry generates over $300 million annually providing nearly 4,000 jobs. Improving equine welfare, particularly within Thoroughbred racing, is within the state’s best interest since Thoroughbreds typically have the highest number of Equine Protozoal Myloencephalitis (EPM) cases. EPM, a neurological condition, can develop in horses exposed to the parasite Sarcocystis neurona (S. neurona). Equines contact S.neurona, when grazing or drinking from areas contaminated by opossum feces leading to the development of symptoms or debilitating complications in the equine host. Antemortem diagnosis is difficult due to the parasite’s ability to infect any part of the central nervous system, the wide variation within an individual’s immune response, and current diagnostic tests only being able to confirm exposure to S. neurona. Approved drug treatments have less than a 50% success rate and may only leave the horse pasture sound. Stress is considered a risk factor as increased cortisol production can suppress the immune system, leaving horses more vulnerable to pathogen accumulation. The equine stress response is commonly assessed by salivary or serum cortisol analysis. Previous studies have indicated that the racetrack environment causes stress among equines due to the intense exercise routines, increased stall confinement, and lack of social interaction. Limited research has been conducted to understand if this increased stress in racehorses has had any negative health consequences. This literature review aims to determine whether the stress related to racetrack environments inhibit a Thoroughbred’s ability to produce S. neurona antibodies, leaving them more susceptible to EPM development. (9)

Barnhart, Brooke*, Adam Cooke, and Kathryn Sarachan Wilson College, Chambersburg, PA 17201. The potential use of Cymbopogon citratus (lemongrass) essential oil to modulate the contents of the gut microbiome.- The gut microbiome plays a significant role in nutrient metabolism, maintenance of the structural integrity of the gut mucosal barrier, and protection against pathogens. There is a great quantity of different species that contribute to the importance of the gut microbiota in mammals. While research into the composition and role of the gut microbiota is ongoing, there are known links between the gut microbiome and mood, mental health, autoimmune diseases, endocrine disorders, skin conditions, and cancer. The microbiome can be disrupted by parturition, breast feeding, medications (including antibiotics), and diet. Certain dietary supplements may be used to support a balanced gut microbiome and improve general health. Lemongrass essential oil is known to have antibacterial properties, to prevent wound infection, and to provide protection against damage in the stomach. While essential oils have been growing in popularity, little research has been done to investigate the effects of essential oils, including lemongrass essential oil, on the gut microbiome. Research in this area is required to determine whether essential oils can be used as a dietary supplement. A dietary supplement of lemongrass essential oil may impact gut microbiome by possibly decreasing certain concentrations of bacteria more significantly than others with its antibacterial and antifungal properties. (97)

Barrales, Luis*, John Martinelli, David Kleiner, Ameer Payton, Edward Winter, and Aikaterini Skokotas Rosemont College, Rosemont, PA 19010. Assessing the role of dityrosine in the UV protection of yeast spores at varying wavelengths.- Yeast spores are known to be resistant to many environmental factors including the mutagenic effects of UV light. The presence of dityrosine in their outermost spore wall may play a protective role by absorbing UV light. The DIT1 gene encodes a formyl transferase enzyme required for dityrosine production. The spores of a heterozygous DIT 1 strain of Saccharomyces cerevisiae were exposed to UV light at different time intervals and wavelengths and their ability to survive was examined. Spores present within tetrads (ascus) and free spores produced by glusulase digestion were analyzed. This study compared the UV sensitivity at the mutagenic wavelength, 254 nm and at 302 nm which is within the range of the absorption spectrum for dityrosine. Tetrads exposed to UV radiation at 254 nm were more susceptible than tetrads exposed at 302 nm. Only 3% survival was observed after a 90 sec UV exposure at 254 nm compared to 63% survival at 302 nm. A similar trend was observed for free spores. Only 0.5% survival was observed after a 90 sec UV exposure at 254 nm compared to 46% survival at 302 nm. The results suggest that dityrosine is most protective at 302 nm and tetrads may contribute to UV protection by allowing spores to hide within the ascus. (36)

Bazuk, Austin*, and Derek Straub Susquehanna University, Selinsgrove, PA 17870. Precipitation chemistry: the effects of the covid-19 pandemic.- The COVID-19 pandemic has altered daily life in the United States in numerous ways. One potential consequence of the pandemic is a reduction in the emission of pollutants into the atmosphere. This study is designed to analyze ions in precipitation samples collected at Susquehanna University and sites from the National Atmospheric Deposition Program (NADP). Precipitation can incorporate atmospheric particles and gases, specifically nitrogen and sulfur, that can be harmful for ground surfaces or waterways due to increased nutrient loading or acidification. In addition to precipitation sampling at Susquehanna University, four NADP sites were selected for this study: Penn State, Pittsburgh, Bronx, and Los Angeles.Precipitation samples at Susquehanna University and the NADP sites are collected using an atmospheric deposition sampler and analyzed using ion chromatography to determine anion and cation concentrations. The data is being split into two time periods, a pre-pandemic period and an initial pandemic period, January 1st-March 13th and March 13th – June 1st. This will allow the data to be compared for the same time periods of 2020 versus the past 10 years to determine differences in the concentrations of nitrate, sulfate, calcium, and ammonium.  The statistical test being use is a two-sample t-Test assuming unequal variance and has a critical p-value of 0.05. If the p-value from the test is under 0.05, the data is significantly different. Preliminary analyses have shown statistically significant differences during the pre-pandemic phase for Penn State and for both Penn State and the Bronx during the initial pandemic period. For Penn State, levels of Ca, NO3, and SO4 were found to have statistically significant decreases. The Bronx station showed only a significant decrease in SO4. Relative concentration changes will be used to identify the effects of the pandemic. The additional two sites will be analyzed to expand on the research. (53)

Becker, Halle*, and Jennifer K Ness-Myers, Ph.D. Messiah University, Mechanicsburg, PA 17055. Efficacy analysis of zebrafish, Danio rerio, as a novel demyelination model through exposure to cuprizone-laden food.- 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, where the immune system incorrectly attacks healthy tissue of the CNS. Most research focusing on these biological processes of demyelination have 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. (82)

Bellerose, Michael*, Matthew O’Neil*, Mina Diep, Abhai Tripathi, and Lawrence Mylin Messiah University, Mechanicsburg, PA 17055. Shelf-life study of glycerol-frozen human erythrocytes used to culture Plasmodium falciparum.- 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) [also known as the Malaria Institute at Macha (MIAM)], which is located in Macha, a rural area 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 to permit cryopreservation of erythrocytes obtained from uninfected individuals in the US, with subsequent transport to Zambia.  We have cryopreserved preparations of fresh, leukocyte-depleted erythrocyte suspensions using minimal aqueous volumes of solutions containing the macromolecular starch-based cyro-protectant hydroxyethyl starch (HES) with or without polyvinyl alcohol (PVA), or with glycerol-based solutions.  Ice recrystallization inhibitors (IRIs) prevent formation of ice crystals during the thawing process.  This presentation will describe efforts to determine if cryopreserved RBCs can effectively support asexual propagation or gametocyte formation for the P. falciparum laboratory strain NF54, and how well these qualities are maintained following long-term -80°C storage. (92)

Berlinger, Blythe* Germantown Academy, Fort Washington, PA 19034. Detecting Water Contaminants using the Bioluminescent Bacteria Vibrio fischeri.- The purpose of this experiment was to establish whether the bioluminescent bacteria, Vibrio fischeri, could be used as an indicator for water contamination. While there are many chemical assays that test for the presence of chemicals, they are expensive, time-consuming, and can be harmful to the environments being tested. Using the light emission and absorbance from the bacteria, qualitative and quantitative data were collected to determine the results. The contaminants used in this experiment were Malathion Insecticide, Copper Sulfate, and Lead Nitrate. To conduct this experiment 12 flasks of liquid culture were inoculated using photobacterium broth and a slant of Vibrio fischeri culture. All contaminants were prepared based on their EPA limits and diluted to achieve the appropriate amount for experimentation. An autoclave was used to sterilize the broth and contaminants. Using a Verner Spectrovis Spectrophotometer, the absorbance of light from each sample was calculated and then transferred into an approximate cell population. Qualitative data was obtained by taking photos of each flask culture periodically every five minutes for 45 minutes. The results demonstrated that the bacteria could detect water contamination as all samples except the control experienced over a 50% decrease in absorbance and visible light. The light depreciation most likely stemmed from the bacteria’s inability to undergo Quorum Sensing, a process that activates the bioluminescence when the bacteria is in high cell density. Overall this experiment can be used to detect contamination in any water source and it offers an organic and inexpensive alternative to most chemical assays.  (76)

Bhatt, Kaivalya* Souderton High School, Souderton, PA 18964. Regression analysis of the effect of ideology on the SARS-CoV-2 (COVID-19) pandemic.- The SARS-CoV-2 (COVID-19) pandemic has ravaged the globe both fiscally and socially. However, the true cause of this issue is hard to highlight. The goal of this study was to examine the correlation between ideology and the SARS-CoV-2 (COVID-19) pandemic is to see if a nation’s core belief system affects the overall pandemic. The two ideologies explored were Individualism, in which the individual is emphasized, and Collectivism, in which the collective unit is emphasized. The number of cases and the number of deaths caused by SARS-CoV-2 (COVID-19) were some of the variables used to represent the pandemic. In terms of experimentation, a list of all countries was imputed into Excel as well as each country’s respected ideology score and SARS-CoV-2 (COVID-19) statistics. The Pearson Product Moment Correlation Coefficient (R-squared value) was, then, used to measure the correlation between the variables. A final multivariable regression was done to account for outliers and to control a specific variable. The resulting data comparing Individualism to SARS-CoV-2 (COVID-19) cases and deaths showed a positive linear relationship—cases and deaths increased as a country’s individualistic nature increased. The resulting data comparing Collectivism to SARS-CoV-2 (COVID-19) cases and deaths showed a negative linear relationship—cases and deaths decreased as a country’s collectivistic nature increased. Furthermore, Australasian countries were outliers because even though they are highly individualistic, they have very low cases and deaths. Latin American countries were outliers because even though they are highly collectivistic, they have very high cases and deaths. Both of these outliers drastically affected the Pearson Product Moment Correlation Coefficient (R-squared value) and were, therefore, removed from the dataset to increase accuracy. Overall, my hypothesis was supported by the data because countries with a collectivistic belief system were less affected by the pandemic than countries with an individualistic belief system.  (14)

Bhuyan, Sangeetha* Germantown Academy, Fort Washington, PA 19034. Inhibiting spotted lanternfly reproduction.- The objective of this study was to determine how to inhibit reproduction in the economically endangering spotted lanternfly(SLF), Lycorma delicatula, by eliminating the gram-negative bacterial endosymbionts that they depend on to grow and reproduce. These bacterial endosymbionts in the SLF gut-microbiome allow the lanternfly to convert the sap they ingest into nutrients. Eliminating these bacterial endosymbionts inhibits the growth stage necessary for reproduction. The milkweed bug was used as a substitute insect for the experiment due to its similar feeding habits and the similar bacteria found in its gut-microbiome. Insect growth regulating insecticides (IGR) was the class of insecticides used as the independent variable because they would most effectively inhibit reproduction as they prevent reproduction, egg-hatching, and molting in young insects. An experimental control group with milkweed bugs that had no exposure to insecticides was tested. The insects from every three chambers were exposed over a two-day period. Each day 3 insects from each experimental group and the control group were dissected and their gut-bacteria was plated(18 Trials). Then gram staining was conducted to determine the number of gram-negative bacteria remaining for each trial. The environmentally safe neem oil was most effective at removing gram-negative bacteria in the milkweed bug. The percent of gram-negative bacteria in the experimental group insects, compared to the control insects, determines how effective different IGR insecticides would be at eliminating the gram-negative bacterial endosymbionts within the SLF. This would inhibit the SLF’s ability to reproduce.     (71)

Biju, Irene* Methacton High School, Eagleville, PA 19403. Mental health in the United States amidst the COVID-19 pandemic.- The purpose of this experiment was to determine if there was a correlation between COVID-19 cases and anxiety levels in the United States, and correlations were analyzed nationally, by age, by sex, by education, by race, and by region. The hypothesis was that there would be a significant positive correlation between COVID cases and symptoms of anxiety because stress is a contributing factor to anxiety. The independent variable in this experiment was the increase in COVID cases (as a percentage of the U.S. population), and the dependent variable was the percentage of the population with significant symptoms of anxiety. To conduct this experiment, data was taken from the Census Bureau’s Household Pulse Survey, which tracked mental health over two time periods in 2020, and from The COVID Tracking Project, which contained accurate case data from state and territory health authorities. STATA, a statistical analysis software, was used to test for correlations, and Pearson’s Regression tests were used to determine significance. The hypothesis was only partially supported. By examining p-values using the Holm-Bonferroni method, it was determined that there was a significant national correlation (p<0.05). However, this correlation was not significant for each of the groups. For instance, while there was a significant correlation between cases and anxiety among men, this was not true for females in the second phase. The results in this experiment would be beneficial for the medical field because the research highlighted specific groups of people who may require more attention in the future as we prepare for any similar situations. (16)

Brown, Thomas*, Hannah Laughner, Joseph Colosi, and Lara Goudsouzian DeSales University, Center Valley, PA 18034. Comparison of bacterialcommunities in different soil environments using next-generation sequencing.- Soil bacterial communities can vary greatly depending on the environment of the soil. We are investigating the differences between the bacterial communities of lawn and meadow environments that reside within five meters of each other. We predicted that soil from the meadow environment will contain a much more diverse bacterial community, as a natural environment should promote more diversity than a groomed one. We collected soil samples from each environment and then extracted the mixed bacterial DNA from each sample. We then amplified the 16S region of the DNA. Once amplified, we mixed the amplicons with specific index primers added together according to their respective environments, forming two mixed samples. The specific index primers allowed us to identify individual sequences from the group sample. These samples were sequenced by massive parallel sequencing (next-generation sequencing). We analyzed the sequence data using the BaseSpace platform (Illumina). We discovered more operational taxonomic units (OTUs) in the meadow environment in comparison to the lawn, but both were largely similar when looking at their more abundant species. (77)

Buckwalter, Silas*, and John Harms Messiah University, Mechanicsburg, PA 17055. Quantifying in vitro matrix deposition to elucidate the target of proglumide-mediated fibrotic inhibition in pancreatic cancer.- 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. (41)

Bullen, Alex *, Alex Grahe*, Penka Kassolis*, and Dr. David Singleton York College of Pennsylvania, York, PA 17405. “Genome annotation of a novel Hafnia alvei bacteriophage isolated from sewage”.- Hafnia alvei ​is a Gram negative bacterium that is commonly found in mammalian gastrointestinal tracts. With reported resistance to B-lactam antibiotics and cephalosporins, phage therapy would be a viable alternative for patients infected with ​H. alvei​; however, a bacteriophage for​ ​H. alvei​ has yet to be reported​. An unknown phage was isolated from a wastewater sample by screening for host specificity. The phage infects ​Hafnia alvei preferentially, but also infects ​Klebsiella pneumoniae​. The DNA was isolated and sequenced separated into 4 contigs for annotation. Likely peptide sequences were designated as Open Reading Frames (ORFs) using GeneMark and GLIMMER. Putative protein functions were assigned to ORFs in DNA Master using the protein blastX function in the NCBI database. The genome of the Hafnia bacteriophage aligned closely to other cataloged bacteriophages for Erwinia,​ ​Salmonella​, and ​Enterobacter​. The identity of each ORF must be confirmed with PCR, but annotation is the first step towards identification of the H.alvei bacteriophage. Having the annotations available in the database will allow for easier identification of the phage by other researchers. Comparison to other bacteriophage genomes will reveal whether the fragments can be joined to each other to create a supercontig. Once the identity of the phage is confirmed, this will allow development of phage therapy to combat antibiotic resistant Hafnia alvei infections. Phage therapy is an advantageous alternative once bacteria develop resistance to antibiotic treatment. (94)

Caroland, Kailey*, John Hanson, and Jane Cavender Elizabethtown College, Elizabethtown, PA 17022. The Effect of T-antigen expression and increase SAM68 levels on the BCL-X isoform selection.- The BCL-2 protein-family are regulators of programed cell death, or apoptosis. Apoptosis is a coordinated set of biochemical reactions that aid in growth and development. Regulatory proteins interact with inhibitors of apoptosis, while others interact with inducers. BCL-X is a specific protein whose mRNA can be spliced to produce products that are either anti-apoptotic (the long form, BCL-XL) or pro-apoptotic (the short form, BCL-Xs). The control of the isoform selection has been ascribed to the splicing factor SAM68.  Our lab is investigating the phenomena that when immortalized-human diploid fibroblasts (HDF(tert)) are transformed with the viral oncoprotein SV40 T-antigen (HDF(tert)+T), the level of SAM68 increases dramatically. To determine if this increased accumulation is correlated to the aggressive growth characteristics, we questioned whether the BCL-X isoform selection was altered. Since BCL-XL has been found to be overexpressed in some human cancers, it was hypothesized that the increase in SAM68 caused by T-antigen would consequently cause an increase in BCL-XL levels. RTPCR was used to compare the BCL-X isoforms in HDF(tert) and HDF(tert)+T cells. Surprisingly, RTPCR data shows that the transformed cells with increased SAM68 protein show no quantitative difference in the BCL-XL to BCL-Xs isoform ratios. Since it has been reported that BCL-XL can accumulate in the cytoplasm, mitochondria, endoplasmic reticulum and nuclear margins, immunofluorescence is currently underway to determine if there are cellular location alterations in the virally-transformed cells. If no differences are found, then it can be concluded that SAM68 does not affect cellular growth via BCL-X expression or location. (25)

Carver, Tiffany*, Deborah Austin, and Abigail Maley Wilson College, Chambersburg, PA 17201. The effects of taurine supplementation with a grain-free diet on cardio health in canines.- Dilated cardiomyopathy (DCM) is a type of heart disease which affects the muscle of the heart causing it to thin and resulting in the enlargement of the left ventricle. If DCM is left unresolved, it can result in congestive heart failure. In the past decade, cases of DCM in canines have increased dramatically. The growing popularity of grain-free canine diets has been suggested as a possible contributor to this increased prevalence of DCM. These diets typically contain very low levels of the amino acid taurine. Taurine is a sulfur amino acid that plays a role in the calcium pools within the cardiac cells that are responsible for proper contractions of the heart. Grain-free diets typically have a reduced amount of animal by-product which is the main source of taurine in conventional canine diets and are generally rich in legumes, such a lentils and peas, which are low in sulfur amino. Legumes also contain a high content of fermentable carbohydrates which leads to a gastrointestinal loss of taurine. Although some evidence links grain-free diets and taurine deficiency, results are contradictory and more research is needed to investigate the potential mechanism by which grain-free diets may contribute to DCM. Additionally, switching from a grain-free to a taurine-rich diet has the potential to reverse symptoms of DCM. Understanding the role diet and taurine may play in DCM in canines is important for maintaining the health of dogs in their roles as companions, therapy, and service animals. (57)

Chew, Ana*, and Wendy Boehmler York College of Pennsylvania, York, PA 17405. The effects of di-(2-propylheptyl) phthalate on developing zebrafish (Danio rerio) embryos.- Phthalates are esters of the chemical phthalic acid and are widely used in plastic manufacturing today. They serve as plastic softeners and are added to things like plastic bags, shower curtains, and medical tubing to make them more flexible. Di-(2-proplyheptyl) phthalate (DPHP) is a commonly used phthalate and was brought into the industry to replace the much more toxic chemical, di-2-ethylhexyl phthalate (DEHP). Studies have shown that these phthalate pollutants are able to detach from the main plastic polymer and leach out into the waterways. These phthalates have been linked to human health complications by effecting endocrine and reproductive systems. Even though many phthalates have been evaluated for developmental toxicity in zebrafish embryos, no study has evaluated DPHP in development. At 24 hours post fertilization (hpf), zebrafish embryos were chronically exposed to environmentally relevant concentrations of DPHP for 4 days (until 5dpf). While there was no significant effects on survival rate, body length was significantly reduced over time and heart rate was significantly increased over time. DPHP also had a profound effect on locomotor activity by 5dpf. Zebrafish embryos chronically exposed to DPHP were significantly hypoactive in comparison to controls. These findings suggest that DPHP may not be a safer alternative to DEHP and future studies will need to elucidate the mechanism of toxicity of these phthalates. (64)

Chiang, Angelika*, Samantha Valaitis, and Quyen Aoh Gannon University, Erie, PA 16541. Role of SCAMP3 Regulating CXCR4 Trafficking.- The CXC-Chemokine Receptor Type 4 is a G-protein coupled receptor. The functions of CXCR4 is to regulate the growth, division, differentiation, and migration of cells. The overexpression of CXCR4 is connected to metastasis in over twenty-three types of cancer along with promoting HIV infection. Under normal conditions, CXCR4 binds to its agonist CXCL12 and is endocytosed into the cell and transported to the early endosomes. At the early endosomes, the CXCR4 is ubiquitinated by the ubiquitin ligase ITCH and is sorted into multivesicular bodies (MVBs) by Endosomal Sorting Complexes Required for Transport (ESCRTs). The MVBs buds off the early endosomes and are targeted for degradation in the lysosomes. The disruption of protein functions during this pathway can inhibit lysosomal degradation and lead to CXCR4 being overexpressed. In this research, we will investigate the role of SCAMP3, a Secretory Carrier Membrane Protein, which regulates CXCR4 trafficking and is known to interact with the ESCRT protein Hs and ubiquitin ligase ITCH. For our experiments, Hela cells, an immortal cell line derived from cervical cancer, were used. We will first do a knockdown of SCAMP3 to silence the SCAMP3 gene and then use RNA interference to examine CXCR4 trafficking in the absence of SCAMP3 while using a well-characterized immunofluorescence assay. The immunofluorescence used to determine the colocalization of CXCR4 in the lysosomes are the lysosomal markers LAMP1/2 and CXCR4 antibodies and in the early endosomes are the early endosomal markers EEA1 and CXCR4 antibodies. ImageJ colocalization program will be used to determine the colocalization between CXCR4 and organelle markers. A trafficking time course will be used to ensure that CXCR4 has arrived in the early endosomes between 30-60 minutes and in the lysosomes at 180 minutes. We anticipate that SCAMP3 could either inhibit or promote the degradation of CXCR4 transportation to the lysosomes. (22)

Chobanoff, Anna*, and Jennifer Elick Susquehanna University, Selinsgrove, PA 17870. Providing an interpretative guide for geology displays: Everhart Natural History Museum, Scranton, PA.- A new guidebook for docents volunteering at the Everhart Museum of Natural History, Science, and Art has been produced to help interpret displays for the tours they give. The Everhart Museum is located in the Wyoming Valley, in Scranton, PA. It’s modest collection contains mostly invertebrate and plant fossils from the Paleozoic Era, predominantly from the Carboniferous Period, which is the bedrock underlying much of the region. Other fossils in the museum include several vertebrates including Mesosaurus, a fish, two Dinosaur reproductions, and a few miscellaneous bird- relate fossils. Interviews with docents and an integration of some 4th Grade level Academic State Science Standards provided the context for the development of this guidebook. Though the docents had educational backgrounds, they lacked a general geologic understanding of concepts and principles related to geologic time, fossil preservation and the importance of the fossils in the Everhart Museum collection. They were also unaware of the education standards they could include their tours. This guidebook may help docents give improved, informed tours while possibly achieving some of the Pennsylvania/National Academic Standards for Environment and Ecology.   (55)

Clements, Caleb*, Catherine Falkenstein*, and Jane Cavender Elizabethtown College, Elizabethtown, PA 17022. Effect of SV40 T-antigen Oncoprotein Expression in Human Diploid Fibroblasts on the Production of SAM68 and subsequent Cyclin D1 Isoform Selection.- Virally induced tumorigenesis accounts for 20% of cancers, totaling 2.8 million new cases yearly. To study viral oncoprotein disruptions, our laboratory employs the simian virus 40 (SV40) T-antigen to induce cellular transformation. In this study, immortalized-human diploid fibroblast cells were transformed with the T-antigen gene.  It was found that the level of the alternative splicing factor, SAM68, increased dramatically in the presence of the oncoprotein. SAM68 RNA-binding protein functions as a splicer, vital to cell proliferation and regulation. This investigation sought to determine if this increased SAM68 was the variant that excludes the KH region (SAMΔKH) and does not exhibit splicing activity; or, if the wild-type isoform was amplified.  To determine if the splicing activity was affected, the downstream target cyclin D was assessed. Cyclin D is an early G1 cell cycle regulator, following cellular division. Two cyclin D isoforms (D1a, D1b) have been correlated to various forms of cancer; thus, it was hypothesized that the viral oncoprotein induced high levels of SAM68 which in-turn created a more active cyclin D, promoting the cell cycle. Literature has shown that increased SAM68 in tumor cells select for the non-spliced variant, and thus similar results are expected. Cyclin D isoform selection following SAM68-induced splicing is yet to be determined in T-antigen expressing cell lines, however other studies have shown D1b variant selection in tumor cells. These studies are essential for viral-induced cancer research due to their insight on preferential splicing behavior, and the resulting impact on downstream proteins involved in oncogenesis. (45)

Cole, Adam *, Michael Shin, and Scott Kieffer Messiah University, Mechanicsburg, PA 17055. Effects of caffeine and CYP1A2, ADORA2A polymorphisms on exhaustive anaerobic performance.- 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. This research examined the effect of caffeine and CYP1A2 and ADORA2A polymorphisms on anaerobic power during exhaustive exercise. 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 discrimination 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. (34)

Croucher, Kristina*, and Michael Foulk Mercyhurst University, Erie, PA 16546. Identification and characterization of the Sciara coprophila cold tolerance gene, Frost (ScFst).- Organisms in the wild routinely encounter extreme environmental conditions.  Seasonal cold temperatures pose a threat to insects in particular; thus, they have developed mechanisms to recover from cold shock. Recently, the effects of cold on the fruit fly Drosophila melanogaster have been studied and a number of genes responsible for cold tolerance have been identified. One of these genes is Frost (Fst). While the specific function of Frost has yet to be determined, expression of the gene has been shown to rise during the period of recovery after exposure to cold temperatures. In this study, we have identified and  begun to characterize a potential Frost ortholog in the fungus gnat, Sciara coprophila. The D. melanogaster Frost sequence was collected from NCBI and BLAST was used to identify a potential Frost ortholog in the S. coprophila genome. Several candidates were initially identified and a reciprocal BLAST search with each of the candidates was completed to identify a potential S. coprophila Frost ortholog, which was named the putative S. coprophila Frost (ScFst) gene. Total RNA was extracted from adult female flies subjected to a three-hour cold stress then allowed to recover for 0, 1, 2, and 3 hours.  Quantitative Real-Time Polymerase Chain Reaction (qPCR) was used to analyze putative ScFst expression at each of these time points. Expression increased for at least the first hour time point. Future plans are to analyze ScFst expression during recovery from cold stress in other developmental stages, such as embryo, larvae, and pupae. We are also in the process of cloning the putative ScFst mRNA into a topocloning vector known as pCR2.1 and then subclone the sequence in frame with GFP. (31)

Crowther, Kallon*, and Wendy Boehmler York College of Pennsylvania, York, PA 17405. The temporal expression pattern of a neuropsychiatric risk gene, complement component 4, during zebrafish (Danio rerio) development.- Schizophrenia is a debilitating psychiatric disorder. There is evidence that its pathogenesis has links to neurodevelopment and genetic variability of the complement system. A recent genome wide association study strongly implicated an increased copy number and increased expression of the complement component 4 (C4) gene in patients with schizophrenia. During development, C4 has been shown to play a role in synaptic pruning and it has been suggested that the overexpression of C4 may cause excessive synaptic elimination potentially underlying the cause of the disorder. To determine the temporal expression pattern of C4 in zebrafish, RNA was extracted from 24, 48, 72 hours post fertilization (hpf) and 5 days post fertilization (dpf) in zebrafish embryos.  Using a reverse transcriptase polymerase chain reaction (RT-PCR) strategy, we found C4 was expressed throughout all developmental timepoints tested. It will be of interest to determine the spatial expression pattern by using whole mount in situ hybridization in this model system. The identification of the C4 gene in zebrafish will allow for future studies that can use the genetic tools available in zebrafish to further elucidate the functional role C4 may have in developmental synaptic refinement. (85)

Cruzan, Annelyse*, Michael Shin, and Richard Schaeffer Messiah University, Mechanicsburg, PA 17055. Altered heavy metal stress response of mutant Arabidopsis thaliana plants in the presence of zinc and nickel heavy metal ions.- 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 and zinc tolerance assays 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 and zinc. 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 heavy metal stress response of the plant. (66)

Culver, Melissa*, and André Walther Cedar Crest College, Allentown, PA 18104. Identifying potential contamination of beer yeast through quantitative PCR with High Resolution Melt analysis.- Beer making involves the fermentation of sugars extracted from malted grains using different species of brewing yeasts.  There are many species of brewing yeasts used in beer production, but two of the most commonly used are the top fermenting yeasts species Saccharomyces cerevisiae used in the brewing of ales, and the bottom fermenting yeasts species Saccharomyces pastorianus used in the brewing of lagers.  Different yeast strains lead to different flavors, alcohol contents, and color profiles, so it is important to use monocultures of the proper yeast strain to obtain the desired beer.  Large breweries reuse the yeast multiple times (re-pitching), however the yeast can become contaminated leading to unusable product. Current tests for contamination are strenuous and take a long time to obtain results, which wastes time and money.  We have focused on creating novel molecular tools that large beer companies could use to test for contamination in their batches of beer faster and more effectively. Our research has focused on using quantitative Polymerase Chain Reaction (qPCR) with High Resolution Melt (HRM) analysis of short tandem repeat loci that have been identified in different yeast species. After qPCR amplification of STR loci, HRM was used to identify the melting point of loci and to identify unique melting temperature signatures of different beer yeasts.  We have been able to generate genetic melting point fingerprints that distinguish between different species of beer yeasts (lager vs ale), and have also been able to distinguish between different strains within the same species of beer yeast.  We are in the process of examining over 20 different strains of beer yeasts to generate genetic identity library, that will allow large brewing companies to rapidly identify the presence and identity of contaminating yeasts in their fermentation tanks. (7)

DeGroot, Jack*, Cody Dosch*, Heather Kwolek*, and Craig Johnson Marywood University, Scranton, PA 18509. A Mathematical Model for Forecasting the Spread of Covid-19 in Pennsylvania.- In an expansion of a completed SIR mathematical Model research project, students Cody Dosch, Heather Kwolek, Jack DeGroot, and faculty advisor Dr.Craig Johnson create a SVEIR mathematical model for the spread of Covid-19. The SIR model is a simple mathematical model of epidemics that utilizes three compartments of Susceptible, Infected, and Removed. The SVEIR mathematical model is an advanced version of the SIR model with additional compartments of Vaccinated and Exposed. The SVEIR model will provide a more precise outlook on how Covid-19 will spread from person to person in Pennsylvania due to additional parameters. These parameters consist of rates per day that pertain to the vaccine coverage rate over Pennsylvania, the vaccine administered into individuals, symptoms occurring for an infected individual, as well as the recovery rate and death rate of Covid-19. The creation of the SVEIR model was done by expanding our original 3 x 3 system of first-order differential equations to include additional compartments consequently creating a 5 x 5 system of first-order differential equations. We wrote and utilized a C# program that uses the numerical method Runge Kutta to forecast the number of Covid-19 cases within a certain time period t within the population of the state of Pennsylvania. We will utilize our SVEIR model to predict the number of covid cases that will occur within the months of May, June, and July. (13)

Dias, Kathlyn*, and Andre Walther Cedar Crest College, Allentown, PA 18104. Identification of Replication Protein A phosphorylation dependent protein-protein interactions via Yeast Two Hybrid system.- The heterotrimeric ssDNA binding protein Replication Protein A is crucial to the study of Cancer because due to its involvement in DNA replication, repair, and recombination. Dysfunctional proteins involved in those pathways have been directly linked to cancer. This study attempts to further understand RPA by identifying the proteins it interacts with when its 32kDA subunit is phosphorylated or dephosphorylated. Discovering proteins that have phosphorylation-dependent interactions with RPA will reveal other proteins that can be used for cancer research and provide a clearer image of the players that could be part of the processes RPA is involved in. Since RPA is highly conserved in eukaryotic organisms a Yeast Two Hybrid assay was used to determine interactions through Saccharomyces cerevisiae. Five RPA variants have been mated with random cDNA yeast library proteins, one of the RPA variants is a hyperphosphorylated 32kDa subunit and another is dephosphorylated 32kDa subunit while the rest are wild type 70kDa, 32kDa, and 14kDa subunits. We have identified a number of candidate proteins that  have displayed phosphorylation-dependent interactions with RPA through yeast two hybrid. (43)

Diep, Mina*, Matthew O’Neil, Michael Bellerose, Abhai Tripathi, and Lawrence Mylin Messiah University, Mechanicsburg, PA 17055. Use of cryopreserved human erythrocytes to culture Plasmodium falciparum.- 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) [also known as the Malaria Institute at Macha (MIAM)], which is located in Macha, a rural area 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 to permit cryopreservation of erythrocytes obtained from uninfected individuals in the US, with subsequent transport to Zambia.  We have cryopreserved preparations of fresh, leukocyte-depleted erythrocyte suspensions using minimal aqueous volumes of solutions containing the macromolecular starch-based cyro-protectant hydroxyethyl starch (HES) with or without polyvinyl alcohol (PVA), or with glycerol-based solutions.  Ice recrystallization inhibitors (IRIs) prevent formation of ice crystals during the thawing process.  This presentation will describe efforts to determine if cryopreserved RBCs can effectively support asexual propagation or gametocyte formation for the P. falciparum laboratory strain NF54, and how well these qualities are maintained following long-term -80°C storage. (4)

Divers, Hayden*, and Sean Georgi York College of Pennsylvania, York, PA 17405. Exploring Expression Level Variations of RERE and CircRERE in Human Cells.- Circular RNAs (circRNAs) have become increasingly relevant in recent years of scientific research, as a result of their identified functions in microRNA sponging, controlling gene expression, influencing cell growth, and acting as disease biomarkers. However, the function and expression of most circRNAs have not yet been fully characterized. Our previous research identified circRERE as a circRNA that is dynamically expressed in developing chicken (Gallus gallus) embryonic retinas, suggesting that its expression levels may be intentionally controlled across cell types during development and beyond. This study aimed to identify human cell types that express circRERE and characterize its expression. Using PCR and sequencing, our research identified two distinct forms of circRERE, which we call circRERE5 and circRERE6, in human melanoma (MCF-7), breast cancer (MDA), glioblastoma (U87), and embryonic kidney (HEK-293) cells. Using qPCR, we were able to verify that circRERE5, circRERE6, and RERE were expressed differentially across each cell type. CircRERE5 showed the greatest expression levels in HEK293 cells, over twice as high as its second highest levels found in U87 cells, and nearly eight times greater than its lowest expression levels found in MDA cells. Furthermore, circRERE6 was also found to show its highest expression levels in HEK-293 cells, around three times greater than its expression levels in U87 and MCF-7 cells, and approximately six times greater than its levels that were measured in MDA cells. These findings could possibly suggest that circRERE5 and circRERE6 are playing key roles in development and/or the progression of human cancers. (21)

Doyle, Chloe*, and Jessica Nolan York College of Pennsylvania, York, PA 17405. Ingestion of microplastics by macroinvertebrates in streams in York County, PA as an indication of pollution.- Microplastics are very small particles that have broken down from larger pieces of plastic. They can also be anthropogenic and used in beauty products and cleaning products. Because of their size, they can infiltrate soil, waterways, and the bodies of organisms. Microplastics can even be ingested by tiny organisms, such as macroinvertebrates, and work their way up the food chain as other organisms consume them. There is no information regarding the environmental influence and prevalence of microplastics in York County, PA. This study looks at macroplastic (plastic litter) pollution in rural and urban streams in York County and if microplastics were ingested by macroinvertebrates. The relative abundance of macroplastics found in and around rural and urban streams was determined. Microplastics in the bodies of macroinvertebrates from the same streams were also quantified by digesting their bodies in a hydrogen peroxide and hypersaline solution. It was found that there were significantly more macroplastic items were present on average in urban streams (48.33) than rural streams (1.33). More microplastics were ingested by macroinvertebrates on average in urban streams (5.66) compared to rural streams (1.66) as well, but the trend was not significant. Pollution sensitive macroinvertebrates tended to ingest more microplastics than tolerant species. No relationship was found between the size of the macroinvertebrate and the amount of microplastics ingested. The results from this study help show what areas of York County are most at risk to being affected by microplastic pollution. (54)

Empfield, Keirstyn*, Patricia Tadley*, Alex Abouafech, Joseph Colosi, and Lara Goudsouzian DeSales University, Center Valley, PA 18034. Discovering antibiotic producing bacteria from soil.- Antibiotic resistant bacteria are on the rise, resulting in a shortage of antibiotics that can be used to treat many types of bacterial infections.  Some bacteria, especially soil bacteria, naturally produce antibiotics effective against other species of bacteria to compete for nutrient resources.  We can take advantage of this survival mechanism to discover new antibiotics.  We created a master plate of 40 distinct bacterial colonies taken from serial dilutions of a soil sample collected from a lawn located on the DeSales University campus. 24 of these colonies grew on our master plate.  Using replica plating, we are testing these isolates for antibiotic production against 10 safe relatives of the ESKAPE pathogens.  One isolate from our master plate has produced zones of inhibition in lawns of both Staphylococcus epidermidis and Bacillus subtilis, indicating susceptibility of these two strains to the antibiotic produced by the novel isolate.  The antibiotic producer is a Gram-positive rod.  We are performing PCR amplification and sequencing of the 16S genomic region of this isolate to identify it.  We are also testing other ESKAPE relatives for sensitivity to the antibiotic. (98)

Evans, Arianna *, and Bridgette Hagerty York College of Pennsylvania, York, PA 17405. The relative activity of the domestic cat (Felis catus) in two county parks in south central Pennsylvania.- Domestic indoor/outdoor hunting cats (IOHC) (Felis catus) are currently estimated to have a population of 95.6 million in the United States with a range of 40-80% of the population being allowed to venture outside without supervision for varying amounts of time. It is estimated that domestic cats are responsible for killing 6.7-20.7 billion small mammals annually as well as driving several island bound species to extinction. The goal of this study was to identify activity levels of domestic cats at varying distances into the forest edge from residential areas in Richard Nixon County Park and John Rudy County Park located in York County, Pennsylvania. Wildlife cameras were used to accomplish this by being placed along the forest edge at both parks. Eight cameras were placed in Nixon Park and three cameras were placed in John Rudy Park where they were adjacent to residential property. Images were analyzed for 30 minute trap events from August to November 2020. During this time there were a total of three trap sessions. Capture success rate was compared between the two parks and distance from the forest edge. While activity did not differ among distances, the capture success rate was much higher at John Rudy Park, as Nixon Park had no cat activity. In John Rudy Park, capture success of IOHC was higher, but capture success of mesopredators, such as red fox and raccoon, were considerably lower. A mark-recapture study could be completed using wildlife cameras at John Rudy Park to successfully identify individual cats in the park and to record their activity. In Nixon Park, other locations that are adjacent to the residential and agricultural land should be included.   (61)

Gallant, Kayla*, and Cynthia Keler Delaware Valley University, Doylestown, PA 18901. The effect of sugar alternatives on the Drosophila microbiome.- Gaining a better understanding of the gut microbiome and how it influences the overall health of an organism has become a major focus of research studies. Tests involving sugar additives are explored in this study to determine the impact of the popular substances Splenda, Stevia, and Sweet n’ Low on the microbiome of Drosophila melanogaster, specifically Lactobacillus species and Acetobacter species levels. Populations of Drosophila were fed diets containing these substances and allowed to feed on the compounds for a week. The populations were fed hydrogen peroxide as a negative control which will kill the gut bacteria. Cornmeal which has been proven to increase Lactobacillus sp. populations and was used as a positive control, and fresh fruit was used as a positive control for the presence of Acetobacter sp. in the Drosophila microbiome. After a week, the flies were collected and stored at -70˚C until bacterial populations could be enumerated. Enumerations were performed using MRS media, selective for Lactobacillus sp., and Ethanol Media, selective for Acetobacter sp. This was repeated so that 3 samples of each population were tested, the average was then taken. Flies fed fruit contained a level of bacteria fivefold less than that of the control and were the only group to contain Acetobacter sp. All three sugar alternative substances tested had reduced levels of Lactobacillus sp. bacteria fivefold less than that of the control sample. Upon comparison of the Lactobacillus sp. CFU (bacteria/mL) in each sample it was determined that all three tested sugar alternatives negatively affected Lactobacillus sp. bacteria in the gut microbiome of the fruit fly. (91)

Goodwill, Gabriella*, Russell Minton, Holly Mihaly, and Riccardo Fiorillo Gannon University, Erie, PA 16541. Phenotypic plasticity in Lithasia geniculata includes shell density.- Pleurocerid snails exhibit shell variation that generally varies with the environment and predation; shells are thicker, more conic, and harder to crush upstream than downstream. Little is known, however, about whether the density of shell material varies in a similar fashion and how it correlates with other shell characteristics and the environment. Using eight populations of Duck River Lithasia geniculata, we measured shell material density as a function of X-ray radiopacity and shell thickness and correlated it with river mile and crushing strength. Populations differed in their density which was positively correlated with river mile whether adjusted for shell thickness or not. Regression indicated that shell density showed positive correlation with thickness and negative correlation with crushing strength. Our results in L. geniculata are the first to show variation in shell material density in pleurocerids, and our data suggest adaptive trade-offs in response to hydrology and predation pressures. (49)

Hancock, Carter* Souderton High School, Souderton, PA 18964. Comparison of computer neural networks’ ability to solve mazes with and without a dedicated memory circuit.- Computer neural networks were inspired by the adaptability of neuron connections in the human brain, and thus it stands to reason that other useful machine learning algorithms can be derived from the workings of the human brain, more specifically, the human mind’s ability to store and retrieve information for later use. Neural networks should be able to gain experience and develop instinct just like humans. This experiment explored whether neural networks with spatial memory could be made and whether they would fare better at solving a maze than neural networks without dedicated memory systems. The results showed that the neural networks with spatial memory had a significantly lower ratio of revisited maze tiles to total visited maze tiles, indicating that the memory circuit did help the neural network complete the maze more efficiently. (40)

Hane, Kimberly*, Andre Walther, and K. Joy Karnas Cedar Crest College, Allentown, PA 18104. Quantitative PCR for forensic bodily fluid identification.- Crime scene analysts face the challenge of reconstructing the series of events that occurred in commission of a crime; this includes identifying bodily fluids and their method for being deposited in order to contextualize the scene. Current analyses tend to be based on enzymatic-based color change tests or qualitative methods involving simple observation of the stains themselves, both of which can generate misleading results, altering the interpretation of the incident. For example, stains generated from sweat, semen, or saliva may all look similar, and while each may yield a DNA profile, the nature of that stain may tell a completely different story for how it came to be at that crime scene. Menstrual and peripheral blood are similar in appearance, as are the spatter patterns that result from expiration and an impact; however, a blood stain that results from a victim coughing up blood carries a completely different meaning than one that was deposited following a blow. To enhance the robustness of stain identification, this study uses quantitative PCR to help automate forensic analysis. The methodology begins with the isolation of DNA from the deposited stain, allowing the sample to be simultaneously analyzed through human STR typing. To differentiate between expirated and impact blood spatter, qPCR was used to identify the oral cavity-specific bacterium, Streptococcus salivarius in blood that was expirated. To distinguish saliva from other fluids, the methylation status of the MDFI gene was determined using bisulfite conversion followed by High Resolution Melt analysis in a real-time qPCR assay. Not only was this method able to accurately classify saliva, but it also differentiated between menstrual and peripheral blood, a feat not achieved by current blood identification protocols. This project demonstrates the utility of molecular methods for characterizing bodily fluids at crime scenes, suggesting a novel approach to forensic analysis. (6)

Holliday, Emma*, M. Dana Harriger, Abigail Maley, and Budhan Pukazhenthi Wilson College, Chambersburg, PA 17201. L-Selectins and their ligands: potential applications for improving fertility in domestic and endangered ungulates.- Rising global extinction rates have prompted conservationists to seek out new strategies to combat declining populations of endangered species.  Improving the reproduction of captive populations is a critical area of research that has led to the development of assisted reproductive technologies (ART) such as in vitro fertilization (IVF) and embryo transfer.  While ART has many benefits for endangered species, it is an expensive and labor-intensive process that can yield poor results.  The ability to pre-determine which individuals are most likely to be receptive of ART may improve the success rates of establishing a pregnancy.  L-Selectin, a biomarker for fertility, may provide important insights about the implantation process, repeat implantation failure, and be used to predict an individual’s fertility status to determine the best candidates for ART.  Studies in humans demonstrate both the role L-selectin and their ligands play in the implantation process as well as the interactions that occur between the blastocyst and endometrium during implantation.  Studies in domestic livestock and non-domestic ungulate species reveal the current success rate of IVF and embryo transfer, and summarizes the incidence of early embryonic loss in ungulate species.  There is a lack of information, however, regarding the efficacy of L-selectin ligand screening for fertility status in equine species.  These findings demonstrate the need to construct a methodology for determining the best candidates for embryo transfer that can be applied to the domestic horse (Equus ferus caballus), and, ultimately, the endangered Przewalski’s horse (Equus ferus przewalskii).  Future applications of L-selectin ligand screening in other endangered species will also be discussed. (12)

Igatpuriwala, Nadia*, and Bernado Mesa-Cruz Elizabethtown College, Elizabethtown, PA 17022. Comparing immobilization performance of two anesthetic protocols (ketamine-xylazine and ketamine-xylazine-Telazol®) in chemically immobilized Ursus americanus (American black bears).- Optimal wildlife anesthetic protocols should induce rapid immobilization, allow for rapid recovery, and provide a wide margin of safety. The ketamine and xylazine (KX) anesthetic protocol is commonly used in chemical immobilizations of Ursus americanus (black bear); however, some biologists report unreliable due to inconsistent recovery times, side effects such as convulsions and hyperthermia induced by required doses of ketamine. In recent years, some biologists have employed the ketamine, xylazine and Telazol® (TKX) anesthetic protocol, which requires relatively lower dose of ketamine and telazol when administered together. Black bears drastically decrease their vital and metabolic rates during hibernation as compared to the active and hyperphagic states. This scenario could alter the efficacy of anesthetic agents during different bear physiological stages. Unfortunately, there is no evidence on whether TKX performs better than the KX protocol under any bear physiological stage. Thus, our objective was to compare the chemical immobilization performance of both KX and TKX protocols in bears housed at Virginia Tech’s Black Bear Research Center. Protocol performance was assessed through vital sign frequencies, induction time, and recovery time in 15 bears. We used linear models to compare variables across different metabolic states. We found more consistent vitals using TKX compared to the KX protocol. Induction times remained similar in pre-hibernation ~10min, yet TKX produced longer inductions (8min) than KX during hibernation, and TKX produced shorter inductions (21min) than KX post-hibernation. Route of reversal agent administration influenced recovery times regardless anesthetic protocol or physiological stage, where intravenous was 21min faster than intramuscular, and 29min faster than per rectum. Both TKX and KX produced immobilization lasting at least 100min. We recommend using KX if immobilizing bears during hibernation and administering TKX during the pre and post hibernation states, as it produces shorter induction times and less variable bear vital signs. (8)

Jaber, Khoula *, and K. Joy Karnas Cedar Crest College, Allentown, PA 18104. fabI gene alteration as a mechanism of triclosan resistance in Enterobacter cloacae.- In response to increased exposure to triclosan, Enterobacter cloacae has developed resistance to the compound. The underlying changes to the genome were investigated focusing on the  fabI gene, which codes for an enzyme involved in the lipid biosynthesis pathway in En. cloacae and is the target for triclosan. Triclosan binds to the target, blocking lipid biosynthesis in En. cloacae and therefore blocking growth of the bacteria. We found that with continued exposure to triclosan, En. cloacae became increasingly insensitive to the compound, and eventually developed complete resistance. We analyzed the specific genetic changes, focusing on the fabI gene in different strains with different levels of resistance. In the completely resistant strain, a known fabI mutation had occurred, but in the moderately resistant strain, there were no signs of the same mutation. There must be another mechanism causing partial resistance in that strain. Further research in this strain suggests exploration in other genetic mutations and possible gene duplication occurrences in the fabI gene.   (90)

Kabonick, Seth *, Noah Smith , Anne Reeve, and Sarah Codd Messiah University, Mechanicsburg, PA 17055. Synthesis of novel hydroxychalcone derivatives as potential inhibitiors of protein tyrosine phosphatase 1B.- 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. (1)

Kalchthaler, Brady*, and Christopher Dolanc Mercyhurst University, Erie, PA 16546. How will the EAB impact Ash trees of different sized diameters?– The emerald ash borer (EAB), Agrilus planipennis, is an invasive wood-boring beetle native to eastern and southeastern Asia. Mature EABs feed on the foliage of ash trees (spp. Fraxinus) causing little damage, while larvae feed on the phloem. Because adult beetles move around, flying from treetop to treetop, tree size would seem to impact the spread. Additionally, evidence suggests that females are limited to laying eggs on trees with prominent bark, or larger, more mature individuals. In this paper, we aim to distinguish which size classes of ash are most susceptible to an EAB infestation. We hypothesize that ash health decline and ash mortality rates will be lower among smaller ash trees. As part of an ongoing census starting in 2016, two forested plots were sampled in Asbury Woods near Erie, PA dominated mostly by pumpkin ash (Fraxinus profunda). Trees in both plots were identified, given an ash-rating on a scale of 1-5 (5 being the worst), and an ash break-up rating (1-5). Ashes were subdivided into three categories (<10 dbh, 10-19.9 dbh, and ≥ 20 dbh) for analysis. Results indicated the EAB prefers the categories of 10-19.9 dbh, and ≥ 20 dbh. The 10-19.9 dbh category exhibited an average ash-rating of 4.21 in the year 2020, and the ≥ 20 dbh category exhibited an average ash-rating of 4.66. The average ash-rating in the <10 category for the year 2020, was 3.43. Lower average ash-rating correlates to healthier trees, suggesting that trees >10 dbh have a higher mortality rate due to their greater decline in health from years 2016-2020. Results do support our hypothesis, however, the average ash-rating for trees in the <10 category are still declining in health, suggesting that ashes in each category are susceptible to infestations and the future of the species remains in question. (63)

Kavanagh, Katie *, and Megan Rothenberger Lafayette College, Easton, PA 18042. Invasive Asian shore crabs and Chinese mitten crabs: current response, best management practices, and competition dynamics with native blue crabs.- The Asian shore crab (Hemigrapsus sanguineus) and Chinese mitten crab (Eriocheir sinensis) are two invasive species with emerging populations on the eastern coast of the U.S. that have raised concern among scientists and conservation managers because of the environmental and economic harm they have caused in aquatic ecosystems in Western Europe and along western coast of the U.S. The objective of this research study is to use the Asian shore crab and Chinese mitten crab as case studies to explore what is being done, what is missing, and what ought to be done to improve management responses to marine bioinvasions.  To address this overall goal, we performed a competition experiment between the Asian shore crab and the blue crab (Callinectes sapidus), an economically important native species. We also performed interviews with conservation managers to understand where gaps between published management plans and on-the-ground management of the crabs may occur and to generate recommendations for their improved management. Preliminary results from laboratory experiments indicate the potential of the Asian shore crabs to compete with blue crabs for food, signaling the importance in managing the growth and spread of already-established Asian shore crab populations. Management responses to the Asian shore crab and Chinese mitten crab, however, have been limited due to constraints in coordination, funding, time, and human resources. (72)

Kenney, Kyle*, Mary Pistack, and Angela Asirvatham Misericordia University, Dallas, PA 18612. The Effects of Rolipram, a Selective Phosphodiesterase Inhibitor, on Immortalized Schwann Cell Proliferation, AKAP-95 and Cyclin D3 Expression.- Schwann cells are a vital component of the Peripheral Nervous System and aid in the repair of axons following injury. The regulation of Schwann cell growth in vitro is facilitated by heregulin, a neuron-secreted growth factor, and an unknown mitogen that activates the cyclic adenosine monophosphate (cAMP) pathway. The abundance of intracellular cAMP is regulated by a family of enzymes called phosphodiesterases (PDEs). PDE inhibitors such as rolipram have therapeutic potential in various disorders and function by increasing the levels of intracellular cAMP. A-Kinase anchoring proteins (AKAPs), a family of scaffolding proteins that belong to the cAMP/Protein Kinase A (PKA) pathway are known to bind both PDE and PKA to regulate cAMP concentration in cardiac myocytes. Previous studies have shown that AKAP95, a nuclear AKAP, known for scaffolding cyclins, is essential for Schwann cell growth. Based on these reports, it was hypothesized that increasing the concentration of rolipram would elicit a dose-dependent increase in Schwann cell proliferation by augmenting the expression of AKAP95 and cyclin D3.  Immortalized Schwann cells were cultured with no mitogens, 12.5 ng/mL heregulin, 1 µM of forskolin (a pharmacological activator of cAMP), heregulin + forskolin, and various doses of rolipram at 0, 1, 5, 10, 25, and 50µM for 12 or 24 hours. Using the MTT assay, preliminary results indicate that cells incubated for 12 hours and 24 hours exhibited the highest rate of proliferation at a dose of 5µM and 10 µM rolipram respectively. Meanwhile, immunoblot analysis revealed that in cells treated with heregulin + foskolin, the expression of cyclin D3 and AKAP95 was highest when incubated with 25 µM and 50 µM of rolipram respectively. These results suggest that increasing the concentration of cAMP by inhibiting phosphodiesterases augments Schwann cell proliferation by amplifying the expression of proteins regulating cell division. (81)

Kimbel, Kylee*, Andrew Resh, and Jennifer Ness-Myers Messiah University, Mechanicsburg, PA 17055. Inhibition of muscarinic receptor subtypes and effects on oligodendrocyte differentiation.- Oligodendrocytes are the myelinating cells of the central nervous system, and these cells and their myelin sheaths are the autoimmune target 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.  Combinatorial treatment of cultured oligodendrocyte progenitors increased myelin-specific gene expression and increased the percentage of mature OLs in the cultures compared to control.  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. Effects of muscarinic antagonists were evident in an incomplete maturation model (cultured OLs), but do not appear to enhance the vigorous myelination program of zebrafish larvae. (83)

King, Jacob*, and Daniel Widzowski Indiana University of Pennsylvania, Indiana, PA 15705. More conditioning and extinction sessions improve conditioned place preference test outcomes in mice.- Methamphetamine (METH) is a highly addictive central nervous stimulant for which there are no approved pharmacological treatment options and significant problems with relapse years after patients stop using. Conditioned place preference (CPP) is a neurobehavioral test used to study drug dependence in many species. In CPP, mice learn to associate METH experiences with one chamber (striped walls) and placebo experiences with another chamber (gray walls) and then show a preference for the METH chamber by spending more time there when allowed to explore both. The purpose of this study was to test an improved method (more & shorter training sessions) for CPP in mice based on a recently published procedure for rats. This CPP protocol involves habituation, conditioning, extinction, and reinstatement phases to determine if certain medication treatments during extinction can reduce METH-seeking behaviors. Within the phases there is a pre-conditioning test, post-conditioning test, extinction test, and reinstatement test. Previous experiments using a quick-training procedure for CPP did not show strong METH-conditioning (55 to 60% average preference). After conditioning with the improved method, mice spent significantly more time in the METH-paired chamber (70% average preference). During the 21-day extinction phase the average preference decreased, although there was day to day variability (some good and bad preference days). By day 14 the preference decreased to 65% and at day 21 it further decreased to 58%. There was evidence of reinstatement on the test day (65% preference) although it was not to the same extent as the post-conditioning phase. Results for extinction and relapse assessment and the effect of a potential treatment (venlafaxine) will also be presented. There was no evidence of the venlafaxine reducing the reinstatement. Overall, the new protocol significantly improved the conditioning phase although further optimization of the extinction methods is needed. (56)

Kowalewski, Matthew*, Jackson Dardis, and Jane Cavender Elizabethtown College, Elizabethtown, PA 17022. Effect of Diquat Dibromide on the morphology, reflex response and viablity of Hydra.- Diquat dibromide (DD) is widely used in the US as an algicide and herbicide targeting broadleaf and grassy weeds.  DD works by causing calcium dysregulation in cells leading to an increase in ATP production in mitochondria, leading to ROS accumulation causing oxidative stress and signaled apoptosis. The use of DD has recently been suspended in the EU; thus, experiments were undertaken to investigate its toxicity using Hydra as a model organism. It was hypothesized that DD calcium dysregulation, will result in degeneration of tentacles and nerves of Hydra leading to decreasing feeding behavior, poke-response and viability. Three hydras were placed into triplicate wells containing 0, 0.025, 0.05, 0.1, or 0.5µg/mL of DD, and two species of Hydra, oligatus and vulgaris, were used. For both species, tentacle length showed a statistical decline at all concentrations. After 2-4 days of exposure to DD, both species exhibited a statistically significant diminished poke and feeding response at all concentrations, eventually leading to the absence of response on day 5. It was found that animals in concentrations 0.025, 0.5, 0.1 and 0.05 µg/mL, experienced steady progression of degradation eventually leading to death. The overall hydra viability was also significantly affected at 0.05, 0.1 and 0.5 µg/mL concentrations, although for 0.025µg/mL, overall hydra viability was extending a day longer. These results show evidence of diquat dibromide causing cellular stress leading to tentacle loss, reduction in feeding response, slower poke reactions, and death in both hydra species left exposed to diquat dibromide. (74)

Labrosciano, Darian*, Ariana Rivera, and Vinayak Mathur Cabrini University, Radnor, PA 19087. Evidence of horizontal gene transfer between Wolbachia and Aedes aegypti.- Horizontal gene transfer (HGT) is the non-vertical movement of genetic material among organisms, and is believed to be an important factor driving evolution. A key question regarding these horizontally transferred genes is the functional role they play in the organism. In the past, HGT studies have focused on prokaryotes, but now there are a number of studies documenting functional HGT in a wide range of eukaryotes. In this study we focused on Aedes aegypti, the yellow fever mosquito, which is a known vector for several diseases. Using information from the NCBI database and the BLAST search tool we found evidence of horizontal gene transfer between Wolbachia and Aedes aegypti. The Wolbachia protein identified in our study is an uncharacterized conserved RhaS protein that has 28 RHS repeats. Using comparative genomics tools and a phylogenetics approach we were able to create the evolutionary relationship between these two organisms. The ProgressiveMauve software was used to generate a synteny analysis of this genomic region where we compared several Wolbachia, and mosquito species to understand the pattern of HGT. Based on our findings, we want to further investigate the functional role of the horizontally transferred genomic regions and expand our study to other mosquito species.  (88)

Lipschitz, Dena*, and Sean Georgi York College of Pennsylvania, York, PA 17405. Effects of Strattera (atomoxetine) on the regeneration and behavior of planaria Dugesia dorotocephala.- Strattera (atomoxetine) is a non-stimulant drug used to treat individuals with Attention Deficit Hyperactivity Disorder (ADHD), which is associated with low levels of norepinephrine (NE) in the brain. This drug is much less commonly used than stimulants, like Adderall or Ritalin, and works as a selective norepinephrine reuptake inhibitor, which increases the amount of synaptic NE in the body. Planaria are commonly used model organisms to study regeneration patterns of the external body as well as the central nervous system, and extensive toxicology reports have been performed using them. Studies have been performed to observe the effects of stimulants on planarian regeneration, however, research is lacking on how non-stimulants affect these flatworms. To answer this question, we observed behavior of the planarian species Dugesia dorotocephala before and after transverse dissections in various concentrations of Strattera. After dissections, the tail fragments were observed to record regeneration time of a new head, and immunohistochemistry procedures were used to compare the newly regenerated brain and neoblasts to the controls. Our results showed that planaria treated with a 10µM Strattera concentration overall regenerated much slower than untreated controls. Results from immunohistochemistry revealed that atomoxetine treated planaria regenerated a smaller head and less distinct nerve cords compared to the controls and showed higher amounts of mitotically active neoblasts closer to the head than in the controls. (47)

Maley, Abigail* Wilson College, Chambersburg, PA 17201. Antennae movements and visual signaling in the Ecuadorian hermit crab, Coenobita compressus.- Although they lack the extreme cooperation and colony building behaviors which characterize eusocial species, many non-eusocial invertebrates, such as the land hermit crabs of the genus Coenobita, exhibit a complex suite of social behaviors. The ecological significance of these behaviors and the variables which mediate social interactions remain poorly understood. Coenobita occupy habitats characterized by change where the distribution of resources, especially food and shells, is in a continual state of flux. Because of the unpredictable and ephemeral nature of these resources, frequent social interactions and intense intraspecific competition are expected to lead to natural selection for signaling systems or communication. Although crabs respond readily to visual stimuli, the potential for vision as a mode of communication in Coenobita has received relatively little attention. Specifically, antennae movements may provide a mechanism for visual signaling in this taxon. I investigated this possibility via behavioral experiments using the Ecuadorian hermit crab, Coenobita compressus. I compared the duration and frequency of antennae movements of individuals when alone and without a stimulus vs when exposed to various conspecific stimuli, including a recently vacated shell, a conspecific removed from its shell, and a conspecific in its shell. Time spent moving antennae and frequency of antennae movements was greater when individuals were exposed to a conspecific stimulus than when exposed to no stimulus and varied between the types of conspecific stimuli. While the results support the hypothesis of visual communication via antennae movement in Coenobita, there are other possible explanations for these observations which remain to be explored. Further work is needed to examine the mode and role of communication in this taxon. (69)

Maria, Vincent*, Francis Krug, and Joshua Slee DeSales University, Center Valley, PA 18034. Investigating the anti-inflammatory effect of curcumin and piperine in preventing biomaterial rejection in vitro.- The inflammatory response to foreign biomaterials is a significant concern in various medical procedures. Chemokines and cytokines recruit monocyte-derived macrophages to the site of the implanted biomaterial, at which they adhere to and attempt isolate it from the body. This typically results in device failure and often requires replacement later in life. Natural products have garnered attention recently as potential therapies that could have anti-inflammatory properties. Curcumin, a yellow hydrophobic polyphenol extract from turmeric, has been shown to have anti-inflammatory characteristics. However, it has a poor bioavailability as it is quickly metabolized upon consumption. Piperine, an alkaloid component in black pepper, has been shown to increase the bioavailability of other nutrients. Through in vitro THP-1 cell adhesion assays, our data show that curcumin alone and in combination with piperine inhibit inflammatory cell attachment to polyurethane films, a common biomaterial. Ongoing experiments with an ex vivo Chandler Loop apparatus are being conducted to confirm the in vitro findings. These results indicate that curcumin and piperine have potential anti-inflammatory properties and warrant further investigation as potential therapies for biomaterial rejection. (30)

Mauro, Steven, Austin Hertel *, Madison Heeter , Olivia Wirfel , and Mara Bestram Gannon University, Erie, PA 16541. Factors influencing COVID-19 spread on a university campus setting: Insights from an epidemiologic and metagenomic surveillance analysis.- COVID-19 has negatively impacted nearly every industry in some manner, and higher education is no exception. Nearly every institution of higher learning has issued responses to mitigate the spread of the virus to its campus community. These methods include but are not limited to wellness surveys, temperature checks, antibody or PCR based surveillance testing, contact tracing, and isolation/quarantine protocols. Gannon University, located in Erie, PA., was one of the first institutions to develop their own in-house PCR based COVID-19 testing programs. In the past year, we have tested over 19,000 samples in this surveillance program, which has been instrumental in allowing Gannon to continue to offer in-person classes throughout both the fall, 2020 and spring, 2021 semesters. It has also been a vital part of continuing athletic competition for the majority of sports offered on campus. This study presents results of our rapid COVID-19 PCR surveillance program in a university-wide setting during the 2020-2021 academic year. Our results indicate a low overall rate of infectivity across campus, most of which were asymptomatic and were at rates below the reported regional averages. However, athletic team type, geographic areas, and size of social gatherings were risk factors that contributed to the spread of COVID-19. This suggests individual practices, not institutional safety procedures, drive COVID-19 presence within campus populations. To understand how localized microbial populations might contribute to or be impacted by COVID-19 infection, we also conducted a nasal and fecal bacterial metagenomic study and viral qPCR analysis of COVID-19 infected individuals. These results coupled with our characterization of routes of COVID-19 exposure reveal insights that can be used for the development of risk aversion strategies that can be applied to other higher education institutions and industrial workplaces. (15)

McKinney, Miranda*, Brooke Mazzotta*, Sarah Sejour*, Kevin Long*, Rachel Wynings, George James, and William Biggers Wilkes University, Wilkes-Barre, PA 18766. Effects of Microcystis aeruginosa on the Metamorphosis and Growth of the Polychaete Annelid Capitella teleta.- Cyanobacterial Harmful Algal Blooms (CHABs) that occur in freshwater and marine ecosystems are well known in causing toxicity to both marine invertebrates and vertebrates.  We have investigated the effects of Microcystis aeruginosa, a cyanobacteria that causes CHABs and produces toxic microcystins, on the metamorphosis and growth of the polychaete annelid Capitella teleta.    This polychaete is viewed to be opportunistic since it tolerates organic pollution loading, high hydrogen sulfide levels, and low oxygen levels very well and therefore becomes a dominant species.  Our laboratory results indicate that the presence of Microcystis aeruginosa in the water surprisingly increases settlement and metamorphosis of C. teleta trochophore larvae.  Juvenile worms of C. teleta also were found to consume the Microcystis cyanobacteria without apparent toxicity as evidenced by fecal pellets, and also exhibited an increased growth rate compared with control cultures grown without MIcrocystis.  These results indicate that Microcystis algal blooms may be beneficial to the growth and development of this hardy polychaete species in the marine environment instead of being toxic. (48)

Mills, Avery*, Jeanne Berk, and Thomas Pritchett Cedar Crest College, Allentown, PA 18104. Analysis of taurine content in traditional and BEG commercial dog foods.- Taurine is an amino acid that is utilized throughout the body and is important for heart functions. In some animals, like felines, taurine is an essential amino acid, whereas it is nonessential in other animals. They can obtain it from diet and from biosynthesis using other compounds. Because felines cannot synthesize their own taurine, taurine deficiency in felines was readily accepted as a cause of dilated cardiomyopathy (DCM) (Kaplan et al. 2018). In 1987, this link between food and feline DCM was established and commercial cat food companies responded to this research by adding taurine to their foods – as a result, taurine deficiency and DCM in felines is now very rare (Pion et al. 1992). The link between canine taurine deficiency, DCM, and dog food has not been as easy to determine because of canines’ ability to biosynthesize taurine. However, the rate of DCM in canines seems to be increasing and leads veterinary professionals to question if taurine deficiency is at fault (Freeman, L.M. 2018). Some foods are more of a concern than others, diets known as BEG diets, which are either boutique brands, contain exotic ingredients, or are grain free, are most concerning. These diets may be associated with taurine deficiency due to a lack of research of the effects of these newer food formulas in canines. This study will examine the content of taurine in dry commercial dog foods, both BEG and traditional diets, using mass spectroscopy techniques. Results will be compared to determine if BEG diets have less taurine than traditional diets. The results from this study will increase knowledge and allow for further research on the possible connection between DCM and diet. (18)

Monahan, Lauren*, M. Dana Harriger, and Shifra Goldenberg Wilson College, Chambersburg, PA 17201. The social behavior of translocated African elephants, Loxodonta africana, into the Sera Wildlife Conservancy in Northern Kenya.- Conservation translocation, the deliberate movement of animals from one location to another, is an approach used in conservation for a number of conservation objectives. Translocation can affect the social relationship of a species and is an essential factor for many species ranging from learning maternal or family bonds to establishing territorial relationships between neighbors. Research indicates that African elephants, Loxodonta Africana, rely on social bonds for survival. Elephants are often translocated to help with their fight against drought and pouching. It is unknown if translocation is affecting these social bonds which is then leading to an unsuccessful reintroduction and negatively effecting that animals’ survival. This study will determine if orphaned calves cooccur with wild elephants after being released into a wildlife sanctuary in northern Kenya. The occurrences are measured through camera traps and observing when orphaned elephants are present with wild elephants at watering holes. Each elephant is individually identified to also determine who the orphans recreated bonds with if at all. Camera traps allow for limited human presence which could impact any elephant interactions. Each image and video are scored to determine which of the ten orphans are cooccurring with the wild elephants. Preliminary analysis suggests that female elephants are able to recreate social bonds when a family member dies, but it is unknown if the human interaction and removal of the orphans from the sanctuary effect this ability. Results of this study will help determine whether and which reintroduced orphaned elephants are cooccurring with wild elephants to fulfill the social relationships they lack. Future studies could compare the released animals’ social interactions over a more extended period of time. This approach will show if the released elephants are becoming more socially integrated over time. (10)

Morley, Nichole*, and Megan Rothenberger Lafayette College, Easton, PA 18042. Fragmentation by roads on microhabitat and abundance of terrestrial salamanders.- The microclimate conditions of forest edges are altered by habitat fragmentation, threatening the regularity of many forest processes and native species.  Conservation biologists use bioindicator species as tools to evaluate forest health because of their relation to many forest processes. Due to their relative abundance in forest habitats, sensitivity to environmental disturbance, and mid-level position in the food web, Plethodon cinereus was evaluated as a sensitive indicator of habitat fragmentation by roads using a multiyear monitoring study in a state park (2014 and ongoing).  Monitoring takes place at an unfragmented and road-fragmented site with biweekly collection of soil pH, canopy cover, leaf litter depth, soil moisture, soil conductivity and salamander abundance from March – November using the artificial cover object (ACO) method. The abundance of P. cinereus, is significantly higher at the unfragmented site where the nearest road is greater than 1000 m away. At the fragmented site, which is approximately 100 m from two roads, salamander abundance increases significantly with distance from the road.  Our results also indicate that P. cinereus is sensitive to microhabitat change, and their abundance is significantly correlated with higher soil pH, lower leaf litter depth, and lower soil conductivity. (11)

Morningwake, Megan*, Brad Engle, Adam Cooke, and Kathryn Sarachan Wilson College, Chambersburg, PA 17201. The overexpression of the eyes absent gene, EYA1, in retinoblastoma, a potential therapeutic target.- Retinoblastoma is one of the most common and deadly cancers in children under three years of age. Worldwide, 9,000 children are diagnosed annually with hereditary retinoblastoma, with a 70% mortality rate in the middle- and low-income countries. The treatments for retinoblastoma (chemotherapy, radiation, and removing the eye) all have serious side effects. At any stage of diagnosis, the goals of treatment should be saving the child’s life, preserving as much vision as possible, and minimizing the damage to noncancerous cells. This could potentially be accomplished by targeting specific genes or gene products that are known to be overexpressed in many cancers. Some genes are upregulated during development; however, they are also upregulated in cancer cells leading to tumor progression and carcinogenesis. The EYA1 gene is a critical developmental gene and its gene product functions as a protein phosphatase and as a coactivator of the SIX/EYA transcriptional complex; it has been shown to be overexpressed in several cancers. In this literature review, the upregulation of the EYA1 gene and its gene product are explored as possible therapeutic targets specific to retinoblastoma cancer cells, opening the door to the possibility of developing targeted therapies that could prove a viable option that will be less invasive to young children and be more effective in treating retinoblastoma. (28)

Nawrocki, Zachary *, and Jane Cavender Elizabethtown College, Elizabethtown, PA 17022. Elucidation of the Isoform and Role of SRSF1 in Simian Virus 40 T-antigen Transformed Human Diploid Fibroblasts.- Cancer results from the accumulation of DNA mutations within one cell, resulting in the production of hyper- or non-functional proteins. Additionally, DNA can remain wild type and the resulting mRNA transcripts can be alternatively spliced to create proteins which contribute to tumorigenesis. To study the mechanisms by which tumor viruses induce this alternative splicing, our laboratory utilized the Simian Virus 40 (SV40) T-antigen oncoprotein. SV40 has been shown to cooperate with oncoproteins to promote transformation of human cells. To determine if alternative splicing plays a role in T-ag-transformation this project is investigated the splicing proteins SAM68 and SRSF1 in immortalized and transformed human diploid fibroblasts HDF(tert) and HDF(tert)+T, respectively. T-ag-transformed cells exhibited higher levels of the splicing factor SAM68. A target of SMA68 is SRSF1, which is also a splicing factor. SRSF1 is involved in pre-mRNA splicing and post-splicing activities like mRNA nuclear export and translation. SRSF1 contains two functional domains: an arginine-serine rich regulatory region, and two RNA-recognition-motifs. Three known SRSF1 isoforms are created by alternative splicing. SRSF1 retaining intron 4, has been shown to cause cells to transition from an epithelial-to-mesenchymal state (EMT) leading to a more aggressive and invasive tumor. It has been shown that SAM68 interacts with SRSF1 to allow for the inclusion of intron 4. This study aimed to use HDF(tert) and HDF(tert)+T to analyze the different SRSF1 isoforms and correlate that to the EMT. It was postulated that cells expressing higher levels of SAM68 will induce more SRSF1-containing intron 4 isoforms. (46)

Nettles, Erica*, and Jennifer Hayden Cedar Crest College, Allentown, PA 18104. Impact of lysine acetylation on Mycobacterium smegmatis survival in the presence of isoniazid.- Tuberculosis (TB) is one of the top ten leading causes of death worldwide, partly due to its ability to become antibiotic resistant. According to the World Health Organization, drug-resistant TB was diagnosed in approximately 450,000 patients in 2019. TB not only affect’s an individual’s health; TB can also pose a finical burden. Drug treatment regimens can cost patients $16,000-$500,000 for drug susceptible and drug resistant infections, respectively. Mycobacterium tuberculosis has become increasingly resistant to the commonly prescribed frontline prodrugs isoniazid (INH). The mechanism of action of INH is well known, where KatG plays a vital role in INH activation. Activated INH then binds to InhA, which results in decreased mycolic acid synthesis and causes cell death. Mutations in katG can lead to unsuccessful activation of INH and consequently, M. tuberculosis becomes resistant to INH. Here, we investigate the role of lysine acetylation in INH resistance using the model organism Mycobacterium smegmatis. Lysine acetylation involves the addition of an acetyl group to a lysine residue, while deacetylation, an NAD-dependent process, involves removing the acetyl group. We determined the effects of lysine acetylation on resistance to INH using Kirby-Bauer methodology, cell survivorship quantification, and Resazurin MIC assays. Understanding the role of lysine acetylation in regulating INH activation and metabolism could potentially contribute to better treatment regimens for TB patients. (5)

Ogrodniczuk, Marcin*, and René Fuanta East Stroudsburg University, East Stroudsburg, PA 18301. Shikimate kinase; searching for potential novel anti-Tubercular agents.- Tuberculosis is a respiratory infection with over 10 million reported cases each year. This infection rate is responsible for over two million deaths annually, second only to HIV in fatalities among infectious diseases. Since 1960 tuberculosis rates have steadily declined worldwide as medical technology has advanced to produce more effective antibiotics. However, studies in the last decade have shown that tuberculosis rates continue to increase. This observation suggests that there is an alarming increase in the prevalence of drug‐resistant strains of tuberculosis, thus the need for the discovery of novel anti‐tubercular agents. When searching for potential enzymatic pathways for drug discovery it its vital that the pathway possesses minimal overlap with the host. The shikimate pathway is a seven‐step metabolic route that produces aromatic amino acids and other cellular metabolites. This pathway is found in microorganisms and has no mammalian counterpart, making any of the enzymes in this pathway suitable targets for screening of potential anti‐tubercular agents. The target enzyme in this project, Mycobacterium tuberculosis Shikimate Kinase (MtSK), catalyzes the 5th step of this pathway. MtSK converts shikimate to shikimate‐3‐phosphate. The overall goal of this project is to express and characterize MtSK in order to screen for potential anti‐tubercular agents. Initial methods included a bacterial transformation of XL‐1 blue competent E.coli cells. This preliminary transformation was performed using a pET-21b plasmid with an aroK gene inserted into the multiple cloning site. Successfully transformed XL-1 blue cells were cloned. A second transformation of  BL21 DE3 competent cells was performed. Initial small scale expression showed the presence of a band around 20 kDa. The theoretical mass of the enzyme is 19.6 kDa which suggests MtSK was successfully expressed within the transformed cells. Expression analysis for large-scale supported data from small scale as a band of 20 kDa once again appeared in the verification SDS-PAGE gel. Purification of MtSK was carried out through nickel affinity chromatography. Subsequently, kinetic characterization and inhibitors studies will be performed using inhibitors like avarone and hymenidin.   (24)

Patel , Mira * Souderton High School, Souderton, PA 18964. Different Temperatures of Water v. Plant Growth.- Problem: How will the temperature of water affect plant growth?   Hypothesis: If the plants are grown with ice-cold water they will grow larger, than plants that are grown in boiling hot water, because plants are prone to cold water due to the natural endothermic energy of the sun. They are most susceptible to cold water because plants use precipitation to carry out their photosynthetic processes.   Independent Variable: the temp of the water  Dependent Variable: height of each plant (cm)  Control Variables: type of soil, type of seed, number of seeds temp of room, placement of plant, amount of water, (etc.)  Control Group: plant sample w/ room temperature water    Procedure: In this experiment, the student will test what temp(s) of water will have the most effect on the growth of a Fenugreek plant. The student will be placing 10 seeds in each flowering pot. Every plant will be watered with its desired temperature of the water. The student will be testing the growth of the plant under ice cold and boiling hot water. The water temperature will be measured by using a thermometer, for accurate data collection. The plants will be watered and measured every other day. The student will also have a control group, she will be growing a Fenugreek plant that is only given room temperature water. At the end of each observation, the student will record all the data into a log book.  (68)

Patel, Megha*, and Christopher Brey Marywood University, Scranton, PA 18509. Cloning and Analysis of Caenorhabditis elegans Krüppel Like Transcription Factor-1 promoter.- Krüppel Like Transcription Factor (KLF) is found in many different species including Homo sapiens. Caenorhabditis elegans is a suitable model organism to study KLFs role in mammalian metabolic regulation as the nematode has only 3 klf members whereas humans have 17. Klf-1 influences cellular apoptosis and lipid metabolism and encodes for a Zinc finger protein. Ce-Klf-1 promoter is 9.5Kbp long. A 500 bp sequence was successfully excised at the ATG 5’ end from 9.5Kbp promoter using PCR and restriction digestion techniques. In this project, we want to excise a 250-300 bp region of Ce-Klf-1 promoter (within 500 bp starting at ATG) with the help of PCR and clone that particular segment using cloning vector. The purpose is to introduce the 250-300 bp promoter region into a cloning and expression vector, upstream to a GFP (Green Fluorescent Protein) encoding start site. This project focuses to determine the expression of GFP protein using the 250-300 bp region of Ce-Klf-1 promoter using an expression vector which will be introduced in C. elegans using microinjection. (89)

Pericak, Olivia*, and Rajinikanth Mohan Mercyhurst University, Erie, PA 16546. Exploring the function of TEM1 within the ethylene pathway of senescence in Arabidopsis thaliana.- Leaf senescence is a degenerative process that occurs among plants undergoing environmental or age-related stress. During senescence, the plant hormone ethylene activates a molecular pathway that collectively relocates nutrients to other areas of the plant that may be capable of regrowing. Despite a recent gain in knowledge of the senescence pathway, the molecular mechanism remain to be elucidated. The research presented here aims to explore the function of a gene known as TEM1 and its function in the ethylene-dependent pathway. Previous research has suggested TEM1 to be a regulatory gene that represses flowering in plants and identifies as an ethylene-inducible gene. This has led us to believe TEM1 represses senescence. To test if TEM1 plays a role in the ethylene-dependent senescence pathway, genetic analysis was performed with tem1 mutants in comparison with wildtype Arabidopsis thaliana plants. By treating leaves of both plants with 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor for ethylene biosynthesis in plants, senescence was induced.  The tem1 mutant showed higher ACC-induced senescence, with the mutant leaves having considerably less chlorophyll, indicating that TEM1 is a possible transcriptional repressor of ACC- and ethylene-induced senescence. Gene expression using qPCR revealed higher ethylene pathway and senescence gene expression in the tem1 mutant, consistent with the phenotype. The results implicate TEM1 as a novel transcriptional regulator of the ethylene pathway regulating senescence and a possible connecting link between flowering and senescence in plants. (67)

Petrick, Hayden*, and Christopher Dolanc Mercyhurst University, Erie, PA 16546. Testing the resource dilution and the resource concentration hypotheses regarding infestation rate of Agrilus planipennis (emerald ash borer) in stands of Fraxinus profunda (pumpkin ash) of differing densities in Northwestern Pennsylvania.- The emerald ash borer (Agrilus planipennis; EAB) is an invasive jewel beetle species from Asia whose larvae mature and consume the cambium of host ash trees (Fraxinus spp.). Because the infestation of EAB is rapidly spreading, determining the validity of two exclusive hypotheses on resources and host density will afford conservation agencies insight into EAB maintenance. The resource dilution hypothesis proposes that a stand with a less dense host spp. leads to a rapid mortality of host individuals, but the resource concentration hypothesis contests this by instead proposing that a stand with a high density of host species leads to a rapid mortality of host individuals. The objective of this study is to produce data that tests these two competing hypotheses. In 2016, two permanent plots with differing densities of the host species, pumpkin ash (Fraxinus profunda), were established before any signs of EAB infestation were observable and ongoing annual censuses have occurred every summer since. The canopy dieback, number of EAB exit holes, DBH and other standard measurements were recorded for individuals in both plots. After four years, the less dense stand of pumpkin ash had an average ash rating of 4.3 while the stand that was denser had an average of 3.98, and the less dense stand had 4.81 more holes per tree than the denser stand despite the stands being less than 100 m apart. The plot with greater host density had a 22.2% mortality rate while the less dense plot had a 52.4% mortality rate. These findings provide preliminary support for the resource dilution hypothesis. Future work is needed to further these findings. If true, land managers will likely want to cease thinning of pumpkin ash stands as a way of preventing infestation as our data suggest this may be to their detriment. (73)

Pikal, Claire *, and Nicholas Sizemore University of Scranton, Scranton, PA 18510. Computational analysis of intermolecular Diels-Alder reactions of α-amido acrylates with furan and 2-(trimethylsiloxy)-1,3-cyclohexadiene.- The Diels–Alder reactions are an important organic transformation that have found utility in a variety of fields such as medicinal chemistry and material design. The intermolecular version of this reaction can bring together cyclic dienes and dienophiles to create bridged bicyclic products. While a number of different diene/dienophile reaction pairs exist, α-amido acrylates dienophiles remain an underused substrate. When paired with cyclic dienes, like furan or 2-(trimethylsiloxy)-1,3-cyclohexadiene, the resulting structures resemble molecules which are known to have interesting biological activity. A computational study to specifically look at the cyclization between these species was done, in an effort to increase understanding about the Diels–Alder cyclization of α-amido acrylates dienophiles, and hopefully, expand the utility of this class of substrates. Initial conformational analysis in Spartan’18 was carried out on starting materials, transition state estimates, and cyclizations products using Merck molecular force field (MMFF) calculations. These conformations were then subjected to optimization and frequency calculations using density functional theory (DFT) at the B3LYP 6-31G+(d) level of theory in Gaussian18. The results of this computational study, including endo/exo selectivity analysis, will be described herein. (19)

Rivera, Ariana*, Darian Labrosciano, and Vinayak Mathur Cabrini University, Radnor, PA 19087. Identification of instances of horizontal gene transfer between Drosophila ananassae and its endosymbiont bacteria Wolbachia.- Horizontal gene transfer (HGT) is the transmission of genetic material among organisms, other than by vertical transmission from parent to offspring. HGT is thought to be an important factor driving evolution. While extensively studied among prokaryotes, there is growing evidence that HGT is also prevalent in eukaryotes. In this study, we focused on the endosymbiont bacteria Wolbachia, and the Drosophila species that it infects. Based on a literature review we identified certain proteins in Wolbachia that have shown evidence of HGT and tested them using the Community Science Project pipeline. We found a particular Wolbachia protein, functionally annotated as DDE-type integrase/transposase/recombinase, which was also present in Drosophila ananassae. This protein in Drosophila ananassae was defined as a putative gag-pol protein based on functional annotation in the UniProt database. Using phylogenetic analysis, we were able to observe the evolutionary relationship between these two organisms and confirm the instance of HGT. Using synteny analysis we were able to observe the arrangement of the gene in several Wolbachia and Drosophila species. Further investigations in this system will focus on understanding the mechanism of HGT and expanding our study to other genes, to discover the extent of horizontal gene exchange. (87)

Roberts, Taylor*, Julia Danko, Mughiara Qadeer, Allison Ahl, Matt Gacura, Prasad Dalvi, and Gary Vanderlaan Gannon University, Erie, PA 16541. Examining Putative Mantel Correlations Between Granular COVID-19 Burden and Election Polling Data, Jan-Oct 2020.- It is well established that war, famine, and disease are tumultuous forces that have driven great changes in a nation’s history.  Here we seek to measure the cost of the COVID-19 pandemic as reflected in the public perception of elected U.S. officials at all levels of government, with a particular emphasis on incumbent leaders.  Using the R-programming language, we plan to perform Mantel Tests to examine if any correlations exist between elevated COVID-19 burden as measured via morbidity & mortality datasets to that of meta-polling data regarding all gubernatorial, house, senate, and presidential races.  Our data structure comprises a total of 266 consecutive days of COVID-19 morbidity and mortality in the year 2020 extracted from public-use databases made available from the Centers of Disease Control (CDC).  Likewise, our polling data is an aggregate dataset derived from a total of 435 governor-level, 784 House-level, 1,873 Senator-level, and 11,282 presidential-level polling results for the same ten-month timeframe across the country.  For all contested races, we seek to test a null hypothesis in which an elevated COVID-19 burden is inversely related to incumbent poll approvals in the sampled window. (96)

Safforld, Keyanna*, Serena Heagney*, Kristina Guth, Monika Girgis*, Hellen Ghanem*, and Christopher Kavanau East Stroudsburg University, East Stroudsburg, PA 18301. Decoding brain signals in the primary visual cortex by simulating brain activity.- One of the great unsolved problems of visual neuroscience is finding a technology to decode complex mixtures of visual signals found in the brain and display these on a display monitor according to what is perceived in the “mind’s eye.” Science and technology are moving closer to these goals through advances in computational neuroscience, a relatively recent discipline. Recent work (Korikawa and Kamitani, 2017) demonstrated that generic object shapes, but not color, may be displayed from brain activity. Here we show that spatial activation patterns in primate striate cortex (area V1) corresponding to color stimuli may be predicted using novel cylindrical and circular arrays of artificial neurons. Our preliminary results suggest we will be able accurately predict, and display, stimulus hues given to test subjects from brain activity. After much trial and error this experiment yielded near-perfect results. In the future, this could potentially lead to new medical technologies to help many of the blind see through artificial brain stimulation, as well as the ability to see what a person is dreaming as they sleep. (39)

Scheppelmann, Erika*, and Bridgette Hagerty York College of Pennsylvania, York, PA 17405. The effect of human disturbance on habitat use of mesopredators in a county park in south central Pennsylvania.- Understanding wildlife and urban relationships is necessary as the encroachment of human development increases in wild spaces. Human disturbance can potentially alter a species behavior and physiology, which could lead to a potential loss in biodiversity, decrease in abundance, and a decline in fitness. Mesopredators are an important component of the wildlife community because they contribute to population control of prey species. We used non-invasive camera trapping methods to examine mesopredator habitat use in a local county park (Nixon County Park near Jacobus, PA). Currently, park managers and naturalists have reported seeing raccoon (Procyon lotor), coyote (Canis latrans), red fox (Vulpes vulpes), and long-tailed weasel (Mustela frenata) within park boundaries, but have limited funds to conduct research. Our goal was to quantify how these mesopredators use the available habitat in the park, and to investigate if human presence in the park is altering habitat use. We strategically placed 12 wildlife cameras in hardwood mixed forests, open fields, and wetlands to monitor how habitat use differs among species within the park boundaries. Additionally, we paired locations with low and high human activity based on distance to trails to compare habitat use to evaluate human impacts on each species. We analyzed images within 30-minute trap events between February and December 2019 and compared the capture success rate (trap events per 76 nights) between locations 20m and 60-80m from trails. Activity for the mesopredators was lower in areas in close proximity to hiking trails throughout the study. We also found that the mesopredators displayed higher activity at night than during the day in all habitat types. We recommend that park naturalists continue to improve the habitat with less human disturbance (e.g., invasive species removal) and educate the public about staying on trails to provide wildlife with areas to avoid human disturbance. (60)

Scholl, Noah*, and John Harms Messiah University, Mechanicsburg, PA 17055. Utilizing a plasma membrane enrichment protocol to optimize western blot detection of the CCK2 and CCK2i4sv receptors.- 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 (CCK2i4svR) 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 CCK2i4svR-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. (32)

Schwab, Solomaya F.*, and Amy Faivre Cedar Crest College, Allentown, PA 18104. Bird activity patterns on the Cedar Crest College campus in Allentown, PA.- Over the course of a season different birds species visit feeders relative to migration patterns, weather, temperature and other factors.  Having baseline data on these patterns is especially important as changes in the climate have been documented to shift migration and feeding patterns. In the fall of 2020 a feeder on the Cedar Crest College campus was monitored daily to record the number of visits and species at the feeder.  Patterns for several species were recorded and compared week to week including visits by tufted titmice (Baeolophus bicolor), mourning doves (Zenaida macroura), Northern cardinals (Cardinalis cardinalis), downy woodpeckers (Picoides pubescens), red-bellied woodpeckers (Melanerpes carolinus), dark-eyed juncos (Junco hyemalis) and white-breasted nuthatches (Sitta carolinensis). Some species were present in fairly consistent, low numbers and other species fluctuated from week to week in number of visits. These data will be compared to visits recorded at a similarly placed feeder on the campus in previous years to determine any shifts in bird species patterns. (62)

Schwenk, Bailey*, and Robert Smith Lycoming College, Williamsport, PA 17701. GIS-based prioritization system for ms4 compliance projects in small municipalities.- The Clean Water Act regulates discharges of pollutants into streams and rivers, which includes point source discharges. Thus, local governments and other entities that manage municipal stormwater systems must meet certain requirements for mitigating stormwater through the MS4 program. This research aims to determine a framework for prioritizing best management practices (BMPs) and locations in small urbanizing areas to fulfill the MS4 requirements. This work is part of a broader initiative to build a college-community partnership and improve local water quality. A list of criteria for BMP selection and placement was generated and GIS data consistent with the criteria were created to generate a spatial model identifying ideal BMP locations. The criteria chosen included the areas outside of combined sewer systems, land outside the floodway, areas of existing BMPs, land parcel size, public ownership of land, impervious and pervious surfaces, and land within MS4 urban areas. We discovered that ideal locations for BMPs are limited in small river-towns such as Williamsport, PA, and that prioritization systems can be a useful tool for small municipalities with limited capacity. (79)

Shah, Raivat*, Carly Shaffer, and Giancarlo Cuadra Muhlenberg College, Allentown, PA 18104. Effects of E-liquids on M1 Macrophage Phagocytosis of Aggregatibacter Actinomycetemcomitans.- A 2020 CDC study revealed that 19.6% of US high school students are self-reported vapers, or users of aerosol releasing electronic-cigarette devices (ECIGs). ECIGs vaporize e-liquids containing various flavors into the oral cavity – the entry point of the vapor. The oral cavity houses a microbiome of bacterial species that may be implicated in oral disease. Aggregatibacter actinomycetemcomitans, a gram-negative pathogenic species of bacteria, occurs in 90% of juvenile periodontitis, a chronic inflammatory disease characterized by the destruction of tooth-supporting tissues. M1 Macrophages are host white blood cells responsible for phagocytosis – a cell ingesting process – of pathogenic microbes. The purpose of this study is to determine the immunological effects of e-liquids -/+ flavors on M1 Macrophage phagocytosis of A. actinomycetemcomitans. THP-1 human monocyte cell lines were first grown in RPMI, stimulated with 200nM phorbol 12-myristate 13-acetate at 37℃, 5% CO₂ (standard conditions) and later differentiated to M1 macrophages with 100ng/mL lipopolysaccharide and 20ng/mL interferon γ. Then, M1 macrophages were exposed to both flavorless and flavored (cinnamon, menthol, strawberry, and tobacco) e-liquids at a final concentration of 1% (v/v) in RPMI (standard conditions) as well as untreated control cells. Treated and control M1 macrophages were then exposed to A. actinomycetemcomitans at an MOI of 100, allowing macrophage phagocytosis to take place. Excess bacteria were removed by washing and M1 cells were incubated with RPMI antibiotic media to kill extracellular bacteria. M1 cells were then washed and lysed with sterile water. Lysates from each well were serially diluted and plated on agar for CFU counts. We expect that -/+ flavored e-liquids will decrease M1 Macrophage phagocytosis of A. actinomycetemcomitans compared to control. This study may indicate that e-liquids and vaping increase the risk of developing periodontal disease via modulation of host M1 macrophage phagocytosis efficacy, particularly among children and adolescents. (95)

Shebby, John*, John Minora, and Christopher Brey Marywood University, Scranton, PA 18509. Chemotaxis behavior reinforces KLF mutation characteristics in C. elegans.- Diabetes is a common and life-threatening illness that is pervasive in our society today. The onset of diabetes and obesity has been associated with mutations in the transcription factor protein family of KLF, which has been identified in metabolic processes in both humans and the model organism C. elegans. Research suggests that in C. elegans, obesity due to KLF mutations is the result of the inability of the organism to metabolize intestinal adipose, which instead accumulates in the intestine. Until now there has been no conclusive evidence verifying that the intestinal fat is completely utilized by the worm, rather this was speculated. In this experiment, it is demonstrated through the vehicle of chemotaxis behavior, that C. elegans with KLF-3 mutations do not metabolize intestinal fat. This experiment examines food-seeking behavior in C. elegans KLF-3 mutants when compared to wild-type (N2) organisms after each groups had been subjected to both normal conditions (well-fed) and stress conditions (starved). Upon comparison, it is expected that the KLF-3 mutants that are well-fed will display the same heightened food-seeking behavior as wild-type organisms that are starved. This behavior is due to the incapability of the mutants to effectively utilize their intestinal fat, prompting the organism to continue to seek out food aggressively despite being in a satiated state. (86)

Silva, Nicole*, Zahaira Velasco*, and Andre Walther Cedar Crest College, Allentown, PA 18104. Examination of growth and capsule formation in mutant strains of the oleaginous Yeast, Cryptococcus neoformans to be used in biodiesel production.- The burning of fossil fuels is a main cause of global climate change due to the release trapped carbon in the form of CO₂ generated in combustion engines.  There is a need to find more carbon neutral and renewable fuels that can be used to power combustion engines in airplanes, trains, and automobiles, since for the power of the combustion engine in unlikely to be replaced in the foreseeable future.  Biodiesel has the potential to be a renewable source of cleaner burning energy that can be used to replace the finite resource of fossil fuels. Biodiesels can be made from biological matter with high concentrations of long carbon chains in the form of fatty acids found in fats and oils.  Biodiesels have been successfully generated from the fatty acids in animal fats, and plant-based oils, but these sources are ethically, and logistically challenging sources that result in minimal savings on CO2  release.  Our lab has explored the used of oleaginous yeasts that can converting plant sugars to high concentrations of fatty acids bound in fats and phospholipids.   One possible system is the encapsulated yeast Cryptococcus neoformans that has been thoroughly studied due to its ability to cause illness in immunocompromised individuals. Our preliminary results show that a strain of C. neoformans containing a mutation causing a defect in capsule formation that is avirulent, is able to produce high levels of lipids that can be chemically converted to multiple fatty acid methyl esters that make up biodiesel. In this study, we have been striving to optimize biodiesel production by examining capsule formation in multiple mutant strains and examining the effect media with different Carbon to Nitrogen ratios has on biomass and biodiesel production.  We hope that our results will provide a new, renewable system to generate biodiesel on a large-scale. (99)

Sollenberger, John*, M. Dana Harriger, and Abigail Maley Wilson College, Chambersburg, PA 17201. Cannabidiol as a therapeutic alternative for treating Systemic Lupus Erythematosus.- Systemic Lupus Erythematosus (SLE) is a chronic inflammatory disease that currently has no cure and current treatment options include many harmful side effects. SLE is generally hard to diagnose and causes chronic inflammation and pain throughout the body. Signs of SLE range from a butterfly-like redness around one’s face to elevated peripheral cell counts, and swelling of affected organs. Individuals diagnosed with SLE are most commonly prescribed the steroidal drug Prednisone which can cause adverse side effects. Cannabidol (CBD) is a chemical extracted from the cannabis plant that has no known psychoactive side effects and has reported anti-inflammatory effects with no harmful side effects when used by humans. CBD is available as an over-the-counter grade and through a prescription for medical grade. Over-the-counter grade CBD is typically infused with oil which can persist after distillation. Medical grade CBD involves winterization followed by a distillation, leaving more CBD and less chemicals that were used in the extraction process. Current findings indicate both forms of CBD having anti-inflammatory properties but the difference in purity has not been tested. Understanding if differences in purity have an effect on suppressing inflammation is important to ensure lower quality CBD are safe treatment options and ensure the medical grade CBD is more effective in its healing properties. Results could suggest using CBD as a treatment option could supplement and/or minimize the reliance on medications like Prednisone with fewer adverse side effects. (33)

Tabaku, Elsa*, Kathryn Sarachan, and Brad Engle Wilson College, Chambersburg, PA 17201. Ceramide and azacytidine in combination as an alternative treatment for triple-negative breast cancer.- Triple-negative breast cancer (TNBC) is a subtype of breast cancer defined by the lack of expression of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) proteins. It represents approximately 10–15% of breast cancers and has a poor prognosis and very limited treatment options. Chemotherapy is one of the most common treatment options being used to treat breast cancer, including TNBC, but the effective dosages often cause damage to healthy cells and tissues. Azacytidine (AzaC) is one commonly used chemotherapeutic drug which functions as a demethylation agent to reduce growth and proliferation of cancer cells. Ceramides (CER) are bioactive lipids which function as tumor suppressors, regulating cell proliferation, differentiation, senescence and apoptosis. Both of these agents have shown to be effective treatment when used on other subtypes of breast cancer. In this literature review, the potential efficacy of a combination of these chemotherapeutic drugs on TNBC is explored. It is proposed that AzaC and CER combined may induce higher rates of cancer cell apoptosis than either agent alone while having no effect or minimal negative effect on the healthy cells and the combined approach would suggest an improvement to current treatment for TNBC. (27)

Tan, Kally*, Lawrence Mylin, and John Harms Messiah University, Mechanicsburg, PA 17055. Differential efficacy of antifibrotic and immunotherapy on T-cell infiltration in murine pancreatic adenocarcinoma.- 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 CCK2i4sv 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 (p<0.02) and significantly less fibrosis. To assess the efficacy of combination therapy, mice (C57Bl/6) were immunized weekly with CCK2i4sv or control peptides starting three weeks prior to orthotopic injection of syngeneic cancer cells (Panc02) engineered to express human CCK2i4svR. 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 was significantly higher in these proglumide-treated mice compared to the previous untreated pilot controls. However, no difference in infiltration was observed in tumors from CCK2i4svR-immunized mice compared to control immunization (p>0.4). 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. (44)

Tasker, Jenna *, Bailey Schwenk, Ruric Bowman, and Robert Smith Lycoming College, Williamsport, PA 17701. Effect of climate on adult Stonefly and Caddisfly activity and abundance.- Climate change is expected to alter the timing of life cycle processes of aquatic organisms. Stream insects have a complex life cycle, and the duration of and transition among life stages is dependent on weather and climate. Thus, local weather should be related to adult insect activity and abundance. We examined the relationship of daily weather patterns with the abundance of adult Plecoptera and Trichoptera. Malaise and canopy traps were deployed at four streams in the Mosquito Creek Watershed (Lycoming County, PA) during a 14-day period in July. Traps were collected daily, and all Plecoptera and Trichoptera were identified to order. Water temperature, air temperature, and wind speed and direction sensors were deployed at each sample site, and a single rain gauge was deployed at the most central site. Abundances of adult insects were not related to terrestrial weather variables, but the abundances of Trichoptera and Plecoptera at ground level were related to water temperature. This indicated that terrestrial weather may not affect adult activity, and patterns of emergence may control adult abundance. Additionally, this result may contradict previous work suggesting that adult insects disperse through stream corridors. (52)

Tews, Veronika*, and Austen Barnett DeSales University, Center Valley, PA 18034. Examining the Evolution of Epidermal Growth Factor (EGF) Pathway Ligands in Insects.- The EGF pathway is a highly conserved and ubiquitously used cell signaling cascade in the development of many animals. In the highly studied Drosophila melanogaster, four ligands ( vein, gurken, spitz and keren) are used to activate the pathway while one ligand (argos) is used to repress activation of the EGF pathway. An arthropod centered phylogenetic analysis showed that the genes which encode the ligands Vein and Argos were present in the last common ancestor of all arthropods. However, this analysis showed the genes encoding Gurken evolved in the last common ancestor of the Diptera, the clade which includes flies. In continuation of this research, evidence was provided showing the orthologues of spitz and keren are the result of multiple independent gene duplication events. In an attempt to determine the ancestral role of the spitz/keren gene during insect development, we used RNAi targeting the orthologues of the genes in two separate lineages, Orthoptera and Hemiptera. These lineages are represented by the cricket Gryllus bimaculatus and the milkweed bug Oncopeltus fasciatus respectively (70)

Thomas, Amilyn * Mount St Joseph Academy, Flourtown, PA 19031. Which ingredient in selected acne medications is the most effective?– Have you ever wondered where acne comes from and how you can treat it? One major cause of acne is the colonization and infection of clogged pores with bacteria. In this science project, I tested different acne medications to determine their effectiveness at killing bacteria.This experiment aimed to determine which ingredients (tea tree, benzoyl peroxide or salicylic acid) used in selected acne medicines would be the most effective in killing and preventing the growth of bacteria on the skin. I tested 3 different medications, 1 for each ingredient. I soaked sterile paper disks in the 3 acne medications and then placed the medication soaked disks on a E.coli coated nutrient agar plate. The experiment was conducted over a 3 to 4 day period where the nutrient agar plates were placed in an incubator to allow the bacteria to grow. Measurements of the sizes of the zones of inhibition around each paper disk indicated which acne medication was the most effective in eradicating E.coli. Surprisingly enough, tea tree oil was the most effective ingredient.   (3)

Thorne, Nathan*, and Michael Shin Messiah University, Mechanicsburg, PA 17055. Genetic Control of Metal Nutrient Uptake in Arabidopsis Thaliana.- Plant cells uptake metals from the soil by operating specific cellular pumps to transport metal nutrients. These transporter pumps are crucial for maintaining the healthy concentration of metal elements. Zinc and nickel are essential micronutrients 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 would provide a window into the MTP1 transporter function and metal nutrient uptake pathways. Unfortunately, a flaw found during a preliminary wild-type experiment highlighted problems with the experimental method. After that result, the purpose became to troubleshooting the experimental method. The ultimate conclusion of this semester of research is that something about the metal ion stock solutions or about the seeds was causing the plates to appear hyper-concentrated.  Two preliminary tolerance experiments were conducted on Columbia line wild-type seeds with zinc and nickel contamination to learn the procedures and to ensure everything was working as expected. The first preliminary experiment with zinc and nickel contamination yielded results that unexpectedly appeared hyper-concentrated. The second preliminary tolerance experiment yielded similar results to the first.  The results, or lack of results, pointed to a problem with the media, the metal ion solutions, or the seeds. Following a chemical digestion to burn off organic matter in the media, the media was analyzed with flame atomic absorption spectroscopy. Although many of the plates were damaged by water or contamination, enough plates survived and ran through the atomic absorption spectrometer to get a good picture of the plates. The analysis demonstrated the concentrations were not hyper-concentrated in the media and pointed to the problem being something in the metal ion stock solutions. (65)

Vasiliadis, Sabrina*, Joseph Tetreault, and Rachel Fogle Harrisburg University of Science and Technology, Harrisburg, PA 17101. Restoration of a Recirculating Aquaponics System.- Recirculating aquaponics is an agricultural technique where the nutrient-rich water produced by fish is utilized to fertilize hydroponic plants. It proposes both societal and environmental benefits by requiring significantly less water and land than traditional farming techniques. This gives aquaponics a unique potential to provide for urbanized and drought susceptible locations that are unable to meet local food demands. Combining the simultaneous farming of fish and plants in a single system can be a challenge as both species have ideal nutrient and pH ranges. Strict water quality parameters must be used to ensure optimal production and fish health. A new undergraduate aquaponics internship was approved by Harrisburg University administration to begin at the start of the spring 2021 semester. The small aquaponics system had not been adequately maintained for several months due to the current pandemic and lack of students on-campus. Lack of maintenance led to the water quality parameters of pH and total ammonia nitrogen becoming dangerous for the fish health and limiting nutrient uptake which diminished plant growth. Over the past two months, the team at HU has restored the recirculating system to meet the ideal parameters reported in aquaponics literature. Improvements to plant production and fish behavior/diet have already been observed during this short period. The data and techniques used in this restoration will be used to provide foundational data in the development of a model for the restoration and management of other small-scale aquaponic systems. (51)

Vieira, John, Jenna Sins*, Mike Ganger, and Gary Vanderlaan Gannon University, Erie, PA 16541. Identification of fern orthologous genes implicated in moss rhizoid and grass root hair development.- The evolution of terrestrial plants is an approximately 500-million-year story. Although all land plants execute an alternation of generations between a haploid gametophyte and a diploid sporophyte stage, the amount of time and space spent in each generation varies across clades. For instance, the bryophytes typified by Physcomitrella patens, are gametophyte-dominant and produce gametophyte structures known as rhizoids for anchoring to substrates as well as for nutrient absorption. Moss sporophytes are dependent on the moss gametophyte, and are diminutive in size, entirely lacking roots. In contrast, anthophytes like Arabidopsis thaliana are characterized by sporophyte-dominant stages with an exceptional spatiotemporal reduction in gametophyte structures. Such is the reduction that flowering plants rely on sporophyte root systems for anchorage and nutrient absorption, forgoing any production of rhizoids. In the middle of this plant evolutionary saga lies Ceratopteris richardii, a fern capable of not only producing gametophyte rhizoids but also sporophyte root systems. Great strides have been made in the elucidation of the genes required in the specification and development of sporophyte root hairs in Arabidopsis as well as for the development of Physcomitrella gametophyte rhizoids, but little is known of the specific genes utilized in ferns for similar processes. Recent advances in genomics and transcriptomics for Ceratopteris richardii permits orthologous gene-function searches. Here we showcase novel fern genes that are expressed in gametophyte and sporophyte stages that share predicted-peptide conservation with known moss and cress genes that are key molecular players in rhizoid and root hair development, respectively. (59)

Vyas, Khushali*, Carly Wood*, and Andre Walther Cedar Crest College, Allentown, PA 18104. The effect of Replication Protein A phosphorylation on telomere shortening in Saccharomyces cerevisiae using a PCR amplification-based protocol.- Replication Protein A (RPA) is a highly conserved, heterotrimeric, single-stranded DNA binding protein that is essential for many DNA maintenance pathways such as DNA replication, DNA damage repair, cell cycle regulation, and telomere maintenance. RPA is phosphorylated in a cell-cycle dependent manner and in response to DNA damage, suggesting that phosphorylation may play a vital role in regulating its function. We hypothesized that phosphorylation may regulate RPA functions in telomere synthesis, and our lab has previously shown that mutations in RPA that effect phosphorylation cause changes in telomere length in the baker’s yeast Saccharomyces cerevisiae (S. cerevisiae).  However, the mechanism for RPA phosphorylation’s regulation of telomere synthesis is unclear. As a result, we have been measuring telomere length of various S. cerevisiae mutants, including those with phosphorylated and dephosphorylated states of RPA. After isolating genomic DNA from S. cerevisiae, telomere lengths were amplified using Endpoint Polymerase Chain Reaction (PCR) and measured through gel electrophoresis. We are currently analyzing the telomere lengths of S. cerevisiae mutants and seeing positive and negative effects on telomere length caused by the addition of different mutations. We have confirmed our Endpoint PCR protocol using a control and have been able to verify our protocol in terms of amplifying telomere lengths. By comparing telomere lengths of RPA phosphorylated mutants to known telomere synthesis mutants, we will have a better understanding of the role RPA phosphorylation plays in regulating telomere synthesis.  Since the abnormal regulation of telomere synthesis is required for cancer cell formation and cancer progression, our research may provide insights into novel treatment targets for cancer. (26)

Wagner, Dylan*, and Jeffrey Thompson York College of Pennsylvania, York, PA 17405. The effects of e-cigarette aerosol extract and e-liquid on the JAK/STAT6 signal transduction pathway.- Electronic cigarette (e-cigarette) use has increased drastically over the years, outpacing traditional tobacco products. E-cigarettes heat a solution of propylene glycol, vegetable glycerin, nicotine, and flavor additives (e-liquid) to produce an aerosol. E-cigarette use has been significantly associated with chronic obstructive pulmonary disease (COPD) and worsened asthma symptoms. JAK/STAT6 is an inflammatory signaling pathway implicated in the etiology of these diseases. The objective of this study was to determine whether e-cigarette aerosol and e-liquid affects the signal transduction of this pathway, and if it may be one mechanism through which COPD and asthma symptoms increase. E-liquid aerosol extracts were created similarly to traditional cigarette smoke extract using a JUUL e-cigarette in conjunction with Virginia Tobacco and Menthol e-liquid. HEK293 IL-13/IL-4 Sensor cells are typically used to detect JAK/STAT6 signaling activity after activation by secreting a reporter protein detectable via a colorimetric reagent. Cells were exposed to each extract and their respective e-liquids in varying concentrations along with or without IL-13, ligand for the JAK/STAT6 pathway. The amount of reporter protein, secreted embryonic alkaline phosphatase (SEAP), was recorded after 18 hours via a colorimetric QUANTI-Blue assay to quantify signaling activity. Dunnett’s test was utilized to determine simple effects within groups after a two-way ANOVA. Signaling activity was significantly greater than the control when cells were exposed to 10% and 5% Virginia Tobacco extract with IL-13 (p= 0.0116, 0.0413). Signaling activity was lower than the control when exposed to 2.5% and 1.25% Menthol extract with IL-13 (p= 0.0121, p<0.0001). Cells exposed to Menthol e-liquid at 0.4% had statistically significant greater signaling than the control (p=0.0159). (29)

Wang, Jason* Germantown Academy, Fort Washington, PA 19034. Sleep Deprivation Induces Brain Inflammation Via Peripheral Monocyte Infiltration.- Chronic sleep deprivation (CSD), referring to the consistent lack of sleep or reduced quality of sleep, is a global concern in more than half of adults worldwide. Research has shown many links between CSD and neurological disorders such as Alzheimer’s disease and Parkinson’s disease; however, the underlying mechanisms remain to be elucidated. In my study, I look to uncover a possible underlying mechanism and identify a potential therapeutic target. Since the association between brain inflammation and neurological disorders is well established, I examine whether CSD can induce brain inflammation through peripheral monocyte infiltration and microglia activation within the brain of mice. By utilizing transgenic mice expressing red fluorescent protein (RFP) under peripheral monocyte-specific promoter CCR2, I demonstrated CSD-induced peripheral monocyte infiltration into the brain of mice through fluorescently visualizing RFP positive cells. Further, I showed that peripheral monocyte infiltration led to microglia activation in the brain by fluorescently labeling CD68, a microglia activation marker. In addition, by comparing the brains of sleep-deprived CCR2 heterozygous mice (CCR2+/RFP) to CCR2 knockout mice (CCR2RFP/RFP), I found that depleting the CCR2 receptor reduced peripheral monocyte infiltration and microglia activation in the brain, thus also reducing brain inflammation. Together, my observations indicate that CSD-induced peripheral monocyte infiltration leads to brain inflammation, suggesting that maintaining sleep hygiene and improving sleep quality may be critical for preventing neurological disorders in later life. In addition, I identify the CCR2 receptor as a potential therapeutic target for reducing the risk of developing neurological disorders. (37)

Wilkins, Ashley*, Mercedes Melo, Emily Bianchini, and Jennifer Hayden Cedar Crest College, Allentown, PA 18104. Finding novel antibiotics under our feet.- Medical innovations have been driven by antibiotics, chemical substances that kill bacteria, in the last hundred years. Many of these antibiotics were first isolated from soil microbes where the production of antibiotics allows them to outcompete their neighbors. However, that progress is being threatened by antibiotic resistance. Created by genetic mutations in bacteria, antibiotic resistance is causing bacterial infections to become difficult, if not impossible, to treat leading to increasing mortality rates of bacterial infections. Every year, over 700,000 people die from antibiotic resistant infections, and by 2050, ten million people are predicted to die from antibiotic resistant infections; thus, there is a dire need for novel treatments. One possible solution is to explore soil microbes for previously undiscovered antibiotics. In this study, microbes were isolated from topsoil and screened against known bacteria to test for antibiotic production. The microbes were then characterized with biochemical tests and identified by 16S rRNA sequencing. Antibiotics produced by a subset of microbes were extracted, re-tested for antibiotic activity, and killing activity was confirmed. For one microbe, the extracted antibiotic was able to inhibit a wider range of known bacterial species than was seen with direct microbial competition.  Additionally, antibiotic-producing microbes and the extracted antibiotics were tested for toxicity against eukaryotic cells.  Identification and classification of novel microbes could lead to the discovery of new, clinically useful antibiotics, an important component of the fight against antibiotic resistance. (80)

Wilson, Jordan E., Kiera L. Shellhammer*, and Amy Faivre Cedar Crest College, Allentown, PA 18104. Stream quality and water chemistry of Elk Ridge Run, a tributary to the Jordan Creek at Trexler Nature Preserve in Pennsylvania.- Stream disturbances often occur due to anthropogenic, terrestrial, and environmental influences. Often times, certain changes need to be made to the environment around a stream to reduce disturbances such as excluding animals from stream embankments to prevent stream degradation. Our study took place years after American bison (Bison bison), were fenced out of a small tributary stream to the Jordan Creek in the Little Lehigh River and Jordan Creek Watershed in Eastern Pennsylvania. The bison had been causing excessive erosion into the stream and increasing the amount of nitrogen and fecal bacteria, E. coli., into Elk Run Ridge. We performed water chemistry analysis using a HACH kit as well as conducted macroinvertebrate sampling during the early spring and fall seasons of 2020, and spring of 2021 at four sites along Elk Ridge Run, as well as one site that was a small tributary to Elk Ridge Run. Macroinvertebrate diversity varied between the seasons with high diversity in spring 2020 and low diversity in fall 2020. Levels of nitrate and phosphorus varied significantly depending on the sampling date but not among locations. If this pattern continues, another animal enclosure that spans the stream may not need to be moved, if it does not appear to degrade the stream. (78)

Witmer, Charles* The Haverford School, Haverford, PA 19041. Bone Quality Assessment Using Radiology.- Prostate cancer is a highly metastatic cancer which often spreads to bone tissue. This metastasis can give rise to bone fractures, so finding an effective way to predict incident (occuring in the future) bone fracture is important. Traditionally, Bone Mineral Density obtained through DXA scans is the metric used to determine incident bone fracture risk. This metric may not always  correctly predict fractures, however, so this study aimed to use bone metabolism to measure fracture incidence risk. Prostate cancer patients receive [18]NaF PET-CT scans to check for metastasis. Uptake of radiolabeled NaF is indicative of bone metabolism as it is absorbed by growing bone tissue. Using Standard Uptake Values (SUVs) from 50 patients with a history of prostate cancer, this study shows that femoral neck bone metabolism is not an effective predictor of incident fracture. However, SUVmean and SUVmax were found to correspond with age, weight and BMD. This study indicates that SUVs may not be useful to predict fractures but may be useful for other measurements. (42)

Woodring, Veronica*, Sam Brady, and Jennifer Ness-Myers Messiah University, Mechanicsburg, PA 17055. Effect of lymphoid cell kinase (Lck) inhibition on oligodendrocyte differentiation.- 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. Future studies will investigate the signaling pathways impacted by loss of Lck activity and their role in oligodendrocyte differentiation. (38)

Xu, Christina *, Michelle James, and Giancarlo Cuadra Muhlenberg College, Allentown, PA 18104. Effects of e-liquids on Streptococcus gordonii biofilm growth.- The use of electronic cigarettes, or vaping, has increased in popularity in the past decade among students in high school and college. The e-liquid contains harmful substances such as nicotine, propylene glycol, vegetable glycerol, and many flavors. The mouth is the first site of exposure to the e-cigarette aerosol and is the home of different commensal and pathogenic bacteria. Most of these microorganisms grow as biofilms, commonly known as dental plaque. Changes in the bacterial interactions may lead to oral diseases. There are limited studies regarding the effects of vaping in the oral cavity, and even fewer studies have focused on the effects of vaping on oral bacteria. This project aims to test the effects of e-liquids with multiple flavors on the biofilm growth of oral commensal bacteria using artificial media. We hypothesize that biofilm growth may be altered as a result of the flavorings in e-cigarette liquid. Bacteria biofilms of Streptococcus gordonii were grown in vitro, exposing the microbes to increasing concentrations of e-liquids (1%, 3%, 5% v/v) as well as growing them over different lengths of time (1 day or 2 days). Flavors of e-liquids include blueberry, cinnamon, menthol, strawberry, and tobacco as well as flavorless. Biofilm biomass was analyzed using a crystal violet staining procedure. This project shows that vaping can affect the growth of S. gordonii. Altering the growth of oral commensal bacteria can lead to further understanding of the effects of e-liquids on the implications of oral health. Exploring the effects of vaping in oral biology deserves more attention since oral health impacts systemic health. (93)

Yeager, Ashley*, and Wendy Boehmler York College of Pennsylvania, York, PA 17405. The effects of L-theanine on zebrafish (Danio rerio) development.- Anxiety is a common psychological disorder that is very prevalent in today’s population. This disorder is commonly treated with a mix of therapy and pharmaceutical drug treatments that can come with a variety of detrimental side effects. L-Theanine, an amino acid found in green tea, has been reported to have an array of health benefits including anxiolytic properties. While the mechanism underlying its therapeutic potential for anxiety is still being investigated, it is unclear whether this type of supplement is safe to use during pregnancy. Zebrafish embryos have continued to emerge as model organisms for drug toxicity studies as embryos are transparent, rapidly develop ex utero, and have close genomic homology to humans. To determine whether L-theanine causes any developmental abnormalities, zebrafish were exposed to different dosages of L-theanine and observed at different exposure time points. While there were no significant effects on survival rate or gross morphological deformities, heart rate displayed a decreasing trend with increasing doses of L-theanine. Further research is needed on the mechanism of this potential teratogenic effect on heart rate. (84)

Yoder, Cheyenne*, Adam Cooke, and Abigail Maley Wilson College, Chambersburg, PA 17201. The potential role of freshwater mussels as indicators of bacterial levels in their environment.- Water is an important public resource threatened by pollution such as wastewater contamination, agricultural runoff, and chemical waste. Bioindicators, organisms used to assess the quality of an environment and its changes over time, are an important method of measuring and monitoring water quality. Because they are directly affected by environmental pollution, bioindicators can sometimes serve as a more sensitive means of assessing water quality than chemical or physical tests. Due to their ability to filter large quantities of water, bivalves, such as mussels, are potential candidates for bioindicators. Bivalves may also provide a means of continuous water monitoring because they are sessile organisms and have the potential to filter and retain particles for long periods of time. The use of bivalves as bioindicators of the bacteria Escherichia coli would help with efforts to monitor water quality for public health.  E. coli is found in the lower intestines of warm-blooded organisms and in fecal matter. It is transferred to water bodies via agricultural runoff, waste from sewage overflows and polluted stormwater runoff. E. coli can cause urinary tract infections, respiratory illnesses including pneumonia, and bacterial gastroenteritis. This review suggests bivalves may be a useful tool for assessing bacteria levels and that Elliptio complanata, the Eastern Elliptio mussel, may be an effective bioindicator of potential sources of E. coli contamination in the Conococheague Creek within the Potomac Watershed. (50)

Zarilla, Alexandria*, Daniel Oar*, and Quyen Aoh Gannon University, Erie, PA 16541. Secretory carrier membrane protein 3’s (SCAMP3) role in beta-amyloid production and secretion.- Alzheimer’s disease is a neurodegenerative disease associated with loss of memory and cognitive function. The aggregation of extracellular plaques containing beta-amyloid is related to the processing of the amyloid precursor protein (APP). When endocytosed to the early endosome, APP will either be degraded by the lysosome or sent back to the trans-Golgi network. Degradation of APP is regulated by endosomal sorting complexes required for transport (ESCRTs). Disruption of ESCRT function leads to accumulation of beta-amyloid. The secretory carrier membrane protein 3 (SCAMP3) interacts and opposes the function of the ESCRT proteins. We hypothesize then that SCAMP3 may regulate APP trafficking by promoting or inhibiting trafficking of APP to the lysosome. As shown with our poster, we will test this hypothesis by examining the effects of RNA interference of SCAMP3 in two assays: (1) an immunofluorescence colocalization assay with full length APP and (2) and ELISA assay to measure beta-amyloid levels. Both assays will use H4 neuroglioma cells that have been stably transfected with APP-EGFP. In the immunofluorescence assay, the cells will be fluorescently labeled with antibodies to the lysosomal proteins LAMP1/2 or the early endosomal protein EEA1. Colocalization of EGFP-APP with EEA1 and LAMP1/2 will be observed under the microscope and quantitatively analyzed to determine the degree of colocalization. We then will use a well-established ELISA assay to measure intracellular and extracellular levels of beta-amyloid. We predict that if SCAMP3 promotes APP trafficking to the lysosome, then a knockdown of SCAMP3 will result in decreased colocalization of APP with lysosomal markers in our immunofluorescence assay and increased intracellular accumulation of beta-amyloid in our ELISA assay. On the contrary, if SCAMP3 inhibits APP trafficking to the lysosome, then there will be increased colocalization on the lysosome and decreased beta-amyloid accumulation. (20)