2018 Glassman Symposium Abstracts

Alumni Poster Abstracts

Identification and Regulation of Calcium Activated Chloride Channels (CaCCs) in Mouse Fiber Cells

Pooja Acharya, ’17, Lisa Ebihara, and JunJie Tong

Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Calcium-activated Chloride Channels (CaCCs) are known to be present in the plasma membrane of a number of different types of cells including epithelial cells, neurons and vascular smooth muscle. They display a variety of important physiological and cellular roles, which include volume regulation, regulation of pH, and cell migration. However, there is not much information regarding the molecular identity and properties of CaCCs in the lens. Therefore, we tested the hypothesis that TMEM16A CaCCs are expressed in mouse lens fiber cells. Immunohistochemistry was used to map the distribution of TMEM16A throughout the lens in cryosections from 10 or 11 day mouse lenses. The sections were imaged using confocal microscopy. Membrane currents from differentiating epithelial cells and newly elongating fiber cells were recorded using the whole cell patch-clamp technique. Our results showed that TMEM16A immunoreactivity was present in the cytoplasm of the epithelial cells and in the membrane and cytoplasm of the superficial differentiating fiber cells near the equator of the lens.  Whole cell patch clamp experiments indicated that these cells express a calcium-activated current whose properties resemble those of TMEM16A.

The Contribution of Sleep Disruption in the Physiological Impact of a Spaceflight Analog

Katrina J. Campbell ’14, Peng Jiang, Vance Gao, Fred W. Turek, and Martha H. Vitaterna

Northwestern University, Chicago, IL; Center for Sleep and Circadian Biology


A widely-used simulation of the effects of microgravity for rodent experiments on Earth involves hindlimb unloading (HU). Multiple spaceflight-associated health effects are found in response to HU. Astronaut studies show sleep in space results in many sleep disruptors. Because sleep loss and microbiome perturbations can produce some of the immune, metabolic, and endocrine changes observed with spaceflight and HU, we investigated the impact of HU on sleep and the gut microbiome. This study included three different mouse strains.


Sleep/wake states were determined from continuous EEG/EMG recording, combined with simultaneous HU for the first time. 12 female mice of each of three strains (C57BL/6J, BALB/cJ, and C3H/HeJ) were studied: six/strain underwent HU, while six underwent a sham (tail immobilization) manipulation. After a baseline recording period, mice were subjected to 4 weeks of HU or sham unloading. The gut microbiome was also monitored through fecal collections prior (-1.5 weeks) to and during (1, 3 weeks) manipulation.


Upon implementation of HU, C57BL/6J and BALB/cJ mice had dramatically increased sleep fragmentation (number of state shifts), but no significant difference in total amount of time spent asleep. In contrast, HU C3H/HeJ mice did not show changes in sleep structure. The gut microbiome as reported by the firmicutes/bacteroidetes ratio in the fecal samples was maintained in C57BL/6J and BALB/cJ mice in both control and HU conditions but was perturbed in the C3H/HeJ HU mice.


Sleep and microbiome disruption occurs in some strains of mice during hindlimb unloading and may contribute to some of its effects. We will investigate the effects of the spaceflight environment on sleep, circadian rhythms, and the microbiome in mice which launched to the ISS in June 2018. The mouse strains C57BL/6J and C3H/HeJ were selected for the ISS experiment. These strains offer an interesting comparison because C3H/HeJ mice produce melatonin while C57BL/6J mice do not. This study will investigate the effects of the space environment on microbiome and sleep, crucial aspects of human health.

Pharmacological Blockade of KCC2 in the Subiculum Triggers Synchronized Inter-ictal Like Bursts

Michael Fiske ’10 and G. MacCaferri

Northwestern Univ. Feinberg Sch. of Medicine, Chicago, IL; Dept Physiol, Northwestern Univ., Chicago, IL

A prominent theory regarding the development of epileptic hyper-synchronization in human and animal models of temporal lobe epilepsy proposes a key role for the specific down-regulation of the KCC2 transporter in subicular pyramidal cells. As KCC2 is essential to maintain the low intracellular chloride concentrations required for hyperpolarizing GABAergic signaling, loss of KCC2 expression would impair GABAergic inhibition and trigger a series of events leading to the emergence of subicular-initiated interictal activity. Although this prediction was supported by computational modeling, direct experimental evidence based on the pharmacological block of KCC2 has not yielded definitive results. Here, we have utilized simultaneous triple cell-attached and whole-cell recording to evaluate the role of KCC2 on network activity and synchronization in the isolated subiculum. Our preliminary data show that the application of a highly selective KCC2 antagonist (VU0463271, 10 μm) on isolated mini-slices of the mouse subiculum generates synchronous interictal-like bursting that depends on depolarizing GABAergic signaling, but is not, apparently, sufficient to trigger ictal-interictal transitions. These events are pharmacologically similar to the inter-ictal events seen in patients, as they can be abolished by application of AMPA or GABA receptor antagonists. Additionally, our preliminary data suggest that epileptiform synchronization in the subicular mini-slices requires excitatory input from parvalbumin+ cells. Brief (1-2 ms) photoactivation of PV+ interneurons expressing channelrhodopsin in slices exposed to the KCC2 antagonist triggered interictal-like spikes that were electrophysiologically similar to spontaneously occurring events. In contrast, prolonged (15 sec) light stimulation of PV+ cells suppressed KCC2 antagonist induced bursting. Our results support the hypothesis that the down-regulation of KCC2 is sufficient to elicit epileptiform activity in otherwise healthy subicular networks.

Alcohol Alters Intestinal Bacteria and Exacerbates Colitis in Mice

Paulius V. Kuprys ’12 , Abigail R. Cannon, and Mashkoor A. Choudhry

Alcohol Research Program, Burn & Shock Trauma Research Institute,

Loyola University Chicago Health Sciences Division, Maywood, IL, USA 60153

Ulcerative colitis (UC) is an inflammatory bowel disease that produces shallow ulcers limited to the large intestine and rectum. No one factor is implicated in the etiology of UC, but it appears to be a composite of environmental, genetic, and bacterial elements. Upon development of UC, patients experience recurring cycles of disease reactivation, leading to abdominal discomfort and bloody stools. Certain foods have been correlated with UC disease flares and onset, although the exact mechanism by which this occurs is not understood. One of these foods is alcohol. To understand how alcohol may influence the disease process of UC, we treated mice with dextran sulfate sodium (DSS) to induce aspects of the UC disease, after which the mice were further treated with alcohol. Using this model, we have shown that alcohol increases intestinal inflammation, exacerbates weight loss, and changes bacterial populations in mice. Compared to control mice, DSS treatment produced increases in gram negative bacteria, as well as decreases in beneficial gram positive bacteria. With the addition of alcohol, levels of gram negative bacteria were further increased and gram positive bacteria were unchanged. The increase in gram negative bacteria may increase the burden of intestinal lipopolysaccharide (LPS), promoting increased LPS leakage. Although DSS increased RNA expression of intestinal antimicrobial peptides, their expression was unchanged in the combined DSS and alcohol treatment, suggesting an alternate mechanism for the observed bacterial population shifts. The changes in bacteria observed in the DSS mouse model post alcohol treatment may be one aspect by which alcohol contributes to the enhanced disease process post DSS treatment. Future experiments will examine serum levels of LPS, intestinal bacteria translocation, and how alcohol produces this shift in bacterial populations (Support: R21AA022324, T32AA013527 and F31AA025536).

Autonomic Arousal Fluctuation Predicts Math Performance

Krista Meuli ’18 and Naomi Wentworth

Department of Psychology and Neuroscience Program, Lake Forest College, IL 60045

Pupil dilation, an autonomic arousal response, can measure attention because pupil dilation positively correlates with attention. This study investigated the predictability of mental arithmetic performance from pupil dilation fluctuation patterns of 11 college students. Arithmetic problems consisted of basic addition and varied in difficulty. The mental arithmetic task was administered while recording pupil dilation at 60 Hz with an ISCAN eye tracker. A pupil diameter baseline was measured before problems. Patterns of pupil diameter change from the baseline over time were analyzed by difficulty and performance. A marginal effect of Difficulty, marginal effect of Performance, and significant effect of Time on Pupil Dilation change were observed. Both Time by Difficulty and Time by Performance interacted significantly. However, Difficulty and Performance did not interact. These findings support the conclusion that attention increased over time during mental arithmetic problems. Furthermore, attention over time increased more on difficult problems and on correct problems.

Quantifying Microvascular Abnormalities with Increasing Severity of Diabetic Retinopathy Using Optical Coherence Tomography Angiography

Peter L. Nesper ’15 ,¹ Philipp K. Roberts,¹ ² Alex C. Onishi,¹ Haitao Chai,³Lei Liu,³ Lee M. Jampol,¹ and Amani A. Fawzi¹

¹Department of Ophthalmology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, United States

²Department of Ophthalmology and Optometry, Medical University of Vienna, Vienna, Austria

³Department of Preventive Medicine, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, United States

 Institute for Financial Studies, Shandong University, Jinan, China

Diabetic retinopathy (DR) is a vascular disease that can lead to irreversible vision loss. In this study, we used a noninvasive imaging technique called optical coherence tomography angiography (OCTA) to quantify retinal microvascular changes during the progressive stages of DR. This cross-sectional study included 137 eyes of 86 patients with DR and 44 eyes of 26 healthy age-matched controls. We analyzed the superficial (SCP) and deep (DCP) retinal capillary plexuses for the following parameters: foveal avascular zone, vessel density, percent area of nonperfusion (PAN), and adjusted flow index (AFI). We used statistical models to adjust for age, sex and the correlation between eyes of the same study participant. All OCTA parameters showed a significant linear correlation with DR severity (P < 0.05) in the univariate models except for AFI in the SCP. These correlations remained significant after correcting for covariables. Interestingly, eyes of subjects with diabetes without clinical DR had increased AFI in the SCP compared to healthy subjects (P < 0.05). We concluded that retinal vascular nonperfusion in OCTA is correlated significantly with disease severity in eyes with DR, and increased flow in the SCP may be an early marker of microvascular changes before clinical signs of DR.

Loss of Transcriptional Repression by BCL6 Confers Adipose Tissue Expansion and Insulin Sensitivity

Madhavi D. Senagolage ’121, Krithika Ramachandran1, Meredith A. Sommars1, Yasuhiro Omura1, Christopher R. Futtner1, Amanda L. Allred1, Jianing Wang2, Cynthia Yang3, Daniele Procissi3, Xianlin Han2, Ronald M. Evans4, Ilya R. Bederman5, and Grant D. Barish1, *

1 Division of Endocrinology, Metabolism, and Molecular Medicine, Department of Medicine, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611

2 Barshop Institute for Longevity and Aging Studies, University of Texas Health Science Center at San Antonio, San Antonio, TX 78245

3 Center for Translational Imaging, Departments of Radiology and Biomedical Engineering, Northwestern University, Chicago, IL 60611

4 Howard Hughes Medical Institute, The Salk Institute for Biological Studies, La Jolla, CA 92037

5 Department of Pediatrics, Case Western Reserve University, Cleveland, OH 44106

Regional body fat distribution, regardless of general obesity, plays a critical role in metabolic risk. Excess visceral fat is directly linked to metabolic syndrome, while subcutaneous body fat is considered beneficial or benign. We have identified an unexpected role for B cell lymphoma 6 (BCL6), a transcription factor, in the expansion of subcutaneous adipose tissue. In adipocyte-specific knockout mice (Bcl6AKO), we found that Bcl6 deletion results in strikingly increased inguinal subcutaneous adipocyte size and tissue mass. Regardless of their increased adiposity, hyperinsulinemic euglycemic clamp studies revealed that Bcl6AKO mice are significantly more insulin sensitive compared to littermate controls. Genome-wide RNA expression and DNA binding analyses in adipocytes revealed that BCL6 controls gene networks involved in cell growth and fatty acid biosynthesis. Using stable isotope labeling and comprehensive lipidomics, we discovered that ablation of adipocyte Bcl6 enhances subcutaneous adipocyte lipogenesis to prevent hepatic steatosis and enhance whole body glucose metabolism. Our studies identify BCL6 as a negative regulator of subcutaneous adipose tissue expansion and metabolic health.

Wnt7a Induces a Unique Phenotype of Monocyte-derived Macrophages: Relevance to HIV Associated Neurocognitive Disorders (HAND)

Jennillee Wallace ’09 1, Victoria Lutgen1, Sreedevi Avasarala2, Brad St. Croix3,Robert A. Winn2, and Lena Al-Harthi1

1 Department of Immunology/Microbiology, Rush University Medical Center Chicago IL, 60612,

2 University of Illinois Cancer Center, University of Illinois at Chicago, Chicago IL, 60612,

3 Center for Cancer Research (CCR), National Cancer Institute (NCI), Frederick, MD 21702.

Macrophages originating from circulating blood monocytes migrate to tissue in response to homeostatic or “danger” signals and undergo substantial morphological and functional modifications based on the dominant signals in the microenvironment. Wnts are secreted glycoproteins that play a significant role in cell differentiation, yet their impact on monocyte differentiation is not clear. In this study, we assessed the role of Wnts1 and 7a on the differentiation of monocytes and subsequent phenotype and function of monocyte-derived macrophages (MDMs). We show that Wnt7a decreased the expression of CD14, CD11b, CD163 and CD206 whereas Wnt1 had no effect. Wnt7a’s effect on CD11b was also observed in the brain and spleen of Wnt7a -/- adult brain mouse tissue and in embryonic Wnt7a -/- tissue. Wnt7a reduced the phagocytic capacity of M-MDM and M1-MDMs, decreased IL-10 and IL-12 secretion and increased IL-6 secretion. Collectively, these findings demonstrate that Wnt7a-MDMs possess both pro-inflammatory and alternative MDM cytokine profiles and reduced phagocytic capacity. Ongoing studies are assessing Wnt7a-MDMs in HIV associated neurocognitive disorders (HAND) using post mortem brain tissue and a humanized animal model.

Course-Based Abstracts

FIYS106 Medical Mysteries of the Mind

Medical Mysteries of Language

Brett Grossman ’22, Danielle Herreweyers ’22, Kealey Humphrey ’22, Josephine Klein ’22, Jaleesa Lalani ’22, Samantha Stubbs ’22, and Megan Truman ’22, First-Year Studies Program and Biology Department, Lake Forest College, Lake Forest, IL 60045

Students of FIYS106 Medical Mysteries of the Mind will present the neuroscience underlying how we communicate through language and they will delve into several current medical mysteries that are the cutting of scientific research. They will educate the audience using posters, models and interactive activities.    

Medical Mysteries of Cognition

Nia Alfaro ’22, Sabina Baitemirova ’22, Carolynn Boatfield ’22, Victoria Cruz ’22, Neeva Shrestha ’22, Kyle Wallis ’22, and Graeme Witte ’22, First-Year Studies Program and Biology Department, Lake Forest College, Lake Forest, IL 60045

Students of FIYS106 Medical Mysteries of the Mind will present the neuroscience underlying how we engage in thought processes and they will delve into several current medical mysteries that are the cutting of scientific research. They will educate the audience using posters, models and interactive activities.    

Medical Mysteries of Sleep

Ayman Atcha ’22, Connor Busch ’22, Bilal Khan ’22, Max Larkin ’22, and Cassandra Majewski ’22, First-Year Studies Program and Biology Department, Lake Forest College, Lake Forest, IL 60045

Students of FIYS106 Medical Mysteries of the Mind will present the neuroscience underlying how we sleep and stay awake and they will delve into several current medical mysteries that are the cutting of scientific research. They will educate the audience using posters, models and interactive activities.

BIOL 130 Deadly Shapes, Hostage Brains

Medical Mysteries of Frontotemporal Dementias

Marissa Burch ’22, Augustana Houcek ’20, Daisy Lazcon ’22, Grace Michel ’22, and Desire Nalukwago Uwera ’22, Biology Department and Neuroscience Program, Lake Forest College, Lake Forest, IL 60045

Students of BIOL 130 Deadly Shapes, Hostage Brains will present the neuroscience underlying the molecular and genetic basis of frontotemporal dementias and they will delve into several current medical mysteries that are the cutting edge of scientific research. They will educate the audience using posters, models and interactive activities.    

Medical Mysteries of Alzheimer’s Disease

Sydni Foley ’22, Aleksandra Kulesza ’22, Bianca Mando ’22, Jose Rivera ’22, Kimberly Tarqueno ’20, and Yesenia Uribe ’22, Biology Department and Neuroscience Program, Lake Forest College, Lake Forest, IL 60045

Students of BIOL 130 Deadly Shapes, Hostage Brains will present the neuroscience underlying the molecular and genetic basis of Alzheimer’s disease and they will delve into several current medical mysteries that are the cutting edge of scientific research. They will educate the audience using posters, models and interactive activities.    

Medical Mysteries of Spinocerebellar Ataxias

Kenna Bailey ’22, Cristina Casado ’21, Jessica Pope ’22, Kylie Rosas ’22, Henry Roman ’21, and Zoe Walts ’21, Biology Department and Neuroscience Program, Lake Forest College, Lake Forest, IL 60045

Students of BIOL 130 Deadly Shapes, Hostage Brains will present the neuroscience underlying the molecular and genetic basis of spinocerebellar ataxias and they will delve into several current medical mysteries that are the cutting edge of scientific research. They will educate the audience using posters, models and interactive activities.

Undergraduate Poster Abstracts

Are Warming Waters Making Us Sick?

Niam M. Abeysiriwardena ’20 a,b, Samuel J. L. Gascoigne ’20 a,c , and Angela Anandappad,e,f

a Neuroscience Department, Lake Forest College, Lake Forest, IL

b Computer Science, Lake Forest College, Lake Forest, IL

c Biology Department, Lake Forest College, Lake Forest IL

d Alliance for Advanced Sanitation, University of Nebraska-Lincoln, NE

e Food Processing Center, University of Nebraska-Lincoln, NE

f Department of Food Science and Technology, University of Nebraska-Lincoln, NE

Global climate change, which has the effect of potentially expanding waters with cyanobacterial populations into higher latitudes, forecasts an increase in the risk of food contamination with toxins. Along with this environmental trend, the eating habits of consumers have been rapidly changing. An increase in the popularity of natural alternatives to pharmaceuticals, herbal remedies and the desire for consuming “super foods” is leading to a change in the dietary patterns of consumers. Similarly, dietary trends are plentiful, with more consumers adopting changes with little medical guidance to dietary plans that are supported by inadequate scientific data. In particular, U.S. consumers are acquiring novel foods that may not be adequately checked for the presence of marine toxins and heat stable toxins in dry or minimally processed foods. Some dry foods are cultivated in or processed in regions that may utilize hypoxic agricultural waters high in cyanobacterial or algal contamination. These may perpetuate increased risks for chronic liver, kidney, and neurodegenerative disorders due to intoxication from preventable foodborne agents. The combination of people favoring less-processed foods and natural sources of remedies, with the increase in the frequency and magnitude of cyanotoxin production in bodies of water, forecasts increasing exposure of people to cyanotoxins that could lead to chronic neurodegenerative disorders in the population.

An Evaluation of Viral Prevalence and Activity in Patients with Multiple Sclerosis

Tariq Aldaas ’21 , Dr. David Everly, PhD., Dr. Joseph Reynolds, PhD., and Dr. Matthew Sorenson, PhD.

Department of Biology, Lake Forest College, Lake Forest, IL 60045, DePaul University, Chicago, IL 60614, Department of Biochemistry and Molecular Biology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

This study is a collaboration between Drs. Sorenson, Everly, and Reynolds. Our long-term goal is to understand the relationship between viral infection, immune activation, and the development of multiple sclerosis (MS) as a means to designing innovative new approaches to treat MS. In our DePaul-RFUMS pilot project, we developed a novel screen to determine the spectrum of viruses in patients with MS using a set of previously characterized samples. Viral nucleic acids were quantified using multiple PCR trials, and viral antigenic reactivity was validated using an autoimmune specific peptide microarray. We leveraged this novel screen in conjunction with Dr. Reynolds’ expertise in autoimmune research to develop a pilot project that was funded by the National Multiple Sclerosis Society (NMSS). The aims of our project are to determine the viral prevalence and viral immune activity in MS patients compared to normal individuals. The results of these studies will provide correlations between viral immune activity and encephalitogenic T cell activation in order to better understand the relationship between viral infection and MS. In addition, these experiments will provide the conceptual framework for future experiments to identify the critical reactive cells in MS patients that are correlated with specific viral exposure. Long-term, determining specific viruses or virus/host interactions that induce MS pathology will help determine the mechanisms that underlie MS development and lead to the development of new and novel personalized therapies for MS patients.

Impact of Several PD-associated Genes on the Toxicity of α-synuclein in a Yeast Model

Ariane H. Balaram ’20 , Paul A. Jones ’18, Alexsandra Biel ’20, Emily Ong ’17, Chisomo Mwale ’19, Maiwase Tembo ’15, and Shubhik K. DebBurman

Neuroscience Program and Biology Department, Lake Forest College, Lake Forest, IL 60045

Parkinson’s disease (PD) is characterized by α-synuclein misfolding and the death of midbrain neurons. PD can be described as familial, or sporadic, both of which are influenced by a multitude of environmental and genetic factors. Familial PD is directly caused by mutations in one of at least ten genes, including SNCA, DJ-1, VPS35, and ATP13A2. SNCA, which encodes α-synuclein, has six identified missense mutations (A30P, E46K, H50Q, G51D, A53E, and A53T) that each cause aggressive PD. Sporadic PD is linked with several risk genes and loci, including VPS13, the Sac I domain of SYNJ1, and the Swa2 domain of DNAJC6. Using our previously established budding yeast model system for α-synuclein, we first show that wild-type (WT), E46K, A53T, H50Q, and A53E α-synuclein are toxic to yeast and show varying degrees of membrane binding and aggregation, while A30P and G51D α-synuclein are relatively non-toxic and show cytoplasmic diffuse localization. What is still not well understood is whether the other PD-causing and risk genes mentioned above can influence toxicity and localization properties of WT α-synuclein and these six familial PD mutants. To test the hypothesis that they do influence α-synuclein, WT and familial mutant forms of α-synuclein were studied in haploid yeast strains that were singly deleted for these six PD-linked genes (all of which are linked to loss-of-function in PD). Results show that some gene deletions increase (Δhsp31) or decrease (Δatp13, Δvps35) α-synuclein toxicity and alter its localization in a highly familial mutant specific way, while others more broadly increase α-synuclein toxicity or aggregation (Δvps13, Δsac1), while still others show no effect (Δswa2). Our findings suggest that WT and each familial mutant of α-synuclein create cellular toxicity and alter localization in distinct ways and that each is likely regulated by different subsets of genes, opening doors for mutant-specific mechanistic insight into the varying modes of α-synuclein toxicity.

Co-localization of Oxytocin Receptors in the Dorsolateral Bed Nucleus of the Stria Terminalis (BNST) with Other Neurotransmitter Systems

Alexsandra Biel ’20, Paulina Lis, and Joanna Dabrowska

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045

Department of Cellular & Molecular Pharmacology, Department of Neuroscience, Chicago Medical School at Rosalind Franklin University of Medicine and Science

The underlying hallmark of stress induced psychiatric disorders like Post Traumatic Stress Disorder (PTSD) is the inability to discriminate between a threat and safety. The Dorsolateral BNST (BNSTdl) has been proposed to regulate the discrimination between signaled and un-signaled threats (Janecek and Dabrowska, 2018) as well as having a role in the translation of stress into sustained anxiety (Dabrowska et al., 2013). The main output neurons of the BNSTdl are neurons producing stress hormone, corticotropin releasing factor (CRF) (Dabrowska et al 2016). In the rat brain, the BNST has one of the highest oxytocin receptor (OTR) expression levels (Dabrowska et al., 2011, Dumais et al. 2013). We recently investigated the role of OTRs in the BNST using a fear-potentiated startle (FPS) paradigm. We demonstrated that blocking OTR in the BNSTdl reduces acquisition of cued fear (Moaddab and Dabrowska, 2017), but the mechanism of this phenomenon is unknown, and the interaction of OTR with local neurons and other neurotransmitter systems still needs to be elucidated. Here, we have employed a double-immunofluorescence technique combined with confocal microscopy (Olympus FV10i) to determine the expression, distribution, and co-localization of OTR with other known neurotransmitter systems in the rat BNSTdl. This was done to examine if OTRs are present on 1) fibers innervating BNSTdl (Serotonin Transporter, 5-HTT; Vesicular Glutamate Transporter, VGLUT2; Oxytocin fibers:, OT) which are extrinsic in nature, or 2) dendrites and/or axons of local BNSTdl neurons, namely neurons producing CRF and/or Ca 2+/calmodulin-dependent protein kinase II alpha (CAMKIIα). To determine possible co-localization of OTR with local neurons, transgenic rats expressing Cre-recombinase (Cre) in CRF neurons of BNSTdl were used and injected with Cre-dependent viral particles expressing mCherry. Co-localizations in the BNSTdl were assessed, and based on the acquired data, our results show that OTR-positive processes are not co-localized with 5-HTT, VGLUT2, or OT. Our results also showed that there is co-localization between CRF neurons and CAMKIIα, but no co-localization of OTR with either of the two.

Hepatitis B Virus Activates Inflammasomes in Human Hepatoma Cells

Michael K. Buabeng ’19, Binod Kumar and Gulam Waris

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045

Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Chronic Hepatitis B virus (HBV) infection is a major health problem across the world. HBV belongs to the family Hepadnavirdae that infects liver cells. Since Hepatitis C Virus (HCV) is known to activate NLRP3-inflammasome for its own benefit and survival in host cells, we aimed to study if HBV also activates the inflammasome complex. We utilized the human hepatoma cell line - HepAD38 that expresses HBV under the control of the inducible tetracycline promoter. To study if HBV production also activates inflammasome complexes, we removed tetracycline from culture media and allowed the production of HBV from day1 to day 10 followed by Western blot analysis of the components of NLRP3 (NLRP3-ASC-Caspase-1) and AIM2 (AIM2-ASC-Caspase-1) inflammasomes. To further confirm the caspase-1 mediated activation, a Western blot analysis was also performed for cleaved caspase-1 and matured IL-1β. The experiments were performed with HepAD38 cells with tetracycline, as control uninfected cells. Our results demonstrated that infection of cells with HBV activated both NLRP3 and AIM2 inflammasome starting at day 2, and the expression of pro- and cleaved-caspase-1 and IL1-β increased gradually till day 10. Thus our data confirmed that HBV infection activates both NLRP3 and AIM2 inflammasomes in human hepatoma cells.

The Role of Nucleolin in Cellular Signaling Pathways for Proliferation of Inflammatory Breast Cancer Cells

Sandra P. Campos ’20, M. Repak, and N. Sharma-Walia

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Inflammatory breast cancer (IBC) is a rare type of breast cancer characterized by redness and swelling of the skin around the breast. Diagnosis of IBC can be difficult, prognosis is poor and further research is needed to clearly understand this disease. Our preliminary studies demonstrated higher expression of nucleolin in the IBC cell lines SUM149PT and SUM190PT as compared to control human mammary epithelial cells (HMECs). Nucleolin is a type of protein found to be involved in cell proliferation and cell death in various cancers such as prostate, colon, gastric, and liver. Previous studies showed that nucleolin is linked to several different types of cellular signaling pathways, such as AKT, PI3K, mitogen activated protein kinases (MAPKs) and mammalian target of rapamycin (mTOR) in other cancers. Therefore, we hypothesized that nucleolin might be regulating proliferation and survival pathways in IBC. The expression and activation of cellular signaling pathways was found to be higher in IBC cell lines SUM149PT, SUM190PT and MDA IBC-3 in comparison to control HMEC cells. By examining the role of nucleolin as a binding partner to cell signaling pathways, we will get a molecular understanding of its function in cell proliferation in inflammatory breast cancer. Understanding the functionality of nucleolin in the signaling pathways of inflammation in the breast cancer scenario will provide new avenues of promising therapeutic targets.

Assessing for Social Deficits in a Mouse Model of Batten Disease

Samuel D. Curry ’19, Jessica L. Centa, and Michelle L. Hastings

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045

Center for Genetic Diseases, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Batten disease is a rare autosomal recessive neurodegenerative disorder that is part of the neuronal ceroid lipofuscinoses class of diseases. The most common form of Batten disease is Juvenile Batten disease, which is caused by mutations in the CLN3 gene (located on chromosome 16). The age of onset is typically around 4 to 7 years of age. Symptoms include learning and behavior problems, dementia, blindness, seizures, dysarthria, and hypokinesia. There is currently no treatment for this disease, leaving patients to typically die between the ages of 15 and 30. The disease is characterized by the accumulation of auto-fluorescent storage material in the lysosomes of neurons in the brain. In the brain of a Batten disease mouse model, we discovered an accumulation of auto-fluorescent storage material in the CA2 region of the hippocampus. The CA2 is a region of the brain that is critical for social memory. Given that there appears to be CA2 dysfunction in mice with Batten disease, we hypothesize that mice with Batten disease will also exhibit social deficits. The present study aimed to identify whether social deficits can be assessed in a mouse model of Batten disease in order to determine whether social deficits can potentially be used as a behavioral diagnostic marker of Batten disease in human patients. In order to test our hypothesis, we created a modified version of a social novelty behavioral paradigm, and compared the social behavior of wild type mice to mutant (Batten disease) mice. Interestingly, we found that the wild type mice appeared to exhibit more social deficits than the mutant mice. A possible reason for why we obtained these results is that the small sample size for both the experimental and control groups may have generated a skewed data set that does not accurately represent the social behavior of either the wild type or mutant mouse genotypes.

The Influence of Predation Risk on Mate Choice in the Waxmoth Achroia grisella

Christopher Edomwande ’19 and Flavia Barbosa

Biology Department, Lake Forest College; Lake Forest, IL 60045

Mating signals are often conspicuous, and therefore, can be eavesdropped on by unintended receivers such as predators. Therefore, we expect that the evolution of female preferences will be shaped by predation risk. However, as predation risk fluctuates in the environment, a female may choose a mate that provides a compromise between her underlying preferences and the threat level. Males of the waxmoth Achroia grisella produce ultrasonic signals to attract females, and females have preference for certain signal traits such as pulse rate. Waxmoths suffer bat predation and can detect bat echolocation signals. Here, we examine how the simulated presence of a predator affects female choice in this species. We presented females with a choice between an attractive and an unattractive mating call (high and low pulse rate respectively), in either the presence and absence of a bat echolocation signal. We investigated how female choice may change under predation risk, despite underlying female preferences for high pulse rate calls: are females more likely to choose a less attractive call when the more attractive one is associated with higher predation risk?

A Genetic Screen for Calcium Channel Localization and Trafficking Mutants in Caenorhabditis elegans

Kendra Fobert ’19, K.H. Oh, and H. Kim.

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045 ; Rosalind

Franklin University of Medicine and Science, North Chicago, IL 60064

The improper function of calcium ion channels has been linked to a variety of nervous system disorders, signifying them for study and experimentation as possible targets for treatment. Calcium channels may be formed by a variety of combinations of several subunits, so a wide variety of these channels with a range of functions exists. The differences between specific channels are important to consider for use with drug therapeutics targeting calcium channels. Because of the diversity of calcium channels, there is still much to be learned about their form and function. The C. elegans unc-2gene encodes a sole channel-forming subunit of the voltage-gated calcium channel homologous to a P/Q type calcium channel found in humans. Like its mammalian homolog, UNC-2 is primarily responsible for neuronal synaptic transmission. In this project, C. elegans was used as a model organism to identify genes that control the localization and trafficking of the voltage-gated ion channel UNC-2. We took advantage of the transgenic line designed to express GFP-marked UNC-2. This transgenic line was put through mutagenesis and examined to find mutants with apparent reduction or loss of the UNC-2 level. The motor capabilities of these worms were severely impacted, causing them to move at much slower speeds than worms of their original line, and in some cases reproduction rates were also reduced. In a few mutants, GFP levels were significantly reduced, suggesting that mutations may reduce UNC-2 levels by influencing UNC-2 stability or trafficking. These results are in alignment with the primary expression of UNC-2 being located in motor neurons and two types of neurons related to egg laying. Further study is currently being conducted with these mutants to determine their specific genetic properties.

The Impact of Phenytoin on preBötzinger Complex Inspiratory Rhythmogenesis

Philip Freund ’19 and Kaiwen Kam, Ph.D.

Neuroscience Program, Department of Biology, Lake Forest College, Lake Forest, IL 60045; Department of Cell Biology and Anatomy, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

The preBötzinger Complex (preBötC) is a rodent medullary neural circuit that generates inspiratory respiratory rhythms in vivo and in vitro. It converts tonic excitatory drive into rhythmic neuronal bursting, which can be detected in both the preBötC itself and the hypoglossal (XII) nerve. Phenytoin is an anti-epileptic drug that selectively binds to Nav1.1 and Nav1.5 voltage-gated sodium ion channels, resulting in a time-dependent block of channel function. Application of phenytoin results in lower neuronal firing rates, especially during high-frequency bursting. Based on this information it was hypothesized that extracellular application of phenytoin to rhythmically active brainstem slices from neonatal mice containing the preBötC would result in a higher interval between preBötC bursts. To test this hypothesis, neonatal mice (P0-P4) were anesthetized, and their brainstems were serially sectioned. A 550 μm slice exposing the preBötC rostrally was extracted from each mouse, and electrophysiological recordings of preBötC and/or XII nerve activity were obtained. Burst duration, interval, and amplitude were measured. Upon application of phenytoin, preBötC bursting was ablated or reduced. This bursting was recovered after the phenytoin was washed off. These results show that phenytoin does have a marked effect on preBötC rhythmogenesis. Future research should consider the how phenytoin affects preBötC rhythmogenesis at the circuit level and how external ionic concentrations might modulate this effect.

Insight into Parkinson’s Disease from Yeasts:Combined Impact of Covalent Modifications and Familial Mutations on alpha-synuclein.

Yoan Ganev ’19, Rosemary Thomas ’18, Chisomo Mwale ’19, Joseph Mountain ’21, Ariane Balaram ’20, Alexandra Roman ’16, Morgan Marshall ’16, and Shubhik DebBurman

Neuroscience Program and Biology Department, Lake Forest College, Lake Forest, IL 60045

Parkinson’s disease (PD) is a neurodegenerative disorder linked to the loss of dopaminergic neurons in the midbrain. A key pathological marker of PD is the presence of Lewy bodies, which are mainly composed of misfolded alpha-synuclein protein. Alpha-synuclein is a highly post-translationally modified protein. While phosphorylation and nitration of alpha-synuclein are well-studied as contributors to PD pathology, less is known about sumoylation, which is proposed to be neuroprotective based on limited studies. The majority of sumoylation takes place on lysine-96 and lysine-102 of alpha-synuclein, and it increases the protein’s solubility. The first goal of this research was to better understand the role of sumoylation in regulating alpha-synuclein toxicity, and we performed four studies towards it.  We evaluated the effects of blocking sumoylation on alpha-synuclein in the well-established budding yeast model for PD and found that alpha-synuclein becomes more aggregated and toxic, losing its membrane localization. Second, we evaluated the effects altering sumoylation pathways by using yeast strains with reduced (ulp1ts) or excessive sumoylation (smt3ts), and found that alpha-synuclein aggregates more with reduced sumoylation, but becomes less toxic with increased sumoylation.  Third, we asked how altering the phosphorylation of alpha-synuclein would alter sumoylation’s protective role and found that blocking phosphorylation reduced alpha-synuclein toxicity in the absence of sumoylation. Finally, we evaluated whether blocking sumoylation on familial PD mutant versions of alpha-synuclein would exacerbate its toxicity, but we have found little evidence to that effect. We also began investigating two more-recently discovered modifications of alpha-synuclein – acetylation and glycation. We found preliminary evidence for the hypotheses that acetylation is protective, glycation is harmful, and both can interact with sumoylation and phosphorylation.

Plasma Membrane Citrate Transporter

Christina Gimondo ’21 and Ronald Kaplan

Department of Biology, Lake Forest College, Lake Forest, IL 60045;Rosalind Franklin University of Medical Science and Lake Forest College.

With new cases of liver cancer on the rise and an overall poor prognosis, the need for drugs targeting the formation of this type of cancer has become increasingly paramount. The plasma membrane citrate transporter, a mammalian ortholog of Drosophila melanogaster’s Indy (I’m Not Dead Yet) gene, facilitates the sodium dependent transport of citrate from circulation into the cell, and is commonly expressed in the liver and brain. Studies have shown that this transporter, encoded by SLC13A5, plays a role in the formation of liver cancer through its supply of the significant metabolic intermediate citrate, and that the inhibition of this gene can stop the growth and proliferation of hepatocarcinoma cells. This research utilizes the HEK293T cell line to study the characteristics of PMCT and test potential inhibitors that could potentially serve as therapeutics using kinetic and protein assays. The study has revealed several promising inhibitory compounds that are effective at therapeutic doses with over 40% inhibition of transport. Further testing will need to be done on these compounds to confirm efficacy as well as explore potential analogs.

Do Modifications in Yeast Cause Parkinson’s Disease? Creating Tools in Yeast

Leslie Gonzales ’21, Zubair Mohammed ’21, Matthew McMahon ’21 , Chisomo Mwale ’19, Yoan Ganev ’19, and Shubhik DebBurman

Neuroscience Program and Biology Department, Lake Forest College, Lake Forest, IL 60045

Parkinson’s disease (PD) is a neurodegenerative disorder linked to the loss of midbrain dopaminergic neurons. A key pathological feature, Lewy bodies, are aggregates composed mainly of misfolded protein α-synuclein. Six familial mutations (A30P, E46K, A53T, H50Q, G51D, A53E) of α-synuclein have been linked with early onset Parkinson’s disease. In addition, evidence shows that α-synuclein undergoes multiple post-translational modifications. Two modifications that we looked at were acetylation and glycation. These affect multiple sites on the α-synuclein protein, but we looked at Lysine-6 and Lysine-10 for our project. Research in our lab has just begun on the effects of acetylation and glycation on the wild type α-synuclein, but no work has been done on the effects of combining the familial mutants of α-synuclein with modifications of acetylation and glycation. We hypothesized that the familial mutations with acetylation enhancement would have protective properties on cells and glycation enhancement would show more toxicity; but the level of that toxicity would depend on the individual familial mutant. To explore this hypothesis, the overall goal of our Richter project was to create each of the six familial mutations on three different versions of α-synuclein template DNA: acetylation-blocking (K6R/K10R/K6-K10R), glycation-blocking and acetylation-blocking (K6A/K10A/K6-K10A), and acetylation mimicking (K6Q/K10Q/K6-K10Q). We used PCR-based site-directed mutagenesis to create these mutations on the three α-synuclein templates. Here, we share our results that we successfully made 18 variants and expressed some of them in budding yeast. Functional studies are already under way to assess the properties of these new variants in yeast.

Analysis of Proliferation Rates of Muscle Stem Cells with 5-Azacytidine Treatment in Cerebral Palsy

Wendy Gross ’20 1,2, Lydia A. Sibley 1,  Austin Menezes 1,3, Richard L. Lieber 1,4,  and Andrea A. Domenighetti 1,4

1 Shirley Ryan Ability Lab, Chicago, IL.

2 Lake Forest College, Lake Forest, IL.

3 University of Cincinnati College of Medicine, OH.

4 Department of Physical Medicine & Rehabilitation, Northwestern University, IL.

Cerebral palsy (CP) is a permanent movement disorder causing activity limitation due to a non-progressive lesion in the developing infant brain. Children with CP develop muscle contractures which are associated with shorter and thinner muscles, smaller muscle fibers, overstretched contractile units called sarcomeres, and a reduced range of motion around the joints. The disability associated with CP spans from limiting the motor function of just one limb to affecting the whole body.

Our lab showed that muscle contractures in children with CP are associated with an epigenetic DNA hypermethylation imprint that prevents MuSC-derived myoblasts from producing new muscle fibers in vitro. We also obtained preliminary evidence that pretreatment with 5-Azacytidine (5-AZA), a FDA-approved DNA hypomethylating agent used to treat blood-related cancers, restored the capacity of CP myoblasts to generate new muscle fibers in vitro, suggesting that 5-AZA may contribute to normalization of the biology of cell division, differentiation, and survival of MuSCs in contractured CP muscle.

The purpose of this study was to investigate the proliferation rates of CP myoblast cultures and compare them to control TD myoblast cultures, and cultures treated with 5-AZA.  We hypothesized that CP myoblasts in culture would proliferate faster than TD, and the drug 5-AZA would ultimately slow down and normalize the cell cycle in CP myoblast cultures, allowing these cells to move to the next step of myogenesis and produce more multinucleated myotubes.

Our results showed possible mechanisms by which 5-AZA improves myogenesis in MuSC-derived myoblasts from contractured CP muscle. Global DNA hypomethylation was associated with decreased proliferation rates, which was more significant in CP myoblasts compared to TD myoblasts. We propose that depletion of the number of MuSCs observed in contractured CP muscle is due to a hyperproliferative phenotype of these stem cells caused by an abnormal epigenetic imprint of the DNA. Repurposing this anti-cancer drug, 5-AZA, to rescue MuSC homeostasis may one day represent an effective way to treat muscle contractures non-surgically.

Selective Serotonin Reuptake Inhibitors versus Multimodal Serotonergic Drugs for the Treatment of L-DOPA Induced Dyskinesias in an Experimental Model of Parkinson’s Disease

Nicole Hedger ’19 , Feras Atwal, Alex Ritger, Janhavi Dhargalkar, Cristian Solis, and Anthony West

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045; Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Parkinson’s disease (PD) is the second most common neurodegenerative disease after Alzheimer’s (Ross, Braithwaite, Farrer, 2008). PD is a devastating hypokinetic movement disorder which affects millions of people per year. The most commonly used treatment for PD is levodopa (L-DOPA), and while it is effective for improving motor dysfunction by regulating corticostriatal transmission and increasing dopamine (DA) presence, it produces debilitating motor side effects termed L-DOPA-induced dyskinesias (LIDs). Recent studies in dyskinetic parkinsonian models have implicated serotonergic raphe-striatal terminals in the uptake and conversion of exogenous L-DOPA to DA, as these neurons possess the enzymatic machinery required to convert L-DOPA to DA and mediate vesicular storage, i.e. vesicular monoamine transporters and tryptophan decarboxylases. Serotonin neurons, however, are unable to detect or regulate synaptic dopamine levels as they lack the feedback mechanism required for the controlled release of the neurotransmitter. As such, a non-physiological release of DA and serotonin (5-HT) appears to occur, suggesting that the serotonergic system might be a key component in the pathophysiology of LIDs. Pre-clinical models and clinical trials have found that by adding to the regimen either selective 5-HT reuptake inhibitors (SSRIs) which block the 5-HT transporter (SERT), or selective 5-HT1A/Breceptor ligands which stimulate 5-HT autoreceptors to potentially suppress DA release from 5-HT terminals, LIDs are markedly reduced. Although promising at first, these drugs are oftentimes associated with reduction in the prokinetic effects of L-DOPA, as well. The goal of the current study was to identify a novel multimodal 5-HT drug which can act to attenuate the expression and severity of LIDs, without interfering with the antiparkinsonian efficacy of L-DOPA. Among these, the multimodal drug Vilazodone is of interest as it is known to exhibit potent SSRI and 5-HT1A partial agonist properties. Unilateral 6-OHDA-lesioned rats modeling PD were treated with either vehicle and L-DOPA (2.5 - 5.0 mg/kg), Vilazodone (10 mg/kg) and L-DOPA, or the SSRI Escitalopram (3 mg/kg). Rats were treated for 5 consecutive days/week, for 2 weeks. On the second day of each week, stepping tests were performed prior to drug administration, and 60 minutes post L-DOPA treatment. Behavioral assessment of LIDs was performed every 30 min for 180 min during the last three treatment days of each week. Vilazodone co-administration significantly reduced LIDs in 6-OHDA lesioned rats when administered 30 min prior to L-DOPA treatment, but had no effects on forelimb akinesia or L-DOPA-induced prokinetic effects. Escitalopram co-administration also lead to a significant reduction in LIDs and did not affect the prokinetic effects of L-DOPA. Additional studies are ongoing to assess the impact of these treatments on corticostriatal transmission and striatal neuronal activity. The current results indicate that together with L-DOPA, multimodal 5-HT drugs such as Vilazodone may be safe and effective co-therapies for reducing side-effects of L-DOPA, such as hyperkinesia and dystonia, in PD patients, allowing for more dosage flexibility in the chronic treatment of PD with L-DOPA.

Evaluating Direct Reprogramming of Human Fibroblasts into Neurons as a New Model for AD

Aiden Houcek ’20 and Dr. Robert Marr

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045; Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Alzheimer’s disease is a neurodegenerative disorder of unknown etiology. Despite substantial investment in AD research, current FDA approved therapies such as memantine have been efficacious in slowing disease progression, but as of now there is no known cure. Although animal models have been valuable for understanding AD pathogenesis, multiple failures of phase 3 clinical trials suggest that new approaches are urgently needed. With the advent of human induced neuron (HiN) technology, we are now able to model AD in human tissue. Here, we employed the use of genetically encoded calcium indicators (GECI’s), which reveal, with extreme sensitivity, intracellular calcium signaling events. Calcium regulation and its relation to AD has recently been shown to preclude many other symptoms of the disease, such as amyloid-b and hyper-phosphorylated tau protein. By using the direct reprogramming method, we can preserve epigenetic markers of AD patient cells, and our work will determine if direct reprogramming is a viable approach for modeling calcium dysregulation in AD, ultimately providing evidence for the benefits of using the disease model to better represent AD pathology.

Chronic Administration of Nicotine Does not Improve Cognitive Flexibility in an Attentional Set-shifting Paradigm

Abagayle L. King ’19 and Jean-Marie Maddux, Ph.D.

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045

A common way to measure cognitive flexibility is through an attentional set-shifting paradigm. Many studies have demonstrated that administering nicotine in rats before the task increases performance. No such studies have been done examining the effect of chronic nicotine administration on attentional set-shifting tasks. Given that nicotine works in the prefrontal cortex to increase cognitive performance, it was predicted that chronic nicotine administration will induce plastic changes in the prefrontal cortex, which would increase performance in a set-shifting task. In the present study, daily nicotine injections were given to rats across a 14-day injection regimen. Rats were tested in an attentional set-shifting paradigm without any nicotine in their system to assess the effect of chronic nicotine administration on cognitive flexibility. Contrary to the hypothesis, nicotine did not have an effect on performance in the set-shifting task. These results pose a challenge to the common finding of acute injections enhancing performance on attentional set-shifting tasks.

Eye-Tracking: Physique, Face, and Distraction

Greg Lammers ’21, Ramona Gyorfi ’20, & Naomi Wentworth

Psychology Program, Lake Forest College; Lake Forest, IL 60045

Physical attractiveness is one of the most powerful distractors for humans. Though, the exact region in which individuals are most distracted by is less than certain. This research aimed to identify whether the physique or face was the more distracting feature of the body. To try and answer this question an initial priming phase was established, showing participants pictures of faces or bodies of the opposite sex. After, in a designed distraction test participants were asked if they could recognize subtle differences in a central stimulus, while distracting stimuli (face and body) were presented on each side of the central stimulus. It was hypothesized that the body stimuli would be more distracting regardless of the priming stimuli presented. To unveil the results, eye-tracking analysis was used to record distraction in three different vulnerabilities.

Crystal Structure Analysis of Gallic Acid Decarboxylase and Pea Pathogenicity Protein

Nebojsa Markovic 1, Cristina V. Iancu 2, and Jun-Yong Choe 2

1 Chemistry Department, Lake Forest College, Lake Forest, IL 60045

2 Department of Biochemistry and Molecular Biology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Tannins are plant metabolites that play a role in plant defense. They are known for their binding and ability to precipitate proteins and thus are linked to different pathologies, including cancer. The hydrolysis of tannins produces Gallic Acids, which are used in the pharmaceutical industries for their anticancer and antioxidant properties. Gallic acids are difficult to degrade and often contribute to pollution. Gallic Acid Decarboxylase (GADC), a protein found in several different types of bacteria and yeast, is an enzyme that decarboxylates gallic acid to form pyrogallol, allowing the natural degradation of gallic acid. This research was conducted to determine the three-dimensional structure of GADC from Arxula adeninivorans (a type of yeast) and also of its homologue, Pea pathogenicity protein (PPP) from Madurella mycetomatis. For protein preparation, we cultured Escherichia coli cells containing the plasmid with GADC or PPP tagged with polyHistidine, and purified the proteins by immobilized metal affinity chromatography (IMAC). We crystallized the purified proteins and collected x-ray diffraction data on the protein crystals at Advanced Photon Source, Argonne National Laboratory. Crystal structures revealed the active site of the enzymes along with several metal binding sites, including cobalt and potassium sites. As expected, mutants of residues in the active site decreased or abolished enzyme activity. We are currently investigating the role of the metal sites in GADC and PPP.

An Examination of the Relationship between Personality and Adherence/Adherence Barriers in Young Adults

Victoria M. Miller ’21, L. Durkin, and R. N. Greene

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045, Rosalind Franklin University of Medicine and Science, North Chicago, IL, 60064


Adherence is the degree to which a person is able to correctly follow medical advice and the most commonly assessed type of adherence is medication adherence. Many factors can affect a person’s medication adherence. Personality factors have previously been linked with predicting adherence levels in adults, such that higher levels of conscientiousness are associated with higher levels of adherence. Little research has been done about the relationship between personality and adherence in young adults. Less research has examined the role of personality in influencing barriers to medication adherence, though there is reason to think that personality may play a role in the nature of barriers experienced.


The aims of this study were to examine the relationship between personality factors and adherence levels and to explore the relationship between personality factors and adherence barriers to see if these predictions were replicable in young adults.


Participants included a sample of 197 between the ages 18 – 24 were asked to complete an anonymous online survey which included measures of personality, medication adherence, and adherence barriers. The factors that can affect a person’s adherence, known as adherence barriers can be classified as: forgetfulness, health and wellness interfering, medication issues, adolescent development, and organizational.


The only personality factor that was significantly associated with overall adherence was conscientiousness, with individuals higher in conscientiousness reporting higher levels of medication adherence. Agreeableness, neuroticism, openness, and conscientiousness were linked to individual adherence barriers.


The knowledge of the relationship between personality and adherence could be helpful when predicting adherence to prescriptions based on personality types. More research could be conducted on the role of personality on adherence to gain a more insightful role in how this could be helpful with understanding if some young adults are more likely to adhere to prescriptions. Future research should continue to investigate these patterns in a larger, more diverse sample.

Identification of Neuronal Composition of the pre-Bötzinger Complex in Neonatal Mice

Grace Monshausen ’21 and Kaiwen Kam

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045, Department of Cell Biology and Anatomy, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

The pre-Bötzinger complex (preBötC) is a neural circuit located in the ventral medulla of the brainstem and the hypothesized circuit for rodent inspiratory respiratory rhythmogenesis. It converts tonic excitatory drive into rhythmic neuronal bursts that can be detected both in the preBötC and the hypoglossal (XIIth) nerve, in vivo and in vitro. Neonatal mice expressing both the EGFP and Dbx1-td tomato genes were used to study the neuronal composition of the preBötC. Dbx1-td tomato strain mice express red fluorescence, marking excitatory glutamatergic neurons, and Glyt2 EGFP strain mice express green fluorescence, marking inhibitory glycinergic neurons. These double fluorescent neonatal mice were perfused to order to fix their brains. Subsequently, their brains were extracted, sliced, and mounted to slides for immunohistochemical staining. The immunohistochemistry performed used anti-ChAT, an antibody that binds to choline acetyltransferase (ChAT) at the terminals of cholinergic motor neurons. DAPI, a DNA stain, was used to identify the nucleus of all neurons in the sample. The immunohistochemical staining as well as the fluorescence of the neurons in the brain slices allow for the neuronal makeup of the slices to be assessed using confocal microscopy. In the future, the fluorescent and stained neurons can then be analyzed using a computer modulated system. The goal of this is to be able to better identify the boundaries of the preBötC and significant collections of neurons surrounding it. Due to the limitations of this work, a larger sample size of mice should be analyzed. Future research should include slicing, performing immunohistochemistry, and analyzing more mice brains to collect a larger sample size of data.

Effects of Prolonged Social Isolation: Sex Differences in Anxiety, Depression, and Sociability Behavior in Male and Female Adolescent Rats

Eliska Mrackova ’19 and Dr. Jeremy A. Rosenkranz

Neuroscience Program, Lake Forest College, Lake Forest, IL 60045, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Clinically, early life stress such as prolonged social isolation is generally regarded as a risk factor for the development of depression and anxiety. Extensive research has found that sex differences in depressive-like behaviors emerge during adolescence, with females more at risk than males. Despite the rising social isolation and loneliness across the world, the causes and effects of social isolation in adolescence remain still poorly understood. Therefore, the purpose of this research project was to determine whether the social isolation duration, sex of the animals or the housing condition would affect the development of depressive-like behaviors. Overall, the Open Field test (OF) revealed no significant differences due to the social isolation condition. Nevertheless, the duration of social isolation (1-week vs 4 weeks), condition (group housed versus socially isolated) and sex (male vs female) had significant effects on social behavior both in the Social Interaction test (SI) as well as the Conditioned Place Preference test (CPP). Appetitive conditioning with anxiogenic bright light showed that after 4 weeks of social isolation both males and females’ number of active nose pokes significantly dropped due to the presence of the bright light stressor and returned to the baseline when the bright light was turned off. Overall, this research adds a new insight into the understanding of sex differences associated with prolonged social isolation and subsequent depressive-like behaviors. Moreover, future research will focus on determining the differences between socially isolated males and females using the deltaFosB staining as an indicator of neuronal activation in response to chronic stress in the Basolateral Amygdala (BLA). 

UNDERSTANDING PARKINSON’S DISEASE IN YEAST MODELS: The Nature of Toxicity Linked with A30P, H50Q and A53E a-Synuclein Mutants

Chisomo Mwale ’19, Emily Ong ’17, Paul Jones 18’, Maiwase Tembo ’15, Charles Alvarado ’16, Michael Buabeng ’19, and Shubhik DebBurman

Department of Biology and Neuroscience Program, Lake Forest College, IL 60045

Parkinson’s disease (PD) is associated with the aggregation and misfolding of alpha-synuclein in midbrain dopaminergic neurons. The gene for alpha-synuclein has six known mutations that cause early-onset familial forms of PD. The pathological determinants of three of these mutants (A30P, E46K, and A53T) are well characterized in diverse model systems and they reveal that each mutant affects cellular toxicity in distinctive ways. The three more recently discovered familial mutants (H50Q, G51D, and A53E) are not extensively studied. We expressed H50Q, G51D, and A53E mutants in budding and fission yeasts model systems and hypothesized that each would generate toxicity by altering their membrane association and aggregation properties, and by disrupting cellular pathways including nitrative stress responses and endocytosis, but each would do so in distinctive ways. First, we found that the H50Q and A53E mutants were toxic to yeast, and bound membranes and aggregated within yeast, while G51D was cytoplasmically diffuse and nontoxic. Secondly, we asked whether the loss of the original amino acid or the gain of the new amino acid in each new familial mutant is responsible for disease. We created four substitution mutations for H50Q, G51D, and A53E in both yeasts models corresponding to the four functional classes of amino acids. We found that H50D was cytoplasmically diffuse and nontoxic, G51A bound membranes and aggregated like WT, G51E was cytoplasmically diffuse and nontoxic like G51D, and A53R was cytoplasmically diffuse and nontoxic, suggesting both the loss of the original amino acid and the gain of the new amino acid are key. Thirdly, we found that some of these new familial mutants had increased toxicity in yeast strains altered for nitrative stress (particularly G51D), sumoylation, and endocytosis. Collectively, this work adds insight into the pathogenicity of different familial PD mutants of alpha-synuclein.

Mutating the Telomere Gene

Brett Palmero ’20, Alexandra Dejneka ’21, Anna Sandler ’20, Junya, Li ’18, and Dr. Karen Kirk

Neuroscience Program and Biology Department, Lake Forest College; Lake Forest, IL 60045

Telomeres are protein-nucleotide sequences at the ends of chromosomes that serve to prevent the loss of genetic information during cell replication. A telomerase ribonucleoprotein complex includes telomerase reverse transcriptase (TERT) and an RNA template (TER) and is responsible for adding telomeric DNA. This holoenzyme is active in cells that divide regularly, like yeast or Aspergillus nidulans, but is repressed in most other cells. Control of telomerase activity is therefore crucial for the maintenance of normal cellular function. Excessive telomere shortening caused by the downregulation of telomerase accelerates aging while upregulation enables cells to replicate limitlessly and become cancerous. Our lab has been studying the biogenesis of telomerase in A. nidulans, a species of fungi that can exhibit a multi-nucleate state known as a heterokaryon, which can help determine if a gene is essential. We assay for the presence of two genetically distinct nuclei via the heterokaryon test, a molecular tool that has allowed us to deduce whether the RNA products for TER and TERT leave the nucleus post synthesis. Researchers in our lab knocked out the genes for TER and TERT, followed by the heterokaryon test, to determine if assembly of the active enzyme occurs in the nucleus or the cytoplasm. Our results suggest assembly in the nucleus, a scenario that is similar to humans but not to yeast, where the telomerase RNA appears to leave the nucleus and potentially be assembled in the cytoplasm. These results further suggest similarities between human and A. nidulans telomerase RNAs, not only structurally as has been shown previously, but now in the confinement of the RNA to the nucleus.

Hepatitis B Virus Infection Leads to Activation of Inflammasome in Human Monocytic THP 1 Cells

Anthony Sullivan ’21 1, Binod Kumar 2, Gulam Waris2

Biology Department, Lake Forest College 1, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064 2

Although Hepatitis B virus (HBV) currently has a vaccine, yet chronic HBV infection is a major clinical problem worldwide. The adverse outcomes of prolonged infection may be liver fibrosis, cirrhosis and hepatocellular carcinoma (HCC). The body’s innate immune system is well equipped to sense the invading RNA and DNA viruses and activate several specific inflammasomes for protection against infections. Although HBV has been reported to induce the activation of AIM2 inflammasome in clinical specimens, very few articles have supported this in laboratory settings in cell line models. The human THP-1 cells have previously been used for showing the induction of inflammasomes by several viruses as well as by LPS-ATP.  Thus, in this study, we aimed to analyze if HBV de novo infection can activate the inflammasomes in THP-1 cells. We used the supernatant of HepAD38 cells as the source of virus as this cell line expresses HBV under the control of the inducible tetracycline promoter. We infected the supernatant collected from HepAD38 Tet+ (HBV-ve), HepAD38 Tet- (HBV+ve) at day 6 and infected the THP-1 cells for 6-8 hours along with relevant control (Mock and LPS-ATP) followed by western blot analysis for the NLRP3 inflammasome complex.  We observed a significant induction of NLRP3 inflammasomes which was supported by increase in the caspase-1 cleavage and matured IL-1β production in Tet- infected cells as compared to Tet+ infected cells. The LPS-ATP treated cells served as positive control for the experiment and showed maximum induction of NLRP3 and cleavage of Caspase-1 and IL-1β. Thus we concluded that HBV infection induces the activation of NLRP3 inflammasome complex in human THP-1 cells.

Investigation of the Neuroinflammatory Response in Hippocampal Subfields After a Single Mild Traumatic Brain Injury (mTBI) in Rats

Danielle J. Sychowski ’19, J. Love, R. Greene, and D. A. Peterson

Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Mild forms of traumatic brain injury (TBI), such as concussions from falls, sports, or combat, are often associated with memory deficits that may or may not be reversible. Even a single TBI can alter the brain in such a way that leaves it vulnerable to further injury. Experiencing multiple TBIs has been shown to increase risk of neurodegenerative disease, particularly chronic traumatic encephalopathy (CTE), which has been increasingly found in the brains of former American football players. We recently described a clinically relevant closed-head model for mild TBI, and are using this model in rats to evaluate mechanisms of neuroinflammation and to explore possible therapeutic strategies. Our analysis is focused upon injury in the hippocampus of rats that underwent mild TBI at a chronic time point. Sagittal sections of the brains were immunostained to detect mature neurons (NeuN), astrocytes (GFAP), and microglia (IBA1), and DAPI was used as a nuclear counterstain. These sections were imaged using confocal microscopy and then analyzed using Stereo-Investigator software to evaluate neuronal loss and inflammation in the CA1 and CA3 hippocampal subfields. Establishing neuronal loss in the hippocampus and understanding the role of inflammation after TBI would provide a useful model for investigating regenerative approaches to replacing neurons through reprogramming induced neurons in the hippocampal CA subfields.

Unlocking Parkinson’s: Which α-Synuclein Modifications Hold the Key to Disease?

Estella Tcaturian ’21, Yoan Ganev ’19, Chisomo Mwale ’19, and Shubhik DebBurman

Neuroscience Program and Biology Department, Lake Forest College, Lake Forest, IL 60045

How proteins maintain their shapes is key to human health and disease and chemical modifications on proteins control their shapes. Parkinson’s Disease is a neurodegenerative disorder in which the loss of motor function results from the death of dopaminergic neurons. In my research project, I focused on evaluating four key modifications (phosphorylation, sumoylation, acetylation, and glycation) on the protein, α-synuclein, whose misfolded shape leads to Parkinson’s disease. Using molecular genetic approaches, we altered α-synuclein and/or yeast (our model system) for these four modifications. Our results demonstrate that α-synuclein’s toxicity is influenced by a combination of these modifications and add insight to this complex and fatal brain disease that has neither cure nor effective treatment. Specifically, we found that acetylation may be protective (and may help reinforce the positive effects of SUMOylation) while glycation is harmful and may interact with phosphorylation. These results are among the first to demonstrate that these modifications work together to contribute to PD pathology. 

Nucleolin’s Role in Angiogenesis/Lymphangiogenesis in Inflammatory Breast Cancer (IBC)

Samantha L. Russell ’20, M. Repak, and N. Sharma-Walia

Biology Department, Lake Forest College, Lake Forest, IL, Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, Chicago, IL

Inflammatory breast cancer (IBC) is an aggressive form of cancer that is usually identified by red swollen breast tissue due to blocked lymph nodes that affects between 1 and 5 percent of the US population and is usually identified at stage three and four. Based on the aggressive nature of IBC, angiogenic factors play a major role in the metastasis and tumor growth of inflammatory breast cancer. Our preliminary studies showed that IBC cell lines (SUM149PT and SUM190PT) expressed higher level of nucleolin as compared to control human mammary epithelial cells (HMECs). Nucleolin is a nucleolar protein involved in cell proliferation and angiogenesis in many cancers. Based on our preliminary results and previous reports on the functionality of nucleolin in aggressive cancers, we hypothesize that nucleolin regulates the expression and secretion of angiogenic factors like vascular endothelial growth factor (VEGF). High levels of pro-angiogenic factors in turn drive the progression of formation of new blood vessels and making tumors accessible to other parts of the body. By using enzyme linked immunosorbent assay (ELISA), Western blot and immunofluorescence assay, we screened for various angiogenic factors and the possible role nucleolin is playing in the regulation of angiogenesis and lymphangiogenesis. Our studies will help understanding the molecular pathways and functions of nucleolin in the IBC progression and strive for possible therapeutic targets.

Examining the Effects of Time Pressure on the Brainwaves

Audrey Taylor ’21 and Naomi Wentworth

Psychology Department, Lake Forest College, Lake Forest, IL 60045

10 subjects (5 male/ 5 female) participated in a word categorization task to study the effects of time pressure on performance in reaction time and accuracy of words correctly categorized. Electrodes recorded the brain activity of participants while performing a word categorization task presented on a computer screen. Time pressure was manipulated by creating two conditions: a fast presentation of the stimulus at (550 ms) and a slow presentation of the stimulus of (800ms). Results revealed a significant main effect for lead (p=.001) demonstrating that the P300 amplitude in the frontal part of the brain (Fz) was greater than in the Parietal part (Pz). Although there were not significant results for gender or conditions, a projected positive slope for the percentage of correct words per their reaction time was apparent.

Barriers to Accessing Opioid Use Disorder Treatment for Pregnant Women and Mothers

Tubanji Walubita ’19, K. Elliott, and P. Corcoran

Lake Forest College, Lake Forest, IL 60045; Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Opioid use disorder (OUD) is a problematic pattern of opioid use that causes clinically significant impairment and distress in an individual. The incidence of OUD in the United States is increasing at an alarming rate, particularly among women of childbearing age. Women are at a high risk of experiencing OUD because at all age groups, women receive more opioid prescriptions than men. OUD has major societal consequences for both women and infants.  The number of deaths due to prescription opioid overdoses among women has more than quadrupled in the past two decades. Furthermore, as more women of childbearing age are diagnosed with OUD, more infants are severely affected by their mother’s opioid use or abuse. Thus, the number of infants diagnosed with neonatal abstinence syndrome (NAS) also continues to increase. Therefore, OUD among women has an immense effect on the most vulnerable members of society – infants. The treatment of pregnant women with OUD is especially complicated because medical providers must consider the effect that treatment could have on the both the mother and the fetus. Moreover, detoxification is not recommended during pregnancy because it is associated with high maternal relapse rates. The issue of treating women with OUD is very complex due to medical and societal concerns. Hence, pregnant women with OUD face numerous barriers to accessing and receiving adequate treatment for OUD. We reviewed current literature to determine an effective form of treatment for pregnant women with OUD and to investigate the barriers that prevent pregnant women and mothers from accessing treatment for OUD. We found that medication assisted treatment (MAT) is the most effective form of treatment for pregnant women and mothers with OUD due to its association with a low maternal relapse rate. Moreover, we found that barriers do exist, and they inhibit these women from seeking treatment. We identified that state policies, stigma, and fear prevent pregnant women and mothers from accessing this effective form of treatment for OUD. Finally, we offer some recommendations for change that can be implemented at the community, state, and federal level to provide better treatment and access to treatment for pregnant women and mothers.