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Parkinson’s Disease (PD) is a common neurodegenerative disease. Although not fully understood, we do know that the alpha-synuclein protein plays a major role in causing PD. α-syn Drosophila flies overexpress the α-syn protein which it normally does not express. This allows it to be used as a model for studying Parkinson’s Disease. Gal4 is a driver that allows for the expression of α-syn. For this project, a cross of GAL4 Drosophila and α-syn Drosophila was compared to two control groups to develop a model. One control group was Gal4 crossed into wild-type background. The second control group was UAS-aSyn crossed into wild-type background. To develop the different variations, female drosophila were collected and crossed with adult male drosophila. Over a period of 8 days, the first generation offspring from the three groups were collected and divided into flies separated by sex. We ran locomotion assays using MWT (Multi-Worm Tracker) to track the movement of the flies. The males appeared to be moving at a slower rate than the females. We recorded survival, tracking the number of flies in each vial twice a week. On the 6th week of survival, there was a dramatic drop in the number of flies. Also, both female and male GAL4 x A-SYN Drosophila had similar survival rates. Once we develop a model we will use the model to run an experiment using PFF (Preformed Fibrils) to essentially test whether or not the overexpression of the α-syn gene protects the drosophila from getting Parkinson’s.

DSU Summer Research Symposium 2021

The Development of a Model to Study Genetic Parkinson’s Disease using Alpha-synuclein overexpressing Drosophila

Welcome to Delaware State University’s Annual Summer Research Symposium, and thank you for attending! Our symposium represents the culmination of students’ summer experiences in research at Delaware State University. This event showcases the work of college students working on faculty-led research projects, as well as middle and high school students experiencing research for the first time working in research groups. Whatever the case, this summer has been a unique experience for the students, and we hope a transformative one. Even for students who are not planning a career in scientific research, their summer experience has given them skills they will draw on for years to come. 
 Please join us on the mainstage and in Gather for the live event, and visit the recorded poster presentations any time.
 The student participants and organizers of the various summer programs at DSU would like to thank all of the research mentors who guided our summer program participants. There were many DSU faculty, graduate students, and other research and program staff who gave their time and energy to help our students have a fun and educational summer experience. Without their generosity and patience these summer programs would not be possible.
 For the research participants, we hope that you will maintain contact with your research mentors and colleagues. Science is a relatively small community made up of disciplinary “neighborhoods”. If you continue to work in a discipline, the other folks in that discipline will be like your neighbors that you run into regularly. Staying in contact with them will help you keep up with the news of the neighborhood, and open doors to educational and career opportunities. Whether you continue in science or take a different career path, for this summer you have been part of the world of research, and you will benefit from that perspective for the rest of your life.

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Welcome to Delaware State University’s Annual Summer Research Symposium, and thank you for attending!

poster

Investigating Environmental Parameters, Total Bacteria and Vibrio spp. in Slaughter Beach, Delaware Devotha Tumushimiyimana1, Rosemary Ebirim2, Michael Watson3, Gary Richards3, Ph.D., Wilbert Long1, Ph.D, and Gulnihal Ozbay1,2, Ph.D. 1Food Science and Biotechnology Graduate Program, Human Ecology Department, Delaware State University, Dover, DE 2Department of Agriculture and Natural Resources, Delaware State University, Dover, DE 3USDA-ARS, Center of Excellence, Delaware State University, Dover, DE Vibrio are members of the Vibrionaceae family are Gram- negative rod -shaped heterotrophic bacteria that are present in diverse array of aquatic environments including oligohaline, and polyhaline water. Vibrio species present in the aquatic environment common to human pathogens are V. vulnificus, V. parahaemolyticus, and V. cholera, and cause wound infections, and gastro- intestinal disease. Other vibrio such as V. coralliilyticus, and V. tubiashii have been known as shellfish pathogens. This research is seeking to know if there are potential risks such as Vibrio spp. for juvenile oysters to grow in Slaughter Beach, if water from Slaughter Beach is suitable to establish an oyster production hatchery, and if Slaughter Beach has potential risks for human pathogenic Vibrio. The four species that are being studied include V. vulnificus, V. parahaemolyticus, V. coralliilyticus, and V. tubiashii. The objectives of this study are to evaluate total Vibrio vulnificus, V. parahaemolyticus, V. coralllilyticus, and V. tubiashi in seawater and oysters collected from Slaughter Beach, Delaware, and to determine relationship between water quality and Vibrio spp. level in seawater, and oysters collected from Slaughter Beach, Delaware. Water samples are collected from two different sites (Cedar Creek and Boat ramp) from June to July 2021 at high and low tides in Slaughter Beach, Delaware. The samples were collected bimonthly. A YSI-556 multiprobe system is used to monitor physical water parameters (Temperature (°C) Salinity (g/L) Conductivity (S/m), pH, Dissolved Oxygen (mg/L), Dissolved Oxygen (%) while chemical water quality parameters are monitored using YSI 9500 Photometer. Turbidity and Total Chlorophyll-a are monitored using a Global Water WQ770 turbidimeter and Turner Design AquaFlor Fluorometer, respectively. Vibrio are monitored by Thiosulfate Citrate Bile Salts Sucrose (TCBS) and Luria Broth (LB). The 100 microliters of seawater are inoculated at 26°C and 37°C, and isolated colonies are re-inoculated on LB agar, and stored for later 16Sr RNA Sequencing. Cedar Creek demonstrated high amounts of Vibrio spp. with high levels of salinity and temperature to both tides from June to July compared to the Boat ramp site. The Boat ramp site demonstrated high turbidity at high and low tides from June to July. This project is currently ongoing and further research is being conducted on seawater and oysters collected on Slaughter Beach, Delaware to have better understanding of the areas for potential oyster culture. Keywords: Vibrio, water quality, Eastern oyster, Vibrio vulnificus, Vibrio parahaemolyticus, Vibrio coralliilyticus, Vibrio tubiashii Acknowledgements: I acknowledge USDA NIFA Capacity Building Grant Award# 2016-06642 for funding my project as part of the student professional growth and my research, Faculty advisor Dr. Gulnihal Ozbay for providing this opportunity to learn from her.

Investigating Environmental Parameters, Total Bacteria and Vibrio spp. in Slaughter Beach, Delaware

Evaluating the Reflective Spectral Profiles of Lipid Membranes in Relation to the Concentration of Cholesterol Minares Ehsani1 and Qi Lu1,2 1Division of Physics, Engineering, Mathematics and Computer Science, Delaware State University, Dover, DE 2Optical Science Center for Applied Research, Delaware State University, Dover, DE Giant Unilamellar Vesicles (GUVs) are excellent model systems for studying physical aspects of biological membranes where the interaction between lipids and proteins takes place. The key advantage of GUVs is that they can be observed directly under the light microscope due to large sizes comparable to mammalian cells. Numerous studies that have shown the effects of cholesterol on the phases and structures of the lipid-cholesterol membranes. The lipid “structures” inside the membrane ocean are known as lipid rafts. Cholesterol (CHOL) increases the rigidity of lipid vesicles by forming raft-like structures. Hence, lipid quantifications are the most important point and serve a key role in lipidomic development. In this project, we focused on the lipid reflective spectral profiles obtained from the hyperspectral dark-field microscopy and we studied their correlations to different concentrations of cholesterol (10, 20, 30, and 40 mol%). Reflective spectral profiles of GUVs with 20 mol% CHOL were found to be distinctly different from the other concentrations in both peak wavelength and FWHM (full width at half maximum). This finding was consistent with other studies, suggesting that the hyperspectral analysis technique can be used to characterize the phase and integrity of lipid membranes.

Evaluating the Reflective Spectral Profiles of Lipid Membranes in Relation to the Concentration of Cholesterol

Parkinson’s Disease (PD) is a central nervous system disorder which causes motor control issues. Current medications help improve the quality of life of those with Parkinson’s by relieving some symptoms, but do not prevent the progression of the disease. A key sign of PD is the formation of Lewy bodies in the striatum and substantia nigra. Lewy bodies are a result of protein aggregation in the brain and a major component of these Lewy bodies is the protein α-synuclein, which is known to undergo many post-translational modifications, one of which being SUMOylation. Small ubiquitin-like modifiers (SUMO) can post-translationally modify proteins affecting their function and stability in a process known as SUMOylation(1). SUMO specific proteases (SENPs) are enzymes which play a role in the de-conjugation of SUMO proteins, making the process of SUMOylation a reversible one(2). In this study, we test the possibility of applying the inhibition of SENPs as a potential treatment for Parkinson’s disease. We tested a natural SENP1 inhibitor, Momordin Ic and a chemical inhibitor, S2A, in PFF treated cells. We have found that both SENP inhibitors are efficacious in reducing cytotoxicity and increasing cell viability after PFF-induced stress. Our results strongly suggest the potential use of the SENP1 inhibitors, S2A and Momordin, as a therapeutic application for Parkinson’s disease.

SUMO Protease1 (SENP1) Inhibitor as a Potential Therapeutic for Parkinson’s Disease

Neurons and Astrocytes are two key components to our brain’s way of relaying information. When several neurons fire action potential at the same time it is called neuronal synchronizing. This is implicated in many neuronal functions that occur during development- synaptic plasticity, establishment of functional neural networks, as well as pathological conditions such as epileptic seizures. Preliminary data gathered by my lab showed that astrocyte-neuron interactions are important for development of neuronal synchronous activity in developing vertebrate brains. We focused primarily on the astrocytic model of G-Protein Coupled Receptors (GPCR)-mediate calcium induced calcium release (CICR). Our model proposes that CICR in astrocytes is controlled by mGluR1, a metabotropic glutamate receptor that leads to the release of glutamate. To demonstrate this we cultured primary astrocytes from optic tectum dissection as well as Human Embryonic Kidney (HEK) Cells . The HEK cells were transfected with melanopsin and GCaMP

An Optogenetic Approach To Understand Molecular Mechanisms of Astrocyte-​Neuron Interactions in the Development of Neuronal Synchrony

The Biopac Student Lab (BSL) is a proprietary educational system, integrating hardware and software for the acquisition of physiological electrical signals from human bodies. In a prior study, we recorded the electroencephalogram (EEG) or brainwaves from a group of volunteers to study how 432 and 440 Hz music tuning may affect responses in human brains. However, the analysis was conducted using the rudimentary built-in functions on BSL with very limited options for EEG analysis and the patterns were not definitive. In this project, our goal is to export data from BSL to a more advanced system AcqKnowledge, which provides specialized EEG analysis capabilities such as approximate entropy, delta power analysis, alpha RMS, etc. We were able to export raw data from BSL to AcqKnowledge of combined EEG signals as well as four channels including alpha, beta, delta, and theta waves using delimited text file with matching sample rate. We will further look into the literature and explore the EEG analysis capabilities on AcqKnowledge in search for an effective strategy to analyze the existing and new data. We anticipate that more advanced analysis of EEG data will shed light on the impacts of music on human brains.

Exporting Data from Biopac Student Lab to AcqKnowledge for Advanced Analysis of Human Brainwaves In Response to Music Tuning

Human bodily functions, e.g., exhaled breath, perspiration, and physiological processes, exhibit several chemical and isotopic tracers that are potential disease signatures. Exhaled human breath analysis can be used for early disease detection and identify and discriminate disease biomarkers while being non-invasive. This project aims to develop rapid, real-time chemical and biological detection using mid-infrared laser systems. We used mid-infrared laser spectroscopy to detect the molecular absorption line transitions. In addition, we designed a long pathlength (30 m) Heriot Laser Optical Cavity to maximize the signal-to-noise ratio of experimental molecular absorption signals. This is a vital part of laser-based precision sensing. Furthermore, we also focused on a volatile organic compound and the detection of Nitrous Oxide (N2O). A rapid laser scan was performed at a frequency of 100 Hz with laser stabilized at a constant 27°C. This allowed us to estimate the current range, 50 mA to 72 mA, and the wavelength range of 4537 nm to 4540 nm of the mid-Infrared absorption from the molecular rotational-vibrational transitions. We also show that mid-infrared sensing is ideal for real-time precision detection of molecules. In addition, molecular density can be precisely estimated by the polynomial fitting of the absorption signal.

Laser-based precision sensing for biomedical applications

Dandelion (Taraxacum officinale) root extract (DRE) is known to have anti- cancer activity against breast cancer, leukemia, prostate cancer, colorectal cancer and melanoma cells (1-5). Even though this plant is sometime seen as a pesky weed in our culture, in other cultures such as Chinese, Middle Eastern, Native Americans, and East Indians, it is appreciated for its medicinal effects against cancer and other diseases ranging from indigestion, heartburn and kidney disease. Based on the reported anti-cancer cell activities of DRE, particularly on aggressive cancer cells but not on normal cells, we decided to study its activity on two breast cancer cell lines, one expressing the cell membrane receptor CD44 (i.e., MCF-7/CD44) and a control cell line negative for CD44 expression (i.e., MCF-7/pLXIN) (3, 6). The MCF-7/CD44 cell line is an aggressive multidrug resistant, very migratory and in vitro invasive while the MCF-7/pLXIN cell line is not. We hypothesize that aqueous DRE will have differential anti-cancer cell activity on these two cell lines based on their CD44 expression status. A cell viability assay (MTS) carried out after 48 hrs of DRE treatment showed that MCF-7/CD44 cells were more sensitive to DRE (when extracted in boiled PBS) compared to MCF-7/pLXIN cells. Treatment of these cell lines with non-boiled PBS extracted DRE showed no cell toxicity effect. This data validated our hypothesis and suggest that DRE might be effective at targeting CD44-expressing aggressive breast cancer cells.

Effects of Dandelion Root Extract (DRE) on CD44(+) and CD44(-) for Cell Viability

We report Raman spectroscopy measurements on samples of Ficoll polysaccharide dissolved in water at different concentrations (up to 1000 mg/ml) and collected at ambient temperature. The goal is to identify and assess Raman spectral signatures associated with the molecular structure of Ficoll PM70 and the concentration effect on these signatures. Raman spectroscopy is used to determine molecular vibrational modes, providing spectral peaks that identify the molecules (e.g Ficoll), especially in crowded and multicomponent matrices. Here, an excitation laser beam is directed onto the solution sample and interact with the structural vibrational modes of the macromolecules, resulting in energy shift of the laser photons that can be readily measured. Ficoll is accepted as a water-soluble, neutral, highly branched polymer and is commonly applied as a macromolecular crowding agent in diverse studies of protein folding and stability as well as cell volume signaling. Further, it is also used to separate blood into its components such as leukocytes and erythrocytes. In our case, we have been using Ficoll as a model crowded system to investigate the complex behavior of diverse nanoparticles. We used a HORIBA MacroRAM spectrometer to measure the Raman spectra of Ficoll solutions. In addition to the commonly reported Raman peaks of water (~1640 cm-1 HOH bend; ~3200 cm-1 OH stretching band) and possibly dissolved O2 (~1556 cm-1) and N2 (~2331 cm-1) we have identified four distinct spectral peaks (~1270 cm-1 ,~1350 cm-1,~1460 cm-1, ~2900 cm-1) likely associated with Ficoll in the 1000 to 3500 cm-1 wavenumber range, a range set to avoid possible spectral contamination from the quartz sample holder. Increase of the Ficoll concentration shows systematic increase of the intensity of the observed Ficoll peaks and systematic decrease in intensity of the two peaks associated with water, the latter effect being due likely to reduction of water content. These observations and a detailed analysis and interpretation of the measured spectral information enclosed in the Raman emission of Ficoll macromolecules may lead to a better understanding of the complexity in chemical analysis of crowded biopolymeric samples.

Raman Spectra of Concentrated Ficoll Solutions

Parkinson’s Diseases (PD) is the second most common neurodegenerative disorder and is characterized by a preferential loss of dopaminergic neurons of the substantia nigra pars compacta. The cause of the disease remains unknown; however, it is thought that environmental factors may contribute to its development. Rotenone is a pesticide that blocks mitochondrial respiration in the Electron Transport Chain by inhibiting Complex I and has been identified as an environmental risk factor for sporadic PD. Here our goal is to develop strategies that can lead to the development of potential preventative measures against PD. Dacarbazine is a potential anti-PD drug that is known for its use in chemotherapy for skin cancer and was identified in a previous screen for neuroprotective agents against the disease. We hypothesize that Dacarbazine can relieve or reverse the effects of rotenone in Drosophila. We set up a study to analyze the effect of dacarbazine against rotenone-induced neurotoxicity using locomotion behavior as a measure of neuronal toxicity. We set up four conditions: rotenone only; rotenone combined with dacarbazine; dacarbazine only; and DMSO only as vehicle only control. Here we present our results in two paradigms: a first one in which we performed a brief chronic exposure to rotenone and/or dacarbazine for three weeks and a second paradigm in which the chemical treated flies were allowed to recover on normal food for two additional weeks. Together, these studies could help determine whether dacarbazine confers protection against rotenone-induced neurotoxicity and could inform future efforts to establish a neuroprotective strategy against sporadic PD.

Ameliorating the effects of an environmental toxin in a Drosophila model of Parkinson’s Disease

Different studies show that TDP-43 plays a major role in association with other neurological diseases, however, we want to test why TDP-43 is a major factor. We injected a mouse with TDP-43 and injected another mouse with Alzheimer's Disease. The two mice were watched for more than over a month now. We used the mice's pellets to conduct the different tests we ran. We sonicatied the different mice pellets for a 1 min 20 minutes with a 5 sec pulse and 5 sec off cycle. Once the sonication was done, we transferred the sonicated pellets to the centrifuge machine which ran for about 35 minutes. Other graduate students began to compare results from these batches of pellets to the others that they tested to see the difference or similarities. These results showed us that the Alzheimer's disease mouse model that we developed over expresses pathological tau and TDP-43.

The Development of a Model to Study Genetic Parkinson’s Disease using Alpha-synuclein overexpressing Drosophila

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