bluuebunny/crossref_metadata_2025 huggingface dataset:
Net Acidity Titration and Water Quality Results for Acid Mine Drainage Water Samples from Morris Creek
Authors: Mosteller Jacob , Serafin Juliana
Published: May 2022

The Morris Creek Watershed Association maintains a passive limestone-bed treatment for acid mine drainage entering Morris Creek near Montgomery, WV.  Water was collected and analyzed from 14 sites in the Upper and Lower Main Stem treatment areas.  Analyses included net acidity and standard water quality tests: pH, turbidity, temperature, total dissolved solids, biochemical oxygen demand, phosphates, total nitrates, and dissolved oxygen (Water Quality with Vernier, 2007), as well as total flow measurements.     The net acidity or “hot acidity” titration is a measure of the highest acidity the water will reach due to dissolved components that undergo redox reactions to produce acidity. Water samples are acidified, boiled with hydrogen peroxide, and titrated with base according a standard method (Standard Methods for the Examination of Water and Wastewater, 2012), This study shows that the results for net acidity are correlated with the water quality tests for biochemical oxygen demand and dissolved solids, while net acidity is inversely correlated with the fecal coliform count, dissolved oxygen, and overall water quality index.  A general decrease in net acidity was found as the water progressed through the two treatment areas.   This study demonstrates the utility of the net acidity titration analysis of water samples and the water quality tests that are most relevant to the titration.  Related and future work includes the determination of dissolved iron, manganese and other metals by atomic absorption analysis.
Using maximum entropy modeling to predict suitable habitat locations for the Cutshin Crayfish (Cambarus taylori)
Authors: Tidmore Eric , Loughman Zachary
Published: May 2022

The Cutshin Crayfish (Cambarus taylori) is a recently described species which is endemic to the Middle Fork of the Kentucky River basin. The Middle Fork of the Kentucky River lies within the anthracite coal fields of Eastern Kentucky, and thus a conservation assessment of C. taylori is warranted. The goal of this study was to predict suitable habitat locations for C. taylori through use of maximum entropy modeling (MaxEnt). The Middle Fork of the Kentucky River’s crayfish fauna was surveyed during the summer of 2014. The occurrence data from this study coupled with landscape scale environmental variables—such as stream order and land use data—was used to create the model. The mean area under the receiver operating characteristic curve (AUC) value was 0.898, showing the model had high predictive accuracy. The model indicated high probability of occurrence in streams at the southern end of the basin, and a low probability of occurrence at the northern end of the basin. Stream order and stream sinuosity had the highest contribution to the model showing that C. taylori prefers 3rd and 4th order streams with low sinuosity. Although the AUC value showed high predictive accuracy for the model, predicted sites will need to be ground validated to confirm the model’s predictive accuracy.
Reassessment of the Crayfishes of the Upper Ohio River Basin in Pennsylvania.
Authors: Khan Tanya N. , Lieb David , McKinney Matthew , Loughman Zachary
Published: May 2022

Crayfishes are keystone species and ecosystem engineers in freshwater systems and are considered the third most imperiled taxa on the planet. In 1906, Arnold Ortmann published one of the most complete historic crayfish surveys ever conducted, which has allowed Pennsylvania to assess changes to crayfish fauna over the past century. More recently, waterways have been impacted by anthropogenic stressors such as urbanization and extractive industry. Our primary goal is to determine changes in species distributions and relative abundances of crayfishes in western Pennsylvania by visiting historical and new sites. Surveys in 2015-16 focused on collecting primary burrowers by trapping and burrow excavation. In 2017, efforts shifted to epigean species in the Upper Ohio River drainage, using standardized sampling of 10 seine hauls/site to assess 265 sites. Populations of primary burrowing crayfishes were found at 20% of 61 historical sites and 72% of 57 new collection sites. Of epigean crayfishes, 79% were found at 102 historical sites and 97% of 163 new sites. These collections represent approximately 60% of the survey, with remaining collections to occur in 2018. Important findings were three nonnative species: Faxonius rusticus (Rusty Crayfish), Procambarus clarkii (Red Swamp Crayfish), and Procambarus acutus (White River Crayfish). Preliminary data suggests that crayfish fauna in western Pennsylvania has remained moderately stable over the last century, though the presence of nonnative crayfishes indicates the need for continued monitoring. Efforts in Pennsylvania must focus on prevention and management of the spread of nonnative species to preserve the native crayfish populations that remain.
Aralia spinosa Extract Produces an Immunostimulotory Response Diminishing Infection by Francisella tularensis
Authors: Myers Jennifer , Collins Elliot , Seeber Roger , Horzempa Joseph
Published: May 2022

Francisella tularensis is a highly infectious bacterium that causes the disease, tularemia. Due to the highly infectious nature of F. tularensis, the Centers for Disease Control and Prevention classified this bacterium as a category A bioterrorism agent. To combat this threat, novel treatments are highly desired.  F. tularensis replicates in macrophages (phagocytic cells of the immune system derived from monocytes) during infection.  In a previous study, we sought to identify extracts from natural products capable of inhibiting intracellular growth of F. tularensis during infection.  From this study, we identified that Aralia spinosa (devil’s walking stick) extract (ethanol extraction of stem) inhibits replication within cells of the THP-1 monocyte line, but does not alter bacterial growth in culture. The Aralia spinosa extract was separated by chromatography and bio-assay guided fractionation was used to isolate the compounds responsible for the aforementioned activity.  Here, Galleria mellonella (waxworm) larvae as well as chicken embryos were infected with F. tularensis LVS (an attenuated type B strain) and were treated with fractions of the Aralia spinosa extract.  To determine whether fractions of Aralia spinosa extract were directly antibacterial, an antibiotic disk-diffusion assay was used. None of the fractions tested inhibited the growth of bacteria outside the context of infection, indicating that this extract does not contain traditional antibiotic compounds. Investigations are ongoing to identify and isolate the active compound of the Aralia spinosa Extract (Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence and funding from the WV Research Challenge Fund [HEPC.dsr.14.13]).
Characterizing the Production of Extracellular Vesicles over Time by LLC1, B16F0, and WISP1 KO B16F0 Cell Lines
Authors: Lanham Benjamin Thomas
Published: May 2022

An emerging mode of intercellular communication is through the release of extracellular vesicles, which can transfer proteins and RNA between cells. Characterizing the role that extracellular vesicles play is accomplished using vesicles isolated from in vitro cell culture. However extracellular vesicles isolated from cell-conditioned media can be contaminated with cellular debris produced during apoptosis. The purpose of this experiment was to determine the rate by which specific cell lines produce extracellular vesicles and how long before cell viability limits vesicle production. Here, we collected sampled from media conditioned by three different cancer cell lines: Lewis Lung Carcinoma (LLC1), melanoma (B16F0), and modified B16F0 (WISP1 KO B16F0). Cell-conditioned media samples were obtained every 12 hours for 72 hours. To characterize the size distribution of the collected vesicles, each sample was analyzed on a NanoSight NS300. Results indicate that extracellular vesicles are primarily produced during the first twenty-four hours without interferences of cell debris. The cells cultured in PBS produced the most extracellular vesicles particles with-in the size range of exosomes. Interestingly, the WISP1 KO B16F0 cell line produced few vesicles, many of which were larger than 200nm after 48-hours. A possible reason is the WISP1 KO B16F0 cell line lacks WISP1, a gene that is suspected of promoting epithelial-mesenchymal transition and suppressing apoptosis. In summary, cell lines should be used to produce vesicles for at most 36 hours, but optimally for 24 hours. As for WISP1, more research is needed identify the role that WISP1 plays on extracellular vesicle production.
Comparison of commercial touch DNA extraction systems utilizing the polymerase chain reaction technique
Authors: Nelson Hannah Dawn
Published: May 2022

Forensic scientists often generate genetic profiles from old blood stains, seminal stains, and hair, but an individual's genetic profile has been accomplished from surfaces touched by human hands. This “Touch DNA” potentially allows for the sensitive identification of DNA from crime scenes where fingerprints are found. The experiment was designed to determine the sensitivity of DNA isolation from fingerprints placed on glass. The two kits used in this experiment were the Invitrogen ChargeSwitch® Forensic DNA Purification Kit and the Promega DNA IQ™ Kit.  The kits provided protocols that claimed to facilitate isolation of genomic DNA from fingerprints.  After touching cleaned glassware with varying numbers of fingerprints, DNA isolation was performed using each of the above listed kits. A sterile swab was moistened with sterile water and used to extract deposited material from the glassware. Isolated DNA was amplified with the highly sensitive polymerase chain reaction (PCR) technique and separated by gel electrophoresis. A NanoDrop One was used to obtain DNA concentrations for all touch DNA samples and dilutions. The two kits successfully isolated sufficient DNA from cheek swabs and dilutions, and the Promega DNA IQ™ system was found to be more sensitive to DNA contamination.  However, the two commercial kits did not successfully extract a sufficient quantity of Touch DNA from a non-porous surface coated with fingerprints. Future experiments should be performed using Touch DNA samples on different surfaces to determine if DNA isolation efficiency can be improved. The project was supported by the NASA West Virginia Space Grant Consortium.
Antimicrobial Effects of Arctostaphylos uva ursi Extract against Acinetobacter baumannii. Dicey Stewart*, Elliot Collins*, James Healy*, Caleb Martin*, Francisco Leon#, Joseph Horzempa*. *Department of Natural Sciences and Mathematics, West Liberty University, West Liberty, WV. #Department of BioMolecular Sciences, University of Mississippi School of Pharmacy.
Authors: Stewart Dicey
Published: May 2022

Acinetobacter baumannii is a Gram-negative, opportunistic pathogen associated with a high mortality rate in those infected with this bacterium. There are few classes of antibiotics that can successfully eliminate A. baumannii infections; therefore, it is imperative to find novel therapeutics to reduce the mortality rate of those infected with these drug-resistant bacteria. Preliminary studies from our laboratory have shown that Arctostaphylos uva ursi (kinnikinnick) leaf extracts (AUU) inhibit the growth of A. baumannii. The AUU compounds were separated by chromatography, and the resulting fractions were tested for antimicrobial activity using an agar-based diffusion assay. This indicated that two fractions exhibited substantial antimicrobial activity. These results were verified using a Galleria mellonella larvae infection model. The G. mellonella model also indicated that active fractions were not toxic to these insects. The fraction with the most robust antimicrobial activity was further separated via chromatography and bio-assay guided fractionation (agar-based diffusion assays and G. mellonella infections) was used to identify fractions containing the active compound. Once the active compound has been isolated via a final round of chromatography using a sephadex column, this molecule will be elucidated via NMR and mass spectrometry.(Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence and through the WV Research Challenge Fund [HEPC.dsr.14.13])
Investigating the effects of summer 2016 floods on mosquito density in West Virginia and the potential relationship between flooding and West Nile Virus transmission in mosquito pools
Authors: Niyogusaba Tim , Watson Mark Brian , Dotseth Eric
Published: May 2022

A major flooding event occurred in West Virginia in June 2016. Multiple counties in central and southern WV (classified in this study as Group I) were hit by this disaster while several others were unaffected (Group II). Given these conditions, we decided to conduct an observational field study aimed at identifying the potential effects of flooding on mosquito abundance and West Nile Virus (WNV) transmission rate. The counties selected for this study have consistently monitored and reported mosquito surveillance data to the WV Department of Health over that past four years, which gave us the unique opportunity to evaluate this data before and after the flood in order to identify potential trends at the county and collection site level. We used descriptive statistics and a G-test of independence to decide on our null hypothesis. Our findings suggest that the flooding event is strongly associated with a significant variation in average annual mosquito abundances pre- and post-flooding for our affected counties, with Kanawha county showing a sharp increase in mosquito activity and WNV transmission rate (MIR) over this period. However, we were unable to identify a linear relationship in all Group I counties and Cabell County, which was unaffected (Group II) also presented significant variations pre- and post-flooding
Prevalence of orthopedic complaints in a rural population in West Virginia.
Authors: Workman Marissa Chase , Popovich Greg E
Published: May 2022

There are numerous medical conditions that impact rural populations. These conditions affect many body regions and systems to varying degrees. Some research suggests that West Virginians have especially high rates of certain orthopedic complaints.   The purpose of this study is to describe the most common conditions/affected body areas of a selected rural population. Over the course of one month, eighty-one patients (72.8% female, 27.2% male) from an out-patient orthopedic physical therapy clinic with a rural clientele were evaluated for their reported condition. During each patient’s evaluation, his or her diagnosis and affected body area and other identifying characteristics such as age, gender, pain level, and duration of the condition were recorded as routine medical documentation. Data were then compiled while omitting the patient’s name to provide anonymity.    For the total population, the following affected areas were reported: back/spine, shoulder, knee, neck, hip, ankle/foot, and general body pain. The collected data reveal spine conditions were the most common complaint for both men and women; 37% of total patients suffered from a condition that involved the spine or adjacent tissues, and 73.3% of those patients were female. Shoulder region conditions were the second most prevalent of total patients (23.5%) where 68.4% of those were female. Only 6.2% of the total patient population suffered from hip conditions, where 100% of those patients were female.    The patients within this rural population presented several different condition areas, most commonly spine, with notable differences between genders.
Nanopore Sensors And Signal Processing
Authors: Bowers Melvin L.
Published: May 2022

Melvin L Bowers III, Department of CME, Shepherd University, Shepherdstown, WV 25443, and Uday Shankar Shanthamallu, School of Engineering, Arizona State University, Tempe, AZ 85281, and Michael Goryll, School of Engineering, Arizona State University, Tempe, AZ 85281, and Dr.Andreas Spanias, School of Engineering, Arizona State University, Tempe, AZ 85281. Nanopore Sensors And Signal Processing With the help of Ion Channels as well as filtering and, signal processing DNA sequencing can become a more efficient process. To see wavelet filtering would be the best for ion channels we imported ion channel data into Qub. Then used MATLAB to show graphically what the signal looked like. We then utilized MATLAB’s toolbox of wavelets to see which wavelet could denoise the noisy ion channel signal, the best. We were able to objectively show the improvement of the signal utilizing specific wavelets to help with the white noise. The maximum observed increase in SNR was 5.42.
Steric-Induced Fluorescence via Methyl Substitution in Chalcone Dyes
Authors: DeRosa Christopher A. , Wilkins Garrett
Published: May 2022

Chemical research of luminescent dyes can have potential benefits in cell microscopy, optoelectronics and textiles. Luminescent chalcones have been studied extensively for their florescence properties and sensing capabilities, such as halochromism (pH), solvatochromism (polarity), and viscochromism (viscosity). Properties such as viscochromism can be utilized to sense the membrane viscosity in cells. To probe the viscochromic behavior of chalcones, bulky methyl-substituents were incorporated into the design of the chalcone to probe planarity, brightness and fluorescence turn-on based of environmental viscosity. This was achieved via Aldol condensation of aromatic ketones and para-(dimethylamino) benzaldehyde to incorporate a bulky methyl groups at an aromatic or vinyl positions. Synthesis of the derivatives was confirmed by 1H NMR, TLC and IR spectroscopy. Preliminary results indicate the dyes have yellow-green florescence in dichromomethane (Q.Y. ~12%), and aggregation induced emission (AIE) properties. This presentation will discuss the optical properties in solution, future directions and possible applications.
A Piperidine-Substituted Chalcone as a Molecular Rotor for Luminescent Viscosity Sensing
Authors: DeRosa Christopher A. , Inman Michael
Published: May 2022

Modern applications make use of the luminescent materials and dyes for optoelectronics, textiles and fluorescence microscopy. Recently, organic-based fluorophores have garnered interest in fluorescent microscopy as brighter and non-toxic alternatives to metal complexes used in cell biology. While some organic fluorophores have emerged as commonly-used imaging agents, such as fluorescein and rhodamine, these lack the ability sense the microenvironment properties with spatial and temporal resolution. As a solution, we proposed a simple chalcone-based molecular rotor to sense cellular viscosity. A bulky piperidine ring is incorporated to undergo twisted intramolecular charge transfer (TICT) producing a dark, non-emissive state in fluid solution (e.g. water), but a rigid, emissive structure in a viscous environment (e.g.  cellular membranes). Dyes were prepared via aldol condensation of the appropriate aldehyde and ketone pairs in low to moderate yields. Synthesis was confirmed by TLC, 1H NMR, and IR. The optical properties of the piperidine-substituted dye and a less bulky dimethylamino derivative were compared for molecular rotor properties and TICT. Preliminary results in dichloromethane reveal green fluorescence and modest quantum yields (~22%). Viscochromic properties in solution, future directions and possible applications will be presented.
The role of FTL_0893 during Erythrocyte Invasion by Francisella tularensis
Authors: Yauch Katelyn , Nachtwey Lisa , Horzempa Joseph
Published: May 2022

Francisella tularensis is a highly virulent Gram-negative bacterium, and the causative agent of the disease, tularemia.  During infection, this bacterium inhabits numerous cell types of its mammalian host including phagocytes of the immune system and erythrocytes – cells incapable of endocytosis.  We previously determined that several F. tularensis genes upregulated in the presence of erythrocytes were also important for invasion of these host cells.  Another such upregulated gene, FTL_0893, has not yet been tested for its involvement in erythrocyte invasion.   In this current study, we sought to generate both a disruption and deletion mutation of FTL_0893.  These mutant strains are currently being tested for their ability to invade erythrocytes.  We are also investigating the contribution of FTL_0893 in the virulence of F. tularensis using various in vitro and in vivo infection models.
GPU Computing: The Future of Computing
Authors: Anthony Parker
Published: May 2022

PARKER ANTHONY, Department of Computer Science, Mathematics, and Engineering, Shepherd University, Shepherdstown, WV, 25443.Computing requirements have grown over the last few years, bringing with it the benefits of complex computing. However, this had led to a bottleneck within the system. Some of the more complex applications, like swarm modelers, need to be able to execute fast and as efficient as possible. Current CPUs slow the process down. The solution is GPU Computing. GPU computing is the latest adaptation of current computer systems to allow for the complex portions of code to be offloaded to the GPU, which has a high rate of parallelism. This allows for the code to be handled in tandem, executing faster. This brings it to the speed of the CPU, which handles the simpler portions of code. The idea of GPU computing is being used in all areas of computing. Whether it be art, medicine, or even society.
Phylogenomics in Asarum section Hexastylis: serial innovations in floral form spawned from two geographically-widespread species.
Authors: Sinn Brandon T. , Daragan Christina , Freudenstein John V. , Barrett Craig F.
Published: May 2022

Asarum section Hexastylis (Aristolochiaceae), is a group of flowering plants endemic to the southeastern United States whose variable floral morphologies and overlapping distributions have long caused taxonomic confusion. Individual species distributions range from the entirety of the southeastern US to only a single gorge. The flowers of section Hexastylis species, to various degrees, appear to have converged on a basidiomycete-like morphology and are putatively pollinated by fungus gnats. Earlier attempts to resolve species boundaries and the evolution of floral traits in the group with commonly-used phylogenetic markers have largely been uninformative at the lowest taxonomic levels.   To uncover species-level relationships, we sequenced thousands of regions dispersed throughout the nuclear genome using Double-Digestion Restriction Enzyme-Associated DNA Sequencing. Data matrices were assembled using the Pyrad pipeline. Phylogenomic inference was conducted under both maximum likelihood and coalescent frameworks. Sequencing was conducted for 97 individuals, representing all taxa in section Hexastylis.   Phylogenomic inference recover two major diversification events within section Hexastylis. Phyletic grades of a geographically-widespread, paraphyletic progenitor species have repeatedly spawned geographically-restricted derivative species with novel floral shapes. Recovered phylogenetic relationships evidence parallel evolution of similar calyx tube shapes, suggesting the operation of geographically partitioned adaptive pressures, most likely associated with fungus gnat pollination.   This work was supported by the National Science Foundation (DEB-1406732) and a William Chambers Coker Fellowship from Highlands Biological Station.
The NSF S-STEM Scholarship Program at Shepherd: major activities and outcomes
Authors: Wang Qing , Wang Zhijun
Published: May 2022

The Scholarships in Science, Technology, Engineering, and Mathematics (S-STEM) Program in the Department of Computer Science, Mathematics, and Engineering (CME) at Shepherd University has supported 57 computer science, mathematics, and engineering majors since June 2013. The overall goal of the program is to increase the number of STEM graduates by providing financial, academic, and social supports, which enable qualified students to overcome certain barriers that prevent them from meeting their educational and career goals. During the 2017-208 academic year, 65 new applications were received and 14 new awards were made. In this poster, major activities and outcomes of supported students are presented. Survey and report results from the last academic year, along with initiatives that increase, retain, and support students in CME fields, are also discussed. The project has been supported by the NSF S-STEM grant DUE - 1259713.
Synthesis and characterization of a cobalt complex bearing a chiral nitrogen-based ligand.
Authors: Keith Seth , Hayashi Kasumi , Webb Joanna
Published: May 2022

Hydrogenation reactions are used to create essential molecules in today’s pharmaceutical, petrochemical, and agricultural industries. Single enantiomer products are desirable and often developed using asymmetric hydrogenation reactions, especially important for drug synthesis in the pharmaceutical industry. Asymmetric hydrogenation reactions currently utilize catalysts made from expensive and toxic metals, such as ruthenium, rhodium, and iridium, and are usually supported by phosphine ligands. Synthesis of an asymmetric catalyst based on a first row transition metal would be advantageous. In addition, chiral nitrogen-based ligands provide an opportunity to investigate a relatively underexplored area of hydrogenation chemistry. Thus, efforts toward a cobalt complex supported by a chiral nitrogen-based ligand, (R,R)-2,6-bis[1-(N-piperidinyl)ethyl]pyridine, are underway. Ligand synthesis, characterized by NMR spectroscopy, and initial reactions with cobalt will be discussed.
Bilateral internal thoracic arteries emerging distal to the anterior scalene muscles: Implications in coronary artery bypass grafting.
Authors: Bertus Brooke E. , ZDILLA MATTHEW J , LAMBERT H. WAYNE
Published: May 2022

The internal thoracic artery (ITA) usually arises from the first-part of the subclavian artery. However, various anatomic origins of this vessel have been reported. In this study, routine cadaveric dissection of a 98-year-old white female who died of congestive heart failure, revealed a bilateral variation of the internal thoracic artery. The ITA, on both the left and right sides, arose at the lateral boundary of the first rib from the terminal portion of the subclavian artery and the initial portion of the axillary artery.  Due to its patency, location, and long-term positive clinical outcomes, the internal thoracic artery is routinely used as a conduit for coronary artery bypass grafting surgery (CABGs).  With more than 400,000 CABGs being performed in the United States each year, knowledge of this vessel's anatomy is immensely important. Recognition of the presented variant may help to avoid confusion during surgery, prolonged procedure times, and downstream complications. The aim of this study is to describe an ITA anatomic variant and to elucidate its clinical significance.
Sexual Dimorphism of the Zygomatic Bone
Authors: Wharton Benjamin Louis
Published: May 2022

This study assessed the posterior contour of left- and right-sided zygomatic bones, hypothesizing that female and male crania would exhibit different contours. The study analyzed lateral cranial views of 252 female and 318 zygomatic bones. The distance from the zygomaticofrontal suture to the jugale was calculated in addition to the length of the posterior border of the frontal process of the zygomatic bone to determine the degree of deviation from a straight line. The contour length between males and females did not differ (t=1.577(593); p=0.12) nor did the degree of deviation from a straight line (t=1.808(593); p=0.07). However, there were significant differences with regard to side (t=5.589(614); p<0.0001 and t=3.742(614); p=0.0002). Discriminant function analysis revealed no significant shape difference between sides or sexes (T2=338.7; p=0.15 and T2=244.3; p=0.94, respectively). However, canonical variate analysis demonstrated statistically significant shape differences among sexes with regard to side. The results of this study reveal morphological variation in the zygomatic bone that may differentiate sex.
Growth of Community-Acquired MRSA strain USA300 Multi-drug Transporter Mutants is inhibited by Extracts of Tyrol Knapweed, Centaurea nigrescens.
Authors: Lasko Kyra P. , Lasko Kyra P. , Robinson Kathryn D. , Robinson Kathryn D.
Published: May 2022

The rates of infection by community-acquired multi-drug resistant Staphylococcus aureus (CA-MRSA) have risen dramatically over fifteen years in the United States.  CA-MRSA is responsible for rapidly progressive diseases, including necrotizing pneumonia, severe sepsis, and necrotizing fasciitis.  Consequently, novel antibacterial strategies are needed to combat the rising antibiotic resistance seen in CA-MRSA strains.  The USA300 CA-MRSA strain has been mutagenized using the Bursa aurealis transposon to create the Nebraska Transposon Mutant Library (NTML). We have screened the 1920 non-essential, defined transposon insertions in the NTML for strains that are either susceptible or resistant to methanol extracts of Centaurea nigrescens (CnME) leaves and flowers. 30 strains containing mutations affecting transporter proteins were identified as having either significant resistance or susceptibility to Centaurea extract.  Insertions in two different drug efflux transporter families have been identified. The EmrB/QacA drug resistance transporter subfamily is a multi-drug efflux pump responsible for the export of toxic molecules from bacteria and yeast. The ABC transporters are involved in drug import and export. These results confirm the effectiveness of the screen as a means for identifying drug-resistance genes affected by the C. nigrescens methanolic extract and suggest a role for drug efflux proteins in the resistance of S. aureus CA-MRSA to antibacterial plant metabolites. Current work is focused on determining the effect of CnME on the expression of selected drug-resistance transporters via qRT-PCR.
Screening of Natural Product Library for the Discovery of Novel Antibiotics that target Monocytic Leukemia Cells.
Authors: Hancher Kayla Michelle , Schmitt Deanna , Horzempa Joseph
Published: May 2022

Leukemia is the leading cause of death in children. The discovery of effective cancer therapies is imperative to combat this deadly disease. The National Center for Natural Products Research (NCNPR) has comprised a library of over 4,000 organic extracts derived from plants, fungi, marine, and algae species with potential anti-cancer properties. In partnership with NCNPR, we have developed a rapid and effective method for screening large quantities of these organic compounds for anti-cancer efficacy against THP-1 cells (a monocytic leukemia cell line). A microtiter-based AlamarBlue Assay was used to identify extracts that reduced the viability of the THP-1 cells.  For this assay, THP-1 cells were treated with an extract from the NCNPR library and were incubated at 37˚C, 5% CO2 for 24 hours. Subsequently, resazurin was incubated for an additional 24 hours.  Change to a red color was an indicator of viability.  Cells treated with extracts that produced little to no color change were identified as leads.  149 lead extracts have been further evaluated for toxicity to HeLa cells (a cervical cancer cell line) to determine whether these compounds may be more broadly applicable. Further testing among Human Embryonic Kidney (HEK) cells to narrow down those extracts cytotoxic to non-cancerous mammalian cells.
Bioinformatics Software for the Detection of Allele-specific Gene Expression
Authors: Miller Jason Rafe , Grzadzielewski Grayson , Sprecher Eric , Hornslien Karina Stensland , Grini Paul E
Published: May 2022

Maternal and paternal gene copies are not always expressed in a 50:50 ratio. Transcriptional imbalance, called allele-specific expression (ASE), has been demonstrated in model organisms and may have evolutionary importance. We are developing a bioinformatics pipeline to detect ASE in RNAseq reads from model plants. The pipeline requires RNAseq reads from homozygous parents, plus RNAseq reads from heterozygous offspring. The pipeline uses software for reference-based assembly, transcript alignment, and differential expression analysis. The pipeline outputs lists of genes whose expression ratios differ significantly from the 50:50 null model prediction. We tested the pipeline on genes commonly expressed in seed endosperm tissue using three strains of Arabidopsis thaliana. Previous studies of this system had relied on counting RNAseq reads that contain known, isolated SNPs. On our data, the two methods had correlations between 0.84 and 0.98 across four experiments. In ongoing work, while developing a version in Python, we aim to demonstrate that the new pipeline is actually more accurate than the isolated SNPs method.
Biologically Inspired Optimization Solutions for Homeostatic Maintenance
Authors: Liao Weidong , Ratledge Anthony
Published: May 2022

Anthony Ratledge, Weidong Liao (Faculty Advisor), Department of Computer Science, Mathematics and Engineering, School of Natural Sciences and Mathematics, Shepherd University, Shepherdstown, WV 25443. Biologically Inspired Optimization Solutions for Homeostatic Maintenance Optimization when applied in computer science refers to implementation of methods to solve problems more efficiently: to realize the most cost effective or highest achievable performance from a computing system. Traditional computation is done primarily through serial processing; one process after another is done, and so on. It is highly inefficient when dealing with complex factors, such as nonlinear computation, multi-objective functions, or nondeterministic variables. Researchers and scientists have found inspiration for such optimization problems in nature and developed an entire design philosophy called biologically inspired computing.   This poster discusses optimization with an emphasis on homeostatic maintenance, which involves adapting internal systems to regulate a consistent state in response to the external stimuli of the environment. In nature, this is an automatic, ongoing process accomplished by the peripheral nervous system through reflexes. This presentation will explore several different biologically inspired optimization software solutions in two rather broad categories: non-swarm intelligence-based algorithms and swarm intelligence-based algorithms. It will then delve into the construction of artificial neural networks as a proposed optimization solution, which features both hardware and software considerations, ultimately to argue in favor of one swarm intelligence implementation for homeostatic maintenance, with experimental support. Finally, this poster presentation will propose a swarm optimization implementation into an artificial neural network to achieve homeostatic maintenance.
Population Density of Crayfish between Two Macro habitats
Authors: Dotson Whitney , Walters Elizabeth , Walters James William
Published: May 2022

Crayfish are significantly important to our streams and ecosystems and more work remains to be done to sample or evaluate the consistency of sampling methods of crayfish in sections of the Pather State Park stream in West Virginia. Crayfish serve as food source, they help control nuisance water weeds, and break down decaying matter. In order to better understand crayfish population distribution and help conserve their habitat it is important to sample and map these habitats. The hypothesis of this experiment is that crayfish will prefer the macro habitat runs over riffles. The experiment will be conducted by choosing three runs and three riffles while measuring substrate size, stream velocity, and stream cover. We have identified two methods for classifying runs and riffles; the empirical method and the Froude number. We welcome the opportunity meet experts at WVAS to help us determine the most efficient method for our sampling. The study is limited to primary habitat of crayfish which consists of substrate larger than 16mm, because areas of sand, silt, and substrate smaller than 16mm would yield little to no crayfish. Crayfish will be caught using a Quadrat sampler and the crayfish will be counted to record densities. Crayfish morphology will be measured with pictures to further identify the species. Anticipated findings of the experiment would be to determine densities of cray fish in macro habitats, measure, and identify crayfish. Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence
. Evaluation of Chlamydia muridarum lung infection in beta-2 adrenergic receptor knock out in a male mouse model.
Authors: Phillips Kiersten , Belay Tesfaye
Published: May 2022

Chlamydia genital infection is the most frequent bacterial STD across the world caused by Chlamydia trachomatis.  Studies on Chlmaydia muridarum infection of male mice with through pulmonary routes have been reported.  Growing evidence shows chronic stress suppresses the immune that leads to infection. Stimulation of beta2-adrenergic receptor (β2-AR) is known to create an immune response through its known ligand, norepinephrine. We have shown that norepinephrine is released when mice are exposed to cold water stress, but its interaction with β2-AR during chlamydia lung infection is not known.  The purpose of this project was to determine the susceptibility of β2-AR knock out male mice to Chlamydial muridarum lung infection. We hypothesized that deficiency in 2-AR leads i) a decreased C. muridarum shedding in the lung of male mice; ii) increased gene expression and production of protective cytokines during lung infection. In our preliminary experiments, we seeded 70 mM-filtered lung lysate to monolayers of McCoy tissue culture following standard methods. After 32 h of infection and staining with fluorescein isothiocyanate-labeled, anti-chlamydial antibody. Inclusion forming units/ml were not determined because cell debris covered monolayers we were able to visualize and count inclusion bodies in monolayers. To improve results, 30 mM-filtered lung lysate will be seeded to monolayers of McCoy tissue culture following standard methods. Gene expression and production of protective cytokines during lung infection is underway. We expect that stressed and non-stressed β2-AR knockout male mice will have about the same IFUs/ml in the lungs because of the lack of β2-AR gene expression. This work was supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence and NIH Grant P20GM103434 awarded to Bluefield State College.
Deployment of E-Learning modules to foster entrepreneurial mindset among engineering students at WVU Tech
Authors: Hatipoglu Kenan , Panta Yogendra
Published: May 2022

“E-Learning Module- Applying Systems Thinking to Solve Complex Problems” were deployed at West Virginia University Institute of Technology (WVU Tech) to foster an entrepreneurial mindset among engineering students. This E-Learning Module were applied to an elective electrical engineering course “EE 493X Alternative Energy Resources” in fall of 2017 to develop a skill set to solve complex problems by applying system thinking. The implementation of Applying Systems Thinking to Solve Complex Problems help realize students to think out of the engineering classroom box and be mindful of the entrepreneurial environment.  The module is bundled up with a number of learning outcomes provided by the KEEN. The modules’ supplementary resources were guided by the instructor in the class and then made accessible to the students through blackboard.  Students reviewed the online contents of the module and then spent their time to complete module activities. Instructor debriefed the classwork activity to highlight on the purpose and the summary of that specific activity. These activities created a very interactive and an effective learning environment advancing student engagements, curiosities, and maximize the learning. Overall, students effectively utilized the module materials for defining system, systems architecture, and system engineering, identify alternative distributed power generation system architectures, decomposing alternative distributed power generation system hierarchy to at least four levels and defining/addressing future integrated power systems from various perspectives, including technical feasibility, value, risk, and societal impact. In the future, the module is planned to be deployed in other courses.
Localization of the key cell division determinant FtsZ in Francisella tularensis.
Authors: Cantlay Stuart , Horzempa Joseph
Published: May 2022

Francisella tularensis, the causative agent of tularemia, is a highly infectious gram-negative bacterium capable of replicating within macrophages, leukocytes, dendritic cells and epithelial cells.  In addition, F. tularensis also invades erythrocytes.    FtsZ, a homologue of eukaryotic tubulin, is an essential cell division protein in almost all bacteria. Polymerization of FtsZ into a contractile ring is a critical first step in division and the FtsZ ring acts in recruiting the cell wall synthesis machinery to the nascent septum.    We have generated a recombinant fusion of ftsZftto emgfp (Emerald Green Fluorescent Protein) which can be expressed, in trans, under its presumed native promotor or a strong Francisella promoter (pGRP).       As expected, FtsZft-Emgfp can be seen localizing as rings in F. tularensis.   Our aim is to utilize fluorescently tagged FtsZ to investigate replication of F. tularensis in a range of in vitro host cell invasion assays and as a marker for viability to study the morphological and physiological differentiation of F. tularensis as it transitions into a viable but non-culturable (VBNC) state.  (This research was made possible by NASA West Virginia Space Grant Consortium Training Grant #NNX15A101H and by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence).
Bioinformatics in the Sequencing Era
Authors: Miller Jason Rafe
Published: May 2022

Transcriptomics, or the analysis of gene expression, is enabled by DNA sequencing machines that decode the nucleotide sequences of RNA molecules from living cells. The large volumes of data require analysis by computer. For organisms with a known genome sequence, computers align each RNA sequence to the genome sequence to identify the gene responsible. The aggregate counts per gene are used to rank the genes by expression. The computational challenge becomes harder when any of the genes under study are similar to each other. The challenge is hardest when the study is focused solely on pairs of genes that are nearly identical: the maternal and paternal copies of the same gene. Nevertheless, there is evidence from plant science that flowering plant genes (maternal) compete with pollen genes (paternal) to reduce each other’s gene expression in their seeds. As computer scientists, we built a computational pipeline capable of detecting this phenomenon. We benefited from plant science collaborators who provided insight, validation, and three billion RNA sequences. We exploited bioinformatics technology including noise reduction algorithms, sequence alignment software, parallel computing environments, and statistical analysis software. We believe our pipeline is less sensitive to machine error, and more tolerant of biological variation, than processes employed to date. Interestingly, our project is focused on a question that was not even posed until the era of the Human Genome Project. Thus, as DNA sequencing technology becomes ubiquitous, and more questions become addressable through sequencing, students of computer science and biology can expect to have growing opportunities to collaborate for scientific discovery.
Aggregation Patterns of Sensory Sensillae in the Food Canal and Cibarium of Glossina morsitans morsitans (Diptera: Glossinidae)
Authors: Joy James E. , Rio Rita V.M. , Setser Emily A. , Humbert Austin A.
Published: May 2022

Mouthparts of hematophagous vectors serve as intermediaries, enabling the transfer of blood and pathogens from and to their hosts. We describe aggregation patterns of basiconic and setiform sensillae in the food canal and cibarium of the medically significant tsetse fly, Glossina morsitans morsitans Westwood. Mean body length of females was significantly greater than males (n = 20 for each sex). Mean lengths of food canal and cibarium were also significantly greater in females, even when correcting for the greater body lengths of females, but there was no significant difference in total number of sensillae in the food canal or cibarium between the sexes. A pair of basiconic (campaniform) sensillae was consistently present in the food canal of every individual, but numbers of setiform sensillae in the canal of both females and males varied from 53 to 74. No basiconic sensillae were observed in the cibarium proper of any individual, but four minute conical basicones embedded in a sclerotized plate at the posterior edge of the cibarial wall were observed. Number of setiform sensillae in the cibarium varied from 5 to 12 in females and 7 to 11 in males. Setiform and basiconic sensillae were significantly aggregated in the proximal-most (i.e., nearest the head) food canal region of both sexes, whereas setiforms were significantly aggregated in the mid regions of the cibarium. Sensilla aggregation patterns in tsetse flies are very different from those documented for tabanid flies indicating potential differences in monitoring blood flow between these two groups of hematophagous feeders.
Using Dermestid Beetles to Enhance Forensic Science Curriculum
Authors: Lowdermilk Kiersten Marie , Henson Kristy
Published: May 2022

KIERSTEN LOWDERMILK Dept of Natural Sciences, Forensic Science Program, Fairmont State University, Fairmont, WV 26554 and KRISTY HENSON Dept of Natural Sciences, Forensic Science Program, Fairmont State University, Fairmont, WV 26554 Using dermestid beetles to enhance a forensic science curriculum In order to conduct accurate, expedient skeletal analysis, scientists rely upon dermestid beetles (Dermestidae). These flesh-eating beetles speed up the decomposition processing leaving behind only skeletal remains and ligamental connections. Dermestid beetles are common in museums and scientific research. These beetles perfectly clean the bones minimizing biohazard waste and damage caused by flensing or carrion-eating animals. For this project, I assisted in the colony creation, maintenance, and the incorporation of a dermestid beetle colony in the Fairmont State Forensic Science Program. Courses that regularly utilize dermestid beetle work include forensic anthropology, forensic biology, and forensic taphonomy. Beetles also aid students and faculty in conducting taphonomic research. To prepare specimens for use in forensic science courses I thaw, flense, and place the specimen in the beetle tank. When bones are finished removed from the specimen from the tank and degreased in a warm water and ammonia bath. After degreasing, I prepare the skeletons for the needs of the specific forensic science course. Skeletal prep has included inflicting trauma, whitening and articulating skeletons for morphological comparisons, cleaning up taphonomic projects, and prepping skeletal material for various undergraduate research projects. Dermestid beetles have advanced laboratory and individual research activities in the forensic science program.