Undergraduate Research Posters 2017

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  Improving the Development of the I-Chart For Use in Biopharmaceutical Manufacturing Operation

  Kimberly Schveder

  The Shewhart control chart is a statistical tool used by pharmaceutical companies, as well as chemical and other batch manufacturers, to help detect errors in the manufacturing process and ensure control of product quality. One particular type of control chart is the I-chart. The average run length (ARL) statistic of the I-chart can easily be determined when output from the manufacturing process is normally distributed with known population parameters. This paper investigates the impact on the ARL statistic when the I-chart is based on mean and standard deviation estimates obtained from small sample sizes of less than 50 batches. The methodology of Quesenberry (1993) is employed to ascertain the impact of small sample estimation on I-chart performance and provide recommendations for how I-charts should be constructed to account for the uncertainty of using a small number of batches to construct them.

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  P1: Assessment of Metabolism-Induced Hepatotoxicity on a 384-Pillar Plate

  Yana Sickar, Soo-Yeon Kang, Pranav Joshi, and Kyeong-Nam Yu

  Microarray bioprinting technology has been explored to create miniaturized 3D cell cultures on a 384-pillar plate, which were combined with drug metabolizing enzymes (DMEs) and test compounds in a 384-well plate for metabolism-induced toxicity assays. Our goal in this study was to demonstrate rapid assessment of metabolism- induced toxicity on the 384-pillar plate and obtain reliable and highly predictive information on compound's hepatotoxicity in vivo. Briefly, human cells including Hep3B human hepatoma cell line as well as human embryonic kidney 293 (HEK 293) cell were encapsulated in alginate-Matrigel on the 384-pillar plate. Test compounds and six different DMEs including cytochromes P450 (CYP450) and UDP-glucuronosyltransferase (UGT) were dispensed in the 384-well plate. By sandwiching the 384-pillar plate onto the 384-well plate, human cells were exposed to the compounds and their metabolites generated by DMEs. The cells were stained with luminescent and fluorescent dyes and IC50 values were calculated using the luminescence and fluorescence obtained. In summary, our approach allowed us to assess mechanisms of metabolism-induced toxicity in high throughput. Thus, the 384- pillar plate could be used as a high-throughput, early stage, microscale alternative to conventional in vitro multi-well plate platforms and provide a rapid and inexpensive assessment of metabolism-induced toxicity at early phases of drug development.

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  P2: Construction of Weather Station for Measuring Wind using a Hovering Drone Network

  Adam Stead, Mark Travis, and Rishi Maheshwari

  Our research aims to build and test the effectiveness of the Sparkfun® Weather Station in its ability to record meaningful data while streaming to a remote device in real-time. This is motivated by our plans to utilize a hovering-drone system to collect meteorological data, such as wind speed, wind direction, humidity, pressure, and temperature. This data is essential to evaluate the wind profiles of a given area and assess available wind resource. The weather station was first calibrated by comparing measurements of the cup anemometer against a standard pitot tube in the lab. We found that the measured wind speed from the cup anemometer needs to be corrected with a factor of 1.4 as wind speed is below 4.5 m/s, but no correction is needed beyond. Overall, the easily-upgradable Sparkfun® Weather Station has been found to be an effective and inexpensive way to collect meteorological data wirelessly. The next step of this research is to incorporate the weather station into the hovering drone to enable measuring wind using a hovering drone network.

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  P1: How High Does the Lower Atmosphere Go?

  Vladimir Sworski and Justin Flaherty

  The Atmospheric Boundary Layer (ABL), consisting of the bottom few kilometers of the troposphere, is a region with strong mixing of moisture and winds. This region's activity has a large impact on weather and climate models. In this study, we use a high resolution computer model: Large Eddy Simulation (LES). Statistics produced require a strong understanding of the height of the ABL. The purpose of this study was to create a method for determining this height accurately and consistently, as previous models demonstrated significant error.

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  P2: Developing Methodologies for Wet-Sample Electron Microscopy Imaging

  Daniel Terrano and Petru Fodor

  Scanning Electron Microscopy (SEM) is widely used to analyze the size, shape, and composition of material systems. However, using this tool for analyzing systems such as particles suspended in solution requires drastic sample alterations, such as precipitation and fixation. Besides altering their environment, this exposes the particles to the harsh conditions within an electron microscope, such as high vacuum and electron beam exposure. To this end, the first goal of this study was to develop methodologies for imaging wet samples using electron microscopy. This is realized by creating a sandwich structure containing the solution of interest between a partially electron transparent window and the aluminum stub. The ability of the developed imaging cells to provide good imaging conditions is demonstrated with a variety of samples including polystyrene spheres, polymeric microgels, and spindleshaped nanoparticles. As some of the systems investigated are temperature sensitive, the second goal of the project was to develop a temperature controlled stage that can be integrated with the SEM. In the future, this heating stage will be used alongside the wet samples to image microgels above and below their critical solution temperature.

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  P1: Synthesis Optimization and Characterization of Polymeric Microgels

  Samantha Tietjen and Samantha Hudson

  Microgels are spherical particles suspended in solution, comprised of crosslinked polymer chains. Due to the amphiphilic property of the parent polymer, microgels display a temperature dependent de-swelling property, and therefore have the potential to be used for drug delivery. In this case, microgels were synthesized using hydroxypropyl cellulose (HPC) polymer and divinyl sulfone (DVS) cross-linker, as well as dodecyltrimethylammonium bromide (DTAB) surfactant to decrease particle size and promote microgel monodispersity. Synthesized particles were then characterized using dynamic light scattering (DLS) for both temperature and angle dependence to determine hydrodynamic radius, Rh, at a range of temperatures showing a transition from the swollen to de-swollen states. Previous studies suggest that increasing the concentrations of either the chemical cross-linker or the surfactant reduce Rh. Primary experiments focused on the variation of DVS and DTAB concentrations. Increasing the DVS:HPC ratio from 1 to 30 results in microgels that decrease in swollen size from 190 to 150nm and deswollen size from 95 to 65nm. However, at higher DVS:HPC ratio synthesized particles grow rather than shrink with increasing temperature. Surprisingly, increasing the surfactant concentration resulted in an increase in Rh; this might be related to DTAB effect on polymer transition temperature. Additionally, DLS experiements revealed a dependence of Rh on microgel concentration in samples. Rh at infinite dilution was extrapolated from the concentration dependence. Continued work with the synthesis procedure also revealed the importance of a meticulous synthesis procedure; most notably in regards to polymer stock preparation, pH and temperature control, and consistent stirring.

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  P2: Is it in the eyes? A pupillometry study of stress reactivity and Borderline Personality Disorder

  Zachary Tokar

  Borderline Personality Disorder (BPD) is characterized by unstable mood states, chaotic interpersonal relationships, and behavioral dysregulation in the form of selfinjurious acts that results in notable functional impairment. Emotion dysregulation, marked by strong shifts in emotional states away from baseline levels across subjective and physiological substrates, is believed to reflect one mechanism in the relationship between BPD and functional impairment. However, it remains unclear whether emotion dysregulation represents a general tendency to experience both positive and negative emotions keenly, or to specifically be sensitized to negative mood states. The present study examined the relationship between BPD symptoms and emotion dysregulation across neutral, negative, and positive valenced emotional states in a sample of twenty-two community dwelling adults with histories of psychiatric disorders. Emotion dysregulation was measured via subjective affect ratings and pupillary responses that index sympathetic nervous system reactivity when participants recalled neutral, stressful, and pleasant events that occurred during the prior 3 months. Results and clinical implications are discussed.

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  P2: Light Scattering Study of the Size and Shape of Mixed Elastin-Like Polypeptide Micelles

  Ilona Tsuper, Daniel Terrano, and Bryce Noe

  Elastin-Like Polypeptides (ELP) can be used to form thermoreversible vehicles for drug delivery systems. The ELP nanoparticles are composed of three-armed star polypeptides. Each of the three arms extending from the negatively charged foldon domain include 20 repeats of the (GVGVP) amino acid sequence. In addition, linear constructs composed of 40 repeats of the same (GVGVP) sequence are introduced into the system. The mixed ELP polymer system is soluble at room temperature and becomes insoluble at the transition temperature (~ 50°C) forming micelles with the foldons on the exterior and linear constructs at the core. Above the transition, the size and shape of the mixed micelles are dependent on the pH of the solution, concentration of the PBS solvent, and the ratio of the linear to foldon concentration. The technique of Depolarized Dynamic Light Scattering (DDLS) was employed to study the structure and dynamics of the mixed micelles at 62oC and maintained at an approximate pH level of 7.3 - 7.5. The ELP foldon micelles have a radius of 10 nm; the introduction of the linear concentration leads to a growth of mixed micelles at a linear rate, when the PBS and foldon concentrations are fixed. A model explaining this linear growth was developed utilizing the molar volumes of the mixed system. Static Light Scattering results seemingly support this model. However, the apparent VH signal found can indicate elongation in the geometry of the particles or anisotropic properties of the core of the mixed micelle.

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  Spurious grain formation due to convection at cross-section-changes during directional solidification

  Noah Weber and Masoud Ghods

  Turbine blades are a critical component in high powered gas turbine engines. These components are directionally solidified to have a single grain orientation, which allows them to operate under high temperature and stress conditions. Spurious grain formation is a major concern when forming these turbine blades. The purpose of this study was to study the effect convection has on forming these defects within turbine blades. Two alloys, Pb-5.8%Sb (solutally unstable) and Al-19%Cu (solutally stable) were directionally solidified upward in a positive thermal gradient (thermally stable) in a graphite crucible having abrupt cross-sectional area change from 3.2 mm diameter to 9 mm diameter. In the Lead alloy after the cross-section-expansion there is no observable new grain formation. However, in the Aluminum alloy there is extensive new grain formation after the expansion.

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  Expression Validation of miR-149-5p targets in prostate cancer

  Grant Wethington and Savita Singh

  Prostate cancer (PCa) is one of the leading causes of mortality in men. Current therapeutics for PCa are androgen depletion by castration or/and anti-androgen based treatments. Androgens are necessary for Androgen receptor (AR) to function as a transcription factor, AR then regulates the expression of genes which promote cancer cell proliferation. However, despite the therapeutic interventions recurred AR signaling, which is facilitated by the acquisition of mutations in AR and its amplification, cholesterol biosynthesis and alterations in the steroidogenesis continue promoting PCa carcinogenesis. Our research is focused on regulatory small RNA molecules know as microRNAs (miRNA) in PCa. Numerous miRNAs fine-tune the expression of multiple genes involved in posttranslational modification, cell proliferation, organogenesis, energy balancing and developmental regulation thus affecting cell proliferation, differentiation, and cell death. miRNA “replacement therapy” is highly promising as it can target the expression of carcinogenesis promoting AR and androgen signaling, the holy grail of PCa therapeutics. In this research project, we have tested the efficiency of miR-149-5p in controlling AR expression using a sensor luciferase reporter assay system in PCa cell culture model. We found a significant downregulation of AR reporter gene in PCa cells indicating the efficacy of miR-149-5p in controlling the growth of AR based tumors. Our result indicates miRNA mediated regulation of genes involved in PCa and therapeutic potential.

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  An Evaluation of Unionidae diversity in the Rocky River and how Population density has Changed Over Time in comparison to Observations made over 15 Years ago

  Jamil Wilson

  This study was undertaken to determine the changes in freshwater bivalve population since the survey conducted in 2001 in the northern region of the Rocky River, Ohio, USA. During our study, we discovered 69 live specimens and 58 shells most of which were found in two distinct locations near the top and bottom of the area surveyed. Our finds consisted of eight species in comparison to the nine that were seen in the same area of the 2001 study. Our results showed that Leptodea fragilis populations have decreased in the main stream of the Rocky River since the observation 2001, others species populations such as Lasmigona costata have slightly increased. However, the overall population of unionid mussels in this area have decreased and seeing as mostly older specimens were found, it appears as if they have not been reproducing despite the improvement of the water quality that has been taking place over the past 15 to 16 years.

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  P2: Dispersion Stability of DNA-Wrapped Carbon Nanotubes in Biological Media

  Fjorela Xhyliu

  Single-wall carbon nanotubes (SWCNTs) are among the most widely studied artificial nanomaterials. Their dispersion stability in biological media is a prerequisite for applications development in biomedical imaging and sensing. This project investigates the dispersion stability of DNA-wrapped SWCNTs in RPMI cell culture media with and without fetal bovine serum (FBS). The synthetic SWCNT mixtures were purified into semiconducting enriched fractions and various single species including (11,1), (9,4), and (7,3) using recognition DNA sequences by a polymer aqueous two-phase (ATP) separation method. The near-infrared (NIR) fluorescence, vis-NIR absorption, and pH of DNA-SWCNT dispersions in different biological media were monitored over time. According to optical properties of SWCNTs, we found that while DNA-SWCNTs were mostly stable over a period of 4 weeks in water, aggregations of SWCNTs occurred in media after 3 days, and FBS containing media after 2 hours. Interestingly, the kinetic measurements of single species in media with FBS showed a monotonic decrease in NIR photoluminescence (PL) intensity for (11,1), (7,3), and (6,4), and relatively stable PL intensity for (6,5) enantiomers within experimental time period. However, increase in PL intensity was observed for (9,4), (7,6), and (7,5), indicating different mechanisms for interactions between DNA-SWCNTs and biological media.