Proceedings of the West Virginia Academy of Science

In defense of information literacy across the disciplines

Nicholas M. Gardner — 2024-04-18

Information literacy, coined in the 1970s, covers a wide range of skills needed to navigate the information landscape to find relevant and useful information to solve problems. The teaching of information literacy is inconsistent across state standards, and West Virginia students entering college are often ill-equipped to find credible, useful information. Information literacy’s core skills include identifying individual information needs, finding relevant sources to solve those needs, and evaluating those sources for credibility. In the pre-Internet, mass media age, it was necessary primarily to evaluate and consider the bias of sources when assessing credibility. Then, mass disinformation/misinformation became a critical issue for information literacy, particularly in the mid 2010s. In the last two years, the now wide scale availability of generative AI tools now requires an even more substantial toolbox for information literacy. Unfortunately, discussions around equipping users with the appropriate skills have become increasingly siloed, with parallel developments in news literacy, digital literacy, media literacy, AI literacy, and reappropriation of science literacy to effectively encompass the same skill set. Here, I show the cross-disciplinary nature and value of information literacy, arguing for its superseding nature against these other literacies, and give recommendations for greater integration of information literacy instruction across college curriculum. Solutions for ILI implementation are readily found in the library and information science literature, and professional staff and faculty are in many institutions prepared or trainable to deliver ILI in both generalized and discipline-specific contexts to empower users to be more information literate and capable of navigating increasingly complex information landscapes.

Integration of Energy Conservation Principles Across Mechanical Engineering Curriculum: A Work-in-Progress Project

Yogendra M Panta — 2024-04-18

The conservation of energy, mass, and momentum stands as fundamental laws in physics, resonating deeply within engineering education. This ongoing project, now in its third year of implementation, aims to seamlessly integrate energy conservation principles across the mechanical engineering undergraduate curriculum. Across various courses, including dynamics, fluid dynamics, and thermodynamics, students delve into the diverse forms of mechanical energy. From kinetic and potential energy in dynamics to fluid flow energies and thermodynamic principles in fluid dynamics and thermodynamics courses respectively, the overarching principle remains: energy is neither created nor destroyed but instead transfers between different forms, maintaining a constant total within a fixed domain.

The project emphasizes the energy balance equation introduced in the first Thermodynamics course, laying the groundwork for subsequent exploration. Students are guided through the transition from the first law of thermodynamics to Bernoulli's equation in Fluid Mechanics, bridging theoretical concepts with practical applications. This integration continues into Heat Transfer and elective courses such as Industrial Hydraulics and Aerodynamics. To assess student comprehension, direct and indirect assessments are conducted, measuring understanding through principle and practical examples. Feedback and questionnaire responses indicate enhanced understanding of energy conservation principles through the synchronization of energy balance concepts across multiple courses. This presentation showcases the evolution of our work-in-progress project, initially presented at the 2022 and 2023 WVAS meetings. By fostering a holistic understanding of energy conservation principles, our aim is to empower students with a comprehensive foundation to tackle real-world engineering challenges.

Using historical biocultural variables and skeletal pathology to assist with osteoprofiles

Kristy Henson — 2024-04-18

The biocultural approach uses biological, environmental, and cultural variables to help identify where an individual spent their childhood or their adult life. When a skeleton is identified DNA may not be sufficient in identification and complete osteoprofiles are needed. This research examines historic skeletal, genealogical, and biocultural information to determine if there is a correlation with the presence of vitamin D deficiency or spinal pathology. One hundred seventy-seven individuals living between the 1830s and 1940s were analyzed and traced through the historical record. Preliminary results indicate that there were trends in determining where an individual spent their childhood and the likelihood of vitamin D deficiency (chi-square p = 0.006; Spearman RS 0.92, p = -0.008), year of death (chi-square 0.005; Spearman RS 0.87, p = -0.01), presence of spinal pathology (chi-square p = 0.05; Spearman RS 0.72, p = -0.03), and trending toward age (p = 0.08). More research is warranted but completing osteoprofiles using biocultural variables may help identify where someone is from and how they lived their lives which in turn will assist with identification.

NSF S-STEM Program - Supporting Undergraduate Cohorts of Career-Ready Engineering and Science Scholars (SUCCESS) Project at WVU Tech – Year 3 updates

Yogendra M Panta — 2024-04-18

The SUCCESS project at WVU Tech aims to recruit, retain, and graduate low-income STEM students with high motivation and entrepreneurial mindset, enhancing their likelihood of employment in technology-based businesses or creating start-up ventures. This report provides updates for Year 3, focusing on the recruitment and retention efforts. Currently, there are 20 scholars in the program, with Cohort 1 comprising seven students and Cohort 2 comprising 13 students. The program has seen some changes in Cohort 2 structure, with three Computer Science (CS) students dropping out in Fall 2023, and four new sophomore students including one Computer Engineering (CpE) and three CS students being admitted. Scholars have engaged in various activities, including completing entrepreneurship courses, attending career fairs, and working on research projects with faculty. The project team is actively collecting and analyzing feedback from scholars to ensure necessary improvements are implemented. Major activities during Year 3 included faculty mentor meetings, IEEE/ACM student branch engagements, and industry mentor interactions. Looking ahead, the project aims to continue supporting scholars in their academic and career pursuits, with a focus on enhancing industry mentorship and peer-to-peer relations. Additionally, the project has diligently explored opportunities to expand its impact to all eight existing engineering programs at WVU Tech and has recently submitted a Track 2 proposal to the NSF S-STEM program. The updates conclude with a discussion on the project's impact on student outcomes and the broader STEM community.

Impact of a Networked Improvement Community’s Interventions on STEM Student Persistence. Review of Results and a Call to Action.

Sue Ann Heatherly — 2024-04-18

It is well known that high impact practices such as student learning communities and peer-mentoring, early engagement in research and faculty-student relationship building can have a positive influence on undergraduate student engagement and retention in their chosen field of study. The First2 Network is a Networked Improvement Community comprised of students, faculty, staff and administrators from eight higher education institutions in West Virginia and one in Kentucky, that has, over the past six years, engaged in creating interventions to implement and test these high impact practices with the aim of doubling the graduation rate of first-generation and other underrepresented STEM students. Relying on data collected, mostly from WVU, we see very promising outcomes on the persistence of STEM majors. These results will be shared. However, our data story is incomplete as there is no mechanism to sort data on first-generation status across all universities and colleges in the state. The value of this information and its inclusion in a future state funding formula will be explored.

An Assessment of Movement and Dam Passage of Paddlefish in the Ohio River, West Virginia.

Stuart Welsh — 2024-04-18

Paddlefish (Polydon spathula), a large-bodied and highly migratory species of large river systems, has experienced population declines or extirpation in parts of its range. As an effort to reestablish Paddlefish populations in the Ohio River of West Virginia, the West Virginia Division of Natural Resources has periodically stocked Paddlefish, but post-stocking studies are needed to assess the status of the reintroduced population, including information on movements of Paddlefish or on their passage through navigation dams of the Ohio River. Using telemetry techniques, we monitored seasonal movements and assessed dam passage of five Paddlefish in the Robert C. Byrd pool of the Ohio and Kanawha rivers. Paddlefish were highly mobile with two individuals using the entire R.C. Byrd pool during the 11-month study period. The highest movement rates occurring during the expected spawning period of April and May, where all individuals frequented the upper section of the pool in Kanawha River. Downstream passage was documented for one individual at the R.C. Byrd lock during July 2023, but no evidence of passage was documented for the locks at Racine or Winfield. Tagging of additional Paddlefish is planned, which will increase sample size for future assessments and continue efforts toward the long-term conservation and management of Ohio River Paddlefish.

Screening Resorufin Pentyl Ether Analogs for Enhanced Antimicrobial Activity Against Francisella tularensis and Neisseria gonorrhoeae.

Mackenzie Hall — 2024-04-18

Antibiotic resistance is an urgent public health threat. The CDC estimates there are approximately 2.8 million new cases of antibiotic-resistant infections annually resulting in 35,000 deaths and billions of dollars in health care costs. The development of new drugs is imperative to combat this crisis and prevent the loss of additional lives from once “curable” diseases. Resazomycins, a novel family of antibiotics, have bactericidal activity against Francisella tularensis and Neisseria gonorrhoeae. One resazomycin, resorufin pentyl ether (RPE), significantly reduces vaginal colonization by N. gonorrhoeae in a mouse model of infection. Repeated administration of RPE, however, fails to clear the infection, in contrast to a single dose of ceftriaxone, an antibiotic commonly used to treat gonorrhea, which clears the infection within 24 hours. Further characterization of resazomycins revealed the efficacy of these compounds is limited by interaction with serum albumin and reduced oxygen concentrations found within mammalian tissues. Therefore, we hypothesize that novel resazurin analogs that maintain antimicrobial activity in the presence of serum albumin and low oxygen will have improved therapeutic efficacy in vivo. Resazurin has been chemically modified to generate ether and deoxygenated compound derivatives. Six ether derivatives and two deoxygenated analogs exhibited robust antimicrobial activity against F. tularensis and N. gonorrhoeae. In the presence of serum albumin, however, reduced efficacy was observed in the six ether analogs yet activity was maintained in the deoxygenated analogs. We plan to screen the efficacy of additional ether and deoxygenated analogs against F. tularensis and N. gonorrhoeae.

Metacognitive reporting provides insight on students’ out-of-class study activities geared toward their learning of chemistry

Michelle Richards-Babb — 2024-04-18

Prompting students to practice metacognition, and encouraging the growth of metacognitive strategies improves student success in chemistry coursework. In this study, students were encouraged to submit weekly metacognitive reports providing (i) their time devoted to the course, including hours devoted to out-of-class additional study, and (ii) detailed summaries of additional study activities. This study was administered to students in two different courses, introductory chemistry and general chemistry I. Introductory chemistry students submitted 1,513 metacognitive reports with self-reported overall means of 2.81 hour per week devoted to attending lecture, 4.75 hours per week engaged in additional study, and 7.56 hours per week devoted to the course overall. Weekly patterns indicate that students’ additional study was focused on days of the week that preceded formal assessments. Our expectation was that general chemistry I students would report more time devoted to additional study outside of class than introductory chemistry students because of the preparatory nature of the course. General chemistry I students submitted 3,551 weekly metacognitive reports with self-reported overall means of 2.83 hours per week devoted to attending lecture, 4.83 hours per week engaged in additional study, and 7.66 hours per week devoted to the course overall. We will discuss the seeming equivalence of time spent on out-of-class additional study for the two populations of students. In addition, we will present qualitative analysis of students’ out-of-class study strategies, including classification of study strategies as deep or surface-level. This work is partially supported by the NSF-funded First2 Network.

Continuing Osteological Research with Fairmont State University’s Human Skeletal Collection

Jay Bow — 2024-04-18

Human skeletal remains have been used at some point by most institutions involved in natural science education, though many real remains are slowly being replaced in classroom use by plastic models. Unfortunately, most antique teaching skeletons still in collections today were obtained during an ethically gray era, resulting in a lack of documentation and accountability. Since their purchase, many of these skeletal remains have been disassociated and damaged due to improper storage, the selling of partial skeletons, and mishandling. Projects such as this one at Fairmont State University are being performed to rehumanize and reassociate individuals using a variety of osteometric and macroscopic analysis techniques and create osteobiographies for them. Fairmont State University’s collection has been determined to contain a minimum of 43 individuals, 28 of whom were able to have profile information restored. Current ethical and legal standards related to the treatment of teaching skeletons are nearly non-existent in the US, but projects such as this one can bring attention to the historically immoral treatment of these individuals.

Spatial and temporal analysis of physical and chemical properties of Short Creek

Elizabeth Burkey — 2024-04-18

Freshwater systems are vitally important. Ecologically, rivers and streams are responsible for immense biodiversity, providing habitat for terrestrial and aquatic flora and fauna. Freshwater systems benefit humans in innumerable ways, providing drinking water to millions of people, means of transportation, food sources, and recreation areas. Nevertheless, the degradation of freshwater ecosystems continues to worsen at alarming rates. Long-term water quality monitoring can help determine the best management practices to improve and restore waterways. This study assessed changes in water chemistry in the Short Creek tributary located in Wheeling, WV, comparing historical data to current, focusing on physical and chemical water quality parameters data recorded from single monthly points obtained from historical data and averaged monthly points from current data. Short Creek represents 4 of 11 sites where data is collected weekly year-round. This data collection has been ongoing since 2018 and is part of a more extensive compilation hosted by Three Rivers Quest and the WVU Water Resource Research Institute. Water temperature, dissolved oxygen, chloride, specific conductivity, pH, and turbidity were recorded and analyzed utilizing Excel and RStudio. The increased frequency of monitoring provides a more accurate representation of Short Creek's overall condition. Both historical and current datasets were incomplete but generally did not show declines. A concerted effort in long-term water quality monitoring is necessary for understanding stream health. Current and historical data combined give us more resolution to estimate trends in water chemistry, better determine stream health progress, and assist in implementing best management practices.

Does Eye Color Affect Vision Acuity in a Variety of Light Conditions?

Tailynn Varney — 2024-04-18

Eye color is a physical characteristic that varies from person-to-person. Due to a range ofpigmentation of the iris, people with different eye colors tend to see the world differently. For those with lighter eyes, it may be more difficult to see clearly in environments with certain light conditions or intensities. Throughout a person’s life, they encounter several different forms of light ranging from natural sunlight to artificial lighting. In my research, I investigated whether a person’s eye color influences their vision in artificial lighting compared to natural lighting.Preliminary results show that light did influence visual acuity. However, there was no correlation between eye color and light influenced vision changes. To assess vision, participants were asked to cover each eye and read a Snellen eye chart. This was completed a total of four times. Using two separate eye charts, participants were tested at ten and twenty feet away from the charts. Each test was presented in a room with only artificial light, and outside of the building in natural light. This information can help individuals ensure that the change in vision, if any, caused by different kinds of light does not negatively influence their everyday life. In extreme situations those with eyes more sensitive to certain types of light could cause harm to themselves or others around them. Having this information about oneself is helpful to all people, not just those with a history of eye issues.

Dielectrophoretic Characterization and Computational Analysis of Peripheral Blood Mononuclear Cells from MMTV-PyMT Mammary Carcinoma Models for Late Carcinoma Detection

Raphael Oladokun — 2024-04-18

Breast cancer is the leading cancer in women, with 284,200 new cases and an estimated 43,600 deaths in 2021. Early detection and proper treatment can enhance outcomes. This study probes the dielectric properties of peripheral blood mononuclear cells (PBMCs) from MMTV-PyMT mice at 14+ weeks (stage IV) using a microfluidic platform for early breast cancer detection.

The central hypothesis is that changes in subcellular components like the cytoskeleton, membrane, cytoplasm, and extracellular matrix at carcinoma onset regulate dielectric properties (conductivity, permittivity), affecting bioelectric signals that aid cancer detection. This is based on preliminary data showing unique PBMC dielectric properties from WT and PyMT tumor-bearing mice, identifying bioelectric signals regulating human adenocarcinoma cells.

We hypothesize isolated PBMCs are altered in cancer compared to healthy bodies, evident in the MMTV-PyMT mouse model for analyzing human breast cancer mechanisms. Although specific PBMC change mechanisms are not investigated, we hypothesize these alterations can be detected using dielectrophoresis, a dielectric characterization technique utilizing non-uniform electric fields to manipulate and separate normal and cancer cells based on differing electrophysiological plasma membrane properties.

Our results present dielectric properties of murine PyMT and WT PBMCs exhibiting unique cellular behavior, with numerical simulations validating these results. These unique characteristics can discriminate between cancer and non-cancer cells. This novel, label-free, rapid (~2 min), low-cost cell sorting technology detects and separates early and late breast cancer stages, leading to preclinical development and future clinical trials. The long-term goal is a non-invasive tool to determine breast cancer at earliest stages without false positives/negatives of standard mammography screening

Whole-Body Loss of NUCB2/Nesfatin-1 Alters Enteric Hormone Expression

Naresh Ramesh — 2024-04-18

Objectives: Post-prandial insulin secretion is predominantly regulated by four intestinal hormones: glucagon-like peptide 1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP) [collectively called incretins], Cholecystokinin (CCK) and peptide YY (PYY). Nesfatin-1, an insulinotropic and anorexigenic secreted peptide encoded by nucleobindin-2 (NUCB2) was reported in the mouse hypothalamus. We have earlier reported that nesfatin-1 stimulates incretins and CCK expression and secretion while being a potent inhibitor of PYY expression, both in vitro and vivo. We further this observation in this abstract by testing the incretin, CCK, and PYY expression in a mouse model of global Nesfatin-1/NUCB2 knockout (NKO).

Methods: For the current study 6-8 weeks old male NKO mice and age-matched littermate C57BL/6J mice were used as controls (n=7). Total RNA extraction followed by Real-Time quantitative PCR (RT-qPCR) of small/large intestine samples was carried out. One-way ANOVA followed by Tukey’s multiple comparison test using GraphPad Prism software was used for data analysis. p<0.01 was considered statistically significant.

Results: We found that whole-body knockout of NUCB2/Nesfatin-1 downregulated large and small intestinal expression of incretins, CCK, and increased expression of PYY.

Conclusion: The current data extends our earlier observation in a mouse model lacking nesfatin-1. Lack of nesfatin-1 affects the healthy expression of incretins, CCK, and PYY in the intestine. These results highlight the importance of endogenous nesfatin-1 and its role in energy balance by modulating crucial enteroendocrine hormones known for their role in energy balance.

Source of Support: Dr. Unniappan's lab (University of Saskatchewan) and Canadian Institutes of Health Research (CIHR).

Combinatorial effects of resazomycins and azithromycin against Neisseria gonorrhoeae

Emily Young — 2024-04-18

Gonorrhea is a sexually transmitted disease caused by the bacterium Neisseria gonorrhoeae. The WHO estimates 87 million new gonococcal infections occur each year making it the second most common sexually transmitted bacterial infection. When left untreated, gonorrhea can cause pelvic inflammatory disease and infertility in women. Treatment is further complicated by the fact that N. gonorrhoeae has acquired resistance to all antibiotics approved for treatment including the gold standard ceftriaxone. One way to address the growing problem of antibiotic resistance is to administer drugs in combination in the hope the two are more effective together than alone. Resazomycins, a novel family of resazurin (Rz) based antibiotics, exhibit antimicrobial activity against N. gonorrhoeae and Francisella tularensis. Here, we used E-test assays to determine if there was a synergistic bactericidal effect between resazomycins and azithromycin, which is commonly used to treat gonorrhea. Multiple clinical isolates of N. gonorrhoeae were more sensitive to azithromycin in the presence of Rz. However, additional testing in a checkerboard assay revealed no synergistic effect between azithromycin and Rz or another resazomycin, resorufin pentyl ether (RPE). Upon further investigation, we determined the conflicting results between the E-test and checkerboard method were due to differences in bacterial load. When the E-test assays were repeated using a lower inoculum of N. gonorrhoeae, increased sensitivity to azithromycin was not observed in the presence of Rz. In the future, we plan to test resazomycins in combination with other antibiotics against N. gonorrhoeae, particularly extensively drug-resistant strains. (This research was made possible by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence and the NASA West Virginia Space Grant Consortium, Grant # 80NSSC20M0055)

The Lead and Arsenic Content in Everyday Drinkware

Jennie VanOsdol — 2024-04-18

This research explores the issue of lead and arsenic contamination in everyday drinkware which highlights the profound implications for consumer safety. The ubiquity of glassware in everyday life underscores the urgency of investigating the sources and levels that pose health risks associated with these toxic elements within drinkware products. Through a systematic examination and use of flame atomic absorption spectroscopy (FAAS), identifying the levels of lead and arsenic contamination has been achieved. This research rigorously quantifies the levels of contamination in commonly used drinkware, revealing possible correlations with concentration, condition, and price points. These correlations furthermore emphasize the need for regulatory measures and heightened consumer awareness. This comprehensive exploration serves as a vital contribution to safeguarding consumer safety in the context of drinkware usage, paving the way for informed decisions and targeted interventions in the pursuit of a healthier living environment. This research found that 20% of all tested samples leached an average of 0.84 ppm lead with a standard deviation of 0.32 ppm. 30% of all samples that tested positive for arsenic had an average of 4.10 ppm leached and a standard deviation of 0.88 ppm. No correlation was found between prices and concentration and a highly negative correlation was found between both metals. Low concentrations of arsenic had high concentrations of lead. Likewise, high concentrations of arsenic had low concentrations of lead. Damaged, or internally exposed ceramics tested higher for both lead and arsenic.

The Isolation and Characterization of Antimicrobial Compounds Extracted from Rhus typhina.

Kaitlyn Bailey — 2024-04-18

Rhus typhina, a plant native to eastern North America, has previously been discovered to exhibit antibacterial activity against various ESKAPE pathogens including Staphylococcus aureus and Acinetobacter baumannii. The purpose of this work is to separate and purify compounds from the crude extracts using column chromatography, identify the pure compounds using NMR, and examine the therapeutic potential of these compounds on A.baumannii and S.aureus. A.baumannii is a non-motile, gram-negative, pleomorphic opportunistic pathogen and S.aureus is a non-motile, gram-positive, coccus-shaped, opportunistic pathogen; Both bacteria are major causes of nosocomial infections due to their ability to resist multiple antibiotics. As multi-drug resistance becomes more prevalent, there is a growing demand for research on new therapeutic agents. Supporting previous unpublished research from our laboratory, the data here suggest that ethanolic extracts of R.typhina contain compounds that exhibit antibacterial capabilities against both S. aureus and A. baumannii. Ongoing studies are focusing on determining the structure of the antimicrobial chemicals and the mode of action of these compounds.

Seeding Your Future and the Impact

Raven Phares — 2024-04-18

The study of the impact of the Seeding Your Future Initiative on the students that attended is to show any correlations that may be present between those that attended and if they went on to go into a STEAM major or if they decided to do something entirely else. The goal of this research is to see how the Seeding Your Future Initiative is impacting the students that attend and to see what demographics are attending, such as location, age, schooling, and grade level. Data suggests that at least three students have gone on to go into STEAM in college. The data for those that have not finished high school shows that most of the students keep returning due to a love for STEAM. Overall, the study is to find any comparisons that have occurred in the last ten years of the Seeding Your Future Initiative and how that has impacted the students who have attended. The source of support for this study was the Shepherd Opportunities to Attract Research Students (SOARS) at Shepherd University.

Examining the Density Gradient Along the Rhizophora mangle Propagule

Marissa Mohn — 2024-04-18

Mangroves grow in tropical regions around the globe, but these dense forests have been declining due to agriculture, industry, and climate change. Mangrove propagules are not seeds but the seedlings themselves that grow on the parent tree until they reach maturity and fall. They are water dispersed, floating horizontally in seawater and vertically in brackish water. The purpose of this study was to further investigate whether there is a gradient change in density longitudinally along the Rhizophora mangle propagule that results in this floatation behavior. This study also explored whether there was a difference in density between the medulla and the cortex within the propagule. Propagules were cut into one-centimeter sections and placed in a pycnometer to calculate their density. The average densities of these categories were graphed and had a correlation coefficient of 0.6036 showing a strong positive correlation. The medulla, or inner ring, was removed from the cortex and placed in water to determine if there was a difference in buoyancies. In the plumule end of the propagule, both the medulla and cortex floated. Towards the radicle end of the propagule, the cortex would sink while the medulla would continue to float. The research raises the question if there is a difference between the cortex in the radicle end and the cortex in the plumule end but supports the idea that there is a change in density along the propagule longitudinally that plays a role in the buoyancy of the mangrove propagules.

Lipidomic analysis by LC-MS/MS of wildtype, mutant and VBNC strains of Francisella tularensis LVS.

Rachelle Patterson — 2024-04-18

Francisella tularensis is one of the many bacteria that can enter a viable but nonculturable state (VBNC). The VBNC state acts as an adaptive strategy that allows for long-term survival of bacteria in unfavorable environments. A defining characteristic of VBNC bacteria is a loss of culturability, making VBNC bacteria difficult to detect in the environment using standard plating assays. Failure to identify VBNC bacteria in the food industry or water distribution centers poses a serious risk to human health. Our preliminary data suggests that lipid metabolism plays a role in VBNC adaptation in F. tularensis. To investigate further, genetic deletion mutants have been constructed from two predicted lipid metabolism genes, FTL_1569 (phosphoglycolate phosphatase) and FTL_1570 (phospholipase D family protein). Lipid extracts were collected from wildtype, mutant and VBNC strains of F. tularensis to determine any lipid species associated with the VBNC state. A single-phase lipid extraction method was used to obtain lipids from F. tularensis cultures grown for 24 hours (culturable) and 336 hours (VBNC). Liquid chromatography with tandem mass spectrometry (LC-MS-MS) and the ThermoFisher Compound Discoverer software were utilized for lipid identification, and comparative analyses. Differences in the lipid profile of wildtype, mutant and VBNC strains of F. tularensis have been identified. (Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence)

Computational Insights into the Mechanism of Boyland-Sims Oxidation Reaction

Isaac Hall — 2024-04-18

The Boyland-Sims oxidation is the reaction between peroxydisulfate ions and arylamines to form the aryl amine ortho sulfate with minor para isomer. The long-standing mechanism involves a nucleophilic attack by the amine on peroxide oxygen to form arylhydroxylamine-O-sulfonate intermediate, which subsequently rearranges to the arylamine o-sulfate. Marjanović et al. have challenged this long-standing mechanism, proposing a nitrenium ion intermediate as the reactive species instead of the uncharged amine. The nitrenium ion proposal was based on relatively low-level calculations. We previously computed the energetics of reaction intermediates in both mechanisms for aniline, 2,4-dinitroaniline, and N,N-dimethylaniline using high-level of density functional theory (B3LYP/6-311++G**) calculations. We present here new computations using density functional theory (B3LYP/6-311++G**) to model both arylhydroxylamine-O-sulfonate and nitrenium ion pathways in a series of aromatic amines. The transition state calculations revealed two possible energetically feasible pathways for the rearrangement of arylhydroxylamine-O-sulfonate to arylamine o-sulfate. Additionally, we computationally modeled the feasibility of the reaction of aniline and tetrathionate to yield 2-aminothiophenol product via Boyland-Sims Oxidation reaction. The results indicate tetrathionate ion is less reactive with aniline than the peroxydisulfate ion.

Effect of Menstrual Cycle on Neuromuscular Efficiency and Muscular Strength in Runners

Hannah Kemp — 2024-04-18

PURPOSE: The aim of this study was to examine the influence of the menstrual cycle (MC) stages on neuromuscular efficiency and muscular strength in female runners. METHODS: Three runners (females) underwent testing using a BIODEX isokinetic dynamometer concurrently with a BioPac EMG, during which peak torque, peak torque/body weight, time to peak torque, mean amplitude, median amplitude, minimum amplitude, maximum amplitude, frequency, and the p-p value were collected throughout the test. During the test, the performance variables were assessed on the quadriceps (rectus femoris) muscle and collected at speeds of 60 degrees/second and 180 degrees/second. STATISTICAL ANALYSIS: A paired-sample t-test was used to determine the correlation between neuromuscular efficiency and peak torque of the quadriceps during extension in the two MC phases. A paired-sample t-test was also used to determine the difference in neuromuscular efficiency between the MC phases, as well as peak torque between the MC phases. RESULTS: Muscular strength testing did not reveal statistical significance between neuromuscular efficiency and peak torque of the quadriceps during MC phases. Furthermore, there was no statistical significance between the differences in peak torque of the MC phases. Maximum amplitude at 60 degrees/second on the left leg between the MC phases revealed statistical significance (p = 0.026). EMG wave frequency at 60 degrees/second on the right leg between the MC phases revealed statistical significance (p = 0.007). CONCLUSION: Results show a significant difference in maximum amplitude and wave frequency at 60 degrees/second during knee extension between the MC phases.

Active Hexose Correlated Compound Feeding to Stress Mice Induces Differential Cytokine Production during Chlamydia muridarum Genital Infection

Kristen Rolen — 2024-04-18

Oral supplementation of Active Hexose Correlated Compound (AHCC) mushroom extract is reported to enhance resistance to infection. However, AHCC feeding to stressed mice during Chlamydia muridarum. genital infection and host immune response remain unexplored. This study aimed to explore cytokine production profiles of immune cells of AHCC-fed stressed mice. Wildtype C57BL/6J mice that were stressed and infected with C. muridarum intravaginally. After seven days, splenic T cells and bone marrow-derived dendritic cells (DCs), and macrophages (MØs) were proliferated, and culture supernatants were collected to determine cytokine production by ELISA. Phenotyping of surface markers of immune cells using flow cytometry is underway. ELISA results show that AHCC increases the production of Th1 (IL-12 and IFN-g) and Th2 (IL-10, IL-13, IL.23) cytokines compared to PBS-fed stressed mice. Increased production of TNF-α in LPS-treated DCs and MØs of WT was observed. Overall, AHCC-feeding increases the production of protective and suppressive cytokines.

Evaluating the effects of metal ions on malate dehydrogenase enzyme activity in Watermelon.

Doris AGYEMANG — 2024-04-18

Malate Dehydrogenase (MDH) is an essential enzyme in the cellular respiration pathway known as the Tricarboxylic Acid Cycle. Using NAD⁺ or NADP⁺ as a cofactor, MDH catalyzes the reversible conversion of oxaloacetate to malate. It can be found in plants, bacteria, and animals. However, some metal ions, such as Cu²⁺, Zn²⁺, and Pb²⁺, can decrease MDH activity. In this study, the recombinant MDH protein (His-MDH) of watermelon was purified and quality tested by SDS-PAGE. The average protein concentrations of elution, cell lysate, and wash were 0.37 mg/mL, 2.23 mg/mL, and 0.24 mg/mL, respectively. According to SDS-PAGE, the size of the purified protein was approximately 37 kD, suggesting the purification of the correct protein.

The enzyme activity of the MDH protein will be tested in the presence of metal ions such as MgSO₄, NiSO₄, ZnSO₄, CuSO₄, and PdCl₂, which have been shown to inhibit dehydrogenase enzymes. Our results provide valuable insight on how different metal ions present in the environment can affect the cell respiration of organisms by affecting the malate dehydrogenase enzyme. Further studies on the roles of metal ions in the TCA cycle and related metabolism, as well as the mechanisms by which metal ions inhibit enzyme activity, will be beneficial in understanding ways to improve soil conditions for higher crop production and for the understanding of evolutionary processes.

Identifying and characterizing antibiotic producers found in and on the haustorium of Epifagus virginiana.

Aida E Jimenez Esquilin — 2024-04-18

Epifagus virginiana, a holoparsite of Fagus grandifolia, utilizes haustoria to parasitize its host. Little is known about the microbiome of this plant's parasitic organ. To shed light on these unexplored ecological interactions we collected eight individual E. virginiana specimens from Kanawha State Forest (Charleston, West Virginia) and used culture-based methods and microscopy to assess total counts and morphotypes, revealing a diverse microbial community on the haustorium surface, including 3 bacterial phyla and fungi. The internal haustorium environment was lower in counts and diversity, indicating the haustorium's internal microbiome is likely a subset of the soil microbiome. We explored the capabilities of these microorganisms to produce bioactive compounds. Fifty-eight morphospecies were investigated for cellulose degradation and antibiotic production. Forty-six isolates were challenged in a cross-streak assay against a Micrococcus roseus (Gram-positive) and Escherichia coli K12(Gram-negative) and 35% showed antagonism for at least one type. Seven surface isolates showed a range of antagonism against M. roseus from 0.85 cm ±0.21 cm to 2.25 cm ±0.07 cm, and 2 against E. coli from 0.75 cm ±0.35 cm to 100%. Two of the internal isolates showed a range of antagonism against M. roseus and E. coli from 1.25 cm ±0.07 cm to 2.35 cm ±0.07 cm and 0.4 cm ±0.00 cm to 2.05 cm ±0.21 cm, respectively. Currently, we are testing these isolates against ESKAPE pathogens and using 16S rRNA sequencing and Biolog ® to further characterize them and their potential as a source of novel antibiotics or bioactive compounds.

Thermotolerance of environmental fungi captured from indoor air.

Abigail Collins — 2024-04-18

While more than six million species make up the fungal kingdom, less than 300 cause disease in humans, in part because many fungal species are unable to grow at temperatures at or above human body temperature. However, there is concern that as global temperatures increase, fungal species that do not currently pose a risk to human health may adapt to growth at higher temperatures, opening the door for the emergence of new human fungal pathogens. This study assessed the growth of five fungal isolates captured from indoor air at temperatures up to 38°C. All five isolates grew well at 24-27°C, grew poorly or not at all at 36°C, and none grew at 38°C. We then investigated the growth of three fungal isolates captured from indoor air at 36 - 39°C when the growth temperature was gradually increased from ambient room temperature to 36 - 39°C, compared to growth after a sudden change of temperature from room temperature to 36 - 39°C. All three isolates still exhibit poor to no growth at 39°C. This research was supported by a NASA-WVSGC Undergraduate Research Fellowship (AEC) and a NASA-WVSGC Research Enhancement Award (LSR).

Determination of Concentration of Heavy Metals in Wood Ash

Matthew Scanlon — 2024-04-18

Heavy Metals such as lead, chromium, and arsenic are thought to be present in wood ashes. As many people use wood ash in gardens, we decided to test how easily these metals can be extracted from wood ash. Metals were extracted from 10-gram samples of ash by mixing with 100 ml of 2.0% trace metal nitric acid, gravity filtering, and final filtering with a 0.22 um. nylon syringe. Absorption data for the extracted samples was compared to a calibration curve of absorption versus concentration constructed from data obtained with a Shimadzu AA7000. Initial results indicate that the concentration of the metals are below the lower limit of detection for Cr, As, and Pb which are 0.2 ppm, 2.0 ppm, and 0.5 ppm respectively. However, the laboratory-fortified matrix sample for each metal failed, thus we will need to perform standard addition tests to certify these results.

This work was supported by NASA WV Space Grant Consortium training grant number 80NSSC20M0055.

Peptidoglycan modulation by Francisella tularensis

Hayden Hess — 2024-04-18

HAYDEN HESS, STUART CANTLAY, JOSEPH HORZEMPA, Department of Biomedical Sciences, West Liberty University, West Liberty, WV 26074 and MELANIE SAL, Department of Biology, West Virginia Wesleyan College, Buckhannon, WV 26201. Peptidoglycan modulation by Francisella tularensis.

Peptidoglycan, a crucial component of bacterial cell walls, plays a fundamental role in maintaining cell shape, integrity, and resistance to osmotic pressure. Mycoplasmas are the only bacteria that do not produce peptidoglycan cell walls. Other peptidoglycan-producing bacteria are capable of transiently existing without a peptidoglycan as an “L-form”, but only on solid media. Preliminary observations made by our laboratory suggested that Francisella tularensis bacteria do not produce detectible levels of peptidoglycan while growing in liquid media. However, increasing salt concentration induces the production of peptidoglycan in all bacteria. In this study we confirmed and extended these observations by incubating bacteria with fluorescent D-alanine (which integrates into the peptidoglycan during synthesis). Fluorescence microscopy revealed that very few F. tularensis bacteria produce detectable peptidoglycan in low salt media, while seemingly all individual bacteria produce this layer in high salt media. Interestingly, the genome of F. tularensis encodes two distinct beta-lactamases. Here we show that both the wild-type F. tularensis LVS and beta-lactamase null mutant were both resistant to high levels of beta-lactam antibiotics in low salt liquid media. However, only the wild type and complemented mutant strains were resistant to beta-lactam antibiotics in high salt media while the beta lactamase null mutant was susceptible. These data further support the conclusion that F. tularensis does not produce, nor need, peptidoglycan in low salt liquid media. Future studies will investigate whether peptidoglycan modulation is required to avoid detection by the innate immune system.

Computational Studies Using COMSOL Multiphysics on Dielectrophoretic Characterization and Separation of Mesenchymal Stem Cells Undergoing Tenogenic Differentiation

Raphael Oladokun — 2024-04-18

Stem cells have unique self-renewal and differentiation capacities, which are advantageous for regenerative medicine and tissue engineering applications. Stem cells can recreate cells needed to repair injured tissues and organs in the body, for example, by regenerating connective tissues, which provide support, protection, and a structural framework for various organs and other tissues. However, a significant limitation is the susceptibility of tendons to injury with long-term loss of function.

Mesenchymal Stem Cell (MSC) therapies are promising for healing tendon injuries and tears but controlling stem cell differentiation and generating homogeneous tenogenic stem cell populations remains challenging. Therefore, separating differentiating and non-differentiating MSCs to collect homogenous cells is critical for further tissue repair after transplantation. To substantiate our hypothesis that differentiating and non-differentiating stem cells have unique dielectric properties, we focused on characterizing-obtaining dielectric signatures for differentiating tenogenic and non-tenogenic MSCs on the third day of differentiation using dielectrophoresis (DEP) focused on crossover frequencies [1, 2]. The estimated values for membrane capacitance, conductivity, and permittivity after 3-day treatment of undifferentiated cells differentiating into tenogenic MSCs are 2.46±0.1 pF, 0.82±0.01 S/m, and 1.97±0.05, respectively, and cytoplasm conductivity 0.82±0.02 S/m. The treated stem cells undergo differentiation, but the untreated cells don’t. The results showed a significant difference between the 3-day treatment and no-treatment group.

The estimated properties from the dielectrophoretic characterization studies are crucial in designing a DEP-based enrichment microdevice to collect homogeneous differentiated stem cell populations for tendon repair. Using particle tracing, creeping flow (transport of diluted species model), and electric current physics in COMSOL Multiphysics simulation software, we designed a microdevice that achieves 100% separation of untreated and treated mesenchymal stem cells undergoing differentiation towards tendon.

Applying dielectrophoresis to the microdevice architecture demonstrated high selectivity and yield, processing one million treated and untreated stem cells in 6.4 and 27.8 hours, respectively.

The Effect of Physical Activity and ACE Scores on Psychological vs. Physiological Addiction

Brianna Hendershot — 2024-04-18

Brianna N. Hendershot, Chelsea L. Robertson, Ryan T. Koenig, Jason W. Metz

West Liberty University, West Liberty WV.

Abstract

Addiction is a growing problem in the United States. Rehabilitation facilities have exercise as a component of their program, yet these facilities are not always within reach of every addict. Understanding that addiction is a chronic disease that is treatable with integrated treatment methods, it’s imperative to find the root of addictive tendencies. PURPOSE: To determine if there is a triangular relationship between addiction, physical activity, and adverse childhood experience (ACE) scores, looking specifically at psychological and physiological addiction types. METHODS: Data was collected from 25 females and 6 males, recruited from West Liberty University via email. Subjects completed a self-reported online anonymous 4-section survey including basic demographics, The International Physical Activity Questionnaire-Short Form (IPAQ-SF), The Health-Resiliency-Stress Questionnaire Part D (HRSQ), and addictive tendencies through questions regarding the criteria for psychological and physiological addiction from the DSM. A moderation analysis was used to triangulate the relationship between vigorous exercise, ACE scores, and addiction scores. RESULTS: The moderation variable was vigorous exercise from the past 7 days and ACE scores. Psychological and physiological addiction were correlated at .477, p=0.835, and .590, p=0.097, respectively. Total addiction produced a correlation of .574, p=0.241. CONCLUSION: A moderation analysis revealed no significant data when looking at vigorous exercise, ACE scores, physiological addiction, psychological addiction, and total addiction, showing the need for future research in this field. Looking at specific addictive behaviors objectively may aid in finding the root of addictive tendencies.

Estimating the dosage and feasibility of intranasal administration of creatine.

Greg Edward Popovich — 2024-04-18

Creatine dysregulation is present in many psychiatric and neurocognitive conditions, supporting the notion that oral creatine supplementation could constitute a viable treatment for conditions such as dementia, depression, and concussion. We recently proposed an intranasal route of administration (ROA) of creatine as a means of efficiently dosing creatine to achieve absorption across the blood brain barrier while targeting preferential delivery to the brain. Indeed, a subsequent murine study supports that creatine can be absorbed via the nasal ROA. The objective of this project is to refine this ROA and estimate appropriate dosage for research trials.

The amount of creatine needed to directly supply the brain is but a small portion of the dose that is administered orally for whole-body saturation. A single dose of creatine is typically 5 g, which results in a calculated blood concentration of 80 mg/dL based upon average adult blood volume. Evidence suggests this is sufficient to raise brain creatine levels. Given the adult brain blood volume of 100‒130 mL, we estimate that a minimum of 80 mg of nasally-administered creatine would need to pass from the nasal cavity across the blood-brain barrier. Even if the nasal columnar epithelium or mucus were to pose a barrier, doubling or tripling the dose would still represent a relatively modest amount of nasally-delivered creatine (e.g., 160‒240 mg) in solution. We predict that the amount of nasally-administered creatine to theoretically raise brain blood concentration to a therapeutic level would be modest and practical.

Developing a Retrieval Augmented Generation (RAG) Chatbot App Using Adaptive Large Language Models (LLM) and LangChain Framework

Cara Burgan — 2024-04-18

CARA BURGAN, Dept of Computer Sciences, Mathematics, and Engineering, Shepherd University, Shepherdstown, WV, 25443, JOSIAH KOWALSKI, Dept of Computer Sciences, Mathematics, and Engineering, Shepherd University, Shepherdstown, WV, 25443, and WEIDONG LIAO (Faculty Advisor), Dept of Computer Sciences, Mathematics, and Engineering, Shepherd University, Shepherdstown, WV, 25443. Developing a Retrieval Augmented Generation (RAG) Chatbot App Using Adaptive Large Language Models (LLM) and LangChain Framework

RamChat is an AI chatbot designed to assist Shepherd University students in navigating the student handbook. Developed in Python, it utilizes both API-based and local Large Language Models (LLMs) for natural language processing (NLP), alongside a vector store system. Our aim is to create a high-quality chatbot app tailored for student use.

We began by researching existing chatbot platforms and created a vector store with embeddings from OpenAI's text-embedding-3-small model, trained on the Shepherd University handbook. Testing each LLM helped assess answer types and accuracy.

Development involved debugging and optimizing RamChat's code, including replacing OpenAI's davinci-002 model with gemma, a local LLM based on Google's Gemini model. Ollama framework aids in automatic LLM selection based on user prompts.

Our conference presentation will detail RamChat's development, methodology, challenges, and insights. RamChat represents an innovative application of AI to enhance the Shepherd University student experience.

The Utilization of Double-Immunofluorescence Microscopy to determine the role of FTL_1199 in erythrocyte invasion by Francisella tularensis.

Elio Delatore III — 2024-04-18

Francisella tularensis is an intracellular bacterium that is the causative agent of the zoonotic disease, tularemia. F. tularensis invades host erythrocytes during infection, a phenomenon that leads to increased colonization of ticks after a blood meal. To better understand the mechanism of erythrocyte invasion, an RNA-seq analysis was conducted to identify genes whose transcription was affected by the presence of erythrocytes. A putative transcriptional regulator, FTL_1199 was determined to be modulated in the presence of erythrocytes. We hypothesized that this gene may therefore play a role in erythrocyte invasion. Deletion of FTL_1199 resulted in a significant reduction in the invasion of rabbit erythrocytes, and complementation of FTL_1199 restored this strain’s ability to acquire the cytoplasmic space of erythrocytes. These findings were determined by a gentamicin protection assay to quantify erythrocyte invasion. To validate these findings, we conducted double-immunofluorescence microscopy. F. tularensis strains incubated with rabbit erythrocytes to allow for invasion. Cells were probed with anti-F. tularensis antibodies followed by red fluorescent secondary antibodies. Cells were washed and erythrocytes were permeabilized. Bacteria were probed a second time utilizing a green fluorescent secondary antibody. Therefore, intracellular bacteria were only labeled with a singular secondary antibody, while extracellular bacteria were tagged with two secondary antibodies. Intracellular bacterial enumeration showed that deletion of FTL_1199 resulted in a significant reduction of invasion (p = 0.0253), while FTL_1199 complementation to the deletion mutant restored invasion to that of wild type (p = 0.3258). These data demonstrate that FTL_1199 plays an important role in erythrocyte invasion.

(Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence, R15HL14735 from NHLBI, and funds from the NASA West Virginia Space Grant Consortium).

Effects of Physical Activity and Bibliotherapy on Anxiety in Middle and High Schoolers

MAKAYLA ROLINCE — 2024-04-18

Anxiety is a common problem within adolescents in the United States. There are therapists and group settings available to this population to ease anxiety but it may not be feasible for them. PURPOSE: To create a program to help middle and high school students handle anxiety. METHODS: There is one participant in this study who will go through a four week program including eight different bibliotherapies that were found on Youtube. The participant will complete exercises based solely on their wants. The child will pick what type and the duration of the exercise. Once the exercise is completed, they will read the bibliotherapy for that week. Once the exercise and readings are completed, they will answer questions pertaining to their anxiety. PRACTICAL IMPLICATION: Regardless of whether their anxiety increases, decreases or stays the same, the child is going to exercise more than before. The bibliotherapies and physical activity combination may provide some relief for the student’s anxiety while also getting them more physically active.

Spatial Distribution Analysis of Invasive Pheretimoid Earthworms in West Liberty University’s Campus Woods.

Jason Ake — 2024-04-18

The United States is currently amidst a wave of invasion by East Asian pheretimoid earthworms. Multiple genera, genus Amynthas in particular, are widespread invaders with powerful ecosystem engineering capabilities and an expanding range. Invasive populations can reach extreme densities and fluctuate heavily on weekly-to-yearly timeframes compared to other earthworms. Despite their impacts, pheretimoid genera are poorly understood, under-researched, and uncontrolled. A deeper understanding of population dynamics and their mechanisms may allow effective management practices to be developed. The research was conducted in the summer of 2023 in a forest stand at West Liberty University, a known invaded site. Spatial Analysis by Distance Indices (SADIE) was used to analyze weekly distributions of pheretimoids, showing aggregated distributions in 5 of 7 weeks of surveying, 2 of which were statistically significant (P value < 0.05). Linear regression showed no recorded environmental variables were statistically significant (P value < 0.05) in more than 1 out of 7 weeks and were inconsistent in their impact. Population density was reduced during this survey compared to previous years, likely weakening significance due to sample size. Distributions still showed aggregation, suggesting clustering behavior is the norm for pheretimoid worms even at low population sizes. As regression couldn’t adequately explain clustering patterns, more environmental variables and new data analysis techniques may be needed for future work to determine the underlying mechanisms for pheretimoids.

The role of Cxcl12b in habenular axon pathfinding in zebrafish

Sara Roberson — 2024-04-18

The habenula is a complex region of the forebrain with multiple populations of neurons making different neurotransmitters such as dopamine, serotonin, GABA, norepinephrine, and acetylcholine. The habenula is involved in behavioral response to pain, stress, sleep, anxiety and reward. The habenula provides the fundamental mechanism in both survival and decision making. Axons of the habenula project to the interpeduncular nucleus (IPN) of the midbrain. Chemokine signaling has been shown to help axons reach this target. Chemokine receptor Cxcr4b is made in the habenula, while ligand Cxcl12b is produced anterior and posterior to the developing habenula. Based on this expression pattern, we hypothesize that Cxcl12b is working as a repellant and therefore, without Cxcl12b the habenular axons will project incorrectly in the anterior/posterior direction. We have identified cxcl12b mutants carrying a transgenic line that labels the habenula and its axons with GFP. However, we have not identified any males carrying the cxcl12b mutation and the transgenic line. Due to this, we will be using the gene editing technology CRISPR to cause mutation in the cxcl12b gene injust one generation. This mutation will permit us to explore the role of Cxcl12b on habenular axon pathfinding.

Smart Parking Space Detection with Generative Artificial Intelligence and Large Language Models

Cameron Vu — 2024-04-18

CAMERON VU, Dept of Computer Science and Math & ENGR, Shepherd University, Shepherdstown, WV, 25443, and DARIA PANOVA, Dept of Computer Science and Math & ENGR, Shepherd University, Shepherdstown, WV, 25443, and JOSIAH KOWALSKI, Dept of Computer Science and Math & ENGR, Shepherd University, Shepherdstown, WV, 25443, and Dr. W. LIAO (Faculty Advisor), Dept of Computer Science and Math & ENGR, Shepherd University, Shepherdstown, WV, 25443, and Dr. O. Guzide (Faculty Advisor), Dept of Computer Science and Math & ENGR, Shepherd University, Shepherdstown, WV, 25443. Smart Parking Space Detection with Generative Artificial Intelligence and Large Language Models.

The increasing relevance of generative AI and large language models is reshaping various sectors of modern society. These advancements have spurred notable progress in fields such as healthcare, finance, and education. Yet, the application of AI extends beyond expert domains, offering simplified solutions to everyday tasks for the general populace.

This project harnesses the power of generative artificial intelligence and large language models to develop a practical application: smart parking space detection. By leveraging these technologies, individuals can effortlessly ascertain the availability of parking spots in monitored lots via camera or photographic monitoring, facilitated by a straightforward algorithm. The overarching objective is twofold: to engineer a user-friendly system utilizing generative AI principles and to demonstrate the potential for such technologies to enhance the daily experiences of ordinary individuals.

Tick Biodiversity and Lyme Disease:

M. Christine Scronce — 2024-04-18

Ixodid (Acari: Ixodidae) ticks are widespread, diverse, and known as carriers of several important diseases. Lyme disease is a tick-borne illness affecting humans, livestock, and domestic animals and is caused by the spirochaete Borrelia burgdorferi. It is the most common tick-borne illness reported in West Virginia with more than 100 cases per 100,000 WV residents. Ixodes scapularis ticks obtain B. burgdorferi from rodents and transmit Lyme disease to their next host during feeding. We analyzed overall tick biodiversity in north-central West Virginia from the years 2018 to 2023. In total, 1,684 individual ixodid (hard) ticks have been collected from four genera: Ixodes (58.5%), Dermacentor (31.6%), Haemaphysalis (8.7%), and Amblyomma (0.2%). Our results demonstrate an increase in tick biodiversity from 2018 to 2023. One striking change included the presence of Amblyomma americanum (Lonestar) ticks in areas where their presence had not been recorded previously. We examined all collected Ixodes ticks for the presence of Borrelia DNA which was detected through real-time quantitative PCR analysis. We tested 745 Ixodes scapularis and found a 2% infection rate. The potential rise of Lyme disease and changing tick biodiversity could pose threats towards community health, agricultural revenue, and veterinary resources. Future work should focus on additional tick-borne illnesses like Rickettsia sp. and Theileria sp. within central West Virginia tick species.

Exploring and Synthesis of bio-active natural product coupling reactions

Sierra Lilly — 2024-04-18

Utilizing natural products in drug discovery research are very comment practice and they lead to more than fifty percent of current medicines in the market. However, there are still needed in new therapeutic entities that can improve the efficiency of drugs against the enzymes. We are hoping this and future study lead to combat unmet needs among healthcare. Having said that, we would like to explore the coupling reactions to make bio-active natural products similar to capsaicin natural product and its chemical synthetic optimization before the its kinetic study. Currently, we are exploring two distinctive coupling reaction conditions to optimize the reaction condition and will present current status of our research project of coupling reaction. Furthermore, we would like to carry on enzymatic profile against resinaase HT enzyme to see how stable amide bond is.

Acclimation of key photosynthetic traits drives edible biomass increases in crop species grown in aquaponics

Mikhaela Ferguson — 2024-04-18

Sustainable agriculture practices may contribute to increasing food demand while alleviating issues with fertilizer over-use and water availability in traditional agriculture. Aquaponics systems provide plants with a constant supply of nutrient dense water from fish efflux. Root uptake filters the fish efflux and water is recycled back to fish tanks, reducing the water needed to produce both plants and fish. Plants acclimate their physiology and anatomy to changing environments, yet, the degree of acclimation in aquaponics is unknown. We tested if acclimation of leaf physiology and anatomy to constant water and nutrient supply in aquaponics affected edible yield. We measured acclimation of leaf gas exchange (A_sat, g_sw), photosynthetic capacity (J_max,Vc_max), stomatal density and chlorophyll production by comparing plants grown in aquaponics or soil for three species: Lactuca sativa (Romaine lettuce), Triticum aestivum (Winter Wheat), and Ocimum basilicum (Basil). We hypothesized that non-limiting nitrogen supply in aquaponics would drive increased production of chlorophyll, increasing electron transport rate (J_max) and carboxylation rate (Vc_max). Additionally, constant water supply in aquaponics would remove limitations driving stomatal closure, leading to higher photosynthesis rates. We found aquaponics plants produced more chlorophyll, driving increased photosynthetic capacity through higher J_max and Vc_max. Additionally, aquaponics plants increased photosynthesis rates (A_sat) via higher stomatal conductance (g_sw) of fewer stomata. Acclimation of key photosynthetic traits in aquaponics plants resulted in more edible biomass compared to soil grown plants, often with reduced investment in root production. Overall, aquaponics appears well suited to producing higher yields of certain crop species, while also reducing environmental impacts.

Evidence of Nesfatin-1 in Mouse Intestinal Enteroendocrine Cells

Naresh Ramesh — 2024-04-18

Objectives: Post-prandial insulin secretion is predominantly regulated by four intestinal hormones: glucagon-like peptide 1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP), Cholecystokinin (CCK) and peptide YY (PYY). Nesfatin-1, an insulinotropic and anorexigenic secreted peptide was reported in mouse hypothalamus. Data supporting nesfatin-1 expression in intestine is minimal. Is nesfatin-1 expressed in mouse intestinal cells? Does it affect intestinal hormones known to stimulate or inhibit insulin secretion? We present STC-1 cells as viable in vitro models to study nesfatin-1’s biological action in intestine.

Methods: Immunofluorescence histochemistry (IHC) of intestinal sections from male C57BL/6 mice were carried out. STC-1 cells (n=8 wells/treatment) were dose-dependently treated with nesfatin-1, followed by RT-qPCR and immunoassay (ELISA) respectively. One-Way ANOVA followed by Tukey’s multiple comparison test using GraphPad Prism software was used for data analysis. p<0.01 was considered statistically significant.

Results: IHC showed NUCB2/nesfatin-1 co-localizing CCK, PYY and GLP-1 in the intestinal mucosa of mice. Static incubation of STC-1 cells with nesfatin-1 upregulated both CCK, PYY and GLP-1 mRNA expression (1 and 10nM) and secretion at 1 hr post-incubation (0.1, 1 and 10nM). Conversely, nesfatin-1 downregulated PYY mRNA expression and secretion in STC-1 cells (all doses tested).

Conclusion: The current data shows cell-specific localization of nesfatin-1 in the intestinal mucosa and establishes STC-1 cells as in vitro models to study nesfatin-1. This study adds a novel function for intestinal nesfatin-1 in stimulating insulinotropic intestinal hormones (GLP-1, GIP, CCK) and inhibit PYY which is insulinostatic.

Role of LpnA in Francisella tularensis Susceptibility to Resazurin

Emma Beatty — 2024-04-18

Francisella tularensis is classified by the Centers for Disease Control and Prevention as a Category A bioterrorism agent due to its low infectious dose, ease of aerosolization, and high mortality rate. Given there is no licensed tularemia vaccine and the possibility of antibiotic-resistant F. tularensis strains, there is an urgent need to develop new treatments against this bacterium. Resazurin (Rz) has bactericidal activity against F. tularensis and other gram-negative bacteria including the human pathogens Neisseria gonorrhoeae and Helicobacter pylori. However, the mechanism of action for Rz has yet to be determined. To identify targets of Rz, we screened for Rz-resistant (Rzr) mutants of F. tularensis LVS. Whole genome sequencing was performed on 42 Rzr isolates and all contained mutations within the coding regions of FTL_0421 and FTL_1504. FTL_0421 encodes for LpnA, the major lipoprotein in F. tularensis. The FTL_0421 mutation in Rzr1 resulted in loss of LpnA expression which correlated with increased sensitivity to osmotic stress. To investigate the individual role of lpnA in Rz-resistance, a deletion mutant (ΔlpnA) was generated using standard molecular genetics techniques. An agar dilution assay was then performed to determine the Rz susceptibility of the ΔlpnA strain. A minimum inhibitory concentration (MIC) of 2.75μg/mL was achieved for the ΔlpnA and the LVS wild-type strain. This data suggests mutation of lpnA alone is not sufficient to confer resistance to resazurin. In the future, we will investigate whether lpnA in combination with other genes like FTL_1504 play a role in resazurin susceptibility in F. tularensis. (Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence & the NASA Space Grant Consortium)

Evaluation of antibacterial effects of stem extracts from Coscinium fenestratum.

Gabriella Morris — 2024-04-18

An increasing number of pathogens are becoming antimicrobial resistant, among them are bacteria Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Enterobacter species, Pseudomonas aeruginosa, known as ESKAPE pathogens. These ESKAPE pathogens have been cited as one of the leading causes of nosocomial infections around the world and have shown higher antimicrobial resistance posing a great clinical challenge and the need of new therapeutic agents. Various plant extracts have been used in traditional medicine since ancient times. Coscinium fenestratum, commonly called yellow vine is a vine native to South Asia and has been used to treat common cold, tetanus, fever, inflammation, wounds, and diabetes especially in countries such as Sri Lanka and India. In this study we extracted the secondary metabolites from dried C. fenestratum stem. The extract was then tested for antimicrobial properties using disc diffusion method. The bacteria assayed include Pseudomonas putida, Mycobacterium smegmatis, and Acinetobacter baylyi which are the safer relatives of P. aeruginosa, M. tuberculosis, and A. baumannii respectively. Methanolic extract of C. fenestratum at 100 µg did not show any antimicrobial activity against M. smegmatis. The effects of the extract will be tested further against P. putida and A. baylyi. If the tests show antimicrobial properties, we will determine the chemical composition of the methanolic extract revealing the chemical agent which is active on the bacteria. Our work will provide valuable information on potential use of C. fenestratum secondary metabolites in treating human pathogens contributing to the drug discovery pipeline.

The effects of nutrient-induced quiescence on neural stem cell proliferation in the Drosophila central brain

Ronaida Ammari — 2024-04-18

In the fruit fly, neural stem cells (neuroblasts) residing within a specialized brain niche divide throughout development to produce the adult CNS. Neuroblasts normally transition between a proliferative or quiescent state during development, and this balance is critical for proper tissue growth and neuron production. NBs enter quiescence at the end of embryogenesis when maternal nutrient stores are depleted and reactivate proliferation soon after freshly hatched larvae consume their first meal. This reactivation is regulated by a nutritional checkpoint that requires dietary amino acids. Neuroblasts in larvae maintained on a diet lacking amino acids halt proliferation and production of neurons for extended periods of time, and this state is reversible after reintroduction of amino acids to their diet. Presumably this developmental plasticity is an adaptation that allows mobile larvae to secure another nutrient source, but the extent to which such a delay affects the structure and function of the adult CNS has not been investigated. We used our previously established nutrient deprivation model to halt neuroblast proliferation in larvae for a prolonged period. Using confocal microscopy, we measured the effect of this treatment on both the number of mitotic neuroblasts and how many daughter cells they produced. Both measures of neuroblast proliferation were reduced in delayed larvae compared to controls, despite having access to dietary nutrients for the same amount of time. These results elucidate an uninvestigated aspect of plasticity in the developing CNS and lay the foundation for further functional study of adult brains.

A Discussion of Year 3 Results of the NSF S-STEM Scholarship Program at Shepherd University

Qing Wang — 2024-04-18

The Track II NSF S-STEM Program in the Computer Sciences, Mathematics, and Engineering Department at Shepherd University started in October 2021 and has thus far supported 22 scholars majoring in computer science, mathematics, data analytics, or engineering. The overall goal of the program is to increase the number and quality of STEM graduates in the STEM workforce by providing financial, academic, and social support, which enables qualified students to overcome certain barriers that prevent them from meeting their educational and career goals. In this poster, the most significant activities and outcomes of the program since its inception are presented. Survey results and outcomes from the last academic year, along with initiatives that increase, retain, and support students in CME fields, are also discussed. This program has been supported by the NSF S-STEM Grant (award No. DUE-2130267).

Enhancing STEM Enrollment, Retention, and Graduation

Johnna Smith — 2024-04-18

JOHNNA SMITH, Dept of Mathematics, Shepherd University, Shepherdstown, WV, 25443, RAINA BURTON, Dept of Mathematics, Shepherd University, Shepherdstown, WV, 25443, QING WANG and KAREN ADAMS, Dept of Computer Sciences, Mathematics, and Engineering, Shepherd University, Shepherdstown, WV, 25443. Enhancing STEM enrollment, retention, and graduation: insights and strategies for student success.

The objective of this study is to synthesize over 20 findings related to STEM success and its measurement to enhance student recruitment, retention, and graduation rates. Surveys were analyzed focusing on various student categories, including underrepresented minorities (URMs), high-achievers, low-income individuals, first-generation students, community college transfers, and gender diversity, among others. Our findings demonstrate that improving students’ self-efficacy and fostering a sense of belonging significantly contribute to their success and persistence in STEM. Consequently, programs such as undergraduate research opportunities, networking, seminars, summer bridge programs, field trips, and workshops are identified as crucial for enhancing students’ confidence and sense of belonging, thereby facilitating their success and perseverance in STEM fields. The project was supported by the Summer Undergraduate Research Experience Grant (SOARS) from West Virginia Higher Education Policy Commission Division of Science and Research and the NSF S-STEM Grant (award No. DUE-2130267).

Characterization of CD4+ t cell subsets during Chlamydia muridarum genital infection

Katrina Davis — 2024-04-18

Characterization of CD4+ T cell subsets in a stressed mouse model during Chlamydia muridarum genital infection. Katrina Davis, Tesfaye Belay. Department of Applied Sciences and Mathematics.

Chlamydia genital infection caused by Chlamydia trachomatis is highly prevalent in low socioeconomic status populations. Whether stress is a risk factor for the high incidence of chlamydia remains to be investigated. CD4+ T cells are the major immune cells that fight chlamydia genital infection; however, the dynamics of CD4+ T cells during stressful conditions and chlamydia infection are unclear. The profiles of CD4+ T cell signature cytokines: T helper 1 (IL-12, Interferon-gamma (IFN-γ), Thelper2 (IL-4, IL-5, IL-13), Thelper17(IL-17), and Treg (IL-10). Mice were stressed for 5 minutes daily for 21 days and infected with 1x 10⁵ IFU/ml of Chlamydia muridarum intravaginally. Non-stressed mice were infected as controls. Mice were sacrificed 7 days post-infection, the spleen and lymph nodes were harvested, and T cells were purified and proliferated for 72 h. Determination of the levels of cytokines in culture supernatants of T cells from stressed and non-stressed mice are underway for poster presentation.

Supported by the NIH grant # 1R15AI124156-01 awarded to BSU

Temperature growth profiles for five fungi isolated from a teaching laboratory in West Virginia.

Elizabeth Rea — 2024-04-18

Every year, fungal infections kill approximately 1.5 million people worldwide. About 90% of these invasive infections are caused by fungi from only four genera. High human body temperature serves as a growth barrier that prevents many fungal species from infecting human hosts. This study assessed the growth of five environmental fungal isolates at temperatures ranging from ambient room temperature to 39°C, above average human body temperature of 37°C. These five isolates were captured from the indoor air of a teaching laboratory at Shepherd University and identified to putative taxon through sequence of the nuclear ribosomal internal transcribed spacer (ITS) region in a previous study. All five fungal isolates grew well at ambient temperature, but did not grow well at 38°C. Specific patterns of growth between room temperature and 38°C were different for each isolate. This research was supported by a NASA-WVSGC Undergraduate Research Fellowship (ER) and a NASA-WVSGC Research Enhancement Award (LSR).

Fingerprint Correlation Between Twins

Cody Barnes — 2024-04-18

CODY BARNES, KRISTY HENSON, MARK FLOOD, Department of Natural Sciences, Fairmont State University, Fairmont, WV, 26201 Fingerprint correlation between identical twins

When teaching about the uniqueness of fingerprints it is commonly said that identical twin fingerprint characteristics are not identical however, limited research has been conducted to determine if there are any correlations between similar print characteristics. In the recent past unrelated individuals have been misidentified based on fingerprint similarities in forensics cases and this may be more likely among twins. This research case study’s objective is to determine if there are any correlations between fingerprint characteristics of identical twins and the probability of misidentification. Identical twin fingerprints were rolled and collected onto 10 print cards then blindly analyzed using Henry Classification, AFIS, and NCIC techniques. Preliminary results indicate a strong relationship between characteristics, but it is not significant (0.49; p = 0.35); fingerprint characteristics between hands were not significantly different (R: p = 0.75, L: p = 0.24). Results indicate there could be a possibility of misidentification, but more twin fingerprints should be analyzed.

DNA Extraction Methods

Katie Long — 2024-04-18

KATIE LONG and MARK FLOOD, Forensic Science Program, Fairmont State University, Fairmont, WV 26554. Evaluation of DNA Extraction Methods.

Three forms of DNA extraction types include solution-based, column-based, and magnetic bead-based methodologies, with the latter being the accepted methodology used in forensic laboratories. Magnetic bead-based DNA extraction uses the negative charge of DNA and the positive charge of the magnetic beads to isolate the DNA strands and produce a purified sample. However, solution and column-based extraction methods use the same techniques of washing to purify DNA and typically have comparable DNA yields. Cheek swabs were obtained from ten individuals and the three extraction methods were performed to determine the quantitation of double-stranded DNA and the quality of the gel electrophoresis profile. Preliminary results indicate that solution-based extraction produces a slightly higher yield of double-stranded DNA having a 3.92% difference between the average quantitation of 26.29 ng/µL from solution-based and 25.28 ng/µL from magnetic bead-based methods. However, it is expected that the magnetic bead-based extraction will produce higher quality gel electrophoresis results. Overall, the methods examined produced similar DNA extraction efficiencies. This research was made possible by NASA West Virginia Space Grant Consortium, Training Grant #80NSSC20M0055 and the STaR Sure Grant.

Exploring mathematic struggles among STEM students at Fairmont State University

Josh George — 2024-04-18

Mathematics serves as the cornerstone of every STEM major, and proficiency in this subject is paramount for achieving academic success, societal integration, and meaningful participation in the workplace. However, numerous students encounter challenges in mastering mathematical concepts. In West Virginia, students face an additional hurdle, poverty, with households in the state ranking among the lowest earners in the nation. This can have significant implications for the funding and resources available to the West Virginia School System. Recognizing the interplay between socioeconomic factors and academic performance is crucial for devising targeted interventions and providing necessary support. In pursuit of this understanding, during the 2023-2024 school year, we administered a survey in 50 STEM courses at Fairmont State University (n = 447) and collected data from the LEAD Center on tutoring. Preliminary results show that math made up 50% and chemistry 37% of requested tutoring subjects in STEM and 69.7% receiving tutoring were in their first two years. Survey demographics are the following: gender identity: 212 male, 212 female, and 4 non-binary; economic status: 82% were middle class, 13% working class, and 4% on assistance; 29.5% are first generation; 62% are within their first two years of study. The most common math completed in high school was algebra (24.8%), trigonometry (34%), and calculus (24.4%). There was a strong correlation between major and enjoying math (0.99) and having a good math teacher in high school (0.62). Through data analysis and working with students directly, our goal is to discover why students struggle with math and how to better prepare future generations.

The Pseudomonas aeruginosa 1244 pilin glycan increases susceptibility to human beta defensin 2 but enhances twitching motility.

Shania Davis — 2024-04-18

SHANIA DAVIS & JOSEPH HORZEMPA, Department of Biomedical Sciences, West Liberty
University, West Liberty, WV USA. The Pseudomonas aeruginosa 1244 pilin glycan increases
susceptibility to human beta defensin 2 but enhances twitching motility.

Pseudomonas aeruginosa is an ESKAPE (highly drug resistant) pathogen that is commonly
acquired during hospital stays. This bacterium produces type IV pili which are adhesins and
motor appendages that mediate a surface motility referred to as “twitching.” These pili are
polymers of protein subunits referred to as pilin. The pilin subunit of P. aeruginosa 1244 is
glycosylated. Previous studies have shown that the pilin glycan affects the efficiency of
twitching motility under certain conditions and the surface polarity of these fibers. Because
modulation of the bacterial surface charge has been shown to mediate defensin resistance in
other organisms, we tested whether pilin glycosylation affected sensitivity to human beta
defensin 2 (HBD-2). Interestingly, the isogenic mutant strain lacking the pilin glycan (1244G7)
showed increased resistance to HBD-2 compared to wild type bacteria and those completely
lacking a type IV pilus (1244.47). This increased resistance to HBD-2 could explain data from a
recent study in which various P. aeruginosa clinical strains were isolated that had pilin
glycosylation defects. We also confirmed and extended previous findings that indicated that the
1244 pilin glycan enhances twitching motility and we showed that this is especially true on
positively charged surfaces (poly L-lysine coated plastic). However, the positively charged
surface was also capable of enhancing twitching of strains producing non-glycosylated pili or pili
with an incomplete glycan (1244.2.1).

Wnt signaling in cranial bones as a proponent of TH-induced craniosynostosis.

Lara Fetty — 2024-04-18

Craniosynostosis (CS) is the premature fusion of the sutures within the skull during embryonic development. Recently, thyrotoxicosis in pregnant women has been shown to increase the risk of craniosynostosis in the developing fetus, but the specific mechanism relating the accelerated bone development in CS to increased thyroid hormone (TH) is unknown. Different cell signaling pathways, such as Wnt, Ihh, BMP, and IGF-1, have been identified in TH regulated bone development. However, the Wnt pathway has been chosen in this study for its extensive involvement in bone development. It is known that Wnt-signaling increases levels of β-catenin which then drives the transcription of Wnt/β-catenin target genes such as Twist1 and Runx2. When upregulated, these genes have been linked to increased osteoblast differentiation and bone formation. In this study, we aimed to determine the level of Wnt signaling activation, in cranial bones of differing embryonic origins (frontal and parietal bones) from an avian model of thyrotoxicosis. Collected bones were processed for assessing β-catenin protein expression by western blotting and Twist1 and Runx2 gene expression by qRT-PCR. Analysis is currently in progress. We predict that differences will be observed between the parietal and frontal bones as the parietal bones derive from the mesoderm whereas the frontal bones derive from the neural crest.

Evaluation of Probable Maximum Precipitation (PMP) for West Virginia to Predict Current and Future PMP

Grace Kerr — 2024-04-18

Probable maximum precipitation (PMP) is essential for dam design criteria. West Virginia Department of Environmental Protection Dam Safety utilizes the 6-hr, 10-mi²(26-km²) PMP established by Hydrometeorological Report 51 based on the analysis of extreme rainfall. These PMP are outdated because there have been unaccounted for extreme rainfall events that have occurred since 1978. In addition, a zone of uncertainty around the Appalachian Mountains exists, including West Virginia (WV). The objective of this research was to determine the PMP throughout WV by studying the area of uncertainty and consider rising dew points to predict future PMP. This study created an updated PMP map of WV, focusing on the maximization of the 1942 Smethport, PA storm. A watershed-specific analysis was conducted at all dams in the state with an approximate watershed area of 10-mi²(26-km²). Current PMP estimations and projections for 2100 were determined. Iso-lines were determined to develop the new updated PMP map for WV that displays current and future PMP estimations. Results suggest that the greatest values of PMP occur in the western region of the state; lowest values of PMP occur in the south-eastern part of the state. The Smethport, PA storm (1942) was considered the governing storm for the entire state; however, it may not apply to the eastern panhandle and Potomac highlands. These regions should be considered for additional analysis. The updated PMP map will be a resource for those involved in dam inspection, repair, design, and modification.

Design and Implementation of Large-Scale Hard Rock AMD Dewatering and REE Production

Ian Bird — 2024-04-18

To develop a domestic Rare Earth Elements (REE) supply chain the Hydraulic Preconcentrate (HPC) produced at the Horseshoe bend treatment plant (HSB) was evaluated as a potential source for feedstock. After the clarifiers were flipped to the desired pH set points, a total of 24 Geobags were repeatedly filled and allowed to passively dewater. The Geobags evaluated were comprised of a mix of Woven, Non-woven, and Capillary Channel Fiber (CCF). The bags were constructed with either a composite of woven and nonwoven or only woven geotextile. Half of the bags of both types also included “fins” made of CCF to evaluate their impact on dewatering. Column Filtration Tests (CFTs) were performed onsite to provide laboratory value for hydraulic conductivity. Throughout their dewatering, the geobags were sampled to determine their rate of dewatering. Influent samples from HSB were also tested to produce a Specific Gravity (G_s), via pycnometer testing and Grain Size Distribution (GSD), via sieve and hydrometer analysis.

The dewatering ability of the geobags was high regardless of bag material with a mean total solids percentage of approximately 40% across all samples after only one month of dewatering. The GSD performed on the HPC is gap graded with most particles having a diameter of approximately 0.5-0.05mm. Hydraulic conductivity values for the filter cakes during Column Filtration Testing ranged from 0.5 to 2.5 meters/day. This data will help to guide development of future large-scale HPC dewatering and the parametric design of an REE oxide production plant at the HSB site.

A molecular biology approach to identify Francisella tularensis proteins that interact with Band 3 of human erythrocytes.

Rori Schreiber — 2024-04-18

Francisella tularensis is a pathogenic gram-negative bacterium that causes the zoonotic disease Tularemia. In addition, F. tularensis is classified as a Class A Bioterrorism agent by the CDC due to the ease of aerosolization and the ability of this bacterium to cause fatal infection in low doses. Previous studies suggest that invasion of mammalian erythrocytes by F. tularensis helps to increase colonization of ticks, an arthropod vector of this pathogen. Our laboratory previously found that the erythrocyte membrane glycoprotein, Band 3 is required by F. tularensis for invasion of red blood cells. Therefore, we predict that bacterial proteins interact with Band 3 to facilitate invasion. To identify these proteins, we will express codon-optimized Band 3 linked to a Glutathione-S transferase tag in F. tularensis LVS. Subsequent pull-down assays may reveal F. tularensis protein binding partners to Band 3 responsible for facilitating erythrocyte invasion.

Genetic and Protein Expression Comparison of Tissues of Differing Embryonic Origin.

Tyler Hill — 2024-04-18

Non-syndromic Craniosynostosis (CS) has an unknown etiology. The overproduction of thyroid hormones (THs) during pregnancy known as maternal hyperthyroidism has been cited by the CDC as a potential cause of CS. Elucidation of the mechanism for TH induced CS may allow for the creation of a novel more suitable treatment option. There are several signaling pathways involved in the differentiation and development of the tissues of the skull such as, BMP, Ihh, and Wnt. Wnt signaling has been linked to the differentiation of both embryonic mesoderm as well as neural crest cells, from which the relevant tissues of the skull originate. The goal of this study is to quantify and compare genetic and protein expression of relevant Wnt signaling molecules and their target genes within the Sagittal and Coronal sutures. Beta-Catenin is a Wnt signaling molecule that acts as a transcription factor modulating the expression of genes such as Runx2 and Twist1. These genes were selected as they are known to be involved in cranial development. Sutures were collected from an avian model of induced thyrotoxicosis and analyzed using qRT-PCR and western blotting. Results of the qRT-PCR showed a significant downregulation in Runx2 and Twist1 expression in the coronal sutures with no change in Twist1 and a significant upregulation of Runx2 expression in the sagittal sutures. Western blot results are still pending. While further investigation into these pathways is necessary, our findings conclude that these key pathways react differently to modulation by thyroid hormones in tissues of differing embryonic origin.

Enhancing Academic Success among STEM Undergraduate Students.

Raina Burton — 2024-04-18

We synthesize 20+ findings regarding STEM success and the measure of such with the aim of increasing recruitment, retention, and graduation of high-achieving, low-income undergraduate students. Students are more likely to enter and graduate from a STEM field when they have high self-efficacy, a sense of belonging, and identify themselves as scientists. We discuss methods of fostering these attributes such as providing undergraduate research opportunities, field trips, and summer bridge programs as well as some in-class methods including sending effort-oriented messages and connecting content to real-life.

Role of Cxcl12a on habenular axon projection in Danio rerio

Sara Roberson — 2024-04-18

The habenula is a forebrain region that is responsible for many emotional responses. The habenular axons project posteriorly to the interpeduncular nucleus (IPN), a region in midbrain. Correct projection of these habenular axons depend on chemokine signaling. Components of this signaling pathway include chemokine ligand, Cxcl12a, and chemokine receptor, Cxcr4b. Cxcr4b is made in the habenula, while Cxcl12a is made along the path of the habenular axons. Loss of either Cxcl12a or Cxcr4b results in abnormal anterior growth of habenular axons. However, the exact function of Cxcl12a as a directional cue at the habenula is not known. To explore this, overexpression of cxcl12a is being performed. Overexpression of cxcl12a is induced by heat shock in larvae carrying a transgenic line that produces GFP in the habenula and its axons. Heat shock is performed at 33 hours post fertilization (hpf), several hours prior to initial axon outgrowth. However, there have been challenges confirming overexpression of cxcl12a at this time point. Once overexpression is confirmed we will examine the habenular axons at 5 days post fertilization (dpf). We hypothesize that the overexpression of cxcl12a in larval zebrafish will cause a disruption in the growth of the axons of the habenula, implying that Cxcl12a is an attractant in habenular axon guidance. If no habenular axon disruption is noted, this implies that 33 hpf is not the correct window for chemokine signaling at the habenula or that excess Cxcl12a is insufficient in disrupting habenular axon pathfinding.

Expression of Chemokine Ligands cxcl12a and cxcl12b in the Habenular Region in Danio rerio (zebrafish)

Sara Roberson — 2024-04-18

The habenula (Hb) is a bilateral cluster of neurons in the forebrain. The interpeduncular nucleus (IPN) is an important component of the midbrain. Together, they make up the Hb-IPN conduction system. This system plays an important role in sleep, reward-based decision making, and various other behaviors and processes. These important structures are found in all vertebrates, but the understanding of the development of this pathway is in the preliminary stages. Previous studies in zebrafish show that chemokine signaling plays an important role in Hb axonal outgrowth. Chemokine (c-x-c) motif receptor 4b (Cxcr4b), which is made in the dorsal habenula (dHb), helps direct axons toward the IPN. Cxcl12a and Cxcl12b are the two ligands in this signaling pathway, which offer guidance cues to dHb axons, though their functions are not yet entirely clear. cxcl12a mutants show dysfunctional dHb axonal projections, with many axons projecting anteriorly, in the opposite direction of the IPN. Mutants with ectopic cxcl12b expression show similar results. Dorsal views show cxcl12a and cxcl12b are both transcribed in regions posterior to the dHb; cxcl12b is also transcribed anterior to the dHb. However, the relative expression patterns of these ligands is unknown, and may give us a clearer understanding of their respective functions in directing dHb axons. To do this, we will employ in situ hybridization, which will allow us to simultaneously visualize where cxcl12a and cxcl12b are being transcribed along the path of the developing dHb axons.