Announcement of the 2022 winners of the Spychala and Women in Science Awards

Abstract

The Pennsylvania Academy of Science works to recognize outstanding presentations of graduate and undergraduate students at the annual meeting each year. We present first, second, and third place oral and poster presenters with the Spychala Award, funded through an endowment created by the Anne Spychala Family Charitable Foundation. A second Spychala fund allows us to recognize achievements by women through the Women in Science award. After a two-year hiatus in these awards due to the global Sars- CoV2 pandemic, we are happy to announce the return of the awards with the return of our in-person annual meeting, 25–27 March at DeSales University.At the 2022 annual meeting of the Pennsylvania Academy of Science, 18 students were selected for outstanding presentations for the Spychala Awards and the Women in Science Award (Table 1). These winners represent nine different institutions and worked on projects ranging from gene expression in cancer cells, through chemotherapeutic drugs, to environmental assessment of sediments and invasive species. These projects also demonstrate some of the breadth of the Academy, with representation from the fields of biology, chemistry, and environmental science. Furthermore, the student winners represent an increasing diversity of maturing scientists, something that the Academy is working toward. Following this brief introduction are the abstracts (as submitted with minor edits) for each winning presentation (in alphabetical order by author).Kwabena AcheampongLafayette College | acheampk@lafayette.eduKhadijah MitchellLafayette College | mitcheka@la fayette.eduAbstract: Lung cancer has the second highest incidence rate and the most cancer-related deaths among males and females in the United States. Specifically, African Americans (AAs) have higher mortality rates and lower survival rates than European Americans (EAs) and other demographic groups. Determinants including geographical, sociopolitical, behavioral, and biological factors contribute to understudied lung cancer disparities. The biological determinants that are investigated in this study are telomere-lengthening network mechanisms and pathways and West African Ancestry. It is hypothesized that WAA is associated with clinically relevant noncanonical telomere lengthening and maintenance pathways in lung cancers from AA and EA patients. We categorized non-small-cell lung cancer (NSCLC) patients as high or low expressers based on median gene expression. and compared 5-year disease-specific survival data by telomere length and high and low WAA. We assessed differential gene expression by WAA for all 2093 genes. DAXX is significantly more expressed in low-WAA LUAD patients. There were no other significant expression differences by ancestry. NSCLC patients with high WAA expressed slightly more canonical pathway genes and LUAD patients with low WAA expressed slightly more noncanonical pathway genes. Patients with high WAA may benefit from combinational drug chemotherapy that disrupts the noncanonical telomere-lengthening pathway. Significant differences in gene expression in the LUAD cohort saw patients with low WAA having higher DAXX expression than those with high WAA. NSCLC patients with high WAA had higher median expression of canonical pathway genes. NSCLC patients with low WAA had higher median expression of noncanonical pathway genes. There are six differentially expressed TMN genes by WAA in NSCLC patients. Future research should quantify and compare the impact of standard combination lung cancer chemotherapy drug response on telomere length in LUAD cell lines with high and low WAA and calculate 5-year disease-specific patient survival based on TMN candidate gene expression.Kiyah BellLycoming College | belkiya@lycoming.eduJeff NewmanLycoming College | newman@lycoming.eduAbstract:Pedobacter is a genus in the Bacteroidota that usually produces pink or yellow carotenoids. Carotenoids are the most diverse and ubiquitous pigments found in nature, synthesized by plants, fungi, algae, and bacteria. They are a class of terpenoids with a linear isoprene backbone, and their structure consists of an extended conjugated system. In this work, five previously identified Pedobacter strains were recovered from the Lycoming College Culture Collection (LCCC). The pigments were extracted with methanol to obtain spectroscopic characteristics by high-performance liquid chromatography. Compounds with a three-peaked spectrum, typical of carotenoids, plus a maximum absorbance around 480 nm were commonly found in the pink strains, and 450 nm was the peak wavelength found in the yellow strains. Phylogenetic analysis using the Genome Taxonomy Database (GTDB) and the RNA polymerase b-subunit (rpoB) tree revealed that the yellow Pedobacter strains and the pink Pedobacter strains each shared a common ancestor and formed separate clades on long branches. A computational genomic study, the Average Amino Acid Identity (AAI), suggested that organisms of different colors should be in distinct genera. After mining the genome for carotenoid biosynthetic genes using the Rapid Annotation using Subsystem Technology (RAST) website, we discovered that the pink species encode b-carotene ketolase, which the yellow species lack. The presence or absence of the b-carotene ketolase enzyme should allow the pink species to produce canthaxanthin and astaxanthin, pink carotenoids, while the yellow species should produce b-carotene and zeaxanthin, yellow carotenoids. We conclude that the pink strains should be reclassified into a new genus, which we propose to call Roseopedobacter, due to their synthesis of pink carotenoids. In the future, we plan to optimize our LC-MS conditions to identify the carotenoids based on the molecular weight and fragmentation patterns.Kailey CarolandElizabethtown College | carolandk@etown.eduJohn HansonElizabethtown College | hansonj@etown.eduJane CavenderElizabethtown College | cavender@etown.eduAbstract: Alternative splicing has emerged as a major player in cancer initiation and progression. The alteration of splicing factor profiles and the effect on their subsequent targets have been reported in several cancers. Comparing tumor samples and tumor cell lines to build protein profiles of specific isoforms has elucidated common alterations that may indicate the important drivers of tumorigenesis. However, when primary tumors and tumor cell lines are used, there is often not a matched control to directly compare the results. But employing the simian virus 40 (SV40) DNA tumor virus model, studies can be designed to investigate the isoform profiles before and after cellular transformation. For this study, human diploid fibroblasts immortalized with telomerase (HDF(tert)) were stably transfected with plasmid encoding the early region SV40 (HDF(tert+T)). Two clones were compared with the parental line, and it was found that the level of T-antigen was directly correlated to increased levels of the splicing factor SAM68. Increased levels of SAM68 and the subsequent downstream altered proteins have been implicated as a driver of, and are now a chemotherapeutic target for lung, colon, and breast cancers. To elucidate the possible role of SAM68 in viral tumorigenesis, downstream splicing targets were assessed using RTPCR and Western blotting. We found no difference in the isoform ratios or levels of protein accumulation of the apoptosis regulator BCLx, bridging integrator BIN1, or cyclin D. Interestingly, it was found that the SAM68 target SRSF1 (another splicing factor) showed increased levels of only the specific SRSF1-208 isoform in the transformed lines. Studies are ongoing to investigate the cellular localization and potential phosphorylation alterations of the SRSF1 protein. Additionally, the tyrosine kinase receptor RON implicated in the aggressive tumor phenotype of epithelial to mesenchymal transition (EMT) and a target of SRSF1 splicing is currently under investigation.Kathlyn DiasCedar Crest College | kcdias@cedarcrest.eduAndré WaltherCedar Crest College | awalther@cedarcrest.eduAbstract: Dysfunction in proteins involved in DNA repair, replication, and recombination is linked to human cancers. Human Replication Protein A (RPA) is a highly conserved heterotrimeric ssDNA-binding protein which participates in many of these pathways involving DNA and is homologous with RPA (encoded by the RFA genes) in the highly studied eukaryotic model organism Saccharomyces cerevisiae. The RPA protein is composed of three subunits (70 kDa, 32 kDa, and 14 kDa), encoded in yeast by the RFA1, RFA2, and RFA3 genes, respectively, and the 32-kDa subunit is phosphorylated in a cell-cycle and DNA-damage-dependent manner, suggesting that phosphorylation of RPA may regulate its function. This study attempts to identify protein interactions with RPA’s 32-kDa subunit when either constitutively phosphorylated or dephosphorylated. Since RPA interacts physically with other proteins, a Yeast Two Hybrid Assay will be used to determine protein interactions in Saccharomyces cerevisiae. The system utilizes the ADE2 reporter gene with five RPA variant-Gal4 binding domain fusion proteins matedagainst unknown cDNA yeast library protein–Gal4 activating domain fusion proteins. The RPA variants individually consist of wild-type subunits of 70-kDa, 32-kDa, and 14-kDa along with two genetically modified subunits: hyperphosphorylated 32-kDa and dephosphorylated 32-kDa. Ten phosphorylation- dependent protein interactions with RPA have been identified through Y2H. Proteins found to have a phosphorylation-dependent interaction with RPA can potentially be utilized in cancer research and in further understanding of DNA repair, replication, and recombination.Bridget FinniffBloomsburg University | baf31406@huskies.bloomu.eduAngela HessBloomsburg University | hess2@bloomu.eduAbstract: Cutaneous melanoma may not be the most common skin cancer; however, it is the deadliest. For patients with metastatic melanoma, the survival rates drop to just 15%. Vemurafenib inhibits a specific mutation in B-Raf (V600E) found in the many metastatic melanomas. Although vemurafenib is somewhat effective in treating melanoma, resistance to the drug occurs at a frequent rate. Resistance to vemurafenib has recently been associated with increased EphA2 expression. EphA2 expression is associated with aggressive forms of melanoma and increased melanoma plasticity as characterized by vasculogenic mimicry. This study explored the mechanisms involved in the development of resistance to vemurafenib by investigating changes in EphA2 expression and vasculogenic mimicry in resistant and nonresistant cells. The data collected so far demonstrate that A375P vemurafenib-resistant cell line was partially resistant to vemurafenib and expression levels of EphA2 were slightly higher in the resistant cell line than in the nonresistant cell line. These experiments benefit cancer research because they shed light on the mechanisms associated with vemurafenib resistance and provide insight into the development of new therapeutic approaches for the treatment of melanoma.Katherine FranzoneCedar Crest College | KAFranzo@cedarcrest.eduAndré WaltherCedar Crest College | awalther@cedarcrest.eduAbstract: According to the American Cancer Society, in 2022 there will be around 1.9 million new cancer cases diagnosed and over 609,00 people are expected to die from cancer-related causes in the United States alone. Current chemotherapeutic cancer treatments are no-specific and target actively dividing cells. This damages the patient’s healthy cells, leading to unwanted side effects and decreasing the efficiency of the treatment. An increase in the specificity of chemotherapeutic drugs would greatly reduce the death of healthy cells by treatment. To better understand the impacts of the DNA damage caused by chemotherapeutic drugs and how eukaryotic cells response to this stress, our project focuses on analyzing the effects of chemotherapeutic agents in the baker’s yeast Saccharomyces cerevisiae, which has been used extensively as a model for human cells. Specifically, we have been analyzing the function of Replication Protein A (RPA), which is a highly conserved protein involved in DNA replication and repair in both humans and yeast. To dissect the function of RPA, our lab has previously generated strains that contain various mutations in the yeast RPA homolog, RFA, along with different known repair genes, and we have been examining the sensitivity of these yeast strains to DNA damage using spot assays to look at the survival of yeast on media at various concentrations of the chemotherapeutic agents campothecin and hydroxyurea. The differences in growth and survival rates of the various strains in the spot assays have allowed preliminary conclusions to be drawn about the importance that the various regions in RPA have in the cellular response to DNA damage caused by chemotherapeutic drugs. A clearer picture of RPA’s role in DNA repair and its interactions with other genes will allow a better understanding of the underlying causes of cancer and more specific chemotherapeutic drugs.Samantha GreenbergLafayette College | greenbsf@lafayette.eduKhadijah MitchellLafayette College | mitcheka@lafayette.eduAbstract: Kidney cancer is a top ten cancer in incidence in the United States. Renal cell carcinoma (RCC) is the most common type (85%), while clear cell RCC (ccRCC) is the most common subtype (80%). African Americans (AAs) notably have lower ccRCC survival rates than European Americans (EAs). Certain genomic/transcriptomic factors have been identified as risk factors for ccRCC survival and may drive this disparity. A gene expression profile called ClearCode34 was previously developed to distinguish between two subtypes of ccRCC, ccA and ccB. Patients with the ccB subtype are known to have lower rates of survival, higher rates of metastasis, and increased tumor size/grade. Genetic ancestry has been linked to lower survival for AA patients with other urologic cancers. Here we show that AA and EA ccRCC patients have different ccA/ccB subtype frequencies differentiated by both self-reported race and West African Ancestry (WAA). mRNA expression, survival data, and self-reported race were downloaded from The Cancer Genome Atlas (TCGA) Firehose Legacy cohort on cBioPortal as a discovery cohort. Genetic ancestry was calculated using STRUCTURE from the Cancer Genetic Ancestry Atlas to group patients with >70% and <70% WAA. Prediction Analysis of Microarray (PAM) in R was used to classify the ClearCode34 subtypes. GraphPad Prism analyzed survival data by comparing across subtypes and races or ancestry groupings. AAs and high WAA patients had significantly higher percentages of ccB-typed tumors. ccB-typed patients showed worse 5-year disease-specific survival rates than ccA-typed. However, these differences were not significant between self-reported races and WAA groupings. Future directions include pathway analysis to identify novel targets in AAs and EAs using a precision medicine approach. All significant findings will be validated in a Cooperative Human Tissue Network/Geisinger cohort.Jazzlyn GrenierYork College of Pennsylvania | jgrenier1@ycp.eduSean GeorgiYork College of Pennsylvania | sgeorgi@ycp.eduAbstract: Circular RNA (circRNA) is a recently discovered genetic material where the 5’ and 3’ ends of one or more exons are covalently bound together. CircRNAs have been found to regulate gene expression and are tied to roles in neurodevelopment and retinal function. One circRNA, circPDE4B, is expressed in human retinal cells and regulates cellular proliferation in alveolar epithelial cells. However, the expression patterns of the linear and circular forms of PDE4B have never been studied across retinal development and in many human cell lines. The purpose of this study was to determine the expression of circPDE4B and PDE4B in multiple human cell lines and throughout the development of chicken (Gallus gallus) retina. PCR results showed that circPDE4B was expressed in HEK-293 (human embryonic kidney) cells, but not in U-87 (glioblastoma) cells, MDA-435 (melanoma) cells, or MCF-7 (breast cancer) cells, while the linear form was expressed in each of the cell lines. This indicates that circPDE4B could have a role in HEK-293 cells. qPCR results showed that circPDE4B expression increased across chicken retinal development, indicating it may have a role in retinal development. However, the expression pattern was similar between the linear and circular forms of PDE4B, so there is likely no preferential splicing occurring from the linear to the circular form of the gene. Future research could aim to determine the function of circPDE4B in HEK-293 cells and developing chicken retinal cells.Daniel GuevinMessiah University | dg1323@messiah.eduLawrence MylinMessiah University | lmylin@messiah.eduAbstract: Pancreatic cancer is currently the fourth leading cause of cancer deaths, with a survival rate of only 10%. The tumors grow deep inside the body and do not produce clearly defined symptoms until later stages. Pancreatic cancer is very aggressive due to genetic makeup and local tissue alterations (e.g., fibrosis) that block access by traditional chemotherapeutic agents. One strategy to combat tumor growth has been to activate (or relieve repression of preexisting) host immunity so that host tumor-specific lymphocytes, specifically T lymphocytes, are empowered to detect and destroy tumor cells. CD8+ or CD4+ T lymphocytes may combat tumors upon recognition of altered peptides presented on MHC molecules expressed by the tumor cells, or on phagocytes that have ingested tumor materials, respectively. CD8+ T cells may directly kill tumor cells, while cytokines secreted by CD4+ T cells empower tumoricidal functions of other cellular effectors. Host immunity can be augmented by vaccination using altered tumor-associated targets. We have designed a system in which a unique tumor target has been incorporated into a highly immunogenic viral oncoprotein, the SV40 T antigen. Immortal cells expressing this recombinant protein given as a vaccine are readily destroyed by the host immune system as tumor-specific T cells are generated. We have tested the efficacy of this vaccine by implanting vaccinated mice with murine pancreatic cancer cells engineered to express the human target protein. In a preliminary study, vaccination did reduce tumor growth. Tumor samples will be analyzed for the presence of infiltrating T cells in a future extension of this study.Sarah KlebElizabethtown College | klebs@etown.eduJodi LancasterElizabethtown College | lancasterj@etown.eduAbstract: Activation of dendritic cells (DCs) through toll-like receptor (TLR) agonists causes release of cytokines. DC2.4 cells are immature murine DCs derived from the bone marrow that express TLRs, including TLR3. The response of DC2.4 cells to varying doses and times of the TLR3 agonist poly(I:C) were conducted. In contrast to previously studied MuTu DCs, DC2.4 cells produced little to no interleukin-12 (IL-12) after 12-h exposure to doses from 1 to 1000 µg/mL poly(I:C). This was surprising because the literature suggests that the expression of TLR3 dramatically increases when DCs are maturing, which should occur upon poly(I:C) exposure. Once fully mature, DCs receive inflammatory signals to reduce TLR3 expression. Treatment of DCs with interferon-a (IFN-α) has been found to enhance IL-12 production by increasing TLR3 expression. DC2.4s exposed to IFN-α prior to poly(I:C) treatment also failed to produce IL-12. Experiments using lipopolysaccharide (LPS) confirmed that the cells can make IL-12 through activation of the TLR4 pathway. Ongoing studies are investigating the functionality of the TLR3 pathway in the cells. Luciferase assays with IFN-β plasmids are being conducted to confirm the presence of a functional TLR3. Western blotting and immunofluorescence are also being explored to qualitatively determine whether the DC2.4 cell line expresses TLR3, as reported in the literature. Analysis of TLR3 levels and production of IL-12 by DC2.4s allows for greater understanding of how different laboratory-derived lines of DCs exhibit responses to TLR agonists. Additionally, the maturation status of the laboratory’s DC2.4 cells is being considered.Alianna LandryYork College of Pennsylvania | alandry@ycp.eduSean GeorgiYork College of Pennsylvania | sgeorgi@ycp.eduAbstract: Cancer is a devastating disease that affects many people around the world. Although there are many treatments available, which are dependent on factors such as stage and type of cancer, there is no cure for most forms of the disease. One important way to take a step forward in treatment is to fully understand all the components of the disease, including novel genetic elements, such as circRNAs. circRNAs are a fairly recent discovery, and are generated as a result of joining the 3’ and 5’ ends of an RNA molecule. Although the functions of some circRNAs have been determined, the function of most circRNAs is still unknown. One circRNA, circBANP, has been found in several types of cancers. In order to further understand circBANP expression in other cancers, as well as in normal development, this study analyzed the expression of linear and circBANP throughout embryonic development of the chicken (Gallus gallus) retina and determined the expression of linear and circBANP in various human cancer cell lines. To analyze the change in expression of linear and circBANP in the developing retinal cells, qPCR was used. Additionally, gel electrophoresis of PCR reactions of linear and circBANP in human embryonic kidney cells (HEK-293), glioblastoma (U 87), melanoma (MDA 435), and breast cancer (MCF 7) cell lines was also performed to determine whether the genes were expressed. The results showed that linear BANP decreased as development progressed, whereas circBANP did not have any significant changes in expression. Gel electrophoresis also concluded that linear and circBANP are expressed in HEK-293, U 87, MDA 435, and MCF 7 human cell lines. These results provide promising potential starting points for further research in which the function of circBANP could be determined in chicken retinal development as well as in the tested human cell lines.Tiffany NguyenYork College of Pennsylvania | tnguyen13@ycp.eduBianca CaresosaYork College of Pennsylvania | bcaresosa@ycp.eduRonald KaltreiderYork College of Pennsylvania | rkaltrei@ycp.eduAbstract: The PI3K/AKT/mTOR pathway is one of the most commonly mutated pathways found in cancer. Cyclin-dependent kinase 5 (CDK5), an essential kinase within this pathway that mediates several biological processes, is up-regulated in many cancers. Studies have shown that decreasing CDK5 reduces the aggressiveness of various cancers. However, little is known about the upstream factors that regulate this kinase, such as PI3K and protein kinase B (AKT). Recent studies have shown that combining a CDK5 inhibitor (Roscovitine, RSV) with chemotherapeutic drugs like temozolomide (TMZ) and doxorubicin (DXR) targets cancer in a synergistic manner to induce apoptosis and decrease cellular proliferation. Our study was designed to determine the cancer-suppressing effects of RSV when combined with the antimetabolite (methotrexate, MTX) within three human cancer cell lines (U87, HT29, and MCF-7). Using this in vitro model, we performed a cytotoxicity assay to determine the LD50 of each antimetabolite MTX and 5- fluorouracil (5-FU). Using the calculated LD50 value, we then measured gene expression of PI3K and AKT (qRT-PCR) under combination treatments with RSV. The LD50 for MTX from each cell line was determined for each cell type (U87: 6.038 μM; HT29: 1.001 μM; MCF-7: 1.200 μM). MTX was then combined with a low (1.0 μM) or high (20 μM) dose of RSV and gene expression was determined. We found that there was a decrease in PI3K and AKT expression when treated with MTX/RSV in U87, not HT29 and MCF-7. Our work suggests that RSV improved the therapeutic activity of MTX in glioblastoma. The combination of RSV and MTX is a potential therapeutic possibility in multiple cancers associated with CDK5 overexpression.Giulia RomanoCedar Crest College | gcromano@cedarcrest.eduLindsey WelchCedar Crest College | lawelch@cedarcrest.eduAbstract: Benzodiazepines are often used for treatment of insomnia, convulsions, and many psychiatric disorders. The widespread use of this class of drugs has raised concern about recreational benzodiazepine abuse. This highlights the importance of chemical detection and concentration determination of the benzodiazepine drugs within a human system. Although there are many different approaches to detection, this proposal outlines a novel presumptive testing method using chemiluminescence. The proposed method offers high sensitivity, a broad linear dynamic range, and quick, simple sample analysis. A comparative study was designed to analyze three azepine drugs, carbamazepine, clonazepam, and diazepam. The chemiluminescent reagents to be discussed are tris(2,2’-bipyridyl dichlororuthenium (II) hexahydrate (Ru(bipy)3Cl2•6 H2O) with cerium (IV) sulfate, known as Method A, and n-bromosuccinimide (NBS), known as Method B. Chemiluminescence was detected using a Berthold Technologies Lumat3 tube luminometer. Method A was more sensitive for carbamazepine and clonazepam, while method B was more sensitive toward diazepam. These drugs were detected in concentration ranges of 1.1. μM to 0.275 mM. The results from this work suggest novel presumptive tests for these drugs of concern. These methods require minimal sample preparation, offer a rapid screening process, and facilitate the detection of these compounds in biologically relevant concentrations.Adina ShresthaLafayette College | shrestha@lafayette.eduKhadijah MitchellLafayette College | mitcheka@lafayette.eduAbstract: Lung cancer is the leading cause of cancer-related deaths in the United States. Approximately 85% of lung cancer patients have non-small-cell lung cancer (NSCLC) consisting of lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). African Americans (AAs) have a higher mortality rate than European Americans (EAs), despite smoking less. When AAs do smoke, most select menthol cigarettes. Cytochrome P450 (CYP) and uridine diphosphate (UDP)-glucuronosyltransferase (UGT) break down menthol and other tobacco carcinogens. Not much is known about how population-specific expression of CYPs and UGTs impacts aggressive lung tumor biology. The Broad GDAC Firehose was used to download normalized mRNA-seq and clinical data (LUAD n = 50 AAs, 342 EAs; LUSC n = 24 AAs, 274 EAs in the Cancer Genome Atlas [TCGA]). The TCGA Splicing Variants Database (TSVdb) was used to access UGT and CYP mRNA isoform data by race. We performed differential expression (DE) on UGT and CYP genes and isoforms using Excel (two-sample t-test assuming unequal variances, P = 0.05). We found that10/20 UGTs and 15/55 CYPs are DE by race and 9 UGT and 15 CYP mRNA isoforms are DE by race; 6/14 UGT mRNA isoforms are DE in survival by race; and 11/22 CYP mRNA isoforms are DE in survival by race. AAs have lower expression of UGT and CYP mRNAs and mRNA isoforms that break down carcinogens, which may help to explain the high incidence of lung cancer in AAs. AAs overall had better 5-year survival than EAs among LUAD patients, especially by high expressors of both short and long isoforms. In the future, AA and EA lung cancer cell lines will be exposed to menthol cigarette smoke to determine population-specific drug responses to chemotherapy agents.Lily VelazcoMessiah University | lv1208@messiah.eduHunter ZondoryMessiah University | hz1163@messiah.eduLawrence MylinMessiah University | lmylin@messiah.eduAbstract: Antibodies protect by various mechanisms, but induction of antibodies that block infection by binding to pathogen surface structures is a goal of current vaccines. Past Messiah University microbiology courses included experiences wherein students blocked infection of Escherichia coli B by bacteriophage T4 using polyclonal T4-specific goat antiserum. Unfortunately, the polyclonal serum is no longer available. We have undertaken to generate mouse monoclonal antibodies that neutralize phage T4 to use in the same laboratory experiences. Our strategy was to immunize Balb/c mice with concentrated T4 suspensions from which endotoxin (lipopolysaccharide) had been depleted by gentle organic extraction. Mice were injected twice over a 2-week period with ~1.6–3.5 × 1010 pfu of T4r+ phage. Serum was prepared from blood collected by cheek vein puncture and assessed for T4-neutalizing activity in a 96-well plate scale-screening assay. Small amounts of E. coli B and T4 were combined in each well. Ongoing infection by T4 limited the density of bacterial growth in a well when measured at 6–8 h. The presence of neutralizing antibody prevented infection, and instead allowed the bacteria to grow to near saturation. Use of serially diluted serum samples allowed relative neutralizing titers to be determined. Exponentially growing E. coli B cells were transformed with plasmids encoding ampicillin and either gentamicin or streptomycin resistance. Infection by T4 was confirmed in the presence of the relevant antibiotic, as well as the ability of each transformant to proliferate if T4 was neutralized by goat antisera under similar conditions. Production and screening of hybridomas is in progress.Blanca VilledaWilson College | blanca.villeda@wilson.eduAdam CookeWilson College | adam.cooke@wilson.eduM. Dana HarrigerWilson College | dharriger@wilson.eduDeborah AustinWilson College | daustin@wilson.eduAbstract: Plastics are ubiquitous today due to their wide range of applications and affordability. Microplastics, plastic debris less than 5 mm in length, are known to be ingested by different freshwater organisms. Once ingested, microplastics can obstruct the organism’s digestive system and clog its feeding appendages. This study investigated whether microplastics are present in the Conococheague Creek and whether there is a correlation between primary land usage and microplastic concentrations in the Conococheague. The Conococheague is a tributary of the Potomac River, which flows into the Chesapeake Bay. It is also the source of drinking water for the Borough of Chambersburg. Four areas of study along a 20-mile reach were chosen based on the land use the creek is flowing through: the headwater of the creek, a recreational area, an agricultural area, and an urban area. Three sediment samples were collected from each site. Microplastics were separated from sediment based on density using zinc chloride (specific gravity 1.6 g/ml) stained with rose bengal dye and identified by stereomicroscopy. Observed microplastics were classified based on their shape: fragments, films, fibers, and other. Microplastics were present in all samples, but there was no statistical difference between the four sites in the average number of microplastics per dry weight. The most abundant type of microplastic in all samples was microfibers. However, there is a trend in the composition of microplastics among the samples. The urban site had the most diverse composition of microplastics, followed by the agricultural site, the recreational site, and finally the pristine site. This study indicates that microplastic contamination occurs at multiple locations along this reach of the Conococheague Creek. Further studies could elucidate points of contamination along the creek and whether the microplastics are affecting the organisms in this aquatic habitat.Talia WatsonCedar Crest College | tlwatson@cedarcrest.eduLindsey WelchCedar Crest College | lawelch@cedarcrest.eduAudra BratisCedar Crest College | abratis@cedarcrest.eduAbstract: In a pandemic, it is of grave importance that novel treatments for diseases be found. Naphthoquinones are a promising alternative to some antibiotics, as they have significant pharmacological properties. The typical catalyst used to synthesize naphthoquinones is hexavalent chromium, which has many unfavorable qualities. Metal acetylacetonate catalysts are a safer alternative when composed of earth-abundant metals such as iron, cobalt, and vanadium. The synthesis of 5-hydroxy-1,4- naphthoquinone (juglone) and 1,4-naphthoquinone was achieved via oxidation of 1-naphthol with tris(acetylacetonato) cobalt (III), tris(acetylacetonato) iron (III), and bis(acetylacetonato) oxovanadium (IV) catalysts. Products of oxidation reactions were characterized with infrared spectroscopy and gas chromatography/mass spectrometry. Other starting naphthyl compounds including 1-bromo-2-naphthol and 2,7- and 2,3-dihydroxynaphthalene were tested, but there was no naphthoquinone product formation. In addition to the successful oxidation of 1-naphthol, oxidation of 1,5-dihydroxynaphthalene and 4-chloro-1- naphthol was also investigated, with promising conversion to naphthoquinones. Based on the effect of catalyst on the selectivity toward each product, the synthesis of naphthoquinones using this novel approach with metal acetylacetonate catalysts was successful for a variety of naphthol starting compounds.