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Poster Session Abstracts

Alumni Abstracts

Clonally related, Notch-differentiated spinal neurons integrate into distinct circuits

Saul Bello-Rojas ’16 ¹ and Martha W. Bagnall ^1
1Washington University in St. Louis, Neuroscience, St. Louis, MO, 63110, USA

Shared lineage has diverse effects on patterns of neuronal connectivity. In mammalian cortex, excitatory sister neurons assemble into shared microcircuits, whereas throughout the Drosophila nervous system, Notch-differentiated sister neurons diverge into distinct circuits. Notch-differentiated sister neurons have been observed in vertebrate spinal cord and cerebellum, but whether they integrate into shared or distinct circuits remains unknown. Here we evaluate the connectivity between sister V2a/b neurons in the zebrafish spinal cord. Using an in vivo labeling approach, we identified pairs of sister V2a/b neurons born from individual Vsx1+ progenitors and observed that they have similar axonal trajectories and proximal somata. However, paired whole-cell electrophysiology and optogenetics revealed that sister V2a/b neurons receive input from distinct presynaptic sources, do not communicate with each other, and connect to largely distinct targets. These results resemble the divergent connectivity in Drosophila and represent the first evidence of Notch-differentiated circuit integration in a vertebrate system.

Evaluation of EZ reflectivity normalization across different OCT devices

Heather Heitkotter ’16 ¹, Koua Vang ², Mina Gaffney ³, Joseph Carroll ^1,2
1Cell Biology, Neurobiology & Anatomy, ²Ophthalmology & Visual Sciences, ³Biomedical Engineering, Medical College of Wisconsin, Milwaukee, Wisconsin, United States, & Biomedical Engineering, Marquette University, Milwaukee, Wisconsin, United States

Purpose : Ellipsoid zone (EZ) reflectivity has potential to serve as a biomarker of photoreceptor integrity, but there is variability in the methods used to extract reflectivity values. Here we evaluate normalization methods of EZ reflectivity across four different OCT devices. Methods : Horizontal 6mm line scans (20 averaged B-scans) were acquired in five control eyes using the Cirrus, Bioptigen, Optovue Avanti, and Spectralis OCT devices. Eight longitudinal reflectivity profiles (LRPs; each 7 pixels wide by 250 pixels in depth) were extracted using ImageJ from each OCT between 0.5–2.0mm (0.5mm intervals) from the fovea. Boundaries of the ganglion cell and inner plexiform layers (GCIPL) and the peak and full width half max (FWHM) of the EZ were identified. Intensity values between the GCIPL boundaries were averaged. Intensity values between the EZ FWHM were averaged, and peak EZ intensity was extracted. Values were averaged across LRPs for each scan. The average intensity of a 300x50 pixel vitreous region was also extracted from each scan. Four intensity normalization methods were evaluated with ICC and Friedman’s test: 1) peak EZ, 2) EZ FWHM, 3) GCIPL normalization (EZ FWHM/GCIPL), and 4) vitreous normalization (subtract vitreous from 3 before taking ratio). Results : The ICC for mean peak EZ (-0.11) and mean EZ FWHM (-0.09) intensity across devices were poor. Friedman test revealed significant differences in peak EZ intensity across devices (Q(3)=86.2, p<0.0001). Dunn’s multiple comparisons test indicated peak EZ differed across all devices except between Optovue vs Spectralis. Mean EZ FWHM across devices also significantly differed (Q(3)=66.0, p<0.0001), and Dunn’s test revealed values from the Bioptigen were significantly different from the other devices (p<0.0001). The ICC for GCIPL normalization across devices was poor (-0.21), and Friedman test revealed significant differences across devices (Q(3)=13.6, p=0.0001), including Spectralis vs Bioptigen (p=0.009) and Spectralis vs Optovue (p=0.018). The ICC for vitreous normalization across devices was poor (0.042), with significant differences revealed by Friedman test (Q(3)=11.2, p=0.002), specifically between Bioptigen vs Optovue (p=0.009). Conclusions : Our data demonstrate that the normalization method alters the relationship of EZ reflectivity across devices. Such variability poses challenges to utilization of EZ reflectivity as a biomarker in clinical studies.

Implications of stress exposure on striatal gene expression and consequent running behavior impairment in young adult rats

Trevor J. Buhr ’18 ^1,2,6, Carter H. Reed ^2,3,6, Ella E. Bauer ^1,2,6, Allyse Shoeman ^1,2,6, Rudy J. Valentine ^3,6, Zack V. Johnson ^5,7, Lynna Chu ^4,6, Peter J. Clark ^1,2,6
Department: ¹Interdepartmental Neuroscience Program, ²Department of Food Science and Human Nutrition, ³Department of Kinesiology, ⁴Statistics Program, ⁵Department of Biological Sciences. ⁶Iowa State University, ⁷Georgia Inst. of Technology.

The World Health Organization lists physical inactivity as the 4th leading risk factor for global mortality. Therefore, identifying the factors that contribute to a reduced willingness to be active may be central to the development of new approaches that increase physical activity. While a growing body of evidence implicates psychological stress as a contributing factor to the sedentary lifestyle, little is known about how stress exposure may alter the brain in a manner that makes individuals more prone to physical inactivity, especially long after the stressor is no longer present. We have found that rats exposed to a single episode of uncontrollable tail shocks (acute stress), 36 hrs before being granted access to running wheels, display at least a two-month decrease in motivation to engage in physical activity, as revealed by a potently reduced daily wheel running distance. Running deficits cannot be explained by changes to eating or body weight, or the expression of depression- and anxiety-like behavior, following acute stress. To identify molecular changes and multicellular pathways that may contribute to reduced engagement in physical activity, we performed single nuclei RNA-seq in the striatum area of young adult rats that were or were not exposed to stress and then subsequently given access to running wheels for 42 days (2x2 design, n=6/group). The striatum was targeted because it has recently been shown to be involved in the acquisition and maintenance of wheel running behavior, and is also sensitive to stress exposure. Data collection is preliminary and ongoing, but Seurat single cell software has initially derived 15 distinct clusters of genetically similar cells, which could be further divided into 35 clusters. Across each cell cluster, a considerable number of differentially expressed genes (DEG) from exercise (i.e. 3539) and stress exposure (i.e. 5767) was observed (q<0.05). Next we use CellChat to estimate strength of molecular communication across cluster pairs and found exercise enhanced cell-cell communication in 19 cluster pairs (with none decreasing in strength. Cell-cell communication will next be tested in stressed conditions. Moreover network mapping of genes within and across clusters will be performed to identify biological pathways influenced by exercise and stress in an in effort to understand factors driving differences exercise motivation.

Observed Intersectionality in the Association between Neighborhood Socioeconomic Deprivation and Obesity

Lauren Giurini ’20 ¹, William Blot PhD ², Loren Lipworth PhD ², Harvey J Murff MD ², Wei Zheng MD PhD ², Shaneda Warren Andersen PhD ^1
1University of Wisconsin-Madison School of Medicine and Public Health, ²Vanderbilt University Medical Center

Obesity and low neighborhood-level SES are more common among Black Americans and individuals with low individual-level SES. We examined the association between neighborhood SES and BMI using data from 80,970 participants in the Southern Community Cohort Study, a cohort that oversamples Black and low socioeconomic status participants. Thresholds from the CDC determined BMI categories, and neighborhood SES was measured by a Neighborhood Deprivation Index (NDI) composed of census-tract variables in the domains of education, employment, occupation, housing, and poverty. Overall, participants were more likely to be obese or overweight in lower SES neighborhoods. We found evidence of interactions by race, sex, and smoking status with NDI where white females and former smokers had the highest odds of obese or overweight status, typically in the lowest SES quintile of NDI. These findings suggest that certain groups are more susceptible to the health effects of neighborhood SES and provide

Current Student Abstracts

Defining the Anatomical Structure of the preBötzinger Complex

Katarzyna Barton ‘24 & Dr. Kaiwen Kam, Department of Cell Biology and Anatomy, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

The preBötzinger complex (preBötC) is a cluster of interneurons located in the ventral respiratory group of the medulla that is essential for the generation of the respiratory rhythm in mammals. Despite its well-understood functional role, the anatomical borders of the preBötC remain poorly defined. We hypothesized that the spatial distribution of GABAergic and glycinergic neurons might help map the location of the preBötC. Tissue histology, thionin staining, and immunohistochemistry were used in brainstem sections from neonatal mice to obtain a more detailed map of the preBötC and surrounding regions. We found that the neurons present within the preBötC coexpress for both GABA and glycine and allow for the borders of the preBötC to be identified more accurately.. We conclude that the GABAergic and glycinergic neurons remain critical for determining the borders and location of the preBötC..  Future research will focus on further mapping of the preBötC and further investigation into the interneurons that make up the preBötC.

Identifying art pigments by the usage of Surface-enhanced Raman Spectroscopy

Naina Beishembieva ’25, Anastasija Rudan ’24, Lira Zajmi ’25, and Dr. Nilam Shah, Chemistry Department, 91��Ƭ��, Lake Forest, IL 60045

One of the most significant challenges in the conservation of art is the identification of organic dyes and art pigments. The purpose of this project is to detect art pigments and determine the best experimental procedures for identifying molecules in low concentrationsusing a technique called Surface-enhanced Raman Spectroscopy (SERS).  Surface-enhanced Raman spectroscopy is a promising technique for detecting dyes on historical textiles because it requires very little sample compared to other analytical procedures In order to be able to see SERS spectra, we need to use a surface with nanoparticles. There are many different types of nanoparticles that have been fabricated through research. They all have LSPR (Local Surface Plasmon Resonance).  Depending on their size, nanoparticles change their colors and have different LSPRs.  In our research, we chose silver colloids as our nanoparticles. We then adjusted our laser to the LSPR value of our nanoparticles.  Produced nanoparticles were aggregated in order to have a higher concentration of nanoparticles in the solution.  There are two ways of aggregation that we utilized and examined their differences. The first method uses colloid paste produced by colloid nanoparticles centrifuged 10 times. The colloid paste has a significantly higher concentration of nanoparticles within it. The second method consists of aggregated colloid paste, which makes the concentration of nanoparticles in the paste even higher. The colloid paste is mixed with Potassium Nitrate, and it results in an aggregated colloid paste. We also examined the role of pH in the identification of peaks.It was found that pH 2 is the optimum pH for carminic acid. Indigo data was not conclusive and further studies need to be conducted. In the future, a broader range of pHs and different nanoparticle synthesis methods will be examined. 

Identifying a new component of Tritonia’s distributed memory for sensitization.

Margaret A. Boersma ’24, Viral K. Mistry, Evan S. Hill, Jeffrey W. Brown, William N. Frost, Stanson-Toshok Ctr. for Brain Function and Repair, Rosalind Franklin Univ. of Med. and Sci., North Chicago, IL

The marine mollusk Tritonia diomedea jumps vertically up off the substrate and executes an escape swim in response to an aversive skin stimulus, such as contact with the limb of a predatory sea star, or a concentrated salt solution. An initial stimulus induces sensitization, a form of nonassociative learning, in which the animal displays a faster swim onset and higher jump height the next time the stimulus occurs. Here, we confirmed the prior observation of jump height sensitization, and further found it to reverse and undergo habituation with repeated stimulation. One noteworthy feature was that the jump height learning curve had the same profile as that for sensitization and habituation of swim onset latency, suggesting common underlying mechanisms. To begin characterizing the neural basis of this learning-related change in jump height, we analyzed large-scale optical recordings of neuronal activity in Tritonia’s pedal ganglion during sensitization of the nerve-evoked motor program. We found that the firing of the ventral flexion neurons that generate the jump height increased in sensitized motor programs, identifying a potential cellular locus for this behavioral learning. We then compiled these optical recordings to begin constructing a functional atlas of the pedal ganglion neurons participating in the distributed memory for escape locomotion. Taken together, these results characterize a new component of Tritonia’s escape swim locomotion learning at a behavioral, network, and cellular level, and have potentially identified a new cellular locus of the animal’s distributed memory for sensitization.

Attentional priority map in the posterior inferotemporal cortex (PIT) as a target for PTSD treatment.

Gabriela Chacón Mora  ‘24, Dr. Asante Kamkwalala PhD, Dr. Leah Rubin PhD MPH
Neurology Department, Johns Hopkins University School of Medicine, Baltimore, MD, 21218

Human’s behavioral goals are guided by an input of sensory information, which is carefully selected in order to focus our attention on specific goals. The ability to shift our attention, despite the presence of multiple stimuli, is known as selective attention- a process controlled by the brain frontal and parietal regions. Recent studies found that there are attentional modulating properties in the posterior inferotemporal cortex (PITd); a region usually associated with visual recognition.
Further studies determined that electrical stimulation of neurons in attentional priority maps facilitate the sections of objects with attention.  Furthermore, patients who have been diagnosed with PTSD usually report high levels of attentional deficits (or negative attentional bias) attributed to trauma-related cues. Based on these studies, electrically stimulating neurons in the PITd of patients with PTSD could potentially be used as target treatment to reduce attentional deficits.

Seed Production increases with Proximity between Flowering Lilium philadelphicum

Sophia Chen ’23, Jared Beck, and Stuart Wagenius, Department of Plant Biology and Conservation, Chicago Botanic Garden, Glencoe, IL 60022

Fragmentation in landscape poses a threat to the reproductive success of native prairie plants that are self-incompatible. Lilium philadelphicum, wood lily, is a self-incompatible perennial and requires diverse mates in order to successfully reproduce. Previous studies found that distance between neighboring plants influences reproduction in some prairie species, such as Echinacea angustifolia. This paper seeks to find similar effects by investigating the relationship between seed set and proximity of Lilium philadelphicum individuals. The sample size (n= 85) used in this experiment was located in western Minnesota. Seed pods from each individual were harvested to obtain seed data. The number of fertilized embryos proportion to the total embryos produced by each individual was quantified as seed set. In addition, GPS location of each individual was recorded for spatial analysis. The relationship between seed set and proximity of neighboring plants was analyzed with a generalized liner model. Results show the increase of proximity between individuals correlates with an increase of seed set. Proximity of an individual between its 5th nearest neighbor shows the most significant effect on the outcome of seed set. Furthermore, decrease in proximity of an individual and its neighboring plants correlates with a decline in seed set. In conclusion, isolation between individual flowering L. philadelphicum impedes its reproductive success. This study provides insight on the negative impacts of landscape fragmentation on native prairie plants. This insight can direct conservation efforts to ameliorate the effects of fragmentation in prairie habitats.

The Mechanism of Predation: Exploring the mechanosensory stimuli of Bumblebee Goby Brachygobius Doraie in detecting prey

Shrija Chhetri ’24, Djina Jovanovic '23, Violet Anderson '23, and Margot Schwalbe, Biology Department, 91��Ƭ��, Lake Forest, IL 60045

In animals, sensory systems are known to mediate a wide range of behaviors. For example, the complex mechanosensory lateral line system of teleost fish detects pressure changes, water flow directionality, and other behavioral abilities such as prey detection. The sensory system is composed of a series of mechanoreceptors called neuromasts arranged in a stereotypic pattern on the head and along the body. Here, we studied the Bumblebee Goby ( Brachygobius Doraie) which has a small and slender body size with a reduced lateral line system, and its feeding behavior. The goal of this study was to test the role of vision and the lateral line system during prey detection by performing trials in the light and the dark. This can be achieved by depriving the sensory capabilities, either vision or lateral line system, or both. In light trials, fish had both their vision and lateral line system, while in the dark trial fish no longer had vision and had their lateral line system intact. When conducting trials, individual gobies were placed in a circular tank and brine shrimp were provided as prey. Behavior was recorded with a camera positioned over the tank and the video was analyzed to quantify the behavior. The data from the trials were used to measure the differences in detection distance, reaction distance, detection angle, and reaction angle. The results demonstrated:
(1) gobies eat prey in both light and dark (2) gobies detect and consume prey in the dark using its lateral line system; (3) prey detection behavior is altered when feeding on the same prey under light and dark conditions, for example in light trials the prey is detected from a greater distance while in dark trials prey is detected from a much shorter distance. Therefore, this study suggests that prey detection behavior involves multiple sensory modalities; depriving the organism of a single sensory system would have a consequential impact on its ability for prey detection. For future experiments, the lateral line system will be ablated for both light and dark trials to further examine the role of the mechanosensory system in behavioral functionalities.

Extending the Therapeutic Applications of LAG3 in Multiple Sclerosis via Genomic Expression

Allison Coffell ’23 and Dr. Joseph Reynolds, Department of Cell Biology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, 60045

Multiple Sclerosis (MS) is an auto-immune mediated neurodegenerative disease of the central nervous system (CNS). Particularly characteristic of MS is the presence of demyelinating plaques in the pons, spinal cord, and periventricular area of the CNS. MS is a T-cell-mediated disease, meaning it causes the myelin sheath in the CNS to be attacked by immune system mediated T-cells, due to the body’s interpretation of these CNS cells as being exogenous immunogen, or “pathogenic.” Upon performing a genotyping experiment on patients diagnosed with Multiple Sclerosis, it was discovered that Lymphocyte-activation gene 3 (LAG3) present on chromosome 12p13 is under expressed. LAG3 is a gene discovered in 1990, was determined to be a part of CD223 (cluster of differentiation 223). It is a significantly important immune checkpoint receptor that serves as an antagonist for activated T cells in the immune system. It serves to inhibit lymphocyte activity and energy. We performed an shRNA experiment to examine the effect of decreased LAG3 on interleukin-17A (IL-17a), a proinflammatory cytokine produced by activated T cells. It was discovered that reducing expression of LAG3 increases expression T-cells with IL-17a, which in turn increases the production of inflammatory responses. A cloning experiment was performed on the LAG3 gene to overexpress it in T cells, resulting in an under expression of IL17-a, revealing a potential role of LAG3 in decreasing an overexpressed immune response, improving symptoms of MS.

Proximate Mechanisms of Evolutionary Trade-Offs in Bean Beetles ( Calllosobruchus maculatus)

Beth DeFoe ‘23, Desire U. Nalukwago, Iman Shepard, and Flavia Barbosa, Department of Biology, 91��Ƭ��, Lake Forest, IL, USA

Trade-offs in response to differential resource allocation typically present as negatively correlated traits, but little is known about their underlying developmental mechanisms. Dimorphism in bean beetles is a plastic response to varying larval densities during development, where dispersal morphs develop under high densities and flightless morphs develop under low densities. This is the result of differential resource allocation to wing and gonad size between densities, and thus a tradeoff between dispersal and reproduction. We investigate the possible role of juvenile hormones (JH) in the proximate explanation of these trade-offs, since JH plays a pivotal role in wing development and sexual maturation in other insect species. We treated beans with a JH antagonist (Methoprene) or JH agonist (Precocene), allowed beetles to develop under controlled density conditions and measured a suite of behavioral and morphological traits. We observed significant effects of treatments on behavioral traits as well as morphological traits. JH inhibitor treatment resulted in larger wings and smaller gonads for both sexes, indicating increased investment in dispersal, which was further supported by significant negative covariances between wing length and gonad size. These findings support the hypothesis that juvenile hormones mediate developmental trade-offs between traits in response to environmental pressure.

Functional dcsE Production as a preliminary step towards D-Cycloserine prophylaxis for Tuberculosis.

Stefanie Dejneka ‘25, Peter Pawlowicz ‘25, Laurel Robbins ‘23 and William Conrad, Chemistry Department, 91��Ƭ��, Lake Forest IL 60045

Tuberculosis (TB) is a respiratory disease caused by the bacterium Mycobacterium tuberculosis. According to the WHO, TB is the second leading cause of death by a single infectious disease behind COVID-19. While a vaccine and antibiotic treatments exist, rising problems of antibiotic resistance worldwide and logistical difficulties in distribution means the burden of TB remains significant around the globe. The goal of this project is to develop a genetic prophylaxis for TB using a six-enzyme biosynthetic pathway to produce d-cycloserine. D-cylcoserine (dCS) is a second-line antibiotic for TB that inhibits bacterial cell wall synthesis. The first committed step towards dCS synthesis is catalyzed by the enzyme dcsE which converts L-serine and acetyl-CoA to O-acetyl-L-serine and Coenzyme A respectively. We endeavored to produce dcsE in human cells and test its functionality as a preliminary step to understand if the dCS pathway could be an effective prophylaxis for TB in human cells. DcsE tagged with FLAG and GFP was overexpressed in A549 cells as determined using fluorescence microscopy. Cells were subsequently lysed, and crude lysate was reacted with L-serine and acetyl CoA to produce O-acetyl-L-serine to be detected using HPLC-MS. The research led to an undetermined conclusion due to constrained time limitations, however further research that was completed showed that dcsE synthesized in human cells was a functional enzyme capable of converting L-Serine and Acetyl-CoA to O-acetyl-L-serine.

Using the JAGB Method to Measure the Distance to the Galaxy NGC 6822

Coral Espinoza ‘23, Abby Lee, Wendy Freedman, Department of Astronomy and Astrophysics, University of Chicago, Chicago, IL 60637

The Hubble constant gives the expansion rate of the universe and constrains the neutrino mass.  It has been measured indirectly through three methods in recent decades using the Cosmic Microwave Background (CMB), Cepheid variable stars, and Tip of the Red Giant Branch (TRGB). These three methods give precise distances to galaxies but give slightly different results of the Hubble constant. The ongoing research of my team hopes to prove that the J-Band Asymptotical Giant Branch (JAGB) method is a precise method for calculating distance so that we could eventually use it to calculate the Hubble constant. I will go over my methods of finding the distance to galaxy NGC 6822 using JAGB stars and the distance which I found to be 0.52 Megaparsec (Mpc). I will also cover the implications of using the JAGB method to calculate the Hubble constant, and why it is important to current physics.

Role of Anti-inflammatory Prostaglandin 15d-PGJ2 in Kaposi’s Sarcoma Associated Herpesvirus (KSHV) Infected Primary effusion lymphoma (PEL) cells

Rabia Fatima 23’,  Nicole Rodriguez, Sarah Narula, Neil Mody Deshmukh, Barry Khim, Olivia Powrozek, Neelam Sharma-Walia, Department of Cellular and Molecular Pharmacology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Kaposi’s Sarcoma (KS) is  the most common cancer in HIV-infected untreated patients caused by the human herpesvirus 8 also known as associated herpesvirus (KSHV). KSHV infection is also  etiologically associated with a highly aggressive B cell malignancy called primary effusion lymphoma (PEL).  KSHV exhibits two phases in its life cycle as latent and lytic. Both phases are critical for the development of KS and PEL. KSHV infection creates a unique inflammatory microenvironment conducive to survival in the host. KSHV utilizes polyunsaturated fatty acid called Arachidonic acid (AA) and its downstream metabolites and receptors to maintain their successful life cycle in the host ( George Paul et al., 2010; Paul et al., 2011; Paul et al., 2013; Sharma-Walia et al., 2010; Sharma-Walia et al., 2014). AA metabolizes the pro-inflammatory cyclooxygenase (COX) pathway to produce prostaglandins (PGs). One of the downstream by-products of the AA pathway is 15-deoxy-∆12,14-PGJ2 (15d-PGJ2), also known as a cyclopentenone prostaglandin. 15d-PGJ2 is a natural anti-inflammatory agent with therapeutic properties.  Incidence rate of KS, KSHV virus load is higher in men than in women.  Previous reports showed a striking expression of androgen receptor (AR) in the nuclei of KS spindle cells at a very high level, compared with the sporadic cytoplasm distribution from control tissues ( Ding et al., 2021).  15d-PGJ2 serves as a potent and direct inhibitor of AR signaling in many cancers ( Kaikkonen et al; 2012) suggesting novel possibilities in restricting the AR activity in many cancer cells.  Therefore, we hypothesized that reintroducing 15d-PGJ2 back onto KSHV infected cells will inhibit androgen signaling.  Using Body-cavity-based KSHV-positive lymphoma cell line (BCBL) and KSHV-negative human B-cell line BJAB cells, we sought the role of  15d-PGJ2  treatment on  AR signaling pathway. We observed downregulation in the gene expression of membrane, intracellular and transcription factors of AR signaling pathway in 15d-PGJ2 treated samples. Downregulated genes are involved in tumor formation, cell proliferation and inflammation.  In conclusion, our results  show that  15d-PGJ2  has the potential to act as a potent and direct inhibitor of AR in KSHV infected cells thus providing a novel lead for targeting druggable AR in KSHV infection associated cancer cells.

Murine Hepatitis Virus and COVID-19: A quantitative approach to understanding the connection

Alex Fayn Jr ‘24, Emily Cao and David Everly Ph.D., Department of Micro Biology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

For multiple years, the fatal acute respiratory disease known as COVID-19 (SARS-CoV-2) has grappled scientists for many years. Without a reliable therapeutic model, the exact efforts to mitigate the transmission and pathogenesis of COVID-19 is not complete. Luckily, another coronavirus, Murine Hepatitis Virus Strain 1 (MHV-1), strain 1, produces a similar pneumonia in mice which contains lots of immunological and pathological hallmarks of human infection with COVID-19. The long-term goal of these studies is to use MHV-1 to better understand COVID-19 pathogenesis.  As a first step, reliable methods to quantitate MHV-1 must be established.  The goal of this summer project was to develop tools to quantitate MHV-1 levels by real-time PCR.  First, MHV-1 RNA was reverse transcribed into DNA and the sequences were cloned into a plasmid to be used as a reference.  This reference has been used to establish copy controls from 300,000 to 3 copies that can be reliably detected by real-time PCR assays.  The next step will be to quantitate viral RNA from infected cells and cell supernatant and compare these numbers to traditional plaque assays.  Together these assays will be used to in in vivo and in vitro studies to help understand viral pathogenesis.

The Sex-Specific Effects of Litter-Bearing on Anxiety and Memory Loss in Alzheimer’s Disease

Olivia Godek ‘23 ¹, Kameron Kaplan ^2,3, Lainey Toennies ², Holly C. Hunsberger ^2,3
191��Ƭ��, Lake Forest, IL
²Center for Neurodegenerative Diseases and Therapeutics, Rosalind Franklin University of Medicine and Science, North Chicago, IL
³The Chicago Medical School, North Chicago, IL

Alzheimer’s disease (AD) is a debilitating neurodegenerative disorder that is associated with progressive memory loss and behavioral changes. AD patients commonly display additional neuropsychiatric symptoms including heightened levels of depression and anxiety, with these effects developing at a faster rate in females than males. Moreover, AD disproportionately affects the female population, although the precise influence of biological sex on the development and progression of the disease are unclear. Because hormones such as estrogen are neuroprotective, scientists speculate that menopause and depletion of these hormones contributes to the earlier cognitive decline seen in women. Previous studies have reported that the increase in estrogen associated with pregnancy may protect neurons from amyloid β (Aβ)-induced apoptotic cell death, a characteristic of AD pathology. However, it is not well understood whether pregnancy has an effect on the progression of AD and consequently on one’s behavior, emotions, and ability to learn. To better understand how pregnancy affects the brain, we used retired breeders from the APP/PS1 (AD) mouse line to investigate the effect of litter-bearing on depression, anxiety, and memory using a battery of behavioral tests. We found that litter-bearing mice exhibited increased anxiety levels in open-field and elevated plus maze tasks as well as circadian differences when compared to controls. Interestingly, in a measure of novelty-induced hyponeophagia, paternal males exhibited decreased latency for approaching food, a behavior considered to be a sign of decreased depression. However, fear memory, avoidance behavior, and perseverative behavior appeared similar across both groups. Developing a better understanding of potential risk factors for AD, including the roles of sex and anxiety, allows us to precisely tailor diagnostic techniques and treatments for females suffering from AD.

Characterization of cognitive and motor learning in a novel stimulus-response association task

Ying Han ‘24, Sayli Korde, Ama Owusu-Ofori ‘24, Bowen Lian, Eun Jung Hwang, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064; 91��Ƭ��,  Lake Forest, IL 60045

Most motor skills we perform everyday are acquired through practice and learning (e.g., tying shoelaces, using scissors, and typing on the phone). Here, we asked whether one’s ability to learn new motor skills is correlated with their ability to learn cognitive skills, in a way that suggests a common brain mechanism supporting both types of learning. To answer this question, we trained mice to learn a novel stimulus-response association task for several weeks and compared their course of cognitive learning to motor learning. In this task, mice needed to turn a wheel with their paws leftward or rightward depending on which side of the screen a visual stimulus appeared (i.e., turn leftward for the right side stimulus and vice versa). With training, some mice successfully learned this task rule and chose the correct responses in more than 80% of trials, while others did not. Thus, mice showed varying capacity in learning the cognitive rule. We also characterized how the motor skill to turn the wheel improves with training. We found that regardless of cognitive learning, most mice became faster both initiating and executing a turning movement in response to the visual stimulus. The enhanced reaction time and movement speed are a hallmark of motor skill learning. Therefore, motor skill learning appears to be decoupled from cognitive learning, suggesting that the neural circuits supporting the two kinds of learning are largely independent.

Fluoxetine Potentiates Oral Methylphenidate-induced Gene Regulation in the Rat Striatum

Willow Highfill ‘23 ¹, Connor Moon ², Michael Hrabak ², Panayotis K. Thanos ³, and Heinz Steiner ², ¹91��Ƭ��, Lake Forest, IL, ²Stanson Toshok Center for Brain Function and Repair, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago IL 60064, ³Clinical Research Institute On Addictions, Jacobs School of Medicine and Biomedical Sciences, University At Buffalo, Buffalo, NY 14203

Methylphenidate (MP) and Fluoxetine (FLX) (SSRI antidepressant) are important pharmacological drugs used as treatments for various disorders. MP is commonly used as a treatment for attention-deficit hyperactivity disorder, commonly known as ADHD, as its function is to inhibit the reuptake of dopamine in the brain to increase the dopaminergic activity in the prefrontal cortex. However, MP has been shown to be highly abused as a cognitive enhancement. In this study, groups of rats received treatments of MP or FLX individually or were administered a combination of MP+FLX. A control group received only water. These treatments were administered orally via drinking water 8h/day for 30 days. Dynorphin and Substance P probes were used to measure gene expression in direct pathway neurons of the striatum; Enkephalin measured gene expression in striatal indirect pathway neurons. Our results demonstrated that MP given alone significantly enhanced Dynorphin and Substance P expression, while Enkephalin showed minimal or no changes in gene expression. When administered alone FLX had no significant effect. However, when MP and FLX were administered together, we observed that FLX strongly enhanced MP-induced expression. Thus, FLX potentiated oral MP-induced gene regulation in direct pathway neurons, mimicking cocaine effects. MP+FLX combinations are used to treat ADHD/depression co-morbidity. Our findings suggest that such MP+FLX combinations may enhance addiction-related gene regulation in the brain.

ALL IN THE FAMILY: DEVELOPMENT OF TOOLS FOR EXPRESSION OF SYNUCLEINS IN SACCHAROMYCES CEREVISIAE 

Nilufar Imomdodova ‘25, Isaiah Moonlight ‘25, Osselborn R., Nassuna T., Bertolotti F., Borland C., DebBurman S.K., Neuroscience Program, 91��Ƭ��, Lake Forest, IL 60045  

Synucleinopathies are a group of neurodegenerative diseases that emerge from misfolding and aggregation of the α-synuclein protein. The most well-studied among them is Parkinson's disease which is connected to aggregation of both wild-type and mutant forms of ⍺-synuclein in the midbrain dopaminergic neurons of the substantial nigra.  α-Synuclein is part of the larger Synuclein family that includes β- and ɣ-synuclein and, while ⍺-synuclein is implicated in synucleinopathies like dementia with Lewy bodies (DLB), two toxic mutations (P128H and V70M) in β-synuclein were also found to be linked to the disease. The role of β- and ɣ-synuclein in neurodegeneration, however, is widely understudied in comparison with ⍺-synuclein. Our lab was one of the first labs in the world to use a yeast model to study α-synuclein pathological potential. Our projects contained two overarching goals: 1) create tools to comparatively evaluate the pathological potential of ⍺-, β-, V70M-β-, P123H-β-, and 𝛄-synucleins, and 2) create tools to examine the impact that disease causing mutations in β-synuclein may have in ⍺-synuclein. In order to comparatively study the cytotoxic and aggregation potential of α-, β-, and ɣ-synucleins we set out to create yeast expression vectors for all synucleins (including P128H & V70M) tagged with emerald green fluorescence protein (emGFP). To do this, we performed a three-phase subcloning process previously employed in our lab. Additionally, to evaluate the pathological potential of β-synuclein mutants (V70M/P123H) on the toxicity of ⍺-synuclein, we engineered mutations into α-Synuclein genes at conserved residues (V70M/P128H) using the same subcloning process. In the initial phase, synuclein genes were amplification using polymerase chain reaction (PCR) and subcloned into an entry vector containing site-specific recombination sites. Next, a recombination reaction was performed between the entry vector and destination vector already containing the emGFP gene. Lastly, we used PCR again to amplify these synuclein-emGFP fusion genes from the destination vector and subclone them into the yeast expression vector pYES2.1 allowing for the study of synucleins in budding yeast. Here, we report successful completion of these steps for all intended synucleins. 

Using Convolutional Neural Networks to Predict Memory Loss in Mild Traumatic Brain Injury Patients

Jovana Jovanovska ‘23, Raneem Samman ‘24, and Dr. Sara Jamshidi, Mathematics and Computer Science Department, 91��Ƭ��, Lake Forest IL 60045

Mild traumatic brain injuries (mTBI) are generally undetectable by radiologists; instead, mTBIs are diagnosed using standardized questionnaires of self-reported symptoms. This summer project attempted to ask, given that a patient has been diagnosed with mTBI, is it possible to predict a particular symptom reported in the questionnaire. We identified memory loss as a symptom with a high probability of detection within MRI imaging. The dataset was constructed from MRI images of primarily cis-male American veterans from the Midwest (total=175, men=165, women=10). Three-dimensional MRI scans were reduced to 6 two-dimensional slices. This was achieved by hand-selecting 6 slices from a randomly selected patient. The remaining MRI scans were reduced to six slices by utilizing the Euclidean distance from the benchmark slices. The dataset was then used to train a convolutional neural network (CNN). Against a testing set, the highest accuracy achieved was 67% with a mean absolute error of 1.26 and a mean squared error of 2.75. In comparison, a linear regression achieved only 15% accuracy against the testing set. Further work is warranted with respect to other symptoms within the questionnaire.

What genes regulate unc-2 levels at the presynaptic terminal in C. elegans?

Wambui Kahende ’24 and Dr. Hongkyun Kim, Department of Neuroscience, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Unc-2 is a gene that has been predicted to enable voltage-gated calcium channel activity in a species of worms known as C. elegans. We study the mechanisms that regulate unc-2 levels at the pre-synaptic terminals. Specifically, what genes or proteins regulate their levels. This research is significant because there are many homologs of our genes in C. elegans, so these findings can help us better understand neural functions in human beings and uncover possible causes of diseases like ataxia, epilepsy, and seizures which are caused by calcium channels.

Riboflavin Encapsulated Sulfobutyl Ether Cyclodextrin Complex to Prevent Dental Caries

Shaad Khalil ‘25, and Dr. Kristen Ahlschwede, College of Pharmacy, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Streptococcus mutans is a bacteria found abundantly in the oral cavity, whose acid producing biofilms are known to contribute to the formation of dental caries. Riboflavin-5'-phosphate sodium salt dihydrate, also known as riboflavin or Vitamin B2, has been shown to counteract S. mutans biofilm formation when photoactivated, but lacks solubility for usage in treatment. Here we investigate the combination of riboflavin and sulfobutylether-beta-cyclodextrin (SBE-CD), a mucoadhesive polymer capable of slowing drug release rate, to inhibit the formation of S. mutans biofilm formation. To answer this question, 2 mg/mL final concentration riboflavin was added to SBE-CD and allowed to equilibrate (Ribo-CD). In a 96-well plate, the diluted S. mutans inoculum was coincubated with various concentrations of Ribo-CD. The plate was treated with a blue LED for 5 minutes and placed at 37°. The biofilm mass was analyzed with crystal violet staining. The disruption continued from 24h to 72h without a decrease in effectiveness. The minimum inhibitory concentration was determined using a crystal violet assay. The Ribo-CD formulation significantly decreased the formation of an adherent biofilm for 24, 48, and 72 h. Furthermore, higher concentrations of Ribo-CD were tested, and concentrations above 200 μl/mL showed diminishing returns of biofilm prevention. The minimum inhibitory concentration was determined to be 200 μl/mL. Lower concentrations of Ribo-CD were outperformed by equivalent concentrations of pure riboflavin under shorter time conditions, though higher concentrations of Ribo-CD greatly outperformed equivalent concentrations of pure riboflavin. This study demonstrates how cyclodextrin could significantly enhance the solubility and release rate of riboflavin and has potential to be used as a preventative measure for dental caries.

Repeated TBI leads to less severe acute outcomes, but has worse long-term effects on mortality and locomotor activity than a single, severe TBI 

Kamden T. Kuklinski ‘23,  Daniel Tulchinskiy '25, Dr. Rebecca Delventhal, Department of Biology, 91��Ƭ��, Lake Forest, IL 60045   

Approximately 1.5 million Americans are affected by traumatic brain injury (TBI) each year with outcomes ranging from hospitalization and survival, long-term disability, or death ¹. Despite many clinical studies into the short- and long-term effects of TBI, the unique effects of repeated TBI are still poorly understood. To address this, we used Drosophila melanogaster as a model organism to compare the outcomes of TBI administered over multiple days to a single, severe TBI administered on one day. We examine outcomes such as acute mortality, lifespan, locomotor performance, and expression of genes related to immune signaling in flies following the injury. Acute mortality was significantly higher among flies in the single-day condition, whereas the long-term mortality was significantly higher in the multi-day condition. This overall trend was consistent with effects on locomotor performance: flies given a single, severe TBI performed significantly worse shortly after the injuries were given, while flies given a TBI over multiple days performed significantly worse long-term. To investigate what mechanism may be mediating the difference in short- and long-term outcomes on injury pattern, we are measuring the immune response at multiple timepoints post-TBI in both conditions.  

Jumping Fish: Exploring different stimuli and associated sensory systems in Silver Hatchfish Gasteropelicus sternicla that trigger jumping behavior  

Kyle Lassen ’25, Alyssa Silva ‘23, Jia Zheng ‘25, and Margot Schwalbe, Biology Department, 91��Ƭ��, Lake Forest, IL 60045 

Many different species of fish are capable of propelling themselves out of water, but very few species are known to routinely jump out of the water. The behavioral “why” and the sensory “how” of this behavior varies among such fish. Some fish may leave the water to avoid predators, while others may do so in order to capture prey just above the water surface. Multiple sensory systems in fish can mediate jumping behavior; for example, a fish could jump when it sees or smells a predator, or it could jump when it detects a sudden water pressure change via their lateral line system generated by a prey or a predator. In our experiment, we studied the jumping behavior in freshwater silver hatchetfish ( Gasteropelicus sternicla) to determine what triggers this fish to jump and link the sensory system(s) associated with their jumping behavior. We tested a variety of stimuli that related to prey detection or predator avoidance and recorded via high-speed video (500-600 frames/sec) the fish’s jumping frequency and the height of the jumps. We also examined the biomechanics of the jump itself by observing the caudal and pectoral fin movements after the fish left the water in the high-speed video. Through the course of the experiment, we found that the hatchetfish jumping behavior was best elicited by a sudden, relatively soft, vibrational stimulus that likely initiated a predator-avoidance behavior, as seen in the wild. This response was most common when a 1 kg hammer was dropped from a height of 4 cm near the tank. Finally, we observed that the hatchetfish continue to move their caudal fin for the duration of their time out of water. Our preliminary findings lay out an important procedural method for further study of the sensory systems and biomechanics involved in the silver hatchetfish’s unique jumping behavior. As we investigate further, we expect to find that this jumping behavior is linked to visual and lateral line pathways.  

Towards the Synthesis of (±)-Chaetoxanthone C and D

Emma Law ‘22, Giselle Schiet ‘23, and Dr. Paul T. Gladen, Chemistry Department, 91��Ƭ��, Lake Forest IL 60045

Natural products have proved to be invaluable in treating a multitude of diseases. Many recently discovered compounds have shown bioactivities including anticancer, antiviral, and antimicrobial activity. One promising class of natural products that has shown these bioactivities is the xanthones. Within the xanthone class, a family of xanthone products called chaetoxanthones have demonstrated activity against Chagas disease, which affects about 6-7 million individuals worldwide (WHO). Herein, we report our efforts to develop a synthetic route to access chaetoxanthone C and D beginning with inexpensive and commercially available starting materials. Our strategic approach relies on the development of a novel direct functionalization reaction of the xanthone core to enable rapid access to natural products. This direct synthetic method represents a significant improvement over existing strategies and will allow for further exploration of biological activities.

Police Expertise in Use-of-Force Rapid Decision-Making

Ceanna Loberg ‘23, Wiktoria Pedryc ‘24, Vivian Ta-Johnson, Ph.D., Psychology Department, 91��Ƭ��, Lake Forest, IL, 60045

Use-of-force decisions among police officers typically occur under stressful and fast-paced conditions. Because judgment and decision-making from more experienced individuals tend to be more effective, efficient, and accurate compared to less experienced individuals, researchers have sought to understand the specific skills involved in the selection of appropriate uses-of-force among expert officers compared to novice officers. In the current study, we examined how a sample of expert ( n = 42) and novice ( n = 36) officers differ in their decisions in use-of-force scenarios. Officers observed a series of body-worn camera footage of real-world police-citizen encounters across the US that were temporally occluded at several decision points. At these decision points, officers were prompted to describe the course of action they would take in the next few seconds if they were the officer on scene. Responses were coded for behaviors ( n = 19) that officers indicated they would engage in. A series of linear mixed-effects models revealed that expert and novice officers differed on their decisions to (1) pursue a suspect; (2) deescalate the situation, and (3) subject themselves into a particular scenario. Practical and research implications for expert performance in use-of-force training and decision-making are discussed.

Immunity Tradeoff Differences in Male and Female Bean Beetles

Katina Lucas ’25, Iman Shepard and Flavia Barbosa, Biology Department, 91��Ƭ��, Lake Forest, IL 60045

Bean beetles ( Callosobruchus maculatus), like other species, have tradeoffs between costly life history traits. Depending on density during the larval stage, they develop into either a dispersal morph with larger wings and smaller gonads, or a non-dispersal morph with smaller wings and larger gonads. Past research shows that density has a strong effect in gonad and wing size in males. In females, however, wing size is not significantly affected by density.  We hypothesized that this is due to a sex-specific trade-off between immune function and fecundity (fertility). We predict that immunity will be overall higher in females, and specifically in those reared under high density. To test this hypothesis, we reared beetles under different densities and measured their gonad size and strength of immune response. We found that under high density, females showed higher immunity and smaller ovariole size while under low density, we observed the opposite pattern.  Density did not affect male immune response, possibly because males have weaker immunity in general.  Overall, we found evidence for a sex-specific trade-off between immunity and reproduction in bean beetles. Future studies will focus on whether these different strategies are advantageous in their different density environments.

“It’s never okay to talk to anyone that way.” Imagined Contact as a Pathway to Understanding and Addressing Dis/ability Related Hate Crime

Johnny Monahan‘24, Sian Jones, Ph.D., and Clare Uytman, Ph.D., Department of Psychology, Queen Margaret University, Musselburgh, Scotland, EH21 6UU

Research has indicated that there is strong cross-national variance in attitudes towards dis/ability. It also exemplifies that responses to dis/ability-motivated hate crime maybe attenuated by imagined contact and identification with the targets. However, much less research has systematically varied the type of impairment in hate crime scenarios, nor compared the responses of those with and without an impairment. We surveyed online N = 619 adults, presenting them with a disability-based hate crime scenario. Participants were randomly assigned to either an imagined contact scenario (present or absent) and were assigned to either respond to a mobility impairment (wheelchair) or hearing impairment (deafness) scenario. Thus, the type of scenario and presence of imagined contact were orthogonally manipulated. We accounted for the nationality of our participants across Hungary, Italy, Scandinavia and the U.K. We measured participants’ responses to the scenario, participants’ direct contact with people with an impairment, and whether they identified as having a dis/ability. Additionally, participants were probed as to their reasoning for behaving in the way they’d identified. Analysis indicated cross-national differences aligned with previous research. We also found that active bystanding was predicted by direct contact, and that relationship was mediated by anger, discrimination perceptions, empathy and intergroup/intragroup anxieties. Participants that declared a disability perceived more dis/ability, and had more direct contact with dis/ability leading to a higher likelihood of non-avoidant behavior. These participants also had significantly lower anxiety when approaching a co-worker with dis/ability. Qualitative analysis enabled us to explore the reasons for these helping intentions.  Facets of qualitative responses were defined under eight categories regarding conflict engagement (avoidance & non-avoidance), emotional response (anger, anxiety, sympathy & empathy), perception of disability, and contact (direct & no contact) with similar situations. This research shows the reasons for active bystanding over non-action are nuanced, and vary according to nationality, and whether or not an individual identifies as having an impairment.

Circadian Control of the Bed Nucleus of the Stria Terminalis (BNST) via Hypothalamis Vasopressin (AVP) Projections

Lorena Monroy ‘24, Susan H. Olson, Valentina Olivera-Pasilio, Joanna Dabrowska, Department of Cellular and Molecular Pharmacology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

The bed nucleus of the stria terminalis (BNST) is a medial basal forebrain structure that regulates an array of physiological and behavioral processes. Previous research has shown that the BNST receives input from the hypothalamus, specifically, the suprachiasmatic nucleus (SCN), which is responsible for circadian rhythms. Arginine vasopressin (AVP) is one of the neurotransmitters that the SCN releases to impose its circadian oscillations onto other brain structures. Yet, it is not well understood whether AVP neurons from the SCN project to the BNST and whether AVP imposes circadian control over the BNST. The purpose of this study was to determine whether AVP expression in the BNST is under circadian control and to determine which hypothalamic nuclei send vasopressinergic projections to the BNST. To examine the relationship between AVP expression in the SCN and activity of the BNST neurons, we performed immunofluorescence and measured fluorescent intensity of AVP in the SCN and striatal-enriched protein phosphatase (STEP), which is a marker of neurons expressing corticotropin-releasing factor (CRF) in the BNST and can be used as a proxy of these neurons’ inactivity. We measured AVP/SCN and STEP/BNST intensity at four zeitgeber times from the same rats that were housed in 12:12 light-dark cycles. We found that the SCN exhibited the highest AVP expression at ZT 11 (one hour before lights off) and lower expression at ZT 1 (one hour after lights on), whereas an opposite pattern was observed for STEP in the BNST with the highest STEP levels at ZT 1 and lowest at ZT 11. Then, to examine which hypothalamic nuclei send vasopressinergic projections to the BNST, we virally injected the supraoptic nucleus of the hypothalamus (SON) and SCN of AVP-Cre transgenic rats (Cre-recombinase under the AVP promoter) with Cre-dependent AAV driving mRuby expression and performed immunofluorescence to visualize AVP. We found fibers in the BNST that colocalized the viral fluorescence and AVP peptide, suggesting that the BNST receives AVP inputs from the SCN and SON. Overall, this research suggests that rhythmic oscillations of the SCN via AVP may mediate behavioral and physiological rhythms through synchronous activity with the BNST.

Sex Differences in Behavioral Responses to Stress and Anxiety in Rats

Rose Jasmin Montes ‘24 and J. Amiel Rosenkranz, Department of Cellular and Molecular Pharmacology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Post-traumatic stress disorder (PTSD) is a psychiatric disorder characterized by increased fear and anxiety. While displaying the same symptoms, women are twice as likely than men to develop such psychiatric disorders as PTSD and general anxiety disorder (GAD). Recent studies have provided insight into the neurocircuitry of the basolateral amygdala (BLA) in fear; however, its role in anxiety/stress remains unknown. Additional evidence shows that females extinguish fear quicker than males. The objective of this study was to determine if there was a sex dissimilarity between anxiety and stress, and more specifically, we aim to investigate the BLA activation during these states. In the first part of the study, we aimed to see a difference in the freezing times of males and females during stressful and anxious circumstances. The two behavioral paradigms administered were repeated restraint stress (RRS) and fear conditioning to see the effects of stress and anxiety on the behavior within the sexes. We found that after repeated restraint stress, females had gained the least amount of weight compared to males. After fear conditioning, however, we did not find significance in their freezing patterns. Therefore, we observed other behavior patterns and found that females were on their hind limbs for a more extended time than males were. Overall, females and males displayed different behaviors to stress and anxiety. Neurons in the BLA were located but were not shown to be activated. This research may provide more details on the sex differences in stress and anxiety, and we also offer some insight into what potential immunohistochemistry staining of the BLA would look like.

Assessing Procedural Justice Through Body-Worn Camera Recordings: A Natural Language Processing Approach

Amelie Motzer ‘24, Matías Fonolla ‘23, Vivian Ta-Johnson, Ph.D., Psychology Department, 91��Ƭ��, Lake Forest, IL, 60045

Body-worn camera (BWC) recordings enable the observation and evaluation of officer behavior with community members. Although usage of BWCs has increased at a rapid pace, BWC recordings are currently under-utilized as a tool for improving the quality of police-community interactions. Recent technological advancements in natural language processing (NLP) provide new approaches that may turn BWC recordings into data that can be used to evaluate officer behavior. As such, we used NLP to contribute to the development of automated analyses of  BWC recordings. Specifically, we developed a classifier that is capable of automatically analyzing verbal behaviors in BWC recordings to identify the camera-wearing officer with 95% accuracy. Other classifiers currently in development include classifiers that are capable of automatically analyzing verbal behaviors from BWC recordings to accurately identify the type of call that the camera-wearing officer is responding to (i.e., domestic violence and mental health calls).

Characterization of age-dependant AD pathology in mice

Anait Nalbandyan ’24, Nikki Barrington, and Dr. Beth Stutzmann, Department of Neuroscience, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Alzheimer’s disease is a fatal neurodegenerative disorder that impacts cognitive abilities of humans by destroying neurons, including neurons in the hippocampal area. While the origin of the disease is unknown, there are several late-stage hallmarks of AD that indicate its progression. They include phosphorylated Tau (pTau) proteins and amyloid beta (Aβ) plaques that accumulate in brains of patients with AD.  We characterized the inventory of mouse models to determine the pattern of protein aggregation in triple transgenic mice (3xTg) with AD across ages of 3 and 6 months. Using immunohistochemistry (IHC) staining for pTau and Aβ was performed on preserved mice tissue, which was then imaged using confocal microscopy, in particular the hippocampal area.

ALL IN THE FAMILY: STUDYING TOXIC TENDENCIES OF THREE SYNUCLEINS IN A YEAST MODEL

Tracey Nassuna ‘23, Ryan Osselborn ’23, Bertolotti, F.B., Moonlight, I.I., Imomdodova, N.M., Zabat, B., Borland, C., DebBurman, S.K., Neuroscience Program, 91��Ƭ��, Lake Forest, IL 60045

Synucleinopathies are a group neurodegenerative disorders that arise from the misfolding and aggregation of alpha-synuclein. The most well-known among them is Parkinson’s disease (PD), which is linked with the aggregation of both wild-type or mutant forms of alpha-synuclein in midbrain dopamine neurons. Alpha-synuclein belongs to a larger family of proteins that include beta- and gamma-synuclein. Recently, two mutant forms (V70M and P123H) of beta-synuclein were shown to cause Dementia with Lewy Bodies (DLB), and gamma-synuclein inclusions were discovered in some cases of human neurodegeneration, but beta- and gamma-synuclein toxicity potential and mechanisms linked to neurodegeneration is still highly understudied. A variety of post-translational modifications and altered cellular conditions liked with PD influence the pathological potential of alpha-synuclein, though their effects on beta- and gamma-synuclein toxicity is poorly understood. For our two related senior thesis projects described here, we tagged wild-type and mutant forms of beta- and gamma-synucleins with GFP at the C-terminus, expressed them in Saccharomyces cerevisiae (budding yeast), and evaluated their toxicity, localization, and expression using serial dilution spotting, fluorescence microscopy, and western blotting, respectively. We report that wildtype forms of both are toxic in yeast (beta-synuclein is more toxic than gamma-synuclein), but none are as toxic as alpha-synuclein, and that their comparative extents of toxicity depended on yeast strain background. Beta-synuclein aggregates like alpha-synuclein does, but gamma-synuclein does not. Both V70M and P123H beta-synuclein mutants are also toxic and aggregate, but at comparable levels as wild-type beta-synuclein. With these methods, we are now evaluating whether beta- and gamma-synuclein properties will be altered in the following conditions: 1) in yeast strains genetically altered for levels of SUMOylation, nitration, lipids, glycation, and acetylation; and 2) in yeast strains deleted for key genes whose human homologs cause autosomal recessive forms of PD. To further assess toxicity of the two beta-synuclein mutants, we are specifically testing whether the loss of the original amino acids (valine, proline) or gain of the new ones (methionine, histidine) make the mutants toxic. We have also genetically engineered the two beta-synuclein mutants into alpha-synuclein to determine if that itself can increase alpha-synuclein toxicity. We plan to report progress in these studies as well. In summary, our ongoing studies will extend our understanding of two proteins found within the human nervous system, beta- and gamma-synucleins, whose links to neurodegeneration is increasingly become more relevant.

Glial-specific knockdown of a subunit of the ER membrane complex (EMC) dramatically reduces survival of D. melanogaster

Maria Jose Orozco Fuentes '24, Otoha Tatami '24, Rebecca Delventhal Ph.D, Biology and Neuroscience Department, 91��Ƭ��, Lake Forest, IL, 60045

The endoplasmic reticulum (ER) is involved in the modification, packaging, and insertion of proteins into the membrane. A newly discovered complex called ER membrane protein complex (EMC) is believed to help with many ER functions. This protein complex is highly conserved from yeast to humans and it is composed of 10 subunits that work in tandem. Prior work showed that transcription levels of EMC4 were decreased in glial cells following TBI, which led us to study the effect of an RNAi knockdown of EMC4 in glial. This resulted in severe phenotypes, such as delayed development, strongly impaired locomotion, a shorter lifespan of 5-6 days, and increased protein aggregation. The dramatic effect of that this knockdown has on fruit flies sheds light on the importance of this subunit to organismal health and protein degradation. Future studies on tissue-specific knockdowns other than glial cells would offer additional insight about the protein processing mechanism that specialized cells have and how they are affected by the loss of function of the subunit EMC4.

The role of V1-STR pathway in visually guided action

Ama Owusu-Ofori ’24, Sayli Korde, Ying Han, Bowen Lian, and EunJung Hwang, Rosalind Franklin University of Medicine and Science, Stanson Toshok Center for Brain Function and Repair, North Chicago, IL, 60045.

Many actions in life are guided by sensory stimuli (e.g., the red traffic light and stop). The brain processes underlying such sensory stimulus guided action selection remain largely unknown. The striatum (STR) has been shown to participate in this process for auditory stimulus via the projection from the auditory cortex to STR. Analogously, we hypothesize that the projection from the visual cortex (V1) to STR (V1-STR) supports visually guided action selection. Our hypothesis predicts that inactivating the V1-STR pathway would impair visually guided action selection. To test this prediction, we trained mice to perform a visually guided action choice task and examined how inactivation of the V1-STR pathway affects their task performance. To inactivate the V1-STR pathway, we took a chemogenetic approach; intraperitoneally injected Clozapine-N-Oxide (CNO) exclusively binds to designer receptor hM4Di and then inhibits neurons. To express hM4Di selectively in neurons in the V1-STR pathway, we injected mice with retrograde virus encoding Cre in STR and virus encoding Cre-dependent hM4Di in V1. These mice were trained to expert level in the visually guided action choice task. Then, we alternated intraperitoneal injections between CNO and saline across days and found no significant difference in the task performance between the two conditions (N = 6 mice). The null effect of V1-STR inactivation suggests two alternatives: 1) hM4Di expression in the V1-STR pathway has failed, or 2) V1-STR neurons are not necessary for visually guided action choice. Our future histological analysis that examines the expression of hM4Di in V1-STR neurons will distinguish between the two alternatives.

Preparation and characterization of 7- or 8-Methyl-5-substituted-chromenopyridines

Giorgio Papadatos ’24, Keiffher Rosendo ’25,   June Mayor ², John Buolamwini ¹,  Ronald Kaplan ²,   Shivaputra Patil ¹, ¹Pharmaceutical Sciences Department, College of Pharmacy, and ²Center for Proteomics & Molecular Therapeutics, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

The discovery of small molecules that would selectively inhibit the human plasma membrane citrate transporter (PMCT) will be of great importance because of its recently reported involvement in cancer, along with other diseases such as obesity, diabetes, and non-alcoholic fatty liver disease (NAFLD). We employed a medicinal chemistry approach to discover a selective PMCT inhibitor. Very recently, the chromenopyridine analog (SAP-165) has been identified as a lead PMCT inhibitor. To develop the possible structure-activity relationship (SAR) studies around the SAP-165, we have designed and prepared twenty new chromenopyridines with suitable substitutions at C-5, C-7, and C-8 positions of the chromenopyridine moiety. All the newly prepared chromenopyridine analogs have been characterized using mass and NMR spectroscopy. The PMCT inhibition and anticancer activities of new analogs will be explored in the near future.

Context is key?: A pilot study to investigate the renewal of Pavlovian conditioned approach behaviors using a within-subject design

Keanna Price ’24, Kotryna Andriuskeviciute ’24, and Dr. Jean-Marie Maddux, Psychology Department, 91��Ƭ��, Lake Forest IL 60045 

Previous research has shown that context can modulate Pavlovian conditioned responses. In Pavlovian conditioning paradigms, a conditioned stimulus (CS) signals the delivery of an unconditioned stimulus (US). Depending on the nature of the CS, two types of conditioned responses can develop: approach to the site of US delivery, termed goal-tracking, and approach to the CS itself, termed sign-tracking. The majority of studies have investigated contextual modulation of the goal-tracking response, but less is known about the context-dependence of the sign-tracking response. Moreover, most studies of the renewal effect, the observation that an extinguished conditioned response can return if testing is conducted in a context different than the extinction context, have done so using a between-subject design. Here, we utilized a within-subject design to investigate the renewal of both sign-tracking and goal-tracking in male Long-Evans rats. Rats received two daily training sessions, each in a distinct context. During acquisition training, rats were presented with two CS levers, each assigned to one context, that signaled sucrose US delivery. Rats then received extinction training, in which the CS levers were presented without the US, and importantly, the initial context-CS assignment was switched. Test sessions were then conducted in both contexts, with serial presentations of both CS levers; hence, each CS lever was tested in its acquisition context and its extinction context. Contrary to prior reports, neither the sign-tracking nor the goal-tracking response showed recovery when tested in the acquisition context compared to the extinction context. These null results underscore the likely parameter dependence of the renewal effect, and indicate this effect is more elusive than the literature suggests. This has important implications for translational research that has utilized context-induced renewal as a model of relapse in substance use disorders. 

How does the host defend itself against tuberculosis? The role of l-plastin in a mycobacterium marinum infection in zebrafish.

Ceylin K. Sahin ‘22, Beau Grimes, Emma Remish, and Dr. Will H. Conrad, Biochemistry and Molecular Biology Department, 91��Ƭ��, Lake Forest IL 60045

Mycobacterium tuberculosis is an issue worldwide with multidrug-resistant forms becoming prevalent due to the lack of effective treatments. The goal of this project is to identify if there are host genes that affect the disease outcome. The influence of the Lymphocyte Cytosolic Protein 1 (LCP1) gene in zebrafish larvae infected with mycobacterium marinum was studied. To understand the immune response in the fish, the bacterial burden was observed in fish that had LCP1 (heterozygotes) and in fish that did not (knockout mutants). M. marinum was injected into the pericardial vein of the zebrafish larvae and the fish were infected. Once the fish were fixed and euthanized, images of the bacterial infection were taken using a fluorescent microscope. The bacteria was tagged with tdTomato, a red fluorescent protein that allows for the bacteria to glow red under the microscope. The images were then analyzed, and the bacterial burden was quantified using ImageJ to count the number of glowing pixels. Following the imaging of the infected larvae, the fish were genotyped. The DNA from the larvae was extracted through a hotSHOT PCR protocol, then primers designed specifically to amplify the wildtype and mutant genomes were used in a PCR reaction to identify whether the infected fish was a heterozygote or knockout mutant. The images of the fish were compared to the genotype and fish without LCP1 were found to have a higher bacterial burden. Understanding why fish without LCP1 have a higher bacterial burden could provide an insight to the role LCP1 plays in the human body’s immune response to tuberculosis. The future directions of this project are to observe more fish in order to replicate the findings and to test other phenotypes in order to have a better understanding of LCP1's role.

Educational Sessions Facilitate Lasting Increase in MDMA Knowledge in Health Profession Students

Dagmara Zajac ¹, Belle Tseitlin ², Madison Stevens ‘23², Taylor Macaulay ¹, Samuel Vincent ¹, & Steven A. Miller ^1
1Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
²91��Ƭ��, Lake Forest, IL 60045

The current study tested the effectiveness of the GPF Foundation 3,4 Methylenedioxymethamphetamine (MDMA) educational program on students at a private, higher education healthcare university. Approximately 56 participants completed the program. An 18-item measure of MDMA knowledge was developed and distributed to participants at multiple time points to assess the effectiveness of the educational program. Multilevel models with random intercepts were estimated using restricted maximum likelihood estimation to examine changes between time points. A. waitlist control study demonstrated that without intervention, knowledge did not change over time, and with it, knowledge increased significantly and did not decrease at follow up.

Non-enzymatic nitro reduction in biological media at physiological conditions

Farhan M. Ahmed ‘24, Hridey Kapoor ‘25, Amy Tram ‘25, Karen P. Gomez ‘22, Dr. Erica Schultz​, Department of Chemistry, 91��Ƭ��, Lake Forest, IL 60045

Nitro aromatic compounds, found in some pharmaceuticals, pesticides, and explosives, contaminate groundwater supplies. However, they can be converted into the aniline derivative, which are less environmentally persistent and less toxic to microbial life, through a nitro reduction. The reduction of nitro aromatic compounds using a catalyst (Pd/C) and a hydrogen source in a biological media under physiological conditions is reported for a range of compounds. This creates the potential to offer new remediation strategies to reduce nitro aromatic contaminants by combining synthetic chemistry with metabolism to be non-toxic to microbial organisms.

Optimizing an external cavity diode laser for maximum power and smooth scanning around 670.7 nanometers.

Lionel Whitehead ’23 and Dr. Michael Kash, Physics Department, 91��Ƭ��, Lake Forest, IL 60045

External cavity diode lasers (ECDL) are commonplace in atomic physics labs as they are an inexpensive and effective way to produce a specific wavelength of light and even “scan” over a certain range of wavelengths. Our ECDL is set up in the Littrow configuration which involves a reflective diffraction grating and another mirror to route the laser back to the table. To control an ECDL in the Littrow configuration, the temperature of the diode must be set so the desired wavelength is in a stable mode of the laser. To achieve smooth scanning, the angle of the diffraction grating and the current running through the diode must be changed together. My research focused on how to determine the optimal rates of change for the current and angle, for which I developed a method that measured the power output of the laser. Separately, I made other changes to the ECDL to optimize the scanning and have proposed further changes.

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