OSCAR Celebration of Student Scholarship and Impact
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Cells, Individuals, and Community College of Humanities and Social Science College of Science Honors College OSCAR Winners

Women’s Perception of Safety as Demonstrated in Tiktok Trend Man Vs Bear

Author(s): Devyn Wilson

Mentor(s): Collin Hawley, Honors College

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Abstract

Stemming from the 2024 TikTok trend asking women if they would rather be stuck in a forest with a man or bear, this study seeks to answer what the internal discourse of women is when choosing a man over a bear, in order to assess their values and perceptions of safety. Early research on this subject focused on crime statistics and victimization rates. As feminist theory developed, researchers began to explore broader contexts, including public spaces, urban design, and societal norms that influence women’s feelings of safety. This transition showcased that women’s safety perceptions are often shaped by societal norms, cultural narratives, and lived experiences. However, this proposed research question explores how women assess and investigate the intersection of physical, emotional, and social aspects of safety. To explore the decision-making process of women, mixed methods including, Critical Technocultural Discourse analysis of social media posts (Brock, 2016), bear attack statistics, crime analysis, and survey analysis of women in the DMV area will be used. Critical Technocultural discourse analysis used to analyze social media discourse to find what women are saying regarding their perceived safety in either scenario, a man or a bear appearing. Bear attack analysis and crime analysis can compare the relative risk of damage if a bear or man appears. Surveys of women in the DMV area allow for a conversational component in which women will be able to express safety in their setting. The findings contribute to broader conversations around safety, risk assessment, trust, and gender in contemporary society.

Audio Transcript

Imagine walking on a trail much like this one. While walking you hear leaves rustling, would you whether the rustling be due to a man or a bear. This question was presented to TikTok users in 2024. What was supposed to be a quick, simple, easy-to-flow trend turned into discussions about women’s safety and how they perceived the world around them. It has been a little over a year since this question first appeared, allowing many people to answer it. But what does this question demonstrate about women’s perceptions of safety? What does this question demonstrate about the perceptions of safety of women. What kind of improvement and conclusions can we draw from this trend to make women feel safer. Furthermore, the project wanted to explore the intersections of social media and outlook on safety.

Academic conversations have yet to cover these discussions. Early research on women’s perception of safety focused on crime statistics and victimization rights. As feminist theory developed, researchers began to explore broader contexts, including public spaces, urban design, and societal norms that influence women’s feelings of safety. Furthermore, safety is grouped into other discussions including MeToo, Not all men, LGBTQ+, and family disputes, among others. While these papers provide context to safety, they fail to capture the true fears and realities of women, in their everyday life. The shift from victimization rates to broader contexts marked the understanding that women’s safety perceptions are shaped by societal norms, cultural narratives, and lived experiences. Despite this shift, academic papers have yet to capture the nuanced discourse of women in everyday life. This disconnect raises important questions about whose experiences are prioritized in academic discourse — and why moments like the “man or bear” debate remain outside scholarly engagement. This project seeks to highlight the internal dialogue of women set upon by this trend Man Vs Bear, exploring how women assess and investigate the intersection of physical, emotional, and social aspects of safety.

This semester primarily focused on IRB approval and gathering survey responses.

Before examining the responses, it’s important to note the inclusion criteria. Respondents had to be women or nonbinary, between 18 and 25 years old, and currently attending a university or college in the DMV area. A total of 70 participants met this criteria.

When asked whether they would rather be in a forest with a man or a bear, responses were as shown in the graph:

Most participants felt confident in their choice. Those who were uncertain said they wanted more context—such as the type of bear, the exact setting, or the man’s behavior—because these factors would meaningfully shape their decision.

Since the purpose of the study was to understand why participants made this choice, the explanations are central to the findings. Common themes included:

Fear of men: This appeared on both sides. Many respondents felt they could more easily predict a bear’s behavior compared to a man’s intentions. Participants mentioned strategies for avoiding bear attacks or managing the situation, while expressing concern about men having hidden motives or personal gains. Several respondents noted that some potential outcomes with men—such as assault, or not being believed afterward—felt “worse than death.”

Some pointed out that they see men every day and have survived those encounters, implying that men may not be as dangerous as perceived. Others emphasized that bear attacks are statistically less common than violent crimes committed by men.

Later questions provided broader context for these choices: 93% of respondents felt that women are conditioned to be more alert or afraid in public spaces than men. No one said “no.” 80% reported they have pretended to talk on the phone, changed direction, or taken defensive actions when encountering an unknown man in a quiet area. These behaviors highlight the role of social conditioning and hypervigilance in shaping perceived safety.

Across responses, control emerged as a major influence. Participants expressed more fear of threats whose motives could be hidden—such as a man who appears helpful but becomes dangerous.

Many respondents also said they would trust their instincts over statistics, suggesting that cultural narratives and personal experience strongly shape safety judgments. For some, choosing the bear aligns with worst-case-scenario thinking, especially for those who stated that certain outcomes with men would be worse than death.

Respondents tended to classify their fear of bears as primarily rational—based on a known, identifiable physical threat.

Fear of men, however, was described as both rational and emotional, rooted not only in real statistics but also in lived experience, cultural conditioning, and emotional memory.

indicating that fear is shaped by:

lived experience,

repeated warnings,

cultural narratives, and

observed stories of harm.

The data shows that young women and nonbinary students interpret safety through a blend of emotion, lived experience, social conditioning, and perceived controllability. The surprising preference for being with a bear over a man is not about the bear itself—it is a reflection of deep concerns about male unpredictability, cultural warnings, and threats to autonomy.

The bear becomes a stand-in for a danger they feel they can understand, while men represent dangers they feel they cannot read or control.

Looking into the coming months, Critical Technocultural Discourse analysis, bear attack statistics, and crime analysis will be put into conversation with the survey results. Together, they will uncover more about perceptions of safety and where gaps can be filled through societal shifts. Ultimately, my work contributes to a broader conversation: how gender, safety, and culture shape the way we move through the world.
The “man or bear” question may sound simple — even silly — but it reminds us that women’s fears are not irrational. They are reflections of real, lived experiences in a world that too often asks them to be cautious — even in the forest.

Categories
College of Science OSCAR US, Global, and Beyond

What Makes Dark Matter Halos Dark

Author(s): Omar Aljebrin

Mentor(s): Ferah Munshi, Physics and Astronomy

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Abstract

We investigate the factors that determine whether dark matter halos form galaxies using the Marvel-ous Dwarfs and DC Justice League zoom-in simulations. By analyzing halo occupation fractions, mass accretion histories, halo concentrations, and gas properties, we find that halo mass alone is insufficient to explain why some halos remain dark. Luminous halos grow faster, achieve higher concentrations, and retain dense, cool gas, enabling star formation. Dark halos grow more slowly, remain less concentrated, and have lower gas densities and higher gas temperatures, preventing efficient cooling after reionization and leaving them dark. These results highlight the interplay between dark matter dynamics and baryonic physics in shaping galaxy formation, demonstrating that mass growth, halo concentration, and gas content collectively determine whether a halo hosts a galaxy.​

Audio Transcript

17:37:02 Hello, everyone. Today, I’ll be talking to you about what makes some dark matter halos dark.
17:37:07 So, first of all, what is dark matter? Dark matter is like any other matter, but…
17:37:13 It’s invisible, it cannot interact with normal matter that we see and touch,
17:37:18 Like, it can’t bump into it, or you can’t feel it.
17:37:23 Um,
17:37:25 And it only interacts through gravity.
17:37:28 So, it can only pull things
17:37:30 using gravity.
17:37:32 And…
17:37:34 Dark Matter Halos is just a clump of dark matter that has collected together.
17:37:40 And from now on, I’ll just call it Halos.
17:37:46 And these halos is…
17:37:48 is the environment where galaxies are made.
17:37:52 But… sometimes galaxies aren’t made.
17:37:56 In those halos, and those are called dark halos. Dark Halos. And for the ones…
17:38:02 that galaxies do form in, those are called luminous halos.
17:38:06 So, the goal of my research is to figure out
17:38:09 what leads to these halos?
17:38:12 not forming any galaxies? What properties, what events?
17:38:17 created? Like, why…
17:38:19 Do these halos have galaxies, and these don’t?
17:38:23 And…
17:38:25 my method to look at… to do that is using, um…
17:38:30 large cosmological simulations.
17:38:34 simulations, uh, that are called
17:38:36 marvelous dwarfs and DCGL.
17:38:39 DCJL standing for DC Justice League.
17:38:46 These were made by…
17:38:49 Dr. Munshi and her colleagues.
17:38:52 Um…
17:38:54 using the simulation, it provided various datas about dark matter halos, their properties,
17:39:01 And I was able to utilize this data to track certain, um,
17:39:07 patterns of dark matter halos.
17:39:10 So, for my first plot,
17:39:12 I’ve calculated the halo occupation fraction versus the maximum
17:39:19 mass, a halo has ever had.
17:39:21 So the Halo occupation fraction is basically saying, okay, for this amount of halos,
17:39:26 Um, how many of them are luminous divided by the total
17:39:30 amount of, um, halos.
17:39:33 And from what you see is…
17:39:34 From the right end… from the right end, from the large end of the mass.
17:39:41 It’s 100%, one, at the top, and then…
17:39:46 Uh, some of… they… they begin to decrease.
17:39:50 And the decrease is gradual, but…
17:39:52 you’ll see that all these simulations have different…
17:39:56 point of mass where that halo occupation decreases.
17:40:00 Which tells us that because their masses
17:40:04 vary for that critical point. That means that halo mass, or halo peak mass, isn’t enough to tell us.
17:40:10 About why some dark… some…
17:40:13 halos don’t form galaxies.
17:40:17 So then, uh, I look at the mass assembly history of the halos, how fast they gain mass.
17:40:24 And what I find that luminous halos
17:40:27 were… or the one that made Galaxies gain mass really fast. Faster than dark halos.
17:40:33 And, of course, the Dark Hills.
17:40:35 didn’t gain mass as fast.
17:40:37 Which could explain why
17:40:40 Dark Halos are… don’t make any galaxies, because they couldn’t accumulate…

Categories
Cells, Individuals, and Community College of Science

Ivermectin as a potential therapeutic for Interstitial Pulmonary Fibrosis

Author(s): Diya Chordia

Mentor(s): Geraldine Grant, Biology

Abstract

Interstitial pulmonary fibrosis (IPF) is a progressive and fatal lung disease characterized by fibroblast activation, myofibroblast differentiation, and excessive extracellular matrix deposition. The TGF-β1/SMAD3 pathway is a central driver of this process, yet current antifibrotic therapies only slow disease progression and remain costly. Recent studies suggest that ivermectin, a widely available antiparasitic drug, may inhibit SMAD3 phosphorylation in non-pulmonary fibroblasts, raising the question of whether it could attenuate fibrotic signaling in lung fibroblasts. This study investigates whether ivermectin suppresses fibroblast activation in IPF by reducing α-SMA expression, inhibiting SMAD3 phosphorylation, and limiting fibroblast migration. Human lung fibroblasts will be treated with increasing concentrations of ivermectin (0.1–3.0 µM) for 48 hours in the presence of TGF-β1. α-SMA gene expression will be quantified by RT-qPCR, and α-SMA, SMAD3, and pSMAD3 protein levels will be examined by Western blot. Functional effects will be assessed using a scratch wound-healing assay performed in 6-well plates, and actin organization will be visualized with phalloidin staining. Data will be quantified using ImageJ and analyzed with one-way ANOVA. We expect ivermectin to decrease α-SMA expression, reduce SMAD3 phosphorylation, and slow fibroblast migration in a dose-dependent manner. These findings would support the potential repurposing of ivermectin as a low-cost antifibrotic therapy and provide new insight into modulation of the TGF-β1/SMAD3 pathway in pulmonary fibrosis.

Audio Transcript

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00:00:08.360 –> 00:00:32.910
Diya Chordia: Hello, everyone, and thank you for taking the time to come visit my poster. My project focuses on exploring the potential antifibrotic effects of ivermectin and antiparasitic in interstitial pulmonary fibrosis, or IPF. IPF is a progressive and ultimately fatal lung disease characterized by fibroblast activation and

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00:00:32.910 –> 00:00:36.299
Diya Chordia: Excessive extracellular matrix deposition.

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00:00:37.020 –> 00:00:52.000
Diya Chordia: Although current medications can slow the disease, they cannot reverse fibrosis, and lung transplantation remains the only definitive treatment. Because of this, there is a real need for new, accessible therapeutic options.

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00:00:52.000 –> 00:01:16.800
Diya Chordia: A key driver of fibrosis in IPF is the TGF-beta and SMART3 signaling pathway. When activated, this pathway increases the expression of markers such as alpha smooth muscle actin, and promotes structural and functional changes that make fibroblasts more contractile and migratory.

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00:01:17.910 –> 00:01:31.869
Diya Chordia: Recent work, including a 2021 paper by Zia et al, suggests that ivermectin can reduce SMART3, activation and decrease

6
00:01:31.980 –> 00:01:47.010
Diya Chordia: alpha-sma expression in fibroblasts from other tissues. However, its effects on lung fibrosis have not yet been studied. This gap in literature motivated this central research question of my project.

7
00:01:47.060 –> 00:01:53.750
Diya Chordia: Our hypothesis is that ivermectin will attenuate fibroblast activation in IPF,

8
00:01:53.750 –> 00:02:09.350
Diya Chordia: by suppressing the SMART3 signaling pathway. Specifically, we predict that ivermectin will decrease alpha-smar expression, reduce SMAR3 phosphorylation or activation, and limit fibroblast migration.

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00:02:11.840 –> 00:02:29.490
Diya Chordia: To test our hypothesis, human lung fibroblasts will be treated with increasing concentrations of ivermectin, from 0.1 to 3 micromolar for 48 hours in the presence of, FPS,

10
00:02:29.530 –> 00:02:48.139
Diya Chordia: to induce activation. Following treatment, SMAR… the alpha-sma gene expression will be measured using RT-QPCR with RNA isolated using the Quergen RNA-Z kit for reliable, high-purity extraction.

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00:02:48.180 –> 00:03:06.720
Diya Chordia: Protein expression will be assessed by a Western blot, probing specifically for Alpha SMAR, total SMART3, and phosphorylated SMART3, which is just activated SMART3, using high-specificity antibodies.

12
00:03:07.660 –> 00:03:20.219
Diya Chordia: To evaluate functional changes, a scratch wound healing assay will be performed in 6 well plates to determine the effect of ivermectin on fibroblast migration.

13
00:03:20.630 –> 00:03:29.809
Diya Chordia: Falden staining will be used to visualize alternatives in actin organization and stress fiber formation.

14
00:03:30.130 –> 00:03:34.659
Diya Chordia: All image-based analysis will be quantified using image J.

15
00:03:34.940 –> 00:03:45.750
Diya Chordia: And data will be statistically evaluated using one-way ANOVA with a significant threshold of, P is less than 0.05.

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00:03:47.670 –> 00:03:55.880
Diya Chordia: Although the full results are still being generated, the expected outcomes are that ivermectin will reduce alpha-smar levels.

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00:03:55.880 –> 00:04:14.110
Diya Chordia: suppress SMAR3-phosphorylation and slow fibroblast migration in a dose-dependent manner. If these predictions hold true, it would indicate that ivermectin can directly interrupt prohibprotic signal signaling and fibroblast activation.

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00:04:15.190 –> 00:04:20.690
Diya Chordia: The broader significance of this work relies in its potential impact on global health.

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00:04:21.120 –> 00:04:43.030
Diya Chordia: IPF affects patients worldwide, but access to current drugs is limited by high cost. Because of this, ivermectin is inexpensive, well-studied, and already widely distributed, and identifying antifibrotic activity could make treatment more accessible, especially in low- and middle-income settings.

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00:04:44.070 –> 00:04:56.380
Diya Chordia: This study also contributes new insights into how the SMART3 signaling can be modulated in pulmonary fibrosis, providing a foundation for future therapeutic development.

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00:04:56.630 –> 00:04:58.630
Diya Chordia: Thank you for listening!

Categories
College of Science Honors College

Automation in Detecting and Localizing Diseased Kidneys in MRI Scans using Deep Learning

Author(s): Rushil Johal

Mentor(s): Suraj D. Serai, Department of Radiology, Children’s Hospital of Philadelphia (CHOP)

Autosomal recessive polycystic kidney disease (ARPKD), a rare genetic disorder affecting 1 in 20,000 children, leads to kidney failure by age 15 due to abnormal renal tubules forming fluid-filled cysts. Diagnosing ARPKD in adolescents prematurely is challenging, as they are in the early stages of kidney development. IMPAKT, a current IT database, helps clinicians identify and manage ARPKD patients early. Traditionally, MRI scans are manually analyzed to collect clinical data, a process prone to errors and inconsistencies. This study introduces a deep learning approach using digital image filtering and artificial neural networks to automate ARPKD detection in DICOMs (digital communication standard for MRI scans). The method accurately localizes cysts by applying spatial high-pass filtering and edge detection, overcoming previous manual detection limitations. This technique can integrate with existing medical records software to predict the likelihood of ARPKD and other genetic diseases at its early stages for risk patients, enabling earlier treatment.

Hello! This is a presentation on the Automation in Localizing and Detecting Disease Kidneys in MRI Scans using Deep Learning. My name is Rushil Johal and with the assistance of the University of Pennsylvania Health System or UPenMed and Children’s Hospital of Philadelphia.

Opening the successfully filed USPTO patent application was a tangible symbol of my biggest dreams. My journey began with sleepless nights, manually tracing and scanning MRI silhouettes of inflamed kidneys to gather vital biological data, such as water volume and iron content. This required drawing 10 outlines per MRI scan layer and each kidney having 40 layers. It’s a cumbersome process for physicians conducting genetic disease testing. I pondered: There must be a more efficient way.

Autosomal recessive polycystic kidney disease (ARPKD) is a rare genetic disorder affecting 1 in 20,000 children. It’s caused by fluid-filled cysts in renal tubules, leading to decreased kidney function and eventual failure by mid-40s. Deep Learning (DL) can revolutionize early ARPKD diagnosis and transform the healthcare industry by collecting and analyzing data from MRI scans for various diseases, potentially saving thousands of lives.

I am studying Computational Data Science and have versatile programming experience in R, Java, MATLAB, and Python. Thus, I apply these hands-on skills in scientific research, specifically interning in MRI Radiology at the University of Pennsylvania’s Perelman School of Medicine and Children’s Hospital of Philadelphia in Summer 2022.

There, I evaluated how an image filtering, DL algorithm is controlled to detect and localize early telltale signs of ARPKD in adolescent patients by examining MRI scans, known as DICOMs.

We‘ll cover the current problems with diagnosing ARPKD and its data collection process, how this DL algorithm functions and its limitations, and the algorithm‘s implications and extensions throughout the medical industry.

To first understand why ARPKD is challenging to detect early and its problematic data collection process, we’ll cover its relevance with DL.

Dr. Anwar Padhani, spokesman for the 2016 International Conference on Radiation Medicine, describes DL branching from ML by utilizing neural networks, like a brain neuron, with many interconnected layers to automatically extract patterns from data, enabling high-level abstraction and complex tasks like image recognition.
ARPKD is difficult to diagnose early due to having no cure, no symptoms nor pain, and delayed development of cysts by the time the risk patient typically reaches 12-14 years old, where tissue trauma has already occurred. Furthermore, the symptoms of excess water retention simulate a steep exponential growth between the patient’s age and kidney size, almost indistinguishable from a healthy, linear-size kidney growth as the patient matures.

Current data collection begins by extruding DICOMS from another picture archiving and communications system, PACs, to be mapped into a standard coordinate system. We collect data by free-hand drawing regions of interest (ROIs) as silhouettes of specific tumors within the kidney. According to nephrologist Dr. Charlotte Gimpel’s 2019 PubMed publication, this tedious methodology is time-consuming, susceptible to human error, unable to localize cysts in DICOMs with blurry qualities, and inconsistent among varying DICOM series.

Understanding DL’s relevancy in streamlining ARPKD’s data, we’ll focus on the algorithm’s operation and limitations.

Digital image filtering and artificial neural networks of multiple input and output automate localizing ARPKD in DICOMs. This steerable algorithm applies high radio frequency and edge detection of image pixels to detect abnormal cysts compared to the rest of the kidney structure. The same approach is applied for Snapchat filters detecting your facial features based on lighting. Once multiple DICOMs of the same patient study and date have been uploaded, the algorithm leverages prior pixel data to improve performance for the same repeated set of tasks.

However, the algorithm may initially experience difficulty differentiating ARPKD’s appearance from a healthy kidney in a DICOM or not generalize to different genetic variations of ARPKD. This can lead to misdiagnosis by false positives or negatives for ARPKD. However, Dr. Kouta Ito’s 2021 publication in the National Center for Biotechnology Information’s findings reveals this algorithm can replace the manual method by decreasing medical uncertainty and random error by 18% with an accurate detection success rate of 93%.

After comprehending the algorithm’s method, let’s focus on its implications and extensions in other fields within healthcare.

This algorithm can be implemented in various medical records software, such as Epic Systems, and includes Multi-Modal Fusion, combining information from multiple imaging modalities, such as CT and ultrasound, to improve localization accuracy. Lastly, the method may detect early patient liver and bile duct cancer risks.

For implications: the algorithm minimizes the time required for diagnosis and treatment planning. It also provides quantitative measurements of cyst size, distribution, and progression over time, helping track disease progression and evaluating treatment efficacy in real-time, not possible prior. Thus, we can tailor personalized treatment plans based on each patient’s severity or unique ARPKD characters. However, legal issues can be a barrier, such as using huge amounts of confidential patient data or a risky financial investment, inaccessible to many underfunded healthcare systems nationwide.

Overall, this image filtering, DL method can change the healthcare industry by diagnosing unaware patients at risk, not just in ARPKD, but any disease studied via MRI DICOMS, even if a cure does not yet exist.

We have covered DL‘s importance in solving ARPKD diagnosis and addressing ARPKD‘s concerns, the new methodology and its constraints, and its impact on healthcare.

Evaluate how you may incorporate DL into your everyday life. Allow an algorithm to teach itself and do the complicated, repetitive tasks for you better after each try. After all, ML is just a baby learning how to walk, but sooner than you realize, will drive a car.

Thank you so much for your undivided attention!

Categories
College of Science OSCAR

Fluorescent Probe Development for Detection of Oxidative Stress-Induced Carbonylation in Cells

Author(s): Amira Anwar

Mentor(s): Ozlem Dilek, Chemistry and Biochemistry

There is an urgent need for precise and robust diagnostic tools for monitoring disease progression in biological systems for healthcare and biomedical research. The development of a non-invasive fluorescence probe that is capable of detecting oxidative stress-induced carbonylation in cells is vital to advance our knowledge of oxidative stress-related diseases. Oxidative stress occurs when there is an excessive amount of reactive oxygen species (ROS) in a biological system that can damage crucial components of the cell and increase carbonylation in the cell environment. This chemical process can be associated with several health complications such as cancer, neurodegenerative disorders, and diabetes. The detection of oxidative stress-induced carbonylation with novel fluorophores can therefore be very helpful for monitoring different cell line systems in vitro. In order to introduce fluorophores into the cellular systems, the design of new probes is essential. Probes should have particular chemical and photophysical characteristics to achieve successful imaging: high stability, less toxicity, fast kinetics, good spectral properties such as large Stokes shifts, reasonable quantum yields, drastic changes on absorption and emission spectra. In this work, we focused on synthesizing a few probe systems for targeting carbonyl moieties of biomolecules in living cells using a click reaction to produce a fluorescent product so that we can be able to visualize the carbonylation process in various cancer cell lines. In the near future, the development of non-invasive fluorescent probes that are capable of detecting oxidative stress-induced carbonylation in cells will be pivotal in understanding the mechanisms that govern disease. In addition, we anticipate that it will help immensely improve diagnostics and drug development in biomedical sciences.

Hello, everyone! My name is Amira Anwar and this is my OSCAR URSP Spring 2024 project. My research focuses on the development of fluorescent probes for the detection of oxidative stress-induced carbonylation in cells. I would like to express my sincere gratitude to my mentor Dr. Dilek, for their invaluable guidance and support throughout this research project. I want to extend my thanks to Eva-Maria Rudler for her wonderful support as well. There is a critical need for precise diagnostic tools to accurately assess disease progression in biological systems for healthcare and biomedical research. These tools are important for early disease detection, guiding treatment decisions, and understanding the mechanisms that govern health and disease. Below are some different types of probes. Oxidative stress occurs due to an imbalance between the production of reactive oxygen species and the biological system’s ability to detoxify them. When there is an excessive amount, reactive oxygen species overwhelm the biological system and can cause chaos in important cellular components including lipids, proteins, and nucleic acids. Oxidative stress can further exacerbate this by increasing carbonylation levels within the cell environment potentially triggering apoptosis or programmed cell death. This type of chemical process is linked to various health complications including cancer, metabolic disorders like diabetes, and neurodegenerative disorders like Alzheimer’s or Parkinson’s disease. Here is a diagram of a normal cell undergoing oxidative stress and then resulting in apoptosis. The development of a non-invasive fluorescence probe capable of detecting oxidative stress-induced carbonylation in cells is crucial for advancing our understanding of oxidative stress-related diseases. These probes can provide a means to directly visualize live cells in real-time and offer incredible insights into the molecular mechanisms. This is a picture of a fluorescent probe that is detecting mitochondrial oxidative stress in cells of mice. Using novel fluorophores to detect oxidative stress-induced carbonylation can be highly advantageous for monitoring various cell line systems in vitro. The design of new fluorescent probes is key in order to introduce them into cellular systems. They need very particular chemical and photophysical properties for successful imaging like high stability to ensure their integrity and reliability in harsh cellular environments. It must have low toxicity levels to allow accurate detection while promising the safety of the biological system. Fast kinetics to enable rapid binding and response to target molecules. There also needs to be good spectral properties like significant Stokes shift, favorable quantum yield, and notable alterations in the absorption and emission spectrums. I focused on synthesizing probe systems to target carbonyl groups in biomolecules found in live cells, utilizing click chemistry reactions to produce fluorescent products. This was done to enable the visualization of the carbonylation process across various cell lines. Here is an example of a click chemistry reaction: this click reaction is referred to as the copper-catalyzed alkyne”“azide cycloaddition. The development of non-invasive fluorescent probes capable of detecting OSIC within cells will play a pivotal role in understanding the mechanisms that govern health and disease. I anticipate this development will advance diagnostics and facilitate drug development in biomedical sciences. Looking ahead I plan to continue my research with OSCAR and focus on several key areas to advance in such as cellular fluorescence imaging techniques, applications of cell culture labeling, potentially investigating additional molecules for metal sensing, and computational calculations. Finally, I would like to say thank you to everyone watching and thank you to OSCAR for giving me this opportunity.

Categories
College of Science OSCAR

Kinetic Analysis of Fosmidomycin Analogs

Author(s): Quintin Carroll

Mentor(s): Robin Couch, Department of Chemistry and Biochemistry

According to the World Health Organization, in 2022 approximately 1.3 million people worldwide died from tuberculosis, making the disease the second-leading infectious killer known to man. For new strains of Mycobacterium tuberculosis (Mtb), the bacterium responsible for the disease, multidrug therapy consisting of isoniazid, rifampin, pyrazinamide, and ethambutol has replaced the now-resistant single-drug protocol. However, the emergence of multidrug-resistant (MDR) strains presents an urgent need for new antibiotics. The 2C-methyl-D-erythritol 4-phosphate (MEP) pathway is an outstanding target for novel drug development due to its essentiality in M. tuberculosis and its absence in humans. This pathway is a seven-step mechanism that results in the synthesis of isoprenoids; cell wall components and other biomolecules essential to the pathogen. The first step of the MEP pathway is the reduction and isomerization of 1-deoxy-D-xylulose 5-phosphate (DXP) via the enzyme 1-deoxy-D-xylulose 5-phosphate reductoisomerase (Dxr or IspC). Fosmidomycin is a natural product known to inhibit IspC, but has poor drug-like properties. Thus, rationally designed inhibitors based on the backbone structure of fosmidomycin have been developed. In this study, we analyzed the effectiveness of several fosmidomycin analogs via a kinetic enzymatic assay buffered at either pH 6.5 or 7.8. Results indicate that the fosmidomycin analogs are especially potent in the more acidic environment (pH 6.5), relative to pH 7.8, which is particularly important since the intracellular pH of M. tuberculosis is 6.5.

Hi. My name is Quintin Carroll, and today we’re gonna be talking about my research on the kinetic analysis of those Fosmidomycin analogs. So starting off, let’s talk about what is tuberculosis. Tuberculosis is one of the deadliest infectious diseases known to man. In twenty eighteen, roughly one point five million people died from its infection, and less than sixty percent of patients who undergo treatment are successfully treated. Thus new treatment options are height of heightened urgency to address the disease. Currently, tuberculosis is treated with a multi drug protocol consisting of rifapentine, isoniazid, pyrazinamide and ethambutol. However, the emergence of drug-resistant strains call for new treatment options of the disease. Mycobacteria tuberculosis is the causative agent of tuberculosis. And it is characterized by a fatty acid outer membrane. This outer membrane plays a significant role in antibiotic impermeability. To develop drugs that target Mtb, we look to the methylerythritol phosphate pathway or Mep pathway for short. This is an outstanding target because it does not exist in humans, thus therapies are highly specific to enzymes in this pathway. Fosmidomycin a natural product has been shown to inhibit ISPC in the Mep pathway. However, poor drug like properties such as unfavorable membrane permeability and poor half life make it an unsuitable antibiotic, thus analogs that can that can address these poor drug like properties or paramount interests. For our methodology, we used recombinant protein expression and isolated and purified our target enzyme. And from there, we perform kinetic analysis using an assay mixture that is buffered to six point five pH. From there, we can incubate for ten to fifteen minutes afterwards, we can monitor the oxidation of nadph to produce a time trace that is pictured above or to my left rather. Previously research has used similar methods, but with a more basic assay mixture at pH seven point eight. Our results show that there are much more potent inhibition at pH six point five compared to pH seven point eight. As we can see from the chart, there is much more compounds that are below the dotted line compared to this bar graph over here, which has many above. In terms of compounds, we have two graphs that display the IC fifty value. Or in other words, the concentration that gives us half maximal and enzymatic activity. analyzing the graphs we can see that the n-acyl moiety on the FR nine zero zero nine eight compound proved to be a positive structural change compared to Fosmidomycin In conclusion, results of this research demonstrate that seven compounds exhibit pH dependent activity. Likely from protonation effects influencing inhibitor binding. However, additional investigation is being done to validate this hypothesis. Furthermore, experiments in the form of antimicrobial susceptibility assays are warranted to test these compounds whole cell Mtb to determine their effectiveness against the fatty acid cell wall.

Categories
College of Science OSCAR

Motor Deficit & Brain Remediation Following TBI with Plasma Administration

Author(s): Petra Honainy

Mentor(s): Jane Flinn, Psychology

Young blood plasma administration in mice has shown to be promising as a potential
treatment for symptoms in neurodegenerative diseases such as Alzheimer‘s disease
and Traumatic Brain Injury (TBI). There is limited research on the pathways and the
extent of the mitigation of motor deficits following young blood plasma administration.
Our study inspects whether young blood plasma can improve motor deficits in mice
with rmTBI and whether these improvements can be seen in the motor cortex. The
mice will be divided in four groups, one receiving TBI and plasma injections
immediately after TBI, one receiving TBI and plasma injections 30 days after TBI, one
receiving plasma injections without TBI, and one receiving saline injections without TBI.
The mice will be tested in the Rotarod test to assess their motor function. Then, brain
tissue will be collected and analyzed for neuronal cell death and restoration using cresyl
violet staining.

Hello! Today I will be discussing on the project, Motor Deficit & Brain Remediation Following traumatic brain injury, or TBI, with Plasma Administration in Mice. Traumatic brain injury is the result of a force on the brain strong enough to damage the brain either by hurting brain cells or its supporting structures. A common TBI known to the public is called a concussion.

If you have ever seen or heard about the consequences of a concussion, you can only imagine what can happen to people who gain multiple traumatic brain injuries over their lifetime. One TBI raises one‘s chances of getting another one. Repeated TBIs can lead to cognitive and motor impairments.

Prevention of TBI would be best, but that is not very realistic, especially as we age and our risks for physical accidents increase. Therefore, I propose looking at how blood plasma from young mice can ameliorate the motor deficits seen from repetitive mild TBIs in adult mice.

Young blood plasma has properties that give it its ability to help a damaged brain. Namely, platelet factor 4 has been shown to reduce inflammation and activate immune supporting cells in the brain. More research needs to be done to assess what other qualities of young blood plasma give its ability to rejuvenate the brain.

Plasma administration to TBI mice will either occur 30 days after the last repetitive mild TBI wasreceived or immediately after the last repetitive mild TBI received. Two other groups of mice will not have TBI and will either have just plasma administration or neither a TBI nor plasma administration. A controlled repetitive mild TBI apparatus will be used to administer the TBIs to the mice while they are under anesthesia. The TBIs will be administered every two days for ten days total. The pre prepared young blood plasma will be administered 24 hours after the last TBI.

The Rotarod test, seen here, will then assess the mice‘s motor abilities by encouraging the mice to stay on a rod due to their innate response to avoiding a fall. The Rotarod test is designed by having a long rod rotate at a certain speed and the mice are timed to assess how long they can remain on the rod.
The mice are placed on the Rotarod (show and fast forward if necessary). Usually there are two people adding mice to the rotarod at the same time to ensure the accurate amount of time the mice were able to stay on the rotarod.

The rod will accelerate over time so that the mice are assessed for speed at which they can maintain staying on the rotarod.

The time it takes for the mice to get off the rod is recorded by the apparatus because the platform below the rod will descend from the weight of the mice and the apparatus will detect this.

Once all of the mice are no longer on the rod they are returned to a transport cage and finally to their home cage!

If motor skills show improvement, it brings us closer to recognizing the potential of young blood plasma, which may someday be applicable to humans. The avenue of research opens new possibilities for treating irreversible conditions such as TBI. Once the scientific community has gathered thorough data demonstrating the advantages of utilizing young blood, we could have a potential new treatment for neurodegenerative diseases.

Categories
College of Engineering and Computing College of Science OSCAR

Optimizing Graphene-Metal Composite Production With Laser Modeling

Author(s): Diego Espinoza

Mentor(s): Pilgyu Kang, Department of Mechanical Engineering

Composite materials are engineered materials made from two or more different composite structures which change physical or chemical properties. Covetics are a type of composite material where graphene is integrated into a metal; these materials show improvement in mechanical strength, electrical and thermal conductivity and increase in material durability. Currently, the formation of covetics has been made using an electric arc method, which gives a non-homogenous graphene distribution, making it more difficult to optimize. Laser-Induced Graphene (LIG) presents a promising alternative. This process employs a laser and a polyimide (PI) film to create porous graphene, infusing it into a metal substrate, resulting in covetic material. Unlike traditional methods, LIG allows for a more controlled and homogenous structure. However, the optimal laser parameters, such as power, wavelength, speed, frequency, and fluence, remain unknown, impacting the quality and efficiency of porous graphene formation in covetics. This research addresses the gaps in understanding the laser parameters’ influence on LIG by developing a photothermal processing modeling of the process. The simulation focuses on correlating laser parameters to the heat distribution during the LIG process. By simulating the thermal changes of the material, the project aims to streamline the determination of optimal laser parameters, bypassing the need for repetitive physical experiments. This computational approach offers a more efficient and cost-effective means of exploring the intricate relationship between parameters and covetic material quality.

Hello, my name is Diego Espinoza and I am physics student at George Mason university working with Dr. Pilgyu Kang. Today I will talk about the work I have been doing under him. Optimizing Graphene Metal Composite Production with Laser Modelling.

We are going to start with defining a composite material, which are engineered materials made from two or more different materials. This new material will have new physical or chemical properties.

A good example of one is steel, which is the combination of Iron and Carbon. One process of making steel is by the use of a furnace which reaches extremely high temperatures and the carbon and iron are mixed.

This composite material obtains improvements in properties like strenght and thoughness.

Now that we know what composite materials are, we are going to see a type of composite material which is obtained by integrating graphene, a 2d layer structure of carbon atoms, into a metal. This new materials can show improvements durability, mechanical strenght, thermal and electrical conductivity.

A way of forming this composite material is by the used of a laser, which can be compare to a furnace for the case of steel. This method allows for a more controlled and homogenous structure. However, the optimal laser parameters like its power, the speed, the lasing time or the frequency are not known. Everytime a study is done, the people working with the materials have to start from 0, constantly creating different samples with different parameters and studying them to see which ones are better.

The project we have been working on intends to adress these challenges. By relating the temperature distribution to the formation of the induced graphene in the metal, we are working on computer simulation that shows us the temperature distribution across the material during the lasing process. During the process we study and used the material properties and how some of them can be changed during the temperature changes. Currently we are still working on the modelling and we are improving it week by week. The overall objective is to have a computational model that, by just changing the materials or the laser parameters, a good approximation of the temperature distribution, and therefore the graphene formation can be obtained.

Categories
College of Science OSCAR

Parasite Prevalance in Crab and Shrimp Hosts in Two Rivers of the Chesapeake Bay

Author(s): Amani Khan

Mentor(s): Sarah Goodnight, Environmental Health and Safety

Parasites are critical components of our ecosystems and impact industries such as fisheries and agriculture, yet there is little information known about them. This research project specifically examined a multi-host digenean trematode parasite in white fingered mud crabs (Rhithropanopeus harrisii) and grass shrimp (Palaemonaetes pugio). These hosts are the second in the parasite‘s life cycle and present as cysts called metacercariae (i.e., the juvenile developmental stage). We aimed to find any patterns between cyst prevalence and host traits (e.g., size and sex). The crabs also were infected with other parasites, one of which is the rhizocephalan barnacle Loxothylacus panopaei, and we were also interested the interactions of parasites within the host. Crabs were collected from submerged oyster shells in a crate that were attached to docks in two rivers. Shrimp were collected from the same locations with a dip net. Hosts were collected monthly from October 2023 to March 2024. Data such as size, sex, number of cysts, and other parasitic infections was collected. Preliminary data shows that larger crabs and male crabs had higher cyst infection rates. Crabs infected with L. panopaei had higher cyst infection rates, as well. This study will be part of a broader, year-long study researching parasite prevalence seasonally in these hosts of this estuarine ecosystem. Additional laboratory work will explore the relationships between these parasites within the crab hosts, with the aim to identify the directionality of those relationships.

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Hi, my name is Amani Khan. And today I’m

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going to be talking about the Oscar

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project that I did, which is parasite

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prevalence and crab and shrimp hosts in

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two rivers of the Chesapeake Bay.

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And let’s get started. So this is

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our crab host Rhithropanopeus

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Harrisii, which is a mud

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crab. And this is our shrimp host,

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Polymonaetes pugio, which is a grass

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shrimp. So we noticed a

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couple cases of these entoniscid

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isopods which can release larvae.

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And these are a parasitic. Of course, we

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also had these loxothylacus rhizocephalans

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, which is an external parasite

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that sits on the apron of the crabs. You

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can see the aprons right down there. And

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this large, you know, white yellow

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ball is the. Parasite,

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it reaches into the crab’s

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body and it takes away the reproductive

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autonomy of the

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crabs. And then moving over to the shrimp

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here we also noticed a couple cases of

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the Bopyrid isopod,

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another parasite. And of course there

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just weren’t a ton of cases of these two

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isopods. So we didn’t really focus on

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that too much And our main parasite that

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we focused on were these digenean

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trematodes, if you can see here these.

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Clear little white balls. And there’s

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also this large white cyst, which is

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another type of trematode, another

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species of trematode. And to clarify, a

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trematode is a parasitic worm.

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So why are we studying parasites? Well,

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they’re critical components of the

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ecosystem and they’re very important for

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biodiversity. But there’s really not a

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lot that’s known about their genetic

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sequences or even their life cycle. And

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as you can see here, this trematode cyst,

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it is a multi host parasite. So

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it moves through about 3 hosts to

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complete its life cycle. But there’s not

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too much known about the life cycle.

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There’s really no published data on the

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genetic sequences. So we’re hoping to

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contribute some knowledge, hopefully. And

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fill fill in some gaps.

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OK, so now for my

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methods.

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We went out about one to two

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times a month to collect crabs and

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shrimp. We set up these crab condos,

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which are basically crates full of

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cleaned and dried oyster shells, and

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they’re deployed in the water. Crabs can

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move in and out of them so that they’re

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so they’re not really traps. But we

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pulled them out of the water, dumped it

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onto a sieve, poured it with water to

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clean it, clean off the shells of some

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mud. And then we picked out about 100

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crabs from the two sites that we

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had. Our two sites were June Parker and

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Boathouse Marinas. When we

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got to the lab, we put the crabs

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in these jewelry boxes in their own

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individual cells.

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And we filled them with water. The shrimp

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were just put in a bucket full of water.

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When it was time for dissection, they

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were frozen for about 20

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minutes. And then we took some host

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traits such as tail to rostrum

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length with the shrimp and carapace width

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for the crabs. And we also sexed them.

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This was all recorded in a data sheet.

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Then we dissected them.

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And we counted the cysts and then we put

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the cysts into a tube for

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DNA extraction and PCR. As you can see

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here, we use the CTAB protocol for

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the DNA extraction. And you can see here

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I’m loading a gel for gel

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electrophoresis with some DNA.

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And we sent off our samples to Ps

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omagen for DNA sequencing when

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we receive the samples back.

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We put them intoGeneious to

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sort of clean up the DNA

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sequences.

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And then we loaded them into

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GenBank. GenBank is a software that

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is used where you can, you know, put some

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DNA sequences into it and hopefully match

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it to a species.

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Unfortunately, we couldn’t find any

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specific species, b

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ut that’s the way it goes sometimes.

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And. Now let’s look at the results.

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OK, as you can see here, I’ve created a

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haplotype network, and haplotype

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networks basically tell us about DNA

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variation.

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So in this green box over here, this

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represents our clear

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cysts. The bigger circles

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mean those individuals that have

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it’s bigger circle means more individuals

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had that haplotype and

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the smaller circles mean less

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individuals had them. These tick marks in

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between each of the lines that

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that indicates more nucleotide

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differences in the DNA sequences. So as

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you can see by both of

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these a

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reas thatthe cyst

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s were not clonal, so they are not

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genetically identical. It’s kind of like

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if one cyst or one person, I

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should say, has brown hair and one

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has blonde hair. That doesn’t mean that

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they’re different species. They’re just

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slightly different. They have different

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polymorphisms. As you can see by

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these two large lines here. That’s a lot

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of differences in nucleotides. So

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these are our white cysts.

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And if you can take a look at this red

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box here. This has a

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lot of nucleotide differences from the

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rest of the white cysts, so this

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might actually mean that it is a

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different species of trematode, so

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that we might actually have. We

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believe this is like cryptic diversity

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where, you know, the white cysts they all

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look the same under the microscope, so

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you can’t really tell if they’re

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different species. But this is telling us

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that yeah, you know, like you might

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actually have a different species of

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white cyst on your hand. And

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I should clarify that most

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of the sequences this is about it was a

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small sample size. It was about 19

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individuals, 18 of which were shrimp and

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one was a crab.

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This just reiterates my points from the

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last slide and these are some g

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enetic diversity metrics. Morphotype A is

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our white cyst and morphotype B is our

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clear cyst. As you can see, there’s more

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diversity, bigger numbers on the top than

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on the bottom, so there’s more diversity,

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and that’s probably due to the one white

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cyst that was found that may be of a

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different species.

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And these are some graphs. This is just

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saying that larger shrimp and larger

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crabs had more trematodes,

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which makes sense if you think about it.

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I mean, if they’re bigger, they’re

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probably older and they’ve had more time

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to accrue more infection. So that’s

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probably why they have more cysts. And

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then lastly, this is another picture of

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the loxo, the Loxothylacus

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rhizocephalan. Crabs that

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had the loxothylacus rhizecephalan

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They also had more

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trematode cysts, which is interesting

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because this may speak to a potential

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facilitation mechanism

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going on. One parasite may infect the

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crab and suppress the immune response of

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the crab, allowing the 2nd parasite to

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come in and invade. Or it may

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mean that you know crab was just at the

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wrong place at the wrong time and got hit

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with two parasites at once. So

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those were my results. Thank you to Mason

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and East Carolina University for funding

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this project as well as our lab and field

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assistance. And these are my references.

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Thank you.

Categories
College of Science OSCAR

Reversing Neuronal Cell Damage Through Mitophagy Induction

Author(s): Sofie Strompf

Mentor(s): Ali Andalibi, School of Systems Biology

Familial Dysautonomia (FD) is caused by the single point mutation of the major signaling scaffolding elongator protein IKAP/ELP1. This disease disrupts the autonomic nervous system, leading to decreased sensory perception, poor reflexes, inhibition of vascular and cardiac functions, orthostatic hypotension, and recurrent severe autonomic crises. Patients with FD experience increased incidence of severe adverse reaction and increased neuropathy after anesthesia. Neuropathy is associated with mitochondrial damage and increased cellular reactive oxidative species (ROS). Mitophagy is a cellular process that recycles damaged or dysfunctional mitochondria. We hypothesize that increasing mitophagy or reducing ROS levels, will lead to greater neuronal cell viability. We will employ the dopaminergic LUHMES cell line and apply a mitophagy stimulator known as CCCP. We expect the cellular viability to be increased because CCCP will cull the defective mitochondrial pool as measured by MTT assay and Western blot for mitophagy markers (PINK1) and apoptosis markers (Caspase-3). Future directions of this experimental plan will include the addition of the anesthetic Ketamine. We hypothesize that a greater level of neuronal cell death will occur after Ketamine application. To counter this death, pre-treatment with CCCP or a ROS inhibitor will delay or prevent Ketamine induced cellular apoptosis. We envision patient pre-treatment with a ROS inhibitor or mitophagy stimulator may counteract anesthetic neuronal damage. Implications of this study could elucidate new methods in preventing anesthesia-associated nerve damage in FD patients as well as diabetic- and cancer-induced neuropathic conditions.

I’m Sofie Strompf and this is my OSCAR URSP Spring 2024 project. My research focuses on the role of mitophagy in anesthesia and neuropathy. The goal of this research is to, in the far future, allow us to pretreat patients who are predisposed to experiencing neuropathy following anesthesia with a mitophagy-inducing drug, to prevent or decrease the likelihood of their developing neuropathy. My mentor is Dr. Ali Andalibi, and I want to give special thanks to Drs. Marissa Howard, Lance Liotta, Jessica Roman, Alessandra Luchini, and Karen Lee, as well as Purva Gade, Matthew Gadziala, and Thomas Philipson for their mentorship. This research was funded in part by the George Mason University Office of Student Creative Activities and Research Undergraduate Research Scholars Program.
So, Familial Dysautonomia is a rare genetic neurodegenerative condition caused by the single point mutation of the major signaling scaffolding elongator protein IKAP/ELP1. This disease disrupts the autonomic nervous system, leading to autonomic distress and recurrent severe autonomic crises. These patients also experience increased incidence of severe adverse reaction and increased neuropathy after anesthesia. Anesthesia is known to induce mitochondrial stress. Neuropathy is associated with mitochondrial damage and increased cellular reactive oxidative species (ROS). This implies a correlation between the increased incidence of neuropathy following anesthesia in patients with FD and the known increase in mitochondrial stress during anesthesia. Mitophagy is a cellular process that recycles damaged or dysfunctional mitochondria. From this knowledge, our hypothesis is that increasing mitophagy or reducing ROS levels will lead to greater neuronal cell viability. Exploration of this hypothesis may elucidate new methods in preventing anesthesia-associated nerve damage in Familial Dysautonomia patients as well as diabetic- and cancer-induced neuropathic conditions. To be clear, this research is not only applicable to patients with Familial Dysautonomia””it is applicable to all patients experiencing neuropathy, especially following anesthesia. Familial Dysautonomia patients are a helpful control group for this experiment, as it is extremely likely that they will experience neuropathy directly related to induction of anesthetic; this occurs at far higher rates in this population than any other patient group, as far as is currently known.
This project is a continuation of my Fall 2023 OSCAR URSP Project. Last semester, I specifically focused on learning skills and techniques that I need to work in biological research and gained valuable certifications that I need to work in a laboratory setting safely.
This semester, I utilized my knowledge of laboratory skills to complete experiments. I utilized skills in cell passaging, cell viability, luminescence imaging, drug dosing protocols, and western blots to do so. The experiments mostly utilized the IOMM-Lee cell line, a malignant meningioma cell line derived from the human brain. In my most successful, complete experiment, the cells were maintained in DMEM-FBS 10% media. They were then plated with control, control-DMSO, varying concentrations of heregulin beta 1″”a tumor growth polypeptide””and CCCP, a known mitophagy stimulator. Half of the wells were imaged for cell viability; half were lysed. Cell lysate was employed in a drug dosing procedure which introduced phosphorylated PINK1 with Actin as a control. A Western blot was utilized to measure P-PINK1 and Actin after 24 hours. This procedure was the first of a series of planned procedures measuring mitophagy markers. This semester, I also had the incredible opportunity to present my current research at the National Conference on Undergraduate Research, or NCUR, in Long Beach, California.
In the future, continuing to use the IOMM-Lee cell line and also introducing the dopaminergic LUHMES cell line, we will continue to measure mitochondrial dysfunction following application of CCCP. We will continue to measure mitophagy markers, including PINK1 and Parkin, and the apoptosis marker Caspase-3, utilizing MTT assay and Western blot. We will also attempt to measure markers of cellular stress associated with induction of anesthesia in order to compare this preliminary analysis of mitochondrial stress with data derived from future experimental processes. Future directions of this experimental plan will possibly include the addition of anesthetics to further explore our hypothesis. We expect that a greater level of neuronal cell death will occur after anesthetic application. To explore new methods in prevention and treatment of anesthesia-associated nerve damage, we will attempt to counter this expected increased incidence of cell death. We will pre-treatment with CCCP or a ROS inhibitor will delay or prevent anesthetic induced cellular apoptosis. We envision patient pre-treatment with a ROS inhibitor or mitophagy stimulator, which may counteract anesthetic neuronal damage.
Obviously, getting anywhere near pre-treatment of patients will happen very, very far in the future. So, in the near future, I plan to continue my experiments and present advances in my research at next year‘s NCUR conference and OSCAR celebration. I plan to continue to work in the Institute for Advanced Biomedical Research, in the Center for Applied Proteomics and Molecular Medicine, where I have been completing my research so far. I have applied for additional funding through outside sources and am hoping that I will be able to not only continue my research next year or through the remainder of my undergraduate career, but that I will be able to carry this research into my graduate career. This has been an incredible experience, and I look forward to continuing my research in the years to come.

Categories
College of Science OSCAR

The Effect of Cadmium on α1 Glycine Receptor Function in Xenopus Laevis Oocytes

Author(s): Diya Sabareesh Meethaleveettil

Mentor(s): Greta Ann Herin, Neuroscience

Cadmium is a known toxic metal and carcinogen commonly found in cigarettes (inhalation) and canned food (ingestion). While the national geometric mean blood cadmium level for adults via inhalation was recommended to be close to 0.4 µg/L, a mean blood cadmium level of 1.58 µg/L was found in a study conducted on heavy smokers. It was also found that smoking one pack of cigarettes per day exposed the human body to 1-3 µg of Cd2+/day, where the cadmium levels in tissues doubled for smokers compared to that of non-smokers. Similar data has also been found for cadmium taken orally; whilst the CDC recommends only up to 0.1 ug/kg/day of Cd2+, studies have found that canned foods contain up to 0.5-0.6 mg/kg of Cd2+. Both these routes of transmission cause adverse respiratory, cardiovascular, renal, skeletal, and developmental effects. Even though most of cadmium‘s toxic effects have been studied, much of its mechanism of effect on neuronal receptor function remains unknown. To address this gap, this project aims to determine if cadmium regulates glycine receptor function in a concentration-dependent manner. This will be done by microinjecting Xenopus Laevis oocytes with the RNA of α1 glycine receptor. Post-expression of receptors, two electrode voltage clamps (TEVC) technique will be used to study the transmembrane current caused by Cl- influx through GlyRα1 receptor on exposure to varying concentrations of Cd2+.

Hi everyone! My name is Diya Sabareesh Meethaleveettil, I‘m here to talk about my project ““ the effect of cadmium on glycine receptors of a1 subtype (GlyRα1) in Xenopus Laevis oocytes. I‘m doing this project in Dr. Herin‘s lab. Without further ado, let‘s get right into it! Cadmium is a highly toxic metal and a known carcinogen that is alarmingly present in cigarettes and canned foods. Many compounds of cadmium are highly soluble in water which makes it easy for them to enter the human body and pose health risks. To understand the role of cadmium, we first need to understand the function of glycine receptors. Glycine receptors are essential for inhibitory neurotransmission. They are quite important in the pain pathways and motor function in the body and are predominantly present in the spinal cord. When glycine neurotransmitter binds to the receptor, it opens the channel and allows chloride influx which will hyperpolarize the cell and decrease the propagation of action potentials. The purpose of the project is to investigate how cadmium affects GlyRα1 expressed in Xenopus Laevis oocytes and this is what the oocytes look like. My hypothesis is that cadmium will inhibit the glycine receptor function in a dose-dependent manner which disrupts the inhibitory effect normally provided by the receptor. I say this because cadmium is structurally similar to zinc which is a known allosteric inhibitor of glycine receptor. Zinc allosterically binds to glycine receptor which changes its conformation and prevents glycine from binding to the receptor. This either causes the channel to completely close or open less which will decrease the flow of chloride through the channel and ultimately decrease its inhibitory effects. This is a picture of zinc and cadmium, which are in the same group in the periodic table, and you can see how structurally similar they are except for zinc missing one shell of electrons. The methodology for this project first starts with the replication of GlyRα1 cDNA and then transcribing it into an mRNA using a transcription kit. We then check the quality of mRNA obtained using spectrophotometry and gel electrophoresis before injecting it into an oocyte. Spectrophotometry allows us to quantify nucleic acid concentration and its purity whereas gel electrophoresis allows us to confirm the absence of impurities. 0.5uL of mRNA will then be injected into each oocyte and we will then incubate it at 30 degrees Celsius for 2-3 days. We then use our Two-electrode voltage clamp or TEVC rig to measure the electrical activity in the oocyte. This technique will clamp the voltage using a continuous supply of chloride ions from a solution of KCl by the current electrode, which will allow us to study only the current passing through the oocyte. This is a picture of how transcription occurs, that is reducing our DNA to mRNA. The other pictures are of the microinjection rig in my lab and also the pipettes we use for injection. We will record data for three different solutions: a physiological saline solution which is ND96 for baseline recording, then with ND96 and glycine which will activate GlyRα1, and then with ND96, glycine, and cadmium chloride to assess how cadmium alters receptor activity. For the solutions with glycine, I would take 0.1mM glycine which is the standard concentration, and for cadmium solution concentrations, I would take 1nM, 10nM, 100nM, 1uM, and 10uM. Hypothetically we should be starting to see an effect at 10nM. This will be done on three types of oocytes ““ uninjected oocytes, water-injected oocytes, and mRNA-injected oocytes which will be the only ones expressing the receptor. This is a picture of the TEVC rig I worked on to obtain the data and to analyze them. This is a picture of the data obtained from a water-injected oocyte that was perfused by ND96. These are the same graphs just zoomed in and out. The top left axis is the current across the membrane in uA, the bottom left axis is the cell‘s voltage in mV, and the X-axis is time in minutes. There aren‘t many changes to the peaks shown here as I haven‘t used cells expressing GlyRα1 yet and also have perfused the oocytes with only one solution. I have been able to get data from 4 oocytes but as you can see there has been some noise, which are the bidirectional changes in amplitude, due to some technological errors which has also made me unable to obtain data for the rest of the oocytes. The noise can also be seen in how the voltage changes, that is different peaks, instead of remaining constant. But we have been able to troubleshoot and correct our rig so I will be able to get more data in the next few days. We should also be aware of the broader impact of cadmium on human health. Inhalation of cadmium from tobacco and ingestion through canned foods can lead to adverse effects on the respiratory, renal, cardiovascular, digestive, and skeletal systems. This signifies the importance of understanding how cadmium interacts with biological processes including receptor function. And as you can see in this chart, cadmium‘s effect on our nervous system via neuronal receptors hasn‘t been extensively studied which is what I aim to find out through this project. Here are my references. And finally, I would like to express my gratitude to Dr. Herin for her continuous support and mentorship throughout this project and my time in her lab. I would also like to thank the OSCAR program for providing me with the funding in the form of the URSP grant. Thank you so much!

Categories
College of Science

The Effect of Anesthesia on Signal Transduction Pathways in Cells

Author(s): Sofie Strompf

Mentor(s): Ali Andalibi, Systems Biology

https://youtu.be/bxDXstmAr8o

The NF-kB inflammation pathway, the JNK cell proliferation and apoptosis pathway, andtheMerlin/NF2 cell growth pathways are essential in the regulation of virtually all major cellular functions and processes. However, little is known with regards to how the signaling scaffolding protein IKAP, a subunit of the Elongator complex, regulates these pathways, both with and without oxidative stress. When IKAP is mutated, its regulation of these pathways is dysfunctional, potentially causing neuronal degradation or death. When mutated IKAP iscombined with oxidative stress induced by anesthesia, these pathways are more upregulated, further harming the neuronal tissue. This studywill complete an in vitro investigation of the effects of a commonly employed dissociative anesthetic on cellular functions implicated in several phosphorylation signaling pathways. Data analysis and interpretation may elucidate new methods in preventing anesthesia-associated nerve damage in patients undergoing surgery.

I’m Sofie Strompf and this is my OSCAR URSP fall 2023 project. My research focuses on the effect of anesthesia on signal transduction pathways in cells. My mentor is Dr. Ali Andalibi, and I want to give special thanks to Dr. Marissa Howard, Purva Gade, Matthew Gadziala, Dr. Alessandra Luchini, and Dr. Lance Liotta. So, the Nuclear Factor Kappa-B inflammation pathway, JNK cell proliferation and apoptosis pathway and the merlin/neurofibromatosis-type 2 cell growth pathways are essential in the regulation of virtually all major cellular pathways. However, little is known with regards to how the signaling scaffolding protein IKAP regulates these pathways, both with and without oxidative stress. When IKAP is mutated, its regulation of these pathways is dysfunctional, which potentially causes neuronal degradation or even death. When mutated IKAP is combined with oxidative stress induced by anesthesia, these pathways are more upregulated, which further harms the neuronal tissue. My study will complete an in vitro investigation of the effects of a commonly employed dissociative anesthetic, ketamine, on cellular functions implicated in several phosphorylation signaling pathways. Data analysis and interpretation may elucidate new methods in preventing anesthesia-associated nerve damage in patients undergoing surgery. This semester, I specifically focused on learning skills and techniques that I need in order to work in biological research. I completed my research at the Institute for Advanced Biomedical Research on George Mason University’s Science and Technology campus. I worked in the Luchini Lab, as well as the Center for Applied Proteomics and Molecular Medicine. This semester, I also gained valuable certifications that I need in order to work in a laboratory setting safely. I also learned more, non-laboratory specific skills. I few things that I want to highlight are my learning of writing an IBC protocol, which was necessary in order to obtain the anesthetic that we are using in the next stage of research in the spring. Additionally, my knowledge in writing an abstract is important in grant proposal writing, as well as conference applications, so that I can continue my research, and share my research, in the future. So, what’s next? In the spring, I am going to continue my research with the OSCAR URSP program. Specifically, I will begin working with neuronal cell lines. Currently I am working with IOMM cell lines and next semester I’ll work with more complicated, more difficult cell lines. We’ll also begin working with the anesthetic and doing specific experimental research with that. We’ll complete data analysis and data interpretation, and I will share my research via conferences and a paper.