Easel #1  - Tamia Abbasi - (9-10am; 3-4pm)

Investigating EGR1 downstream targets in osteosarcoma

Tamia Abbasi¹, Syeda Maryam Azeem², Shahana Mahajan PhD^1,2,3,4

¹Department of Medical Laboratory Sciences, Hunter College

²The Graduate Center of City University of New York, Biology

³The Graduate Center of City University of New York, Biochemistry

⁴Weill Cornell Medical College, Brain and Mind Research Institute

Hypothesis/Statement of Problem: Osteosarcoma, a type of malignant bone cancer, has an increase in glutamate receptor activity which aids in cancer survival and proliferation. Despite medical advancements, survival rates have not greatly improved over the past three decades. Riluzole, an FDA approved glutamate blocker, used for treatment of amyotrophic lateral sclerosis, is currently being studied to be repurposed as a potential therapeutic agent for osteosarcoma. Both in-vitro and in-vivo, Riluzole has shown to inhibit osteosarcoma growth and cause cells to undergo apoptosis. EGR1 plays an important role in many physiological processes due to its involvement in cell proliferation and apoptosis. In osteosarcoma, EGR1 expression is highly downregulated but when cells are treated with Riluzole, EGR1 expression increases inducing apoptosis. However, the role of EGR1 interaction with downstream genes in inducing apoptosis has not been studied well in osteosarcoma.

Methods: Total RNA was extracted after 0h, 2h, and 4h treatment with 100 uM Riluzole for U2OS cells and 0h, 4h, and 6h treatment with 100 uM Riluzole for LM7 cells. Total RNA was eluted in 30 µL of RNase-free water and 1 µg of total RNA was reverse transcribed to cDNA. Quantitative PCR reactions were run using Sybr Green and the comparative Ct method was used to determine fold change in expression. Samples were run in triplicates with verified primers and normalized to Actin.

Results: We detect a pattern of increase in pro apoptotic genes. We intend to repeat the qPCR experiments to further validate our current results.

Easel #2 - Spartak Abramov - (1-2pm; 3-4pm)

Impact of AI on Human Thought

Spartak Abramov

Even in the arts and social sciences, where critical thinking, creativity, and empathy are highly valued skills, artificial intelligence (AI) has the potential to replace human thought. AI systems are increasingly capable of carrying out activities that were previously believed to be the sole preserve of human intelligence, such as the analysis of literature, music, and art, or the interpretation of social phenomena.

By displacing human scholars in the humanities and social sciences, AI might render human thought ineffective in these domains. Huge data sets can be analyzed by AI systems, which can also spot patterns and trends and produce insights that are on par with or better than those of human experts. The interpretation and analysis of cultural products like literature, music, and art could thus be taken over by AI systems rather than human researchers.

By impeding human creativity and invention, AI may also render human thought outdated. AI systems are made to function inside predefined boundaries and base their judgments on predetermined rules and algorithms. This may hinder their capacity to produce novel theories and methodologies by preventing them from contemplating novel and innovative ideas.

In the study of social phenomena, AI may eventually render human thought obsolete by substituting emotional intelligence and empathy. AI systems might be able to evaluate social data, spot patterns, and detect trends, but they might find it difficult to comprehend the complex emotional and cultural factors that influence how people behave.

Finally, AI has the potential to replace human intellect in the humanities and social sciences, which would have significant ramifications for human growth and our capacity to comprehend and analyze human culture and behavior. To guarantee that we are utilizing this technology in ways that support, rather than impede, human flourishing, it is crucial to take into account the ethical and societal consequences of its use as we continue to develop and use AI systems in these sectors.

Easel #3 - Hagar Abuzaid - (1-2pm; 3-4pm)

Computational identification of Conotoxin Ligand Specificities Using Machine Learning and Protein Language Model

Hagar Abuzaid¹, Tara Doma Lama¹, Eric Li², Anita Raja², Mande Holford³, Weigang Qiu¹

¹Department of Biological Science, Hunter College

²Department of Computer Science, Hunter College

³Department of Chemistry, Hunter College

Hypothesis/Statement of Problem: Conopeptides are toxins that are released from the venom of cone snails (Canidae) ranging from 10 to 45 amino acids in sequence length. Conotoxins are highly diverse and consist of multiple families driven by co-evolutionary pressure between Canidae and their prey species. Conotoxins show high specificity to target proteins including ligand-gated, voltage-gated, and G-protein coupled receptors (GOCRs). Due to their high degree of specificity and chemical potency, they were considered possible drug candidates for the treatment of neurological disorders and pain. Here, we applied the latest bioinformatic analyses to the discovery of conotoxin target specificity. We first used Conodictor2, a command-line program based on machine-learning with high accuracy and efficiency and able to identify conotoxins from a whole cone snail venom gland transcriptome in a short time. Conodictor2 requires the assembled transcriptome or raw reads file in DNA or amino acids to report its prediction on a protein file. We tested the accuracy of the prediction made by Conodictor2 through running the program on multiple conopeptides sequences and comparing it to some known methods used previously in the analysis of conopeptides. Next, we will develop a new classifier based on the protein language model that are able to classify conotoxins according to their predicted ligand and biochemical specificity. It's been hypothesized that despite conotoxins being hypervariable, the superfamiles and cysteine frameworks are determined based on the conserved sequences, hence, we should be able to find the association between superfamily and framework. It's been statistically hypothesized that there is no association between the cysteine framework and superfamily.

Methods: Sample Collection: The Isolated and sequenced transcriptome used as the data resource was collected from Professor Mande Holford's Lab. The transcriptome is of venom glands collected from multiple marine snail species. The type of sample is an amino acid sequence of 807 sequences in length and the sequencing of format is in FASTA.

Machine learning methods: Conodictor 2 is used as the primary program in identification of conotoxins using raw reads file in DNA or amino acids/Transcriptome. The reported results obtained by the program will be reported on a protein file. Other programming languages were used to conduct the data as R language and UNIX.

Results: On conducting the results, it was found that Conodoctior was able to classify superfamilies accurately. Also, using regular expressions, we were able to identify and classify cystine frames. Our statistical analysis clearly demonstrates that the distribution of superfamily isn't uniform throughout the frameworks and vice versa. Gene determines the primary amino acid sequence. Hence, there must be some inherent molecular mechanism that must have made the M superfamily have predominant cysteine frameworks.

Conclusion: All in all, Conodictor presents a fast and accurate tool for identifying the superfamilies of conopeptides. However, there are still some unanswered question that Conodictor was not able to determine which is why some superfamilies are biased towards some frameworks than other. Finally, that give our lab the idea to develop a machine learning classifier that is based on protein language models.

Easel #4 - Oumayma Agdali - (11am-12pm; 2-3pm)

Effects of Chronic Social Defeat on Pre-Pulse Inhibition in African cichlid fish Astatotilapia burtoni

Oumayma Agdali,^1,2 Sandra Knapczyk,¹ Zoia Choudhry,¹ and Thomas Preuss¹

¹Department of Psychology, Hunter College

²NIH BP-ENDURE Program, City University of New York, Hunter College

Hypothesis/Statement of Problem: African cichlid fish Astatotilapia burtoni have flexible social hierarchies where dominant males display aggressive behaviors toward subordinates. It has been found that the subordinate males show reduced prepulse inhibition (PPI) compared to their dominant conspecifics (Neumeister, 2017). PPI is a sensorimotor gating mechanism that filters out potentially disruptive sensory inputs during information processing. It is categorized as when a weak stimulus (prepulse) can suppress the startle response to a subsequent stronger startle stimulus (pulse). Deficits in PPI are associated with neurological and psychiatric disorders, namely schizophrenia (Parwani, 2000). Interestingly, the social defeat hypothesis for schizophrenia posits that long-term exposure to social defeat increases the risk for the disease (Selten, 2013). We now asked how chronic social defeat during early development will affect PPI in A. burtoni hatchlings.

Methods: We set up a chronic social defeat paradigm in which A. burtoni hatchlings were paired with slightly larger 4 week-old juveniles in small arenas for a 12 week period. During this time the juveniles displayed aggressive behaviors in the form of chasing. Following this 12 week period, the juveniles were removed and the hatchlings remained in the arenas for an additional four weeks without the exposure to a social stressor (the juveniles). Throughout the 16-week course of the study, PPI testing was conducted through the use of an auditory stimulus to elicit startle escape behavior or a c-start response.

Results: Preliminary results showed that social defeat has a transient effect on startle responsiveness between paired hatchlings and individual hatchlings. Furthermore, PPI in hatchling remains modifiable during development despite chronic social defeat.

Conclusions: Together, this implies a resilient plasticity of PPI in A. burtoni which may serve an adaptive role.

Easel #5 - Maxwell Aguilar - (11am-12pm; 12-1pm)

Assessing drug permeability and concentration of (-)-stepholidine derivatives following voluntary oral administration to determine its effectiveness in treating methamphetamine use disorder.

M. Aguilar¹; T.M Rodriguez^1,2; E. Rodriguez ^1,3 ; H.K Namballa⁴ ; K. Grondecki^1,3 ; B. Yoo⁴ ; W.W Harding^2,4; P.A Serrano^1,2,3

¹ Department of Psychology, Hunter College, ² PhD Program in Biochemistry, The Graduate Center of CUNY, New York, NY, 10016, USA

³ M.S Program in Cognitive Neuroscience, The Graduate Center of CUNY, New York, NY, 10016, USA

⁴ Department of Chemistry, Hunter College, City University of New York, NY 10065, USA

Hypothesis/Statement of the Problem: Developing effective pharmacological agents to treat methamphetamine use disorder (MUD) remains a challenge. Chronic use of methamphetamine alters dopamine (DA) levels, potentiating drug-seeking behaviors. Polypharmalogical agents that simultaneously target DA D1, D2 and D3 receptors can be used to mitigate the effects of MUD. These agents must cross the blood-brain barrier (BBB) upon oral administration to be therapeutically viable. (-)-Stepholidine (SPD), is a tetrahydroprotoberberine alkaloid, that displays a D1 agonist /D2 antagonist /D3 antagonist profile and consists of anti-addiction properties. However, SPD consists of poor oral bioavailability, limiting its therapeutic activity. Therefore, we use SPD derivatives, such as prodrugs that improve the delivery of SPD into the brain, following voluntary oral administration (VOA).

Methods: A VOA mouse model was utilized to deliver the drugs - SPD, tetrahydropalmatine (THP) and stepholidine diacetate (SPDD). Liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to assess the permeability and concentration of drugs in male mice brains. A single dose of SPD (10 mg/kg) was administered, and brains assessed at 15- and 30-min post VOA. For THP, a single 20 mg/kg dose was administered, and brains assessed at 15-, 30-, 60-, and 120- min post VOA.

Results: THP was detected at all time-points in the brain; however, no drug detection for SPD. Interestingly, a single dose of SPDD (20mg/kg) led to detection of SPD in the mouse brain, 15min post VOA, indicating that SPDD improves oral bioavailability and BBB permeability of SPD.

Conclusion: We must further evaluate SPDD as a potential pharmacological agent in treating MUD.

Easel MN1 - Ayah Ahmad - (9 - 10am)

Cytomegalovirus-responsive CD4 T cells are predominantly stable and cytotoxic over the first year post-transplant

Ayah A. Ahmad¹†, Lauren E. Higdon²†, Jonathan S. Maltzman^2,3

¹Macaulay Honors College, Hunter College, The City University of New York, USA

²Department of Medicine/Nephrology, Stanford University, Palo Alto, CA, USA

³Geriatric Research Education and Clinical Center, VA Palo Alto Health Care System, Palo Alto, CA, USA

† These authors have contributed equally to this work and share first authorship

Hypothesis: Cytomegalovirus (CMV) infection is a significant complication in humans after solid organ transplantation. Immune surveillance in healthy individuals typically controls primary infection of CMV. However, as a herpesvirus, it establishes latency such that it remains dormant within the body with the potential to periodically reactivate. Transplant immunosuppression impairs the anti-viral response to CMV. To date, anti-viral CD4 T cells have been shown to play an independent role in adaptive immunity through production of inflammatory cytokines and lysis of infected cells. Additionally, previous studies on the effects of transplantation and associated therapies show that CD4 T cell immunity is crucial to immune control of CMV. Further in-depth analysis is required regarding CMV immunity in the context of transplantation.
Methods: Therefore, we investigated how the protective CD4 T cell repertoire is affected by CMV in kidney or heart transplant recipients in the first year after transplant. We analyzed peripheral blood samples from pre-, 3- and 12-months post-transplant through flow cytometry and measured gene expression and clonal expansion via targeted single cell RNA and T-cell
receptor (TCR) sequencing, respectively. Clonal analysis involved sequencing of TCRs, which are protein complexes on the surface of T cells that are responsible for antigen recognition and can be used to define specificity.
Results: Recipients with prior exposure to CMV exhibited an aged phenotype compared to recipients with no exposure to CMV, and maintained phenotypic stability over time. Consistent with previous studies, our findings indicate CMV-responsive CD4 T cells were largely anti-viral and cytotoxic populations which were also predominantly stable over time. We found that expanded CMV-responsive CD4 T cells were also mainly characterized by an aged cytotoxic phenotype.
Conclusions: Overall, we found that transplantation does not significantly affect CMV-responsive CD4 T cells during the first year post-transplant but differentiation to cytotoxic CD4 T cells plays a dominating role in this immune response. Further investigation of the CD4 T cell repertoire in response to CMV within transplant recipients is necessary to better understand how this stability affects homeostasis and how its cytotoxicity may be protective against CMV.

Easel #6 - Rida Ahmed - (11am-12pm; 1-2pm)

Evaluation of FMRP expression in adult Fmr1 knockout mice following neuron-targeted therapy

Rida Ahmed^1,2 and Tatyana Adayev²

¹Macaulay Honors College, Hunter College, City University of New York

²Department of Human Genetics, NYS Institute for Basic Research in Developmental Disabilities

Hypothesis/Statement of Problem: Fragile X syndrome (FXS) is an X-linked neurodevelopmental disorder characterized by a CGG-repeat expansion (>200 repeats) in the FMR1 gene, which results in the absence or reduction of fragile X messenger ribonucleoprotein (FMRP). Previous gene therapy studies have demonstrated that restoring FMRP in the central nervous system (CNS) using adeno-associated viral (AAV) vectors can correct pathological abnormalities in mouse and rat models of FXS.

Methods: The ssAAVphp.eb-hSyn-mFMR1IOS7 vector and an empty vector were delivered to the brain via single peripheral intravenous (IV) administration into the tail vein of adult Fmr1 knockout (KO) mice and wild-type (WT) controls, respectively. Behavioral assessments were conducted four weeks post-injection using the following tests: open field, marble burying, accelerating rotarod, and context fear conditioning. Following sacrifice, cerebral tissue dissections from the brain stem, cerebellum, hippocampus, and neocortex were biochemically analyzed using immunohistochemical (IHC) staining and the quantitative FMRP (qFMRP) assay to determine the distribution and magnitude of FMRP expression throughout the brain.

Results: The qFMRP assay and IHC analyses of AAV-treated mice revealed neuron-specific expression, and unequal FMRP distribution throughout the brain stem, cerebellum, hippocampus, and neocortex. In all tested brain regions, FMRP expression surpassed control FMRP levels in WT mice with varying degree. The gene therapy led to full correction of motor learning and partial correction of associative learning and memory, with no effects on hyperactivity or anxiety-like behavior. Negative correlations were found between FMRP expression in the neocortex of treatment mice and exploratory behavior (measured in the open field test), and between FMRP expression in both the hippocampus and neocortex of treatment mice and motor coordination (measured in the accelerating rotarod test).

Conclusions: Excessive over-expression of FMRP may explain why certain phenotypic behaviors were not significantly altered by Fmr1 delivery. Behavioral correlation data suggest that reducing the delivery of FMRP to the neocortex and hippocampus may furthermore lead to improvement in exploratory behavior and motor coordination in treatment mice. These results demonstrate that peripheral administration of AAVphp.eb may be used to achieve CNS expression of Fmr1 and correct specific behavioral abnormalities in preclinical studies using adult mouse models of FXS.

Easel MN2 - Asia Akperov - (1-2pm; 3-4pm)

Ex vivo expansion of primary acute leukemic blasts in an endothelial cell-based co-culture system

Asia Akperov^1,2, Sonali Persaud¹, Rosemary Neigenfind¹, Wenbin Xiao^1,3

¹ Levine Lab, Human Oncology and Pathogenesis Program, MSKCC

²Hunter College

³Department of Pathology and Laboratory Medicine, MSKCC

Statement of Problem: Acute myeloid leukemia (AML) is a rare group of blood cancer with high mortality rate. Due to limited cells from primary AML samples, researchers have attempted with limited success in developing an ex vivo culture system to expand primary leukemic blasts while maintaining leukemogenic potential.  Meanwhile, several ex vivo culture systems have been established to effectively expand normal hematopoietic stem cells (HSC), a rare type of precursor cells that has the potential to self-renew and differentiate. Feeder cell layers formed by endothelial cells can promote the expansion of human cord blood-derived HSCs in the presence of cytokines. Recently, feeder cell-free and cytokine-free culture conditions have also been established to expand human cord blood-derived HSCs.

Methods: We established an ex vivo culture system with 3 conditions to evaluate whether these conditions may expand primary AML blasts and maintain the leukemogenicity. The conditions include: 1) endothelial cells transduced with an adenovirus gene, early region 4 encoded open reading frame-1 (E4ORF1) (named as AD52) as a feeder layer with a cytokine cocktail (FLT3L 50 ng/ml/TPO ng/ml/SCF 50ng/ml), 2) three-activator (3a) cocktail of UM171 (70 nM), 740Y-P (1 μM), and butyzamide (0.1 μM), 3) AD52 and 3a cocktail.

Results: CD34+ blasts expanded by 10 times in AD52 with cytokine cocktail condition and 5 times in 3a condition. The combination of AD52 and 3a was similar to AD52 and cytokine cocktail. The ex vivo expanded blasts gave rise to AML upon transplantation to immunocompromised mice.

Conclusion: Identifying conditions under which an ex vivo culture system expands primary acute leukemic blasts allows us to expand our study of the pathogenesis and therapeutic vulnerabilities of AML. This is essential for developing novel therapies and improving patient outcomes.

Easel #7 - Sera Aktas - (9-10am; 3-4pm): Combination Therapy of Chemotherapy, Navitoclax, and a Long Non-Coding RNA for Triple Negative Breast Cancer Treatment

Sera Aktas^1,2, Anthony Ramadei^3,4, Amy Yu², Devorah Natelson^3,4, and Frida E. Kleiman^3,4

¹Macaulay Honors College, City University of New York (CUNY)

²Biology Department, Hunter College, CUNY

³Chemistry Department, Hunter College, CUNY

⁴Biology Program, Graduate Center, CUNY

Hypothesis/Statement of Problem: Chemotherapy remains the predominant treatment for triple negative breast cancer (TNBC), despite the frequent recurrence of chemoresistant clones caused by surviving therapy-induced senescent cancer cells, which can return to a proliferative state. There have been attempts to target these dormant cancer cells through the use of senolytics, which are small-molecule drugs designed to target senescent cells. The most studied senolytic, navitoclax, has a strong cell-killing effect by inhibiting pro-apoptotic Bcl-2 and Bcl-xL in combination with DNA damage inducers doxorubicin and paclitaxel, which are chemotherapeutics frequently used in breast cancer. In addition, navitoclax downregulates the cyclin-dependent kinase inhibitor p21, which promotes survival in cancer cells undergoing treatment by inducing senescence. Recently, we identified a long non-coding RNA from the CDKN1A locus, which codes for p21, generated by alternative polyadenylation after DNA damage. We term this transcript Selective Polyadenylation Upon Damage (SPUD), which functions to upregulate p21 translation. We demonstrate that siRNA-mediated knockdown of SPUD (siSPUD) significantly decreases p21 protein levels in stressed and non-stressed cells. My central hypothesis is that downregulation of SPUD prior to chemotherapy treatment will decrease the number of breast cancer cells entering senescence, allowing for navitoclax to eliminate the surviving senescent cancer cells.

Methods: 10 cm 50% confluent tissue culture plates of MCF-7 breast cancer and MDA-MB-231 TNBC cells were treated with 100% optimum containing 12.5uL lipofectamine RNAi MAX and 75 picomolar SPUD siRNA or Control siRNA for three hours. After which the optimum was replaced with full serum cell media. 24 hours later both sets of cells were treated with 0.25uM doxorubicin for 24 hours, after which doxorubicin media was replaced with normal cell media. The cells were left for three days to allow senescence to occur. The cells were fixed and stained with X-gal using the Cell Signaling β-galactosidase staining kit. Senescent cells have elevated lysosomal β-galactosidase activity, even at a pH of 6, and β-galactosidase breaks down X-gal into the blue product 5,5'-dibromo-4,4'-dichloro-indigo. Therefore, cells with a blue stain in pH 6 are considered senescent.

Results:  Representative images of senescence-associated β-galactosidase staining under brightfield at 200X are shown. Blue stained cells are positive for senescence-associated β-galactosidase. Quantification of senescent cell count to non-senescent cell count ratio for MCF-7 and MDA-MB-231 cells shows a statistically significant decrease (p<0.05) in the senescent/non-senescent cell ratio for siSPUD treated MCF-7 and MDA-MB-231 cells compared to siControl treated cell.

Conclusions: My preliminary studies indicate that siSPUD treatment decreases the number of breast cancer and TNBC cells that enter senescence after doxorubicin treatment. Our studies will provide a potential combination therapy for triple negative breast cancer that can shift the balance of cancer cell fate from proliferative/senescent to apoptotic.

Easel MN3 - Malika Alamova - (11am-12pm; 12-1pm)

Identification of a subpopulation of granule cell precursors that is the cell of origin of SHH-Medulloblastoma

Malika Alamova,^1,2 Salsabiel El Nagar,² Alexandra L. Joyner²

¹City University of New York Hunter College, New York, NY

²Developmental Biology Program, Sloan Kettering Institute, MSKCC, New York, NY

Hypothesis/Statement of Problem: Medulloblastoma (MB), a cerebellar tumor, is the most common malignant pediatric tumor. While survival rates for patients are high, there remains an urgent need for less toxic treatments, as current therapeutic modalities (surgery and chemotherapy) result in long-term side effects. Four molecularly distinct subgroups of MB have been identified: WNT, Sonic Hedgehog (SHH), Group 3, and Group 4. The SHH subgroup accounts for ~30% of all cases and originates from granule cell precursors (GCPs) defined by the gene Atoh1. GCPs are the most abundant neurons in the brain, and their proliferation is stimulated by SHH. The GENSAT database, large-scale central nervous system gene expression atlases, and several studies strongly suggest that GCPs are a heterogenous population where each subpopulation presents distinct characteristics and different susceptibilities to tumorigenesis. Our lab demonstrated that constitutive activation of SHH signaling via the expression of SmoM2 at mouse postnatal day 0 (P0) only in Nestin-expressing GCPs, that represent 2% of GCPs, or in all GCPs, results in similar survival rates with tumors restricted to the lateral cerebellum. Thus, we hypothesize that Nestin-expressing GCPs are more susceptible to tumorigenesis than others.
Methodology: To analyze if Nestin-expressing GCPs are solely responsible for SHH-MB, we studied the cerebellum at P12 in Atoh1-SmoM2 and Nes-SmoM2 MB models, where SmoM2 expression is induced in scattered Atoh1+ and Nestin+ cells, respectively.
Results: Nestin-expressing GCPs formed lesions in the posterior and lateral cerebellum whereas Atoh1-expressing cells formed lesions through the cerebellum.
Conclusion: Studies have shown that SHH-MB tumors are prevalent in the posterior and lateral cerebellum. Our findings convey that the Nestin-expressing GCPs also form lesions in the exact same location. These findings should ultimately lead to development of more effective treatments for SHH-MB.

Easel MN4 - Sherelyn Alejandro-Merchan - (10-11am; 1-2pm)

Using JWST to Study Brown Dwarfs and Giant Exoplanets

Sherelyn Alejandro-Merchan^1,2, Jacqueline Faherty², Kelle Cruz^1,2, Genaro Suarez², Yeeun Park², Kay Lee², Keun Kwak², Daniel Jeon², Teawoo Kim²

¹CUNY Hunter College, New York, NY,

²American Museum of Natural History, New York, NY,

Hypothesis/Statement of Problem: Brown dwarfs are a class of astronomical objects that provide a link between stellar and planetary astrophysics.  In recent years, astronomers have focused on atmospheric studies of brown dwarfs as they can be studied directly with small, medium and large class observatories.  While they are far easier to study than extrasolar gas giant planets, brown dwarfs are difficult to characterize because parameters like dust, age, mass, clouds, magnetic activity and rotation drive large diversity in their observable properties. By determining fundamental parameters like mass, gravity, effective temperature, and bolometric luminosity, we can test theoretical atmosphere models as well as theories of substellar and giant planet formation. In order to calculate fundamental parameters, I am working on a project to construct the spectral energy distributions (SED) for a sample of the coolest star-like objects known using archival data and new data from the James Webb Space Telescope.

Methods: In order to compile each SED, I combined parallax measurements with spectral and photometric data from the optical to the mid-infrared. SED construction was done using the Python-based open-source package called SEDkit, and the photometric and spectral data used are collected from the literature but compiled in the SIMPLE Archive (http://simple-bd-archive.org/). I combed the literature and SIMPLE to identify and load the relevant and useful spectral and photometric data into Python. After loading the data, I modified spectra and photometry measurement to be read in properly by SEDkit. Using each SED, I used SEDkit to obtain an empirically calculated bolometric luminosity (𝐿_𝑏𝑜𝑙). Adding an age estimate that I found from the literature, I then again used SEDkit to use evolutionary models to estimate a radius, mass, gravity, and to semi-empirically calculate the effective temperature.

Results: To date, I have compiled 50 SEDs and have derived empirical and model-dependent fundamental parameters such as bolometric luminosity, effective temperature, radius, mass, and surface gravity for them.

Conclusions: These parameters provide us with a further understanding of substellar mass objects in addition to providing the scientific community with a clearer understanding of the atmospheres of these complex worlds which will be of importance to future generations when Earth is no longer habitable.

Easel #8 - Raed Algahim - (9-10am; 10-11am)

Modification of Tetrahydroprotoberberine Template to Synthesize D1 Receptor Ligands

Raed Algahim,¹ Ashok Gudipally,^1,2 Hari K. Namballa¹ and Wayne W. Harding^1,2,3

¹Department of Chemistry, Hunter College

²PhD Program in Chemistry, The Graduate Center, City University of New York

³PhD Program in Biochemistry, The Graduate Center, City University of New York

Hypothesis/Statement of Problem: Selective partial agonists of the dopamine D1 receptor are promising as therapeutics for treating cocaine addiction. (-)-Isocorypalmine, a tetrahydroprotoberberine (THPB) alkaloid, possesses D1 partial agonist activity and has anti-cocaine effects in animal studies. However, the role of the phenolic group present in (-)-isocorypalmine on its D1 partial agonist activity has not been established yet. The goal of this project is to determine the significance of the phenolic group of (-)-isocorypalmine on its D1 partial agonist activity by synthesizing novel (-)-isocorypalmine analogues with modified phenolic groups.

Methods: Starting from commercially available berberine, the benzodioxole group was converted into a 2,3-dihydroxy product in 90% yield. This product was then transformed into (-)-isocorypalmine via sequential methylation and enantioselective reduction. Novel (-)-isocorypalmine analogues were synthesized by functionalizing the C2 phenolic group of (-)-isocorypalmine with ether, amine, and amide groups.

Results: The pharmacological evaluation of the analogues is still in progress. However, the successful synthesis and spectroscopic characterization of the novel (-)-isocorypalmine analogues have been achieved.

Conclusions: The synthesis and pharmacological evaluation of novel (-)-isocorypalmine analogues will provide insights into the role of the phenolic group of (-)-isocorypalmine on its D1 partial agonist activity. These findings will be useful in designing THPB-based D1 partial agonist experimental therapeutics for treating cocaine addiction.

Easel MN5 - Haya Alkiswani - (10-11am; 11am-12pm)

Optimizing Beta-Cyclodextrin Derivatives and Conditions for Clearance of Lysosomal Content

Haya Alkiswani^1,2,3, Patricia A. Fontan, PhD³, Marcelo Nociari, PhD³

¹ Macaulay Honors College, CUNY Hunter College, New York, NY

² Department of Biological Sciences, CUNY Hunter College, New York, NY

³ Weill Cornell Medicine, New York, NY

Hypothesis/Statement of Problem: Lysosomal storage disorders (LSDs) are a group of inherited metabolic disorders due to mutation either in lysosomal hydrolases or membrane transporters. Due to these deficiencies, lysosomes accumulate unprocessed substances, leading to pathological symptoms and neurodegeneration. Neurodegenerative processes, such as age-related macular degeneration (AMD) and juvenile macular degeneration (Stargardt), accumulate lipids such as N-retinylidine-N-ethanolamine (A2E) within the lysosomal system. Recent studies indicate that solubilization of A2E with beta-cyclodextrins has the potential to facilitate their extracellular elimination. βCDs are molecular rings formed by seven D-glucose units. βCD-rings can encapsulate A2E -like lipids in their central nonpolar cavity. Moreover, more recent preliminary data from the lab indicate that Anionic-beta-cyclodextrin derivatives (AβCD), have not only the ability to form soluble complexes with A2E but trigger massive emesis of lysosomal content. Thus, removal of lipofuscin from cells involves both A2E complex formation and exocytosis. The goals of the experiments presented here was to optimize the type and number of anionic moieties in the βCD ring to maximize their formation of soluble complexes with A2E.

Methods: Dilutions of 1 mM, 3 mM, 5 mM, 10 mM, and 20 mM AβCD (succinyl βCD, oxalic βCD) were prepared in both water and 1 M NaCl (to assess the influence of ionic strength). The same concentration dilutions of a neutral methylated beta-cyclodextrin (MBCD) were prepared in water, and all doses were transferred to a 96 well plate. A2E was then prepared and added to all wells, and the treatments were left to incubate. The fluorescence resulting from A2E affinity was recorded for all treatment doses at 590 nm after 24 hours and after 72 hours of incubation. A similar protocol was followed to assess the effect of pH levels on A2E affinity in succinyl- βCD (SUβCD) by comparing A2E fluorescence in treatments of MBCD (control) and AβCD pH 1.5, pH 2.6, pH 4.7, pH 7.3, and pH 9.7 after 24 and 72 hours. Finally, to determine the most optimal carboxylic acid, this protocol was used to compare A2E fluorescence in AβCD, oxalated-cyclodextrin derivatives, and MBCD in various pH conditions after 24 and 72 hours of incubation.

Results: AβCD showed significantly greater fluorescence at 590 nm when compared to the neutral MBCD. Ionization showed a slight impact on affinity in our study. Additionally, our study showed that acidity was directly proportional to the level of A2E fluorescence and affinity. Lower pH levels led to greater A2E affinity when compared to higher pH levels, and treatments of SUβCD in acidic conditions

induced the greatest A2E complex formation when compared to other drug options and conditions.

Easel #11 - Diyorbek Bahromov - (9-10am)

[⁴⁴Sc]Sc-HOPO-octreotate for PET imaging of somatostatin receptors

Diyorbek Bahromov¹, Michael D. Phipps, Lynn C. Francesconi¹

¹Department of Chemistry, Hunter College, the City University of New York, New York, NY

Hypothesis/Statement of Problem: The development of a novel radiopharmaceutical, [⁴⁴Sc]Sc-HOPO-octreotate, using scandium-44 and a bifunctional HOPO ligand conjugated with octreotate peptide, will enhance PET imaging capabilities for SSTR-positive NETs and improve patient outcomes.

Methods: Synthesize a bifunctional HOPO ligand and conjugate it with octreotate peptide to form HOPO-octreotate. Radiolabel the compound with scandium-44 (⁴⁴Sc), a radioisotope suitable for PET imaging. Employing preparative high-performance liquid chromatography (HPLC) for purification of the bifunctional ligand and the HOPO-octreotate product. Use analytical HPLC and NMR spectroscopy for compound characterization. Evaluate the performance of [⁴⁴Sc]Sc-HOPO-octreotate in PET imaging of SSTR-positive NETs in preclinical studies.

Results: The successful development of [⁴⁴Sc]Sc-HOPO-octreotate will provide a reliable and efficient method for producing this novel radiopharmaceutical. The new compound is expected to enhance PET imaging capabilities for SSTR-positive NETs and improve patient outcomes. Additionally, the study will contribute to the effectiveness of radio-scandium in nuclear medicine by demonstrating the potential of scandium-44 for PET imaging applications.

Conclusions: This study represents a significant advancement in the field of cancer imaging and treatment, specifically for neuroendocrine tumors (NETs). The development of a novel radiopharmaceutical, [⁴⁴Sc]Sc-HOPO-octreotate, for PET imaging of NETs that express somatostatin receptors (SSTRs) has the potential to improve diagnostic accuracy and ultimately patient outcomes. The purification process ensures high purity and homogeneity, which is critical for accurate imaging. The use of scandium-44 as a radioisotope in this study is promising and may pave the way for further exploration of its potential in nuclear medicine.

Easel #12 - Mark Bubnovich - (12-1pm; 2-3pm):

Kinetic and thermodynamic effects of sucrose on chymotrypsin-catalyzed peptide synthesis

Mark Bubnovich,^1,3 Maithrey Ramakrishnan,^2,3 Salma Kassem PhD,^2,3 Rein V. Ulijn PhD^1,2,3

¹Department of Chemistry, Hunter College

²Department of Chemistry, The Graduate Center, City University of New York

³Nanoscience Initiative, Advance Science Research Center, City University of New York

Hypothesis/Statement of Problem: Enzymes have solidified their crucial role as biological catalysts in pharmaceutical development, biosensing, and chemical transformations by making otherwise impossible or slow reactions proceed at accelerated rates. Their catalytic activity stems from the fragile three-dimensional structure of the active site that can be compromised through changes in pH, temperature, mechanical forces, and salinity. Therefore, there is a growing demand for methods of protein stabilization and enhancing performance in synthetic reactions. Here-in, we describe how modification of the solvent environment by addition of sucrose reduces the dielectric constant and stabilizes chymotrypsin and improves its catalytic activity in peptide bond formation.

Methods: The extent to which the folding of the protein was affected by sucrose was measured using Circular Dichroism (CD) of 1 mg/ml and 5mg/ml chymotrypsin samples in 3 M sucrose and pH 8 phosphate buffer. Infrared spectroscopy (IR) was used to monitor the shift of the peak around 1600 cm^-1 to a higher wavenumber in the lyophilized samples of 10 mM, 30 mM, 70 mM, 90 mM N-Acetyl-L-Tyrosine. Subsequently, enzymatic peptide coupling of a model reaction between N-BOC-L-Tyrosine and C-NH₂-L-Glycine in up to 3M sucrose was analyzed using Liquid Chromatography - Mass Spectrometry (LCMS). 

Results: Insignificant alteration in the CD spectra of chymotrypsin in 3 M sucrose and in phosphate buffer indicated that improved activity of the protein did not come from the induced change in shape but was motivated by the solvent-solute interaction. Shift in the ketone peak on the IR spectrum upheld the hypothesis that lower dielectric constant in presence of sucrose would change the ionization state of the starting material. Monitoring dipeptide coupling through LCMS revealed an improved kinetic and thermodynamic profile of the enzyme in sucrose as opposed to the phosphate buffer that could be related to sucrose-induced suppression of ionization and thereby enhancing reactivity in peptide coupling.

Conclusions: This study suggests that sugar-rich media improves catalytic activity of chymotrypsin and can be used to enhance peptide-bond formation.

Easel #13 - Zijing Cao - (12-1pm; 2-3pm)

Induction and Secretion of Cathepsin-L in Trypanosoma brucei brucei

Zijing Cao^1,3 , Bernardo Gonzalez Baradat¹ , Daniel Lopes¹ , and Jayne Raper^1,2

¹ Department of Biological Sciences, Hunter College

² PhD Program Biology and Biochemistry, The Graduate Center, The City University of New York

³ The National Institute of General Medical Sciences Research Initiative for Scientific Enhancement Program, Hunter College

Hypothesis/Statement of Problem: Trypanosoma brucei are eukaryotic parasites that cause African Sleeping Sickness Disease in humans and Nagana in mammals. T. brucei have a cysteine protease called Cathepsin-L (TbCatL), an essential protease in the parasite. Elevation of TbCatL levels in trypanosomes has been associated with efficiency in crossing the host blood-brain barrier and evading the hosts' innate immunity. However, the pathway, mechanism, and secretion of TbCatL has been under-investigated. The aim of this project is to induce TbCatL secretion in trypanosomes in different media conditions as a potential marker for the exocytic pathway.

Methods: Parasites (Trypanosoma brucei brucei strain 2T1) were cultured in different media conditions to compare the secretion levels of TbCatL. The control condition parasites were grown in 10% Serum Plus HMI9 Media. The experimental condition parasites were grown in modified 0% Serum Plus HMI9 Media. The supernatant with secretion products were obtained. The presence of TbCatL in all samples was determined by gel electrophoresis and western blot. Cell viability was measured by microscopic live cell counts and live dead Calcein-AM assay.

Results: There was an increased level of TbCatL secretion in the induced experimental group vs uninduced trypanosome control group based on band intensity from western blots.

Conclusions: Reducing trypanosome media supplements may induce the secretion of TbCatL. This is a preliminary study, and the experimental conditions can be utilized to measure TbCatL trafficking and secretion from parasites with genes knocked-out that regulate exocytosis.

Easel MN6 - Sangita Chakraborty - (10-11am; 11am-12pm)

Enhanced Pyruvate Uptake by Naive Embryonic Stem Cells

^1,2Sangita Chakraborty, ²Benjamin T. Jackson, ²Lydia W.S. Finley

¹McNulty Scholars Program, Hunter College, The City University of New York,

²Cell Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center

Hypothesis/Statement of Problem: During early embryonic development both rapid cellular proliferation and the maintenance of pluripotency are tightly regulated to control proper cell fate specification. However, little is known about the metabolic requirements that allow the dramatic cellular changes and unique demands of this critical period. We previously found that mouse embryonic stem cells (ESCs) cultured in the naive 'ground state' of pluripotency have unique amino acid intake strategies, but how naive ESCs obtain other nutrients to sustain their growth remains unknown.

Methods: To study these nutrient acquisition strategies, we utilize pluripotent embryonic stem cells (ESCs) cultured in different media conditions that allow us to effectively model the different stages in embryonic development. ESCs cultured with serum and leukemia inhibitory factor (S/L) represent a heterogeneous and metastable population while addition of MEK and GSK3β inhibitors (S/L+2i) represent the naïve or 'ground state' of pluripotency. Nutrient consumption by S/L or S/L+2i-cultured cells was measured by gas chromatography-mass spectrometry of conditioned media collected over 24 hours from each respective cell state. MCT1-deficient cell lines were generated through CRISPR-Cas9 genome editing and the gene knockout was confirmed by Western blot. Expression of MCT1 was exogenously restored in MCT1-deficient cell lines with the addition of MCT1 cDNA using the piggyBac transposase system. Clonogenic capacity of ESCs was measured using colony formation assays.

Results: Here, we profile the nutrient consumption of naive and differentiated ESCs and found that naive ESCs preferentially consume pyruvate relative to their more differentiated counterparts. Mechanistically, pyruvate uptake was dependent upon the monocarboxylate transporter MCT1, as MCT1-deficient ESCs were unable to increase pyruvate uptake in the naive state. Restoration of MCT1 in MCT1-deficient cell lines was sufficient to rescue pyruvate consumption. Notably, MCT1-deficient ESCs display decreased proliferation and naive colony-forming ability.

Conclusion: Collectively, these results suggest that enhanced pyruvate uptake enables naive ESCs to sustain their cellular fitness and the pluripotent state.

Easel #14 - Nancy Collie-Beard - (10-11am; 12-1pm):

Baseline Anxiety and Metabolism are Associated with Vulnerability to Activity-Based Anorexia in Mice

Nancy Collie-Beard¹, Sadiyah Hanif¹, Nesha Burghardt^1,2

¹Department of Psychology, Hunter College of the City University of New York;

²Psychology Program, Behavioral and Cognitive Neuroscience, The Graduate Center, CUNY, New York, NY

Hypothesis/Statement of Problem: Up to 65% of patients with anorexia nervosa are also diagnosed with an anxiety disorder, but it is unclear if anxiety is a risk factor for anorexia or a consequence of malnutrition. 

Methods: Using the activity-based anorexia (ABA) rodent model, we tested the relationship between pre-existing levels of anxiety and ABA vulnerability in adult C57BL/6 female mice. Anxiety was evaluated in the open field test one day before mice were individually housed with running wheels. Baseline bodyweight, food intake, and wheel running were recorded for 7 days before food was removed. During the following 10 days of food restriction, food access was limited to the first 2 hours of the dark cycle, during which time mice could consume as much as they wanted. Vulnerable mice were removed from the experiment when they lost at least 25% of their baseline weight.

Results: We found a positive correlation between the number of entrances into the center of the open field and day of ABA removal (r = 0.75, p<0.001), indicating that more anxious mice are more vulnerable to ABA. We also found a positive correlation between baseline food intake and ABA removal (r = 0.48, p<0.01). Similarly, baseline food intake divided by bodyweight correlated positively with ABA removal (r = 0.45, p<0.01), suggesting that a lower baseline metabolism may also increase ABA risk. 

Conclusions: Our preliminary results indicate that anxious individuals with a lower baseline metabolism who diet and exercise may be at higher risk of developing anorexia nervosa. Understanding these relationships further may provide much needed insight into the biological substrates of this poorly understood eating disorder.

Easel #15 - Rania Darwish -  (12-1pm; 1-2pm)

Electrophysiological shift in the diverse VTA projecting BNST neurons following chronic social defeat stress in mice

Rania Darwish^1,2, Emine Ustundag¹, Lauren Renzoni^1,2, Otabek Abulkhairov ¹, Yuka Miura^,3 Allyson K. Friedman^1,3

¹Department of Biological Sciences, Hunter College of the City University of New York;

²Macaulay Honors College, Hunter College, City University of New York;

³Graduate Center of the City University of New York

Hypothesis/Statement of Problem: Neuronal activity can be greatly impacted by the presence of chronic stress, oftentimes causing negative changes in emotional states, such as depression and anxiety. The response to stress varies between individuals- some displaying decreased performance and others remaining resilient. In this study, we aimed to investigate the mechanisms by which stress alters the internal channel network that allow for resilience. The BNST uses GABAergic interneurons to regulate the firing of dopaminergic cells within the ventral tegmental area (VTA), a region linked to stress-related susceptible and resilient behavior. VTA-projecting BNST neurons integrate stress input into a reward circuitry. The diverse population of GABAergic neuronal projections to the VTA present a variety of ion channel distributions, which could factor into one's resilience to chronic stress or lack thereof.

Methods: To test our hypothesis in this study, we examined the electrophysiological properties of four sub-types of virally identified VTA-projecting BNST neurons in C57BL/6J adult mice with varying responses to chronic stress. We compared three different groups of mice: those that successfully adapted to chronic stress, those susceptible to chronic stress, and those without exposure to chronic stress.

Results: Based on the emotional response of mice, we hypothesized that of the four subtypes, only some of them will be sensitive to stress.

Conclusions: Our findings will contribute to a better understanding through which VTA-Projecting BNST neurons control resilience to stress and their electrophysiological properties. This knowledge could pave the way for developing novel therapeutic interventions to treat stress-related disorders.

Easel #16 - Moitrayee Dasgupta - (11am-12pm; 1-2pm)

Effects of BRCA1-Mediated Ubiquitination of HuR on RNA Metabolism in Breast Cancer

Moitrayee Dasgupta¹, Devorah M. Natelson^1,2, Gamage Aruggoda^1,3, Anthony Ramadei^1,2, Amy Yu¹, Sera Aktas¹, Frida E. Kleiman^1,2,3

¹Chemistry Department, Hunter College, City University of New York (CUNY)

²Biology Program, Graduate Center, CUNY.

³Biochemistry Program, Graduate Center, CUNY

Hypothesis/ Statement of the Problem: Breast cancer is the most commonly diagnosed cancer type in women. BRCA1 mutations predispose affected individuals to increased risk of breast cancer by 65%. BRCA1 is a gene involved in DNA repair and other cellular processes. Additionally, BRCA1 is known to form a heterodimer with BARD1 and act as an E3 Ub ligase enzyme, but its targets and the implication of this enzymatic activity in breast cancer are not fully understood.

Methods: To assess the BRCA1/BARD1 mediated ubiquitination of HuR and its effects, western blots, sub-cellular fractionations, and RNA-imunoprecipitiation assays were conducted in different cellular conditions such as cell lines expressing functional (MCF7/MDA-MB-231) or non-functional BRCA1 (SUM1315), BRCA1/BARD1 knockdown, and UV treatment.

Results: Our preliminary studies indicate that BRCA1/BARD1 ubiquitinates HuR, modulating intracellular localization of HuR and interaction with target mRNAs. Non-degradative HuR ubiquitination is reduced by BRCA1/BARD1 depletion and after UV treatment. Additionally, SUM1315 cells show decreased levels of ubiquitinated HuR, elevated levels of chromatin-bound HuR, and HuR does not appear to localize to cytoplasm after UV damage, when compared to MCF7 cells with functional BRCA1. Furthermore, HuR binding to mRNA target TP53 increases in BRCA1/BARD1 depleted cells and in SUM1315 compared to MCF7 cells.

Conclusion: Together, these data suggest that HuR ubiquitination by BRCA1/BARD1 affects the sub-cellular localization of HuR and decreases HuR binding to target transcripts. Changes in this controlled pathway by BRCA1 mutation might result in transcriptome dysregulation and homeostatic imbalance, affecting breast cancer outcomes and therapies.

Easel #17 - Arva Demaliaj - (11am-12pm; 1-2pm)

Determining the role of sodium accumulation in TLF-mediated lysis of African Trypanosomes

Arva Demaliaj,¹ Sara Fresard,^1,2 Jayne Raper ^1,2

¹Department of Biological Sciences, Hunter College-CUNY

²Biology Program, The Graduate Center CUNY

Hypothesis/Statement of Problem: African trypanosomes are unicellular eukaryotic parasites. In cattle, trypanosome infection causes nagana, a wasting disease resulting in a major agricultural and economic burden in sub-Saharan Africa. Humans and some non-human primates are protected against most species of trypanosomes due to an immunity complex called Trypanosome Lytic Factor (TLF). TLF is a specialized High-Density Lipoprotein (HDL), that carries a lytic cation channel-forming protein, Apolipoprotein-L1 (APOL1). The mechanism of APOL1-mediated lysis remains controversial. We propose that after receptor-mediated endocytosis, APOL1 is inserted in the endosomal membrane due to the acidic pH. Once the endosome is recycled to the neutral environment of the plasma membrane, the channel opens. Sodium and calcium ions influx, causing an osmotic imbalance, resulting in the trypanosome swelling and bursting. Other models suggest APOL1 is trafficked to the mitochondria, triggering mitochondrial dysfunction and apoptotic-like cell death. We hypothesize that the mitochondrial membrane depolarizes due to the sodium influx through APOL1 at the plasma membrane, which causes sustained plasma membrane depolarization.

Methods:  To test this hypothesis, flow cytometry was used, with fluorescent indicators ING2-AM a sodium-specific dye to measure the timing of sodium ion accumulation, and TMRE to measure mitochondrial membrane depolarization in response to TLF.

Results: Within 30 minutes of TLF treatment, sodium accumulation, and mitochondrial membrane depolarization were detected.

Conclusions: From our results, we cannot conclude that mitochondrial membrane depolarization happens due to the sodium influx through APOL1 at the plasma membrane. To dissect the two events we will evaluate shorter times of TLF treatment, and test if replacing Na+ with TMA+, a larger positive ion that cannot pass through the APOL1 ion channel, prevents sodium influx and thus mitochondrial depolarization.

Easel #18 - Amar Dhanjal - (11am-12pm; 1-2pm):

Generating an Inducible, Transgenic Control to Study the Mechanisms of APOL1 Mediated Lysis in Trypanosoma brucei brucei.

Amar Dhanjal^1,2, Sara Fresard^1,3 and Jayne Raper ^1,3

¹ Department of Biological Sciences, Hunter College-CUNY

² Macaulay Honors College, Hunter College, City University of New York, New York, NY, USA.

³ Biology Program, The Graduate Center CUNY

Hypothesis/Statement of Problem: Trypanosoma brucei spp are parasites that cause African Trypanosomiasis in humans and animals through the bite of tsetse flies. Humans and some baboons are protected against T. b. brucei infection due to an innate immune complex called Trypanosome Lytic Factor (TLF). TLF carries a pH-gated, cation-channel forming protein called Apolipoprotein-L1 (APOL1) that lyses parasites through osmotic swelling. The exact mechanism by which APOL1 lyses trypanosomes is currently disputed. APOL1 requires activation in the parasite's acidic endosomal system, and we hypothesize that it is subsequently recycled to the neutral environment of the plasma membrane, where it forms channels. Two subspecies of T. brucei, T. b. rhodesiense and T. b. gambiense, have evolved genes to evade APOL1-mediated lysis and are thus able to infect humans, but not baboons. T.b. rhodesiense evades lysis due the presence of the Serum Resistance-Associated (SRA) protein. SRA binds directly to human APOL1 in endosomes, blocking activation and channel formation, allowing the parasite to proliferate in humans. To fully understand the mechanism of APOL1-mediated lysis and intracellular trafficking, we generated an inducible transgenic T. b. brucei cell line that expresses SRA, to use as a control system in our experiments.

Methods: We used a cell line with an inducible promoter in a landing pad to generate SRA-expressing cells. To determine whether the SRA gene was successfully integrated into the genome, and when expressed, confers resistance to APOL1, we performed PCR, Western blot analysis, and 24 hr trypanosome lysis assays with recombinant human APOL1 and 8hr assays with purified TLF complexes. Percent lysis was determined using a viability indicator, alamarBlue.

Results: Under induction with doxycycline, a synthetic version of tetracycline, SRA expressing parasites were fully resistant to both human rAPOL1 and purified TLF in comparison to uninduced controls.

Conclusions: SRA expressing parasites are resistant to lysis with human APOL1 and purified TLF complexes. This biological control will be used in further studies to elucidate the mechanism of APOL1 mediated lysis and help us to better understand the differences between human and baboon innate immunity.

Easel #19 - Doobneek - (10-11am; 3-4pm)

Solutions for Directing Human and Vehicle Traffic

Doobneek¹

¹Department of Studio Art, Hunter College. Deposited graduate student at NYU Tandon, Transportation Engineering class of Fall 2023. Other credentials include: four out of five years in school of architecture in Chelyabinsk, Russia.

Hypothesis/Statement of Problem: The intersections of multiple roads often create issues for drivers who have to yield to each other and enter bottlenecks at the highway esplanades. I Create sophisticated solutions for narrow city plans that allow cars to drive using one lane throughout the whole intersection depending on what direction is needed to be achieved. To prevent same-level intersections with human traffic all roads have to be lifted above the ground with hermetic glass covers to prevent air and noise pollution or hidden underground.

Methods: Software is needed to direct drivers to pre-select what direction they need to go after they exit intersection, because several blocks before the entrance the street divide on multi-level 1-2 lane sleeves that have no exists up to the very merge with the next street, same for the cars that are making the same journey on the way back, to prevent the delay of drivers attempting to turn around if they realized they took the wrong exit and remove a risk of face-to-face collisions.

Results: The delays in traffic jams are reduced, accidents' fatalities are reduced due to the only nature of collisions is the same-directional traffic or the borders of the road, the waste of energy and air pollution reduced due to constant flow of traffic and less wait time with a running motor.

Conclusions: Same tactic can be used in directing human traffic in airports, where people need to exit from one long terminal through a hub where all terminals meet into another terminal, their traffic can be directed so they never intersect their paths. It will occupy the same space but reduce the travel time.

Easel #20 - Ahmed Farag - (1-2pm; 2-3 pm)

COL1A1 modulates the invasive and migratory abilities in Osteosarcoma.

Okkeun Jung¹ , Ahmed Farag ², and Shahan Mahajan^,1,2,3,4

¹The Graduate center of City University of New York, Biology, New York, NY

²Department of Medical Laboratory Science, Hunter College, New York, NY

³The Graduate center of City University of New York, Biochemistry, New York, NY

⁴Weill Cornell Medical College, Brain and Mind Research Institute, New York, NY

Hypothesis/Statement of Problem: Metastasized bone cancer is often more aggressive and proliferative than primary bone cancer, leading to higher mortality rates for patients. Despite extensive research, the factors and mechanisms that drive osteosarcoma to become more invasive and metastatic remain unclear. Therefore, we hypothesized that modulating a protein that is highly expressed only in the primary osteosarcoma would reduce the migratory ability of the metastasized osteosarcoma.

Methods: To determine the protein upregulated in the primary osteosarcoma, we analyzed the three RNA-seq datasets of each primary and metastasized osteosarcoma samples. We then performed qPCR to validate this observation in other osteosarcoma cell lines, primary osteosarcoma (MG63, HOS, SAOS2) and metastatic osteosarcoma (MG63, MNNG, 143b and LM7). Moreover, we performed western blot to examine the protein expression level because RNA-seq only shows mRNA expression levels. Finally, we will examine the migratory ability through Boyden chamber invasion assay after suppressing the mRNA expression level using RNA interference in primary osteosarcoma.

Results: A comparison of metastasized osteosarcomas and primary cancers reveals that COL1A1 is highly expressed in primary osteosarcomas. Indeed, the basal mRNA expression levels of COL1A1 in nine osteosarcoma cell lines are only highly expressed in the primary osteosarcoma cell lines. Furthermore, the protein expression levels of COL1A1 are followed by the mRNA expression levels. Finally, the suppression of COL1A1 mRNA might increase invasive abilities in the primary osteosarcoma cell lines.

Conclusions: This finding suggests that COL1A1 may be a key gene involved in regulating the migratory and invasive abilities of metastatic osteosarcoma cells.

Easel #21 - Mariam Farag - (11am-12pm; 1-2pm)

Wheat Germ Eukaryotic Initiation Factor 3d (eIF3d) Binds with High Affinity to 5'-Capped Human cJun mRNA Construct

Mariam Farag,¹ Km Mustaksid,^1, and Dixie Goss¹

¹Department of Chemistry, Hunter College

Hypothesis/Statement of Problem: Contamination of water by dyes is a serious issue for the environment and human health. Hydroxyapatite (HAP) is a powerful, low-cost adsorbent for pollutants through ion exchange. As a naturally occurring part of bones and teeth, it is biocompatible, making it well suited for applications in environmental remediation. Modification of solid supports can change their affinity for substrates in solution and this project aims to measure the effects of metal modified HAPs on the adsorption of the dye, Janus Green B (JGB).

Methods: HAP was synthesized with modifications of Cu, Mn, Fe, Cd, Ni, Zn, Co, and Sr that replaced 10% of the calcium ions. There were additional batches of CuHAP with replacement of 2%, 4%, 6%, and 8% of the calcium ions. Adsorption studies were performed by creating a 0.1 mM stock solution of Janus Green B (JGB), which were serially diluted to create 5 solutions that were 0.01, 0.02, 0.04, 0.06, and 0.08 mM. Nothing was added to control solutions, while 100 mg of HAP was added to test solutions. UV-Vis spectroscopy was then performed to measure amount of adsorption.

Results: The adsorption of JGB from the HAPs was found to follow the Langmuir isotherm, a model used to predict the maximum adsorption. Fe10HAP is shown to increase adsorbance of JGB marginally, while other metal modifications of HAP do not show significant effect on the ability to adsorb JGB. Variation of concentration of the metal also does not show any significant improvements.

Easel #22 - Ruixin Feng - (11am-12pm; 3-4pm)

Improving the yield of designed, Fibril forming Collagen Mimetic peptides. 

Ruixin Feng¹, Jui Chaugule², Yujia Xu*¹

¹City of New York Hunter College, New York, NY

²The Graduate Center, CUNY, New York, NY

Hypothesis/Statement of Problem: Collagen forms the essential structures in connective tissues. Its functions depend on the formation of a complex fibrillar complex where three polypeptide chains first form a triple helix; the triple helices are further packed into fibrils. The super twist of the packed triple helices affects enzyme and cell receptor binding sites. How the fibril formation of collagen modulates the interactions between collagen and the other molecular ligands remains unknown. Our lab has developed fibril-forming collagen mimetic peptides (FCMPs) that are generated by bacterial expression using designed genes. These FCMPs are useful tools to study the effects of fibril packing on ligand binding and other functions of collagen. However, the main obstacle is the low yield of the FCMPs that limited many applications. I worked to optimize the purification process of the FCMPs. We have identified the thrombin cleavage of the expressed fusion protein as the crucial factor limiting the yield of FCMP.

Methods:  The FCMP Col3+ is expressed in E. coli as a fusion protein having a N-terminal Trx- tag. The protein was expressed and purified using standard methods. The Trx-tag is removed by thrombin cleavage. SDS PAGE was used to check the purity and yield of the protein.

Results: By optimizing the affinity chromatography procedures and combining the dialysis with thrombin cleavage we collected purified protein samples in fewer fractions with higher concentrations. The time for thrombin cleavage is also shortened.  The overall outcome is more concentrated purified protein samples with higher purity.

Conclusions: The optimizations of the protocol led to a pure and higher yield of FCMP. The futural goal of this research is to use the FCMPs for fundamental research of the biological functions of collagen and for developing novel biomaterials.

Easel #23 - Dominique Forbes - (10-11am; 2-3pm)

Exogenous Purified MDMX Addition to Triple Negative Breast Cancer Extracts Promotes Increased Cellular Protein Ubiquitination

Dominique Forbes¹, Rusia Lee^1,2, Jill Bargonetti^1,2,3

¹ Department of Biological Sciences, the City University of New York Hunter College, New York, NY

² Biology and Biochemistry Programs, City University of New York Graduate Center, New York, NY

³ Department of Cell and Developmental Biology, Weill Cornell Medical College, New York, NY

Hypothesis/Statement of Problem: MDMX and MDM2 are proto-oncogenes often amplified and overexpressed in breast cancer. MDM2 is an E3 ubiquitin ligase that ubiquitinates p53. MDMX, a homolog of MDM2, enhances the E3 ubiquitin ligase activity of MDM2. MDMX and MDM2 use their RING domains to form a heterodimer that robustly ubiquitinates p53 and MDM2. We found that MDMX promotes circulating tumor cells of human xenograft triple-negative breast cancer cells (TNBCs) in immunocompromised mice. We hypothesize that MDMX activates MDM2 to ubiquitinate multiple proteins to drive p53-independent cancer-promoting metastatic functions.

Methods: To study if MDMX stimulates the ubiquitination of targets other than p53, we generated a plasmid for in-vitro expression of MDMX. We performed an in-vitro ubiquitination assay using purified MDM2 or MDMX and cell extracts from TNBCs with mutant p53, with and without depletion of MDM2 or MDMX.

Results: We successfully confirmed that: 1) purified MDM2 displays E3 ubiquitin ligase activity, 2) purified MDMX enhances the ubiquitination activity of MDM2, and 3) MDMX alone has no E3 ubiquitin ligase activity. Under the conditions tested, the addition of purified MDMX to cell extracts with, or without, depletion of MDMX caused overall increases in cellular protein ubiquitination. Greater upregulation of ubiquitination was observed in MDMX-depleted cell extracts suggesting dynamic regulation of reaction kinetics. We suspect the formation of MDM2-MDMX heterodimers has a rate-limiting role on the ubiquitination of targets and that target modification is dynamically reversed by deubiquitinases (DUBs).

Conclusions: In future experiments, we will use this newly designed system to identify novel targets of MDMX-enhanced ubiquitination.

Easel #24 - Jonathan Gao - (12-1pm; 2-3pm)

The Impact of Fructose on High-Fat Induced Obesity

Jonathan Gao,¹ Mujmmail Ahmed,² Shiri Li,² and Marcus DaSilva Goncalves²

¹Department of Chemistry, Hunter College, City University of New York, New York, NY USA

²Division of Endocrinology, Weill Cornell Medicine, New York, NY USA

Hypothesis/Statement of Problem: Fructose, a sugar found in many processed foods, has been previously shown to elongate intestinal villi, leading to enhanced fat absorption and adiposity in mice. Increased uptake of fructose in intestinal cells, leads to increased fructose-1-phosphate (F1P) levels by ketohexokinase (KHK) and inhibition of pyruvate kinase (PKM2) by elevated F1P leads to cellular metabolic rewiring and elongation of villus structures. ​​We hypothesize that KHK-deficient mice and TEPP-46-treated mice fed with high fat and high fructose will not elongate their villi in response to fructose and will not experience as much weight gain as control mice fed matched diets.

Methods: To investigate the role of KHK and PKM2 in diet-induced obesity, we use mice genetically deficient in KHK or exposed to the PKM2 activator, TEPP-46, to assess weight gain, adiposity, and villus length. There were four groups based on different diet and treatment including high-fat-high-sucrose with TEPP-46 (Group 1), high-fat-high-sucrose (Group 2), high-fat-high-dextrose (Group 3), and control (Group 4). Body weight change, food and water intake, liver and gonadal white adipose tissue (gWAT) weights, and villus length in the small intestine were measured. Mouse body composition was measured through weekly EchoMRI scans in the forms of fat and lean mass. Energy expenditure, physical activity, and food intake were monitored on weeks 3 and 4 with the Promethion metabolic cage system.

Results: Groups 1, 2, and 3 gained more weight than group 4 (28.7 g, 29.1 g, and 27.3 g, vs xxx g). Food intake for Group 4 (1.27 g/day) and Group 2 (1.48 g/day) was higher than that of Group 1 (0.54 g/day) and Group 3 (0.46 g/day). Group 3 consumed the least amount of water (0.88 mL/day) while Group 4 consumed the most (1.56 mL/day). The livers of Groups 1 (1.11 g) and 2 (1.10 g) were heavier than those of Groups 3 (0.93 g) and 4 (1.02 g). The gWATs of Groups 1 (0.34 g), 2 (0.37 g) and 3 (0.37 g) were higher than that of Group 4 (0.26 g). Although an increase in fat mass was observed for Groups 1, 2, and 3 (1.67 g/day, 1.78 g/day, and 2 g/day, respectively), Group 1 was the only group to show an increase in lean mass (0.38 g/day). The villi length of mice in Group 2 was 301 µm and was longer than Group 1 (252 µm), Group 3 (256 µm), and Group 4 (276 µm).

Conclusions: Our experimental data shows that fructose may play a role in high-fat induced obesity, as demonstrated by the differences in body weight. These findings also indicate a potential correlation between an increased consumption of TEPP-46 and a decrease in villi length. Further research will allow us to fully understand fructose's role in obesity and will provide us with important pre-clinical data to support human trials using KHK-inhibiting and PKM2-activating drugs to counter the effects of fructose.

Easel #25 - Alexandra Genovese - (9-10am; 1-2pm)

Contrafreeloading in domestic pet dogs (Canis lupus familiaris)

Alexandra Genovese^a, Liza Rothkoff ^a, Kayla Cohen^a,  Lynna Feng ^b, Sarah-Elizabeth Byosiere^a

^a Thinking Dog Center, Department of Psychology, Hunter College, City University of New York, United States 

^b Guide Dogs for the Blind, United States

Hypothesis/Statement of Problem: Contrafreeloading can be defined as an animals' preference to work for food when freely available food is present. To date, multiple studies have evaluated contrafreeloading in various animal species, observing that many non-domesticated species (e.g. maned wolves, red jungle fowl) and domesticated species (e.g. pigs, goats) demonstrate a preference to work for food when readily available food is present. Most recently, Delgado et al., (2021), observed that domestic cats prefer freely available food over food that requires effort. 

Methods:  In an adaptation of this research, we assessed whether or not another domesticated companion animal, dogs, prefer to work when presented with a food puzzle (snuffle mat) and a tray. Thirty-eight pet dogs participated in the study in which they were presented with ten feeding trials (50% of food in a snuffle mat and 50% of food on the tray). All feedings were video-recorded and behaviorally analyzed. 

Results: We did not find evidence of contrafreeloading behavior in domestic dogs. Dogs, as a group, approached the tray first most often (M = 0.262, SD = 0.228, p = 0.0215), and of the 38 dogs, only one dog demonstrated a significant preference to contrafreeload while 22 demonstrated a preference to freeload. 

Conclusions: Overall, our results suggest that domestic pet dogs do not show a preference for contrafreeloading. Our results could have implications for the future of dog enrichment.

Easel MN7 - Greta Ghita - (9-10am; 12-1pm)

Characterizing EZH2, RBBP4 and their Co-Expressed Genes as Potential Markers of Progenitor-Like Populations in GBM
Greta Ghita¹, Yanhong Yang², Viviane Tabar²

¹John P. McNulty Scholars Program, Hunter College, City University of New York

²Department of Neurosurgery, Memorial Sloan Kettering Cancer Center

Abstract: Identification of effective and specific markers is a vital part of developing new therapeutic treatments for Glioblastoma (GBM), with classifications for different tumor cell subtypes still being determined. The existence of cancer stem cells in particular helps give rise to a highly heterogeneous and invasive tumor microenvironment within the brain, with markers being necessary to help distinguish between cells at all stages of differentiation and their associated proliferative capacities. Recently, the emergence of a highly proliferative "progenitor-like" cell population with expression profiles characteristic of neural progenitor (NPC) and oligodendrocyte progenitor cells (OPC). Our project aims to characterize genes that are associated with characteristics of progenitor-like cells in GBM and could possibly be used as more specific markers for this population. Notably it has recently been found that the PRC2 complex is associated with upregulation of genes in NPC and OPC-like cells, with further analysis finding that the EZH2 and RBBP4 subunits in particular could help regulate proliferation and be associated with oncogenic activity.

Methods: Utilizing computational analysis through R and WebGestalt, sets of genes coexpressed with each of the subunits was found to help identify potential markers and assess the role of EZH2 and RBBP4 in Glioblastoma.

Results:  Using WGCNA, it was found that genes highly co-expressed with EZH2 include PLK1, CDC6, and KIF18B, all genes associated with cell division regulation. Contrastingly, for RBBP4, genes highly co-expressed in GBM RNA-seq datasets include a variety of cellular functions, such as cell differentiation, transcription and degradation of mRNA, as well as cell division and growth. It was also through WebGestalt that the majority of genes highly coexpressed with either EZH2 or RBBP4 were located in the nucleus, and in particular ubiquitin-like protein ligase binding genes were enriched.

Conclusions: RNA-seq analysis supports the idea that the PRC2 complex may help to act as a marker of highly proliferative progenitor-like populations in GBM. In particular, it was found that Polo-like kinase 1 (PLK1) was highly co-expressed with EZH2 in the TCGA glioblastoma bulk RNA-seq datasets. PLK1 remains a highly important gene in studying cancer and metastasis in particular, and whose inhibition results in decrease of tumor growth. Within the enriched group of ubiquitin-like protein ligase binding genes, cyclin B1 (CCNB1) in particular forms the maturation factor with p34 in order to maintain a normal G2/M transition phase in cell division. Data suggests that it may also play a role in metastasis of high-grade gliomas, as a recent study looking at differentially expressed gene networks in glioblastoma, CCNB1 was found to be greatly enriched in the p53 pathway, along with CDK2, which was also highly co-expressed with RBBP4.

Easel #26 - Abdelrhman Gouda - (10-11am; 11am-12pm)

Dimethyl Sulfoxide Combined with Polyethylene Glycol 400 as a Vehicle for Salvinorin A

Abdelrhman Gouda¹, Emma R. Huels^2,3,4, Tiecheng Liu², George A. Mashour^2,3,4, Dinesh Pal^2,3,4

¹Hunter College, New York, NY

²Department of Anesthesiology

³Neuroscience Graduate Program

⁴Center for Consciousness Science, University of Michigan, Ann Arbor, MI

Hypothesis/Statement of Problem: Despite a resurgence in psychedelic research, salvinorin A, has received little attention. One of the major hindrances has been its poor solubility in aqueous solutions, necessitating the use of organic solvents. We investigated different combinations of dimethyl sulfoxide (DMSO) and polyethylene glycol 400 (PEG400) as a vehicle for intravenous delivery of salvinorin A in rats.

Methods: First, we tested the solubility of salvinorin A in different proportions of DMSO and PEG400 and found it to be soluble in a 35:65 DMSO:PEG400 ratio with a maximum solubility of 1 mg/mL. Next, to determine the effect on patency of the microrenathane catheters, we soaked the catheters for 2h in either 35:65 DMSO:PEG400 solution or 35:65 DMSO:PEG400 diluted with saline (14:26:60 DMSO:PEG400:saline). The tubing began to break down after 20 minutes of exposure to the DMSO/PEG400 alone, while the addition of saline preserved patency with only minor blockage occurring. Finally, five male rats were surgically fitted with a catheter for administration of the 14:26:60 DMSO:PEG400:saline solution and electrodes for simultaneous electroencephalogram recordings to quantify power spectral changes (1-180Hz) before and after vehicle administration.

Results: The results showed that a solution of DMSO (35%) and PEG400 (65%) was sufficient to dissolve salvinorin A (1mg/mL), while the addition of saline preserved the catheter tubing.

Conclusions: The solution did not elicit changes in spectral power or behavior, supporting its use as a vehicle for intravenous salvinorin A administration in rats.

Easel MN8 - Leya Groysman - (10-11am; 11am-12pm)

Assessment of Bone Progenitor Cell Proliferation in Response to Exosomes 

Leya Groysman^1,2; Adrianne Nemchik^1,2; Jenn Park,MD¹; Fernando D. Arias, MD¹; Alexandra Verzella, BS¹; Roberto L. Flores, MD¹; Piul S. Rabbani, PhD¹

¹ NYU Langone Health Hansjorg Wyss Department of Plastic Surgery

²  Macaulay Honors College at Hunter College CUNY

Hypothesis/Statement of Problem: Bone tissue engineering is a promising alternative to invasive procedures such as bone graft surgery, particularly for pediatric patients.  Exosomes, secreted vesicles that carry signaling molecules, from bone-derived multipotent stromal cells (MSCs) offer bone tissue growth potential. This is due to the fact that exosomes carry molecular cargo that correspond to the source cell that they are released from. Here, we explore the impact of exosomes on leptin receptor (LepR)-expressing bone progenitor cell growth in vitro.

Methods: We used a CyQUANT assay to assess whether mouse cranial LepR+  proliferate in response to MSC exosomes. We computed the results using GraphPad Prism. 

Results: The proliferation of the LepR+ cells would indicate that they contribute to bone growth. We assessed whether tissue cultured leptin-receptor positive or leptin-receptor negative progenitor cells respond better to the exosome addition. We found that both cell types had a significant fold-change proliferation in response to a lower dose (50 million particles) exosome treatment, in comparison to the initial cell densities. We observed a statistically significant (p = 0.0079 and p=0.0003, respectively) higher proliferation difference between the lower-dose group at higher initial cell densities (3,000 and 5,000 cells) versus the higher-dose group (500 million particles) at lower initial cell densities (1,000 cells). Both exosome dosages resulted in growth saturation at higher initial cell densities compared to cells grown without exosomes, 72 hours post plating the cells.

Conclusions: Cell proliferation in response to exosomes could provide further insight into the impact these nanoscale vesicles have on cell behavior, help identify extracellular vesicle-based cell communication, and contribute to the regenerative process in a bone defect. MSC-derived exosomes could be used to advance therapeutic tissue engineering therapies to alleviate the treatment challenges for pediatric patients with cleft lip and palate disorders.

Easel #27 - Yuval Guetta - (1-2pm; 3-4pm)

Mapping mPFC Projections Involved in Fear and Extinction Memory Retrieval

Yuval Guetta,¹ Carolina Fernandes-Henriques,^2,3 Mia Sclar,³  Rebecca Zhang-Shen,³ Lyubov Yusufova,³ and Ekaterina Likhtik^2,3

¹Department of Psychology, Hunter College

²The Graduate Center, City University of New York

³Department of Biology, Hunter College

Hypothesis/Statement of Problem: Projections from the Prelimbic (PL) and Infralimbic (IL) regions of the medial prefrontal cortex (mPFC) to the Basolateral Amygdala (BLA) are involved in  fear and extinction memory retrieval, respectively. However, there is much debate surrounding the separation vs overlap of function between these two sets of projections.

Methods: To address this question, we used retrograde tracing and functional interrogation via immediate early genes to map activity in PL- and IL- projections to the BLA across cortical layers and the anterior-posterior axis during fear and extinction memory retrieval. Furthermore, for comparison, we used the same approach to map activation of IL and PL projections to the Basal Forebrain (BF), a critical structure for attention processing and learning, which also receives strong input from the mPFC.

Results: Overall, we show that during extinction retrieval, IL activity is upregulated in deep layers (L5) relative to superficial layers (L2/3), and that the posterior IL is more active. Further, we show that IL-BLA and IL-BF projectors differentially localize within mPFC layers, with IL-BLA projections primarily arising from L2/3 at all tested locations on the anterior-posterior axis, whereas IL-BF projections consist of two groups, one arising from L2/3 and one from L5.  Interestingly, we found that IL-BLA projectors are equally active during fear and extinction retrieval, whereas deep-layer IL-BF projectors selectively increase activity during extinction retrieval. 

Conclusions: Together, these findings demonstrate that IL activation shows spatial and projection-specific constraints during emotional memory retrieval.

Easel #28 - Bartlomiej Halibart - (9-10am; 2-3pm)

Characterizing the Expression of Top Genome Wide Association Study Candidate Gene in Mouse and Human Samples in the Setting of Urinary Tract Infections

Bartlomiej Halibart^1,2; Katherine Xu²; Krzystof Kiryluk²

¹Chemistry Department, Hunter College, New York, New York, USA.

²Department of Medicine, Columbia University, New York, New York, USA.

Urinary tract infections (UTIs) are common bacterial infections that typically affect the bladder but can travel up the ureters to affect the kidney. Difficulty in accurate diagnoses of UTIs arises from the overlapping symptoms with sexually transmitted infections (STIs) upon presentation as well as irregular urinalysis results. As a result, these patients with misdiagnosed UTIs were exposed to unnecessary antibiotics, which could possibly lead to bacterial antibiotic resistance. Multiple factors may affect an individual's predisposition to UTIs such as anatomical structure, vaginal or urethral biota, menopause, hormones, diabetes, along with social and behavioral factors. We have recently performed the largest UTI Genome Wide Association Study (GWAS) to date with four patient cohorts and identified a possible candidate. Additionally, we wanted to investigate if the top GWAS gene can be secreted into the urine. Human urine samples were obtained from patients diagnosed with a UTI and patients not diagnosed with a UTI. Western Blots were performed on the urine samples to determine whether the gene protein is secreted into human urine since the protein can be cleaved and whether there is a difference in concentration of the protein between patients with and without UTIs. Additionally, we performed in situ hybridization to localize gene expression in UTI male and female mouse kidney, ureter, and bladder. The analysis performed suggests that the GWAS gene expression potentially varies between patients with and without UTI infections. The field of precision medicine would benefit from this type of insight on a potential gene candidate that demonstrates promise in diagnosing UTIs.

Easel #29 - Mohammed Haque - (2-3pm)

Constructing MUSE: A Simpler Approach to Stellarators

Mohammed Haque,¹ Steven Greenbaum², Michael Zarnstorrf^,3

¹Department of Physics, Hunter College, Undergraduate

²Department of Physics, Hunter College, Professor of Physics

³Princeton Plasma Physics Laboratory, Chief Scientist

Hypothesis/Statement of Problem: Fusion energy has the potential to meet the world's demand for clean energy. The stellarator was first invented in 1951 as a potential device for fusion reactions. It uses external coils to generate a magnetic field and a rotational transform to confine the fusion plasma in a toroidal region. Early stellarators had poor plasma confinement due to large neoclassical loss and Magnetohydrodynamic instabilities. With the development of more advanced design tools, modern stellarators have been optimized to minimize neoclassical loss (and to be MHD stable). Today, Stellarators have complex designs that make maintenance and access difficult. It has been proposed by Mike Zarnstorff, et. al.[1] that using planer coils and permanent magnets to generate a magnetic field can significantly reduce the difficulty in building a stellarator. A permanent magnet stellarator would make construction much easier, cheaper, provide us with more freedom for optimizing the magnetic field, make the design and construction modular, and would enable rapid prototyping. We are building the world's first quasi-axisymmetric permanent magnet stellarator(MUSE)[2] to demonstrate the feasibility of this approach for building optimized stellarators.

[1] 2020 Helander, et. al., Phys. Rev. L., 124, 095001

[2] 2022 Qian, et. al., Nucl. Fusion, 62, 084001

Methods: MUSE makes use of sixteen planar coils as well as neodymium magnets to create our desired magnetic field. The magnetics were held in place by 3D-printed magnet holders made from nylon which provided us with a balance of strength and accuracy. Each magnet well has a specific height for a particular magnet. Magnets were manually inserted and secured with Loctite and epoxy.

Results: We have completed the engineering portion of our experiment and now have begun to turn on our power supplies. In the coming weeks, we will be conducting electron beam mapping experiments to map the flux surfaces of MUSE and verify that we have indeed created a stellarator.

Conclusions: Our research will help us gauge the feasibility of alternative technologies in stellarators. Our results will help us determine if coil simplification could be a lucrative technique in fusion experiments.

Easel #30 - Aisha Haroun - (12-1pm; 2-3pm)

Understanding the Effects of Disease-Causing Mutations on the Self-Assembly of Fibrillar Collagen 

Aisha Haroun^1,2, Sidorela Reci^1,2, Faizunnahar Dewan^1,3, Yujia Xu¹

¹Department of Biochemistry, Hunter College

²Macaulay Honors College, Hunter College, City University of New York, New York, NY, USA

³PhD Program in Biochemistry, The Graduate Center, City University of New York

Hypothesis/Statement of Problem: Collagen plays a crucial role in providing structural support to skin, bones, muscles, and connective tissue. Its basic structure consists of a triple helix composed of three polypeptide chains with (Gly-X-Y)_n repeat sequences, where X and Y represent any amino acid residue. Any amino acid residue replacing Gly on this repeat sequence would decrease the stability of the triple helix. Our focus is on a fibril-forming collagen model that forms mini fibrils from triple helices, contrary to non-fibrillar collagen. Lethal mutations that prevent peptide folding entirely are found nearer to the C terminus of the peptide. Less detrimental mutations are closer to the N terminus meaning that the peptide would already be partially zipped due to the directionality of folding, resulting in a less impactful consequence. A mutation from glycine to valine decreases the stability of the triple helix in our model, which hinders fibril formation, and thus bone strength and density, as seen in Osteogenesis Imperfecta (OI), or Brittle Bone Disease.

Methods: In order to determine the impact of mutations on the self assembly of the triple helix into fibrils, we engineered a mimetic fibril-forming collagen model with a glycine to valine missense mutation at the N-terminus. The mutant was expressed and purified from transformed Escherichia coli cells.

Results: Our preliminary studies indicate triple helix formation of the fibrillar collagen model, which may be confirmed by using Circular Dichroism (CD). We are continuing to characterize the N-mutant through quantitative analysis of protein concentration and the use of electron microscopy to determine if fibrillogenesis has occurred.

Conclusions: We know from previously conducted studies the dire clinical implications of glycine mutations like in the case of OI and our goal is to increase peptide yield and ultimately determine how mutations affect the self association of the triple helix.

Easel #31 - Tasmina Hassan -  (10-11am; 3-4pm)

Novelty Search Promotes Antigenic Diversity in Microbial Pathogens

Tasmina M. Hassan¹, Brandon Ely², Winston Koh¹, Eamen Ho¹, Anh V. Pham¹, Weigang Qiu^1,2,3

¹Department of Biological Sciences, Hunter College, City University of New York, New York, NY 10065, USA

²Department of Biology, Graduate Center, City University of New York, New York, NY 10016, USA

³Department of Physiology and Biophysics, Institute for Computational Biomedicine, Weill Cornell Medical College, New York, NY 10021, USA

Hypothesis/Statement of Problem: Driven by host-pathogen coevolution, cell surface antigens are often the fastest evolving parts of a microbial pathogen. The persistent evolutionary impetus for novel antigen variants suggests the utility of novelty-seeking algorithms in predicting antigen diversification in microbial pathogens. In contrast to traditional genetic algorithms maximizing variant fitness, novelty-seeking algorithms optimize variant novelty. Here, we seek to understand which algorithm promotes antigenic diversity best.

Methods: Using Python and R, we designed and implemented three evolutionary algorithms (fitness-seeking, novelty-seeking, and hybrid) and evaluated their performances in 10 simulated and 2 empirically derived antigen fitness landscapes.

Results: The hybrid walks combining fitness- and novelty-seeking strategies overcame the limitations of each algorithm alone, and consistently reached global fitness peaks. Thus, hybrid walks provide a model for microbial pathogens escaping host immunity without compromising variant fitness. Biological processes facilitating novelty-seeking evolution in natural pathogen populations include hypermutability, recombination, wide dispersal, and immune-compromised hosts. The high efficiency of the hybrid algorithm improves the evolutionary predictability of novel antigen variants.

Conclusions: Facilitated by large and rapid genetic changes brought by recombination, hypermutability, genetic drift, and diversifying natural selection, novelty search promotes antigenic diversity in natural microbial pathogen populations. Simulated evolution with a combination of fitness- and novelty-seeking algorithms provides a way to predict the rise of high-fitness antigen variants given an empirically derived antigen fitness landscape. Evolutionary centroids of high-fitness variants are promising candidates for broadly protective, escape-proof vaccines against a diversifying microbial pathogen.

Easel #32 - Jasmine He - (1-2pm; 2-3pm)

Synthesis of Nickel-modified Hydroxyapatites for Water-Remediation

Jasmine He¹, Spiro Alexandratos, PhD¹

¹Department of Chemistry, Hunter College

Statement of Problem: Toxic, non-biodegradable metal ions are present in water from industrialization processes and are of interest for removal, as metal ions may have carcinogenic properties and accumulation in organisms may result in adverse outcomes^[1]. Hydroxyapatite (HAP) is an inorganic compound derived from natural sources that can eliminate metal ions from water through adsorption^[2]. We examined whether the incorporation of nickel would alter HAP affinity towards metal ions.

Methods: Various concentrations of nickel-modified hydroxyapatite (NiHAP) are synthesized using the procedure in the Alexandratos Lab Manual, and the structure was assessed using Fourier Infrared Spectroscopy (FTIR)^[3]. Adsorption isotherms were performed with Janus Green B (JGB) serving as proxy for metal ions. The data points were fitted to the Langmuir and Hill Equations to determine Q_max and K_d values.

Results: FTIR confirmed incorporation of nickel into the HAP crystal structure. Q_max and K_d are decreased in every concentration of NiHAP tested.

Conclusions: NiHAPs improve affinity for JGB and may have higher potential for removal of metal ions. Future research should modify HAP with other transition metals to see effects on Q_max and K_d.

Easel #33 - Ryan Henry - (12-1pm)

Investigating the Role of Spontaneous Neural Activity in Modulating Mauthner-cell and Decision Making in Fish

Ryan Henry,^1,2 Denis Shor,^1,3 and Thomas Preuss¹

¹Department of Psychology, Hunter College

²NIH BP-ENDURE Program, City University of New York, Hunter College

³CUNY Neuroscience, The Graduate Center, City University of New York

Hypothesis/Statement of Problem: Spontaneous neural activity in the nervous system is thought to reflect the prior history of neural states and plays an important role in the updating of ongoing processing within these networks in response to internal and external stimuli. This suggests that spontaneous activity is not simply random noise but constitutes information relevant for optimal subsequent behavioral responses. Previous research showed that escape directionality is determined by minor asymmetries in the excitatory/balance between the paired Mauthner-cells (M-cells). The exact mechanisms through which spontaneous activity influences neural circuits and decision-making processes however, is largely unknown. Thus, we asked now how this excitatory/balance, i.e., directional decision making is modulated by spontaneous neural activity in the two M-cells.

Methods: Goldfish (N=5)  were anesthetized with a combination of immersion in chilled water and MS-222 (60 mg/l) and immobilized with intramuscular injections of d-tubocurarine (0.15 mg/g BW) and respirated with a steady flow of aerated water containing the anesthetic through the mouth. The M-cells are large neurons in the medulla, which receive multimodal inputs. Due to their large size and an electric signature they are readily accessible for in vivo intracellular M-cell recordings. To characterize spontaneous neural activity, we performed simultaneous recordings from both M-cells. We also used sound pips and spinal cord stimulations to reveal if spontaneous activity can be modulated.  Visual analysis of the recorded neural activity was used  to identify asynchronous and rhythmic activity within and between the two M-cells.

Results: Preliminary results indicate the presence of rhythmic activity within each individual M-cell but also asynchronous activity between the cells, which warrants further analysis to identify potential anti-correlative features or differences within shared frequencies.

Conclusions: Ultimately, a deeper understanding of the role of spontaneous activity in modulating neural circuits and decision-making could have broad implications for our knowledge of the nervous system and its intricate functions. The results of the current study can be used to update a biophysical model of the M-cell used to simulate intrinsic dynamics related to directional decision making. By incorporating the presence of a spatiotemporal structure of intrinsic activity into future models, researchers can gain a better understanding of the nervous system's functionality and responsiveness to internal and external events. This type of insight can contribute to the development of future innovative brain-computer interfaces.

Easel #34 - Destiny Howell - (10-11am)

Brown Dwarf's Variability Kicks Down the Door to Learn about Its Atmosphere

Destiny Howell¹, Yifan  Zhou²

¹Department of Physics and Astronomy, Hunter College CUNY

²Department of Astronomy, The University of Texas Austin

Hypothesis/Statement of Problem: Our main scientific goal is to measure the brown dwarf's variability through time series direct imaging. We want to use variability to study atmospheric properties of brown dwarfs. We want to use a time series direct imaging to understand patchy clouds on brown dwarfs. The brown dwarf's atmosphere will be used as a template to study other planets.

Methods: We conducted time series direct imaging observations to collect raw data using the DIAFI instrument on the 2.7 meter Harlan J. Smith Telescope at the McDonald Observatory. Once the raw data were collected, we performed time series direct imaging. Next, photometry was measured for each targeted source. We constructed empirical corrections to calibrate our light curves. Finally, we created a fitted light curve model to the observed data. This derives the variability's amplitude and period.

Results: The results show both brown dwarfs have similar rotation periods. Both brown dwarfs have a rotation period of roughly 2.4 hours. Furthermore, one of the brown dwarfs has an amplitude of 1.72% while the second one has an amplitude of 0.87%.

Conclusions: From this research, we can make some conclusions on the rotation period, cloud coverage, and patterns on certain hemispheres. Both brown dwarfs are much faster compared to Earth. The rotation rate is ten times the Earth's. Also, one of the brown dwarfs has more complex cloud structures with greater variability.

Easel #35 - Darvin Huang - (12-1pm; 3-4pm)

Modification of hydroxyapatite with transition metal ions for adsorption of Janus Green B: Experimental Analysis on the Langmuir model

Darvin Huang, ¹ Spiro Alexandratos PhD ¹

¹Department of Chemistry, Hunter College

Hypothesis/Statement of Problem: Contamination of water by dyes is a serious issue for the environment and human health. Hydroxyapatite (HAP) is a powerful, low-cost adsorbent for pollutants through ion exchange. As a naturally occurring part of bones and teeth, it is biocompatible, making it well suited for applications in environmental remediation. Modification of solid supports can change their affinity for substrates in solution and this project aims to measure the effects of metal modified HAPs on the adsorption of the dye, Janus Green B (JGB).

Methods: HAP was synthesized with modifications of Cu, Mn, Fe, Cd, Ni, Zn, Co, and Sr that replaced 10% of the calcium ions. There were additional batches of CuHAP with replacement of 2%, 4%, 6%, and 8% of the calcium ions. Adsorption studies were performed by creating a 0.1 mM stock solution of Janus Green B (JGB), which were serially diluted to create 5 solutions that were 0.01, 0.02, 0.04, 0.06, and 0.08 mM. Nothing was added to control solutions, while 100 mg of HAP was added to test solutions. UV-Vis spectroscopy was then performed to measure amount of adsorption.

Results: The adsorption of JGB from the HAPs was found to follow the Langmuir isotherm, a model used to predict the maximum adsorption. The metal modified HAPs showed mild improvements to maximum adsorption, with iron modifications having the the most significant effect. Variance of the concentration of the metal modifications did not significantly impact absorbance.

Conclusions: The findings indicate that metal modifications improve the adsorbant properties of HAP towards dyes, with iron modifications being most effective. Variation of concentration of the metal does not show any significant improvements. This results indicate iron modified HAP will be a promising target for future studies.

Easel #36 - Mohab Idris - (9-10am; 12-1pm)

Developing a VLP System for SARS-CoV-2 and Assessing the Antiviral Potency of Peptides

Mohab Idris^1,2, Jennifer Drew-Bear ^3,4, Anne Moscona ^3,4,5,6

¹Hunter College 695 Park Ave

²SPURS Biomedical Research Program, Columbia University Irving Medical Center

³Department of Pediatrics, Columbia University Vagelos College of Physicians and Surgeons

⁴Center for Host-Pathogen Interaction, Columbia University Vagelos College of Physicians and Surgeons

⁵Department of Physiology and Cellular Biophysics, Columbia University Vagelos College of Physicians and Surgeons

⁶Department of Microbiology and Immunology, Columbia University Vagelos College of Physicians and Surgeons

Hypothesis/Statement of Problem: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus responsible for the COVID-19 disease, and also responsible for nearly 800 million cases and over 6 million deaths worldwide. To fuse to the host cell, SARS-CoV-2 utilizes 4 major proteins, Membrane (M), Envelope (E), Spike (S), and Cytoplasmic Nucleocapsid (N). Since the Moscona Lab is BSL-2, direct research on SARS is prohibited. Thus, by generating a successful VLP system which mimics the natural SARS-COV-2 assembly and cell entry processes, it enables the tracking of viral fusion via Electron Cryotomography. With the help of our collaborators we developed two different inhibitory lipopeptides that will be assessed for their antiviral potency. The most potent lipopeptide will be potentially selected for in vivo testing.
Methodology: To produce our VLPs, we transfected HEK 293 T cells with Plasmids encoding the spike protein of SARS-CoV-2, which were co-transfected with the E, M, and N-HiBiT plasmids. As for our EVs, we transfected HEK 293 T cells with plasmids encoding the hACE-2-LgBiT receptor. To characterize sequential structural transitions in SARS-CoV-2 spike proteins, we used Cryo-Electron Tomography to image interactions between our VLPs and EVs. To visualize expression of the VLP system, two western blots were conducted. One blot was exposed to Anti-HRC Spike solution and the other was exposed to Anti-haCE-2 solution. A fusion assay was conducted to see if our VLP system fuses with and without trypsin in the presence of the lipopeptides. The most potent lipopeptide was administered intranasallay to ferrets to assess its biodistribution in vivo.

Results: When we mix the EVs and VLPs and add Trypsin, through fragment complementation we see light, and this means we are witnessing viral assembly and entry. A drop in Relative Light Luminescence was detected when the lipopeptides were administered in the fusion assay.
Conclusions: We conducted successful research on SARS-CoV-2 by developing a VLP System. Utilizing this VLP system we were able to assess the antiviral potency of our peptides. The most potent lipopeptide was administered dimer intranasally to naive ferrets (which aren't infected with the virus), and it completely prevented SARS-CoV-2 direct contact transmission when these naive ferrets were housed with ferrets infected with the SARS-CoV-2 virus.

Easel MN9 - Nusrat Jahan - (2-3pm; 3-4pm)

Understanding if a Star's Changing Radial Velocity is from an Orbiting Planet or Stellar Activity

Nusrat Jahan¹, Dr. Megan Bedell³, Dr. Lily Zhao³, and Lianys Feliciano²

¹Department of Physics, Hunter College, City University of New York

²Department of Physics, City College of Technology, City University of New York

³Center for Computational Astrophysics at the Flatiron Institute

Hypothesis/Statement of Problem: A star's changing radial velocity can be impacted by an orbiting planet or stellar activity. The radial velocity technique uses the fact that a star is moving because of a gravitational tug from an orbiting planet. The greatest source of error when finding smaller exoplanets comes from mistaking stellar activity for orbiting planets.

Methods: We used the spectra of star HD 26965 from the ultra-stabilized Extreme Precision Spectrograph (EXPRES) to study and compare its different wavelength chunks.

Results: We found that different wavelength chunks vary in behavior, which is unexpected as they should all have the same radial velocity value within the same observation. By finding the correlation between known activity indicators and observations of star HD26965, we can determine if the star is impacted by stellar activity or an exoplanet.

Conclusions: I will present on how the difference in behavior is related to the effects of stellar activity. Studying these high-precision spectra allows us to further our understanding of how to best measure a star's radial velocity and disentangle an orbiting exoplanet from stellar activity.

Easel MN10 - Syeda Jannath - (2-3pm; 3-4pm)

Selection and Characterization of Resistant Variants Against SARS-CoV-2 Inhibitory Compounds

Syeda Jannath¹², Alison Ashbrook², Hans-Heinrich Hoffmann², Cindy Meyer², Aitor Garzia², Michael Miller³, Stacia Kargman³, David Huggins³, Nigel Liverton³, Peter Meinke³, Thomas Tuschl², and Charles M. Rice²

¹Hunter College, The City University of New York

²The Rockefeller University, New York

³Tri-Institutional Therapeutics Discovery Institute, New York

Hypothesis/Statement of Problem: Viruses in several RNA virus families pose a known or suspected risk as drivers of future epidemics and pandemics as observed recently for SARS-CoV-2. One focus of pandemic preparedness efforts is the development of small molecule, direct-acting antivirals that target essential viral functions to prevent or mitigate virus infection and severe disease. Such small molecules need to be identified, moved through the preclinical pipeline, and be poised for clinical testing and use. Data in the lab have shown that the N7-methyltransferase (MTase) of the SARS-CoV-2 nonstructural protein (nsp14) is an important antiviral target that mediates 5' RNA capping of the viral genome, which is important for genome stability, viral protein translation, and subverting host detection. However, it is unclear whether small molecule-mediated blockade of nsp14 MTase activity will select for drug-resistant viral variants. We hypothesize that passage of SARS-CoV-2 in the presence of nsp14-specific compounds will select for resistance mutations among discrete viral loci that can affect enzymatic function and drug interaction.

Methods: To inform the development of compounds that have a high barrier to resistance, we serially passaged SARS-CoV-2 in the presence of subinhibitory concentrations of two nsp14 MTase-targeting compounds. The resultant virus populations were sequenced to identify candidate resistant mutations, and isogenic viruses encoding candidate mutations will be engineered using reverse genetics.

Results: The Nsp14-targeting compounds did not impair cell viability at the concentrations or time points tested. SARS-CoV-2 infection was reduced by ~95% with 300 nM concentration of TDI-626 or 800 nM concentration of TDI-686 and SARS-CoV-2 subjected to 5 passages in the presence of nsp14 compounds induced CPE that was comparable to SARS-CoV-2 CPE induced in the absence of inhibitor. Additionally, approximately 50-300-fold higher concentrations of nsp14-targeting compounds were required to inhibit SARS-CoV-2 serially passaged in cells treated with subinhibitory concentrations of compound.

Conclusions: This work will enhance our understanding of antiviral drug resistance to develop effective therapeutics that are broad in their inhibitory activity to cover current and future threats from virus families of pandemic concern.

Easel #37 - Alison Juray - (9-10am; 11am-12pm)

Site Specific p53 Mutagenesis Through Inducible CRISPR Base Editing in Pancreatic Organoids

Alison Juray,¹ Sujen Rashid,² Adrian Vega, PhD² Andrew Wenger,²  Whitney Diaz,² Maria Paz Zafra, PhD² Alyna Katti, PhD³ Lukas Dow, PhD^2,3 Despina Siolas, MD, PhD^2,3

¹Yalow Honors Scholar Program, Hunter College, City University of New York, New York, NY, USA

²Department of Medicine, Division of Hematology and Oncology, Weill Cornell Medicine, New York, NY, USA

³Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA

Hypothesis/Statement of Problem: Pancreatic cancer, the third-deadliest cancer in the United States, is associated with a poor prognosis. KRAS is the main oncogenic driver in pancreatic adenocarcinoma (PDA) followed by TP53, however, the cooperative phenotypes produced by distinct KRAS and TP53 mutations are not well characterized. TP53 is mutated in up to 70% of PDA, which may be gain-of-function mutations that bestow neomorphic capabilities or loss-of-function mutations silencing its tumor-suppressor activity. Various p53 mutations have been identified in PDA, although the specific effects they have on the tumor microenvironment, immune evasion, and resistance to cancer therapy remain poorly understood.

Methods: Here, we use a novel inducible CRISPR/Cas 9 gene editing system to introduce specific Trp53 mutations into Kras mutated pancreatic organoids to study their impact on the tumor-immune microenvironment. Organoids mimic the function and behavior of 3D tissues in vitro, allowing us to determine the impact of the six most common p53 mutations in a physiological setting.

Results: Kras^G12D mutated pancreatic organoids with a doxycycline inducible CRISPR/Cas9 editing enzyme were nucleofected with different Trp53 mutant oligonucleotides. To remove p53 wildtype cells, pancreatic organoids were selected with nutlin, causing apoptosis. DNA from nutlin surviving cells were extracted, followed by PCR sequencing of the p53 gene to confirm the mutational status. Ongoing experiments include orthotopic implantation of p53 mutated pancreas cell lines into the pancreata of wild type syngeneic mice for tumor formation and histological analysis.

Conclusions: These experiments aim to understand the impact of p53 mutant alleles on pancreatic tumor development and the surrounding immune microenvironment.

Easel #38 - Sindy Kalauch - (9-10am; 1-2pm)

Sleep Quality and Suicide Ideation among Adolescents: Mediating Role of Rumination and Depression

Sindy Kalauch,¹ Christina Rombola,¹ Ana Ortin-Peralta,² and Regina Miranda^1,3

¹Department of Psychology, Hunter College

²Department of Psychology, Yeshiva University

³The Graduate Center, City University of New York

Hypothesis/Statement of Problem: Prior studies have found a correlation between sleep disturbances and suicide ideation (SI) and suicide attempts (SA). However, there are only limited findings among adolescents, a high-risk group for suicide attempts. This present longitudinal study aimed to examine the relationship between sleep disturbances and SI among adolescents ages 12 to 19 and how rumination might mediate this relationship. We hypothesized that low sleep quality would be associated with increased SI severity and that rumination would mediate the relationship between sleep quality and SI.

Methods: We recruited 163 participants, with SI or SA, from four different health facilities in New York but we only included data from 101 participants who completed the baseline and 12-month-follow-up. We evaluated sleep quality via the Sleep Quality Scale SQS, suicide ideation with the Suicide Ideation Questionnaire SIQ-JR, rumination with Children's Response Styles Questionnaire CRSQ, and depression with the Patient Health Questionnaire -9 PHQ-A. The data were statistically collected with the software RedCap and analyzed with SSPS and the Process Macro for SPSS.

Results: Analysis suggested no significant relation between sleep quality and SI but a significant indirect effect of sleep quality on SI via rumination and depressive symptoms, suggesting that rumination and depressive symptoms serially mediated the relationship between sleep quality and suicide ideation.

Conclusions: There was no direct impact of sleep quality on SI, but there was a serial mediation effect that suggested improving sleep quality may reduce rumination, which in turn may reduce depression and lower SI severity. These findings encourage further research to clarify how sleep disturbances affect SI via rumination and depressive symptoms.

Easel #39 - Elizabeth Katanov - (12-1pm; 3-4pm)

Influence of Inhibitory Neuron Spike Timing on Seizure Generation in the Hippocampus

Elizabeth Katanov^1,2, Zoé Christenson Wick, PhD², Paul Philipsberg, MS², Sophia Lamsifer², Cassidy Kohler^2,3, Tristan Shuman, PhD²

¹ Yalow Honors Program, Hunter College, CUNY, New York NY

² Icahn School of Medicine at Mount Sinai, New York NY

³ New York University, New York NY

Hypothesis/Statement of Problem: In the hippocampus, a brain region strongly associated with learning and memory, a rhythmic wave called theta is thought to organize the timing of neuronal firing. This phenomenon, called phase-locking, is hypothesized to balance neuronal excitation and inhibition, and facilitate cognition. We recently found that in healthy mice, hippocampal inhibitory neurons preferentially fire at theta wave troughs, while inhibitory neurons in epileptic mice fire more distributedly. We hypothesize that impaired inhibitory neuron phase-locking may contribute to seizure generation in an epilepsy mouse model, and that restoration of this timing may reduce the neuronal hyperexcitability which leads to seizures.

Methods: We developed PhaSER (phase-locked stimulation to endogenous rhythms), allowing us to specifically stimulate neurons at either the peak or trough of theta waves in real-time. To manipulate neuronal activity, we express light-sensitive ion channels in hippocampal inhibitory neurons which stimulate or suppress firing upon delivery of red or blue light, respectively. To determine if phase-locking controls seizure generation, we induce an acute seizure with injection of kainic acid followed by excitation of hippocampal inhibitory neurons at the peak or trough of theta, while inhibiting firing outside of these windows. The seizure latency, or time for a seizure to occur, is measured.

Results: Our preliminary data suggests that trough stimulation of inhibitory neurons elongates seizure latency in epileptic mice, and that peak stimulation in control mice shortens seizure latency.  

Conclusions: This work suggests that inhibitory theta phase-locking is a causal regulator of seizure generation and may be a target for future epilepsy therapies.

Easel #40 -  Shadman Kazi - (10-11am; 2-3pm)

The Effect of Nanocage Size on their Uptake by Cancer Cells

Shadman Kazi¹, Hiroshi Matsui^2,3

¹Yalow Honors Scholar, Hunter College of The City University of New York

²Department of Chemistry and Biochemistry, Hunter College of the City University of New York

³Department of Biochemistry, Weill Cornell Medicine

Hypothesis/Statement of Problem: Nanoparticles are capable of being effective drug delivery vehicles, but their success/specificity depends on many factors such as size, shape, and chemical composition. In this study, we hypothesized that changes in the relative concentrations of coating agents for cage-shaped iron oxide nanoparticles, IO-nanocages, change their size or shape and the size of these IO-nanocages influences their uptake by cells. Specifically, larger IO-nanocages are expected to have a higher rate of uptake because of their increased surface area as a result of being larger in size and ability to conjugate more cargo onto the cage.

Methods: To test this hypothesis, we first optimize the synthesis protocol to change the size of IO-nanocages by varying the total volume of water used during synthesis, as well as the concentration of iron. A control nanocage sample would have a normal water injection amount of 1 mL and iron concentration of 2.5 M, these amounts were changed to a combination of 0.5 mL, 2.5 mL, 3 mL of water injection volume, and 0.625 M, 1.25 M, and 5 M of iron injection. After these cages were synthesized, they were coated with dihydrocaffeic acid (DHCA). Next, the effect of nanocage size on their uptake by cancer cells by injecting into cancer cell samples and the effectiveness of nanocages depends on how readily functionalized they were, and their ability in drug delivery in therapeutic or diagnostic measures was also analyzed. Uptake was observed via confocal imaging.

Results: We found that with the increase of both water volume and iron concentration in the synthetic procedure used, there is an increase in the size of nanocages (to a certain extent), decreased hollowness of the nanocage, and a more polydisperse (increased variability) sample observed due to the increased introduction of water and iron. FOr the effect of nanocage size on cellular uptake, preliminary results using confocal imaging indicated that larger cages have improved uptake by cancer cells.

Conclusions: The significance of this is the ability to tailor a nanocage for targeted delivery of drugs to specific tumor sites due to the different size constraints. We are also able to utilize the increased surface area in some cages to conjugate more of the desired protein or cargo and potentially transport more into the targeted cell. 

Easel #41 - Imad Khalil - (11am-12pm; 12-1pm)

The Expression of the Short Isoform of armitage Leads to Abnormal Maternal mRNA Regulation During D. melanogaster Oogenesis

Imad Khalil¹, Caroline Casella^1,2, Diana P. Bratu¹ and Livia V. Bayer¹

¹Department of Biological Sciences, Hunter College, CUNY, New York, NY,10065

² Macaulay Honors College, Hunter College, CUNY,New York, NY,10065

Abstract:  Transposable elements called transposons are DNA sequences that jump within a genome and insert themselves into functional gene loci resulting in deleterious mutations. To safeguard the genome, transposons are degraded via the PIWI-interacting RNA (piRNA) pathway. The RNA helicase Armi (armitage) is a crucial player in the piRNA pathway, as well as  being involved in the regulation of maternal mRNAs during Drosophila melanogaster oogenesis. The armi gene encodes for two mRNA isoforms (short and long), which are differentially expressed within the egg chamber. The long isoform is expressed in the 16 germline cells (an oocyte and 15 supporting nurse cells) and in the monolayer of somatic follicle cells, surrounding the germline. The short isoform is only expressed in the follicle cells. We are interested in deciphering the regulation of these isoforms based on previous studies of armi mutants which only affected the long isoform. Using the Gal4-UAS system to induce the knock down in expression of the long or both isoforms in either germ or somatic cells, has enabled us to  characterize the roles of each gene product. We hypothesize that in the absence of the long transcript, the short isoform is upregulated causing major defects in the regulation of the maternal mRNA, oskar. Using distinct sets of single molecule FISH probes which we generated, we visualized the presence of each isoform in wildtype egg chambers. We used these probes in combination with systematic knockdowns of these isoforms in the germline and/or soma to determine the expression and localization of each armi mRNA isoform in the absence of the other. The insight gained is important not only for Drosophila egg development, but also for the implications armi's human ortholog MOV10 might have due to its involvement in cancerous tissue and antiviral activity with Coronaviruses.

Easel #42 - Aysha Khan - (9-10am; 1-2pm)

Relationship Between Adverse Childhood Experiences and Menopause Symptoms

Aysha Khan,^1,2 Clara Law,^1,3 Natalia Provolo,⁴ Kathleen C. Gunthert, PhD,⁴ Evelyn Behar, PhD^1,3

¹Department of Psychology, Hunter College

²Macaulay Honors College

³Department of Psychology, The Graduate Center, City University of New York

⁴Department of Psychology, American University

Hypothesis/Statement of Problem: Most women undergoing the menopause transition report experiencing bothersome physical and emotional symptoms, including vasomotor symptoms (VMS). It is critical to understand the factors that predict VMS severity because VMS are associated with psychological disturbance, and are the primary reason why women seek health care during menopause. A major limitation of studies examining the relationship between ACEs and VMS is that they failed to compare VMS to non-VMS physiological symptoms. The absence of such comparisons precludes our ability to determine the degree to which ACEs predict VMS specifically. The present study examines the degree to which different ACEs differentially predict VMS versus non-VMS physiological symptoms.

Methods: A nationally representative sample of 227 peri- and early post-menopausal women completed the Childhood Trauma Questionnaire (Emotional Abuse, Emotional Neglect, Physical Abuse, Physical Neglect, Sexual Abuse) and the Menopause Symptom Questionnaire (VMS, Physiological Non-VMS).

Results: Results indicated that both the Physical Abuse (B=.110, t=2.47, p<.05) and Physical Neglect (B=.122, t=3.27, p<.01) subscales independently predicted severity of NonVMS physiological symptoms when controlling for other subscales. Additionally, only the Physical Abuse subscale independently predicted VMS severity when controlling for other subscales (B=.132, t=2.47, p<.05).

Conclusions: These results present a challenge to past studies concluding that ACES uniquely predict VMS. Rather, two types of ACEs (physical abuse and physical neglect) predict general physiological symptoms. Although childhood physical abuse does predict VMS, this is not distinct from its prediction of general physical symptoms.

Easel #43 - Zaeem Khan - (10-11am; 3-4pm)

Effect of the distal loop interactions of the hairpin ribozyme on the catalytic rate

Zaeem Khan^1,2 and Nancy L. Greenbaum¹

¹Department of Chemistry, Hunter College of the City University of New York, New York, NY 10065

²Macaulay Honors College at CUNY, Hunter College, New York, NY

Hypothesis/Statement of Problem: Ribozymes are enzymes composed of RNA. Numerous small nucleolytic (RNA-cleaving) ribozymes have important roles in gene expression in bacteria and plants. The single-stranded RNA satellite genome of the tobacco ringspot virus (TRSV) includes a ribozyme called the hairpin ribozyme (TRhp). As part of the replication cycle of the TRSV satellite RNA, the hairpin ribozyme catalyzes the reversible cleavage and ligation of the negative (antisense) strand of the satellite RNA. The ribozyme sequence folds into a three-dimensional conformation comprising four helical loops, with stems A and B interacting to produce the ribozyme's catalytic site. Beyond these catalytic loops, distal loops with complementary sequences may interact via base pair formation to stabilize the loop-loop interaction, thereby stabilizing the active site and enhancing the rate of catalysis. The goal of this study is to determine whether these distal loops of the TRhp interact and impact on the rate of catalysis.

Methods: Two RNA samples representing the ribozyme were transcribed, one with the wild-type (native) sequence and another in which the sequence of the distal loops was mutated to prevent interaction. We measure the rate of catalysis by the appearance of the cleaved substrate strand with respect to time for the two samples using denaturing polyacrylamide gel electrophoresis. 

Results: To date, we have transcribed and purified samples representing the wild type and mutant ribozyme and a separate substrate strand. We are now attempting to pair the two strands. Once pairing is achieved, catalytic assays will be performed.

Conclusion: Results of this study will provide further insight into the role of stabilizing distal elements in activity of the hairpin ribozyme. An understanding of these "design principles" will assist in an understanding, and engineering of, ribozymes catalyzing other reactions.

Easel #44 - Sara Khasib -  (11am-12pm; 12-1pm)

Nuclear pore complex components play an integral role in regulating P-body formation and mRNA localization during Drosophila melanogaster oogenesis

Livia V. Bayer,¹ Sara Khasib,^1,2 Harpreet Kaur,¹ Andrew Baumer,¹ Diana P. Bratu¹

¹ Department of Biological Sciences, Hunter College, CUNY, New York, NY 10065

² Thomas Hunter Honors Program, Hunter College, CUNY, New York, NY 10065

Abstract: The focus of intense research in recent years has been on cytoplasmic membraneless organelles that form via the liquid-liquid phase separation (LLPS) process. One such organelle, a processing body (P-body), is comprised of proteins that contain intrinsically disordered regions (IDRs), which proved to be the driving force behind LLPS. Two P-body proteins, Cup (human eIF4T) and Me31B (human DDX6) are expressed during Drosophila melanogaster oogenesis. Maternal mRNAs such as the post-transcriptionally regulated oskar mRNA, are stored in P-bodies. oskar is required for proper embryonic patterning and germline formation. Here we address the role of some nuclear pore proteins (nucleoporins, Nups) that create a complex (NPC) which also phase separates similarly to P-bodies. We hypothesize that these residents of NPCs participate in the formation of P-bodies and aid in targeting mRNAs to P-bodies. Nups rich in FG-domains form a highly selective hydrogel barrier that requires transport receptors for the nucleo-cytoplasmic transport of large cargoes. Nup154 (human Nup155) is one nucleoporin which has already been identified to interact with Cup. We found that the knockdown of the NPC components altered the cytoplasmic localization of Cup and Me31B. They no longer form condensates, but instead appear diffused in the cytoplasm. In order to assess Nups' role in coupling mRNAs to P-bodies inside the nucleus, we used single molecule fluorescence in situ hybridization (smFISH) to visualize oskar mRNA. Upon knockdown of Nup154, oskar mRNA no longer colocalized with Cup in the cytoplasm, and its export out of the nucleus appeared to be hindered as indicated by the formation of large oskar mRNA aggregates in the nucleus. We show, for the first time, the connection between Nups and proper P-body function, which has tremendous implications in development, especially neurogenesis, as well as numerous diseases.

Easel #45 - Julie Ko - (11am-12pm; 12-1pm)

Suppression of mitotic division during D. melanogaster oogenesis is regulated by Cup

Samantha N. Milano^1,2, Harpreet Kaur¹, Julie Ko¹, Diana P. Bratu^1,2 and Livia V. Bayer¹

¹ Department of Biological Sciences, Hunter College, CUNY, New York, NY 10065

² Program in Molecular, Cellular and Developmental Biology, The Graduate Center, CUNY, New York, NY 10016

Abstract: Maintaining the spatial and temporal regulation of cell division is crucial for not only proper tissue function, but also for its development. We study cell cycle regulation during Drosophila melanogaster egg development where the mitotic signals are highly regulated throughout oogenesis. Oogenesis begins with an initial stem cell division event followed by four rounds of mitotic cell divisions with incomplete cytokinesis, that gives rise to an interconnected chamber of 16 cells. An important protein necessary for mitotic progression is Cyclin B (CycB). After the 4th cell division, CycB along with other mitotic factors which initiate cell division become suppressed. Among The proteins that have been identified as crucial for this repression is the RNA-binding protein Bruno 1. Bruno 1 directly binds cycB mRNA's 3'-UTR, inhibiting its translation. Here, we show that Bruno 1 expression is in turn regulated by Cup, an eIF4E binding protein. In the absence of Cup, Bruno 1 levels decrease, which then allows cycB mRNA to be translated, and CycB protein to accumulate. This accumulation causes unregulated, aberrant mitosis within the egg chamber. This finding underscores the idea that maintaining proper cell cycle division is mediated via numerous co-dependent proteins and therefore the misregulation of any one of them leads to improper initiation of mitosis. Unraveling the mechanism of this type of regulation is of utmost importance for understanding basic tissue development, and largely the progression of abnormal development such as progression of cancer.

Easel #46 - Jonathan Kogan - (9-10am; 12-1pm)

Anxiety and Depression Symptoms among Emerging Adults who Experienced Adverse Childhood Experiences: Do Approach and Avoidance Coping Strategies Help?

Jonathan J. Kogan¹, Melanie Abbondola BA¹, Tracey A. Revenson PhD¹

¹Department of Psychology, Hunter College, City University of New York, New York, USA

Background: Adverse Childhood Experiences (ACEs) have a severe negative impact on individuals long into adulthood. Studies have shown that ACEs are linked to higher rates of anxiety and depressive disorders years later. Emerging adults (18-29 years old) are highly susceptible to depression and anxiety symptoms. Two modes of coping, approach and avoidance coping, have been shown to have different effects on mental health in this population.

Aims: This study examines the relation of ACES to depression and anxiety among this population, as well as the ability of coping to buffer these effects.

Method: Participants were recruited through ResearchMatch (an NIH platform), the undergraduate subject pool at Hunter College, and flyers posted around Hunter. Inclusion criteria were: being ages 18-29; having experienced at least one ACE between the ages of 0-18, and being fluent in English. Participants (n = 241) completed a Qualtrics survey containing a 15-item ACE questionnaire, as well as self-report scales to measure approach and avoidance coping at the time of the ACEs (Brief COPE), and current depressive symptoms (PHQ9) and anxiety (GAD7).

Results: ACEs were strongly correlated with anxiety (r = 0.353, p < 0.001) and depression (r = 0.342, p < 0.001). Approach coping was not significantly related to anxiety or depression, while avoidance coping was significantly correlated with anxiety (r = 0.458, p < 0.001) and depression (r = 0.494, p < 0.001).

Conclusion: ACEs have a significant positive correlation with anxiety and depression symptoms. Avoidance coping styles are correlated with worse anxiety and depression symptoms.

Easel #47 - Christine Kuang - (10-11am; 1-2pm)

Whole-Mouse Circuit Mapping Reveals Stress-Induced Remodeling from the mPFC

Christine Kuang^1,2, David James Estrin², Shane Johnson², Kenneth Johnson², Pooja Suganthan^2,3, Thaira Ahmed², Ashna Singh², Matthew Wright¹ & Conor Liston²

¹Department of Psychology, Hunter College, the City University of New York, NY

²Department of Psychiatry, Weill Cornell Medicine, New York, NY

³Department of Biology, Macaulay Honors College, New York, NY

Hypothesis/Statement of Problem: The medial prefrontal cortex (mPFC) region is responsible for decision-making, processing threat, and retrieving long-term memory. Dysfunction in the mPFC is linked to stress disorders such as PTSD and impairment in mPFC-dependent behaviors. Previous studies have shown that chronic stress exposure drives excessive, branch-specific dendritic spine elimination and circuit reorganization, weakening connectivity in the mPFC region. Understanding the structural-functional brain connectivity of the mPFC is necessary to progress toward treating stress-induced neurological disorders. Given prolonged stress, we hypothesize that brain regions innervating the mPFC modulate their connectivity to the mPFC.

Methods: We injected helper and rabies virus, a retrograde tracer, into the mPFC of the mouse brain. Following 21 days of repeated corticosterone (CORT) exposure, we performed a series of SHIELD tissue clearing and immunostaining to achieve uniform optical transparency and fluorescent labeling. We used light sheet microscopy to visualize 3D circuitry across the intact brain. Recent advancements in deep-learning have enabled large volume analysis to achieve highly accurate and unbiased cell counts.

Results: We found significant differences between CORT and vehicle cell counts across several brain regions, consequentially remodeling connectivity from the mPFC.

Conclusion: These findings suggest that stress-induced circuitry changes from the mPFC may compromise functional synaptic integrity and lead to maladaptive neuronal plasticity, prompting long-term effects on the brain.

Easel #48 - Rudina Kurtaj - (9-10am; 12-1pm)

The Impact of Psychosocial Services on Emotional/ Social Loneliness Among Underserved Breast Cancer Patients in the Bronx

Rudina Kurtaj,^1,2 Tasmia Kabir², Christina Martinez², Johnna Bakalar², MPH, Brittany Miller², PhD, and Alyson Moadel-Robblee²

¹Department of Human Biology, Hunter College

²Bronx Oncology Living Daily Program, Montefiore Einstein Cancer Center

Hypothesis/Statement of Problem: The COVID-19 pandemic has exacerbated a number of psychosocial stressors, including loneliness. The Montefiore Einstein Cancer Center in Bronx, NY provides free psychosocial services to cancer patients who already experience loneliness to a heightened degree and seeks to understand whether uptake of these services mitigates feelings of loneliness.

Methods: Quality of Life surveys were conducted during the pandemic (2020-2022) with a convenience sample of cancer patients at two time points (baseline and 3 months). Both timepoints included the 6-Item Scale for Overall, Emotional, and Social Loneliness (range 0-6). Following the first timepoint, patients were offered the option to engage in psychosocial services.

Results: In the sample of breast cancer patients (n=67), 92.5% identified as non-white and the average age was 56.76 years (range= 28-82 years). Half (n=34) of the sample engaged in psychosocial services.. Based on an independent sample t-test, results show a statistically significant difference in the average change in loneliness score between those who did not receive services (μ=+0.5294) and those who did receive services (μ=-0.9697) across two time points (p=0.017). While those who did not engage in psychosocial services demonstrated a 9% increase in loneliness, those who did engage saw an average 16% decrease in loneliness scores.

Conclusions: Engagement in psychosocial support services after a breast cancer diagnosis shows promise for reducing loneliness in underserved, vulnerable populations. As social connectedness can create other opportunities for advancement in financial, physical, and mental health, our findings suggest referral to these services immediately after diagnosis may be beneficial.

Easel MN11 - Iana Leshchynska - (1-2pm; 3-4pm)

Identifying the Function of Previously Uncharacterized Transcription Factors Insm1, Zbtb18, and Cxxc5 in Granule Cell Development

Iana Leshchynska¹, Kärt Mätlik², Mary Beth Hatten²

¹Yalow Scholars Program and McNulty Scholar Program, Hunter College, The City University of New York, NY

²The Laboratory of Developmental Neurobiology, The Rockefeller University, New York, NY

Hypothesis/Statement of the Problem: Controlling motor behaviors is a hallmark of the cerebellum, although its participation in non-motor behaviors has also been found in recent research studies. Impairments in cerebellar development result in various neurological diseases, such as autism spectrum disorder, ataxia, and medulloblastoma. Granule cells (GC) are the most numerous cells of the cerebellum and heavily affect the formation of cerebellar cortex layers and cerebellar circuitry. Analysis of processes that affect granule cell development, therefore, could provide for more understanding of developmental diseases associated with the cerebellum. While transcription factors (TFs) are known for orchestrating granule cell development, there are many whose mechanisms are unknown. Here, based on preliminary results from the Hatten lab, we selected three candidate transcription factors (Insm1, Zbtb18, and Cxxc5) for subsequent study. Our goal was to overexpress these proteins in cultured granule cells and in cerebellar slice cultures to study how perturbation of these proteins might affect granule cell morphology,  migration, and/or maturation.

Methods: Genes for the three transcription factors were cloned in overexpression vectors co-expressing the Venus fluorophore. Clones were sequenced to confirm the presence of the protein-coding inserts in the overexpression vectors. We were not able to clone Insm1, therefore, the following experiments were performed using constructs overexpressing Zbtb18 and Cxxc5. The constructs were overexpressed in cultured granule cells and in ex vivo organotypic slices of the cerebellar cortex using electroporation. Immunostaining was performed to analyze the morphology of the cells overexpressing Zbtb18 and Cxxc5, and to infer the developmental stage of electroporated cells.

Results: We have identified a few overly large granule cells during the imaging of the cerebellar culture with overexpressed Zbtb18 and Cxxc5, which could suggest apoptosis or other abnormality; however, because they showed similarity in the phenotype we want to confirm if they were just an artifacts.

Conclusions: While the two gene overexpression produced granule cell cultures, we will re perform the genetic manipulation with a new set of starter cells. The knockdown and overexpression experiment is going to be performed with Zbtb18 and Cxxc5 to determine the morphological differences if present and further evaluate the effect of the studied TFs on the GC development. Understanding the functions of the TFs in the cerebellum will help to tailor the therapeutic applications in subsequent studies for various neurological diseases, such as autism spectrum disorder, ataxia, and medulloblastoma.

Easel #49 - Petvy Li - (10-11am; 3-4pm)

Biomarkers of Ischemic Damage Associated with Liver Preservation Injury Prior to Transplant

Petvy Li¹, Guergana G. Panayotova MD MHS², Keri E. Lunsford MD PhD², James V. Guarrera MD²

¹Department of Chemistry, Hunter College

²Department of Surgery, Rutgers New Jersey Medical School

Hypothesis/Statement of Problem: Demand for liver transplant continues to increase while the donor organ pool remains limited. Post-transplant complications, such as early allograft dysfunction (EAD) can occur due to ischemic injury to the donor graft during liver preservation. There is a critical need to improve liver graft preservation methods to optimize post-transplant outcomes. Conventional static cold storage (SCS), or "on ice," preserves livers at 4^oC. Hypothermic Oxygenated Machine Perfusion (HMP-O₂) is a novel preservation technique, during which the liver is continuously preserved in a nutrient-rich oxygenated solution. Clinical data shows that HMP-O₂ improves post-transplant outcomes. The exact molecular pathways ameliorated by HMP-O₂ remain unknown. Herein we explore nuclear factor kappa B (NF𝜅B), a transcription factor critical for the initiation of inflammatory responses, as a potential key player in preservation injury mitigated by HMP-O₂.

Methods: Liver biopsies were obtained following the preservation period, during the transplant procedure (SCS N=6, HMP-O₂ N=6). Total and nuclear NF𝜅B-p65 tissue expression was detected via immunohistochemistry, IHC (CellSignaling, 1:600). Staining density analysis was performed by two independent, blinded investigators, and results averaged between the two. Quantification of results was performed with Fiji ImageJ (NIH). Statistical analysis was performed via GraphPad Prism v9.4. A p<0.05 was considered significant.

Results: There was decreased density of cytoplasmic staining of NF𝜅B-p65 in livers preserved using HMP-O₂ compared to SCS (p=0.04). Similarly, nuclear translocation of NF𝜅B was decreased in the HMP-O₂ cohort (9% vs 12%, p=ns). In subgroup analysis, patients with EAD post-HMP-O₂ showed increased parenchymal and nuclear NF𝜅B staining compared to those without EAD (p<0.0001). Conversely, incidence of EAD did not affect tissue staining density of liver NF𝜅B post-SCS. In this limited patient sample, there was no significant correlation between tissue parenchymal and nuclear staining (r=0.3, p=0.4).

Conclusions: Livers preserved using HMP-O₂ showed decreased parenchymal and nuclear NF𝜅B-p65 compared to SCS. Results did not show clear correlation between NF𝜅B-p65 tissue density and incidence of EAD. Increased NF𝜅B-p65 might represent an early preservation injury mediator. Early downregulation of NF𝜅B might be a central pathway mitigated by HMP-O₂.

Easel #50 - Gigi Lin - (1-2pm)

Genetic polymorphisms/AIMs and its association with HNSCC susceptibility given tobacco exposure

Gigi Lin¹, Elizabeth Blackman², Karthik Devarajan², Cherie Erkmen³, Denise Gibbs², Jeffrey Liu⁴, Thomas J. Ow⁵, Freda Patterson⁶, Aditi Satti³, Yan Zhou², Camille Ragin²

¹ CUNY Hunter College SCRI

² Fox Chase Cancer Center- Temple University Health

³ Lewis Katz School of Medicine, Temple University

⁴ Department of Otolaryngology, Lewis Katz School of Medicine, Temple University

⁵ Albert Einstein College of Medicine

⁶ Temple University

The primary risk factors that are associated with the development of head and neck squamous cell carcinomas (HNSCC) are tobacco use, alcohol consumption and human papillomavirus (HPV) status. The oral cavity and larynx are the predominant cancer sites associated with tobacco use. Among the race groups, the Black male population consistently has one of the highest general incidence rates across the three cancer subsites. The 178 ancestry informative markers (AIMs) will be evaluated within this case-control study to determine whether any of these AIMs is associated with risk of developing HNSCC. This study is pertinent to understanding the racial disparity that exists in cancer incidence by considering the potential relationship with genetic ancestry.

The study (N=718) consists of African American/Black cases (n=305) and controls (n=395). We generated descriptive tables that seek to characterize the conditions of the population according to demographics, primary site, and cancer grade. To investigate independent associations of genetic variants in the development of HNSCC in this Black population, age-adjusted odds ratios, and p-values to test statistical significance were used. A Benjamini-Hochberg adjusted p-value was generated for each SNP overall and individual SNP genotypes based on a false discovery rate (FDR) of 0.1.  110 AIMS were found to be statistically significantly associated with HNSCC risk based on a FDR of 0.1. There are AIMs that are associated with tobacco-metabolizing genes that are significant, in which they play a role in the risk of developing HNSCC.

Easel #51 - JunXian Ma - (10-11am; 12-1pm)

Crucial Forces In Stable Collagen Mimetic Protein (CMP) Trimer Formation

JunXian Ma^1,2, Yujia Xu^1,2, Sophie Xu^1,3

¹Department of Chemistry, Hunter College

²Macaulay Honors College, CUNY Hunter College, New York, NY

³Department of Chemistry, The Graduate Center, City University of New York

Hypothesis/Statement of Problem: Collagen is the most abundant protein found in the human body, of which type I collagen plays a crucial role in biological activity, primarily functioning as an extracellular scaffold but also participating in wound healing, molecular signaling, and organ structure. As such, collagen has been one of the most sought-after biomaterials for studying its assembly, organization, and mutations on their ability to form triple helices. The collagen peptide is composed of a repeating (Gly-X-Y)_n amino acid sequence. Previously, our lab has designed collagen mimetic proteins (CMPs) for studying collagen I triple helix formation and stability. Our mimetic collagen peptide consists of a Histag on the N terminus followed by a Gly-Pro-Pro (GPP) sequence, and then a GPCC and foldon on the N or C terminus. Despite a 70-80% homology between ɑ1(I) and ɑ1(2) collagen monomer chains, ɑ1(I) triple helix homotrimers have been demonstrated to have a much higher thermostability compared to their ɑ2(I) homotrimer counterparts.

Methods: To investigate the residue responsible for ɑ2(I) homotrimer thermostability, an isoleucine to proline mutation was introduced on the ɑ2(I) homotrimer with the GPCC and foldon both on the C terminus. Additional peptides were manufactured with the GPCC on the N or C terminus to corroborate previous studies on CMP homotrimers with different GPCC placement. The thermostability for each CMP homotrimer under different pH and salt concentrations was determined with CD spectroscopy.

Results: Our research demonstrates that isoleucine to proline mutation on the 68^th residue, GPCC sequence positioning at either the N or C terminus of the CMP, and GPP sequence length confers increased thermostability to the ɑ2(I) homotrimers.

Conclusions: In this study, we identified key interactions in collagen triple helix formation that would aid in CMP production for further biomaterial studies and applications.

Easel #52 - Nawshin Maleeha - (12-1pm; 3-4pm)

Acute vs Chronic Cuprizone-Induced Demyelination Differentially Affects Behavior

Nawshin Maleeha¹, L. Emma Denholtz^2,3, Carmen Melendez-Vasquez^2,3, Ekaterina Likhtik^2,3

¹ Psychology Department, Hunter College, CUNY, New York, N.Y.

² Biology Department, Hunter College, CUNY, New York, N.Y.

³ Biology Program, The Graduate Center, CUNY, New York, N.Y.

Hypothesis/Statement of Problem: Cuprizone is a copper-chelator that induces oligodendrocyte apoptosis and drives demyelination. Although this model has been widely used to study the pathophysiology of multiple sclerosis, its effects on behavior at acute and chronic stages of demyelination are unknown.

Methods: In the present study, we tested male C57BL/6J mice exposed to the cuprizone diet for 6 weeks (acute demyelination), and 12 weeks (chronic demyelination) to age-matched controls on three types of memory: object recognition memory, using the Novel Object Recognition (NOR) task, spatial working, and reference memory, using the Y-Maze task.

Results: The NOR task revealed that animals developed significant memory impairments only after chronic demyelination. In contrast, the Y-Maze revealed that spatial working memory was already impaired after acute demyelination, when mice showed significantly less spatial alternation than controls. Acutely demyelinated mice also showed significantly less rearing in the novel arm than controls without differences in overall rearing, suggesting a specific impairment in exploratory behavior in a novel context and increased anxiety. Notably, spatial reference memory was not impaired at any stage of demyelination, as mice visited the novel arm equally above chance regardless of demyelination status. A preliminary analysis of myelin basic protein (MBP) density after chronic demyelination revealed two trends: 1) MBP density increased in the entorhinal cortex, a region implicated in object recognition memory, with NOR performance, and 2) MBP density increased in the dorsal hippocampus, which supports spatial working memory, with Y-Maze spatial alternation.

Conclusions: This work is foundational for developing effective physiology and behavioral treatments for multiple sclerosis.

Easel #53 - Nisha Manahil - (1-2pm)

Identifying Factors Responsible for Sex Disparities in Melanoma Progression

Nisha Manahil¹, Daniel Deegan², Nora Engel²

¹ Department of Chemistry, Hunter College

² Fels Cancer Institute for Personalized Medicine

Hypothesis/Statement of Problem: Melanoma is an aggressive skin cancer that develops in melanocytes. The overall incidence and mortality rates of cutaneous melanoma are substantially higher in men than women. Sex disparities in melanoma have conventionally been attributed to hormones but sex chromosomes may also play a role, given their possession of genes that impact downstream autosomal targets. In this study, we examined the effects of both sex hormones and sex chromosomes in melanoma incidence, progression, and metastasis through the use of the Four Core Genotypes (FCG) model. 

Methods: In this model, the founder male is mated with a wild-type female to obtain the four core genotypes, XXF, XYF, XXM, and XYM, where F and M denote gonadal female and male, respectively. The FCG mice were injected with either XX or XY melanoma cell lines and tumors were allowed to grow until day 40 post-injection. Tumors were extracted and sent out for immune profiling and sequencing to pinpoint sex-biased transcription and epigenetic factors. The FCG mice were monitored until day 80 to determine metastasis to the lungs and then euthanized.

Results: Preliminary trends in tumor growth were observed; For the female cell line, only male mice developed tumors. For the male cell line, all mice grew tumors. Notably, the largest tumor observed was 1.4 cm^3 in an XY female injected with the male cell line.

Conclusions: These trends are suggestive of sex chromosome-linked factors contributing to the sex biases in melanoma growth and progression, and further research may facilitate the development of sex-specific therapeutic strategies.

Easel #54 - Abigail Manoylo - (12-1pm; 3-4pm)

Physician Certification to Enable Patients to Access Medical Cannabis: State Requirements

Abigail Manoylo,^1,2 Deborah Korenstein M.D.,² and Nirupa Raghunathan M.D., M.S.²

¹Department of Human Biology, Hunter College

²Department of Medicine, Memorial Sloan Kettering Cancer Center

Statement of Problem: Cannabis is among the most widely grown drugs in the world. Some US states legalized cannabis use in medical contexts to control symptoms such as pain and nausea most commonly in the setting of illness such as cancer or HIV/AIDS. Physician knowledge about cannabis is limited; it is not traditionally discussed in formal medical education and studies have shown a lack of physician comfort surrounding it. Some states with medical cannabis (MC) programs have specific requirements for physician qualification to certify eligible patients for the drug, but requirements differ by state. Our aim was to evaluate state requirements for physician certification to enable patients to access medical cannabis.

Methods: We searched Google to document requirements for physician certification to enable patients to access MC in states where it is legal. We viewed each state's department of health website and other relevant sites regarding state MC programs. For states with insufficient online information, we called and sent emails to health departments and/or specific state MC programs inquiring about physician certification requirements.

Results: MC is legal in 37 of 50 states (74%), including 19 states in which recreational cannabis is also legal; we found sufficient information online in 29 of the 37 states with legal MC (78%). The other eight states responded to calls and/or emails requesting information. Among the 37 states, ten (27%) require specific physician education to allow MC access for patients. All required courses are online with a median minimum time requirement of 2.875 (range, 1-4) hours. All courses require payment, with median cost $191 (range, $75 - $289). Seven states have specific state-approved courses with a median number of courses available being 2.8 (range, 1-7); three require physicians to take any credible course available online. Of the ten states with educational requirements, we reached nine for information on recertification. Seven require recertification after a median of 1.5 (range, 1-2) years. In 14 (38%) of 37 states with legal MC, an established physician-patient relationship was required for physician certification. Three organizations offer MC educational courses approved by multiple states - The Answer Page, The Medical Cannabis Institute, and Medical Marijuana 411.

Conclusion: Medical cannabis is legal in ¾ of US states, but fewer than ¼ of states with MC programs require physician education to certify the drug.

Easel #55 - Anriela Marte - (2-3pm)

Using SNAP-TAG to Visualize APOL1-mediated Lysis in Trypanosoma brucei brucei
Anriela Marte¹, Kayla Leiss¹, Jayne Raper^1,2
1Department of Biological Sciences, Hunter College-CUNY
²Biology Program, The Graduate Center CUNY

African trypanosomes are flagellated, extracellular parasites that cause African trypanosomiasis in humans and cattle. Trypanosome lytic factor (TLF) is a modified high density lipoprotein that provides innate immunity to Trypanosoma brucei brucei in humans and other higher order primates. Apolioprotein L1 (APOL1) is the lytic component of TLF through the generation of cation channels which cause an ionic imbalance leading to hypotonic lysis. We hypothesize that the mechanism of APOL1 lysis involves insertion into the acidic endosomal membrane with activation channel opening occurring upon recycling back to the neutral plasma membrane. In order to test this hypothesis, we plan to use fluorescent microscopy to assess if APOL1 does recycle to the plasma membrane of TLF treated Trypanosoma b. brucei. APOL1 is 42 kDa and to fluorescently label it, we will use SNAP-tag which is a protein derived from human O6-alkylguanine-DNA-alkyltransferase, an enzyme involved in DNA repair. The SNAP-tag protein reacts with O6-benzylguanine and is able to covalently bind to benzylguanine derivatives tagged with a fluorophore with high specificity. To achieve the visualization of SNAP-tag labeled APOL1, we will use HiFi DNA assembly to tag the APOL1 gene with the SNAP-tag sequence downstream of the APOL1 signal peptide. The PCR products of this reaction will be confirmed by size and sequence. They will be expressed from mammalian and bacterial plasmids to test the functionality of the tagged-channel. We expect that visualization of the trafficking of tagged-APOL1 will test the hypothesis that APOL1 recycles to the plasma membrane to effect lysis of the parasite.

Easel MN12 - Jeslyn Mei - (12-1pm; 3-4pm)

Lineage-Specific Overexpression of LMX1B Elicits Different Responses in the Basal and the Apical Calvaria

Jeslyn Mei,^1,2 Maria Pacheco-Vergara,² Thach-Vu Ho,³ Angel Cabrera Pereira,² Yang Chai,³ Juhee Jeong²

¹Department of Psychology, Hunter College

²Department of Molecular Pathobiology, New York University College of Dentistry

³Center for Craniofacial Molecular Biology, University of Southern California

Hypothesis/Statement of Problem: The goal of our research is to disrupt osteogenesis in the anterior part of the calvaria, the upper part of the skull, and examine the consequences to calvaria formation. This improves our understanding of the genetic mechanism by which sutures of the calvaria develop positional identity during embryonic development. When the sutures fuse prematurely prior to the formation of the brain, this can lead to a birth defect called craniosynostosis.

Methods: We investigated the effect of neural crest-specific overexpression of LMX1B on calvaria formation using a Cre-loxP system on a mouse model. The anti-osteogenic property of LMX1B is crucial to proper calvaria formation. Subsequently, we used ROSA26-YFP Cre Reporter to label the mutant versus non-mutant cells. Finally, we conducted microCT morphometric analysis of mutant and control calvaria.

Results: MicroCT morphometric analysis of mutant and control calvaria revealed different results in the basal and apical coronal suture of the mutants. The apical suture, as expected, shifted to the anterior, but the basal suture shifted to the posterior. Further lineage analysis showed that mesoderm-derived cells contributed to the frontal bone to compensate for the loss of neural crest-derived bone in the basal calvaria. However, the apical calvaria lacked compensation by the mesoderm bone.

Conclusions: The basal calvaria has a compensatory mechanism to resist anterior shift of the coronal suture. The apical calvaria lacks this compensatory mechanism.

Easel MN13 - Edwina Mensah - (9-10am; 3-4pm)

Correlation of collagen fiber alignment with prostate cancer aggressiveness

Edwina Mensah¹, Mariia Aleksandrovych², Chelsea Yu¹, Min Xu²

¹Department of Chemistry, Hunter College

²Department of Physics and Astronomy, Hunter College and The Graduate Center, City University of New York

Hypothesis/Statement of Problem: Cancer prognostic factors are needed to inform treatment regimens and therapeutic targets for patients with tumors. It has been shown that collagen fibers and their structures are affected by tumors. The organization of collagen fiber of tissues rich in collagen, such as stomach tumors and pancreatitis, correlates with the recurrent time of cancer.

Methods: In this work, we analyzed the collagen structure of prostate cancer tissue sections stained with Hematoxylin and Eosin (H&E) that highlights collagen fibers. We used the open-source MATLAB software framework "CurveAlign" to measure fiber alignment on region rich in collagen fibers.

Results: Our findings suggest a positive correlation between the recurrence time and collagen fibers alignment, with aggressive (< 5 years) cancerous tissue exhibiting lower collagen fiber alignment compared to non-aggressive (> 5 years) cancerous tissue.

Conclusions: In summary, qualitative analysis of the orientation of collagen fibers may provide additional, clinically relevant information about tumors and the important time of reappearance.

Easel #56 - Michelle Merav - (1-2pm; 3-4pm)

Repurposing Antidepressant Agomelatine as a Potential Treatment for Alzheimer's Disease

Michelle Merav^1,2,3,4, Grace Terry^2,3,4,5

¹ RISE fellow

² Figueiredo-Pereira Laboratory

³ Department of Biological Sciences

⁴ Hunter College of the City University of New York

⁵ Biochemistry PhD program, City University of New York

Hypothesis/ Statement of Problem: More than 6 million people currently live with Alzheimer's disease (AD) of which two out of three are women. Alzheimer's is a neurodegenerative disease with limited treatment options and no long-term therapies. Using a systems pharmacology approach to identify previously used pharmaceuticals with repurposing potential to treat AD, our lab selected agomelatine (AGO) as a possible AD therapeutic. Agomelatine, a synthetic melatonin derivative, is an antidepressant used to treat depression and anxiety in Europe and Asia. AGO acts as an agonist of MT1/MT2 melatonin receptors and an antagonist of the 5HT2C serotonin receptor. Due to its multitarget properties, AGO has potential to treat AD as it is a complex disease.

Methodology: We used the Fisher transgenic 344-AD rat model of AD, which expresses human mutant "Swedish" amyloid-precursor protein (APPsw) and Δ exon 9 presenilin 1 (PS1ΔE9). We orally treated TgF-344AD rats and their wild type littermates with AGO (~9mg/kg/day in rat chow) starting at 5 months of age. At 11 months of age, we used an active place avoidance task to assess hippocampal-dependent spatial learning and memory.

Results: Preliminary results indicate that AGO-treated transgenic females exhibit improved learning and memory compared to their untreated transgenic counterparts. Moreover, we are analyzing rat hippocampal tissue for amyloid-beta plaque burden and microgliosis by immunohistochemistry.

Conclusion: These ongoing studies will examine if improved cognitive performance in AGO-treated transgenic females correlates with reduced pathology, such as amyloid-beta plaque burden and microgliosis. We expect our pre-clinical studies with agomelatine to be translatable into clinical practice to treat AD.

Easel #57 - Nyhal Metidji - (10-11am; 2-3pm)

Using Inducible CRISPR-Cas9 Systems to Study the Exocytic Cathepsin L Pathway in Trypanosoma brucei 

Nyhal Metidji ¹, Bernardo Gonzalez-Baradat PHD ¹,Daniel Lopes¹, Jayne Raper PHD^1,2

¹Department of Biological Sciences, Hunter College;

²Biology PHD Program, The Graduate Center, The City University of New York, New York, NY

Hypothesis/Statement of Problem: Trypanosoma brucei are eukaryotic parasites transmitted by the tsetse fly, with certain subspecies being shown to cause African sleeping sickness in humans or nagana in livestock. To protect against parasitemia, humans and other primates have Trypanosome Lytic Factors (TLF) that contribute to their innate immunity. The lytic component in TLF, Apolipoprotein L1 (APOL1), forms channels in the parasitic membrane, allowing for osmotic lysis to occur. Trypanosomes also have the essential lysosomal protease, Cathepsin L (CatL), which has been shown to degrade APOL1. It is not well understood how CatL is secreted through the exocytic pathway, where it can encounter APOL1. We hypothesize that the use of an inducible CRISPR-Cas9 system will allow us to explore the trafficking of CatL.

Methods: To test our hypothesis we will implement a CRISPR-Cas9 system in a line of 2T1 T.brucei parasites, which we will transfect with T7 polymerase and Cas9 genes. Integrating sgRNA plasmid with CRISPR-Cas9, we will be able to knockout specific genes and thereby proteins associated with the exocytic pathway. With this we can elucidate the trafficking CatL by monitoring any changes of CatL secretions.

Results: We have transfected the 2T1 trypanosomes with the T7 polymerase regulated by a tet on/off landing pad and Cas9 plasmids, attaining an inducible system. Using doxycycline Cas9 expression was induced. As a control we are currently transfecting these parasites with the Rab11 sgRNA, which should produce a lethal knockout confirming the functionality of our system.

Conclusions:  From our results we will be able to better understand the way in which CatL is secreted through the exocytic pathway in T. brucei 2T1, allowing us to understand where and how CatL and APOL1 interact.

Easel #58 - Alexis Mitelman - (1-2pm; 2-3pm)

Exploring the Contribution of ATP-Producing Metabolic Pathways Towards Axon Survival

Alexis Mitelman,^1,2 Alexander P. Walsh,² David J. Simon²

¹Department of Chemistry, Hunter College, New York, NY

²Department of Biochemistry, Weill Cornell Medicine, New York, NY

Hypothesis/Statement of Problem: Neurons, the principal cells in the nervous system, can send signals across a distance up to a meter in length in humans via a long cable-like structure called an axon. Several neurodegenerative disorders, including Alzheimer's and Parkinson's disease, have been characterized with axonal pathologies and cellular bioenergetic failure.  However, little is known regarding how axonal metabolism relates to its survival. Axons can be independently studied using an injury model to identify axon-intrinsic survival factors, in which physical separation of the axon from its cell body induces the loss of nicotinamide adenine dinucleotide (NAD+), an essential redox cofactor, and is followed by the catastrophic loss of ATP and the initiation of degeneration. Axonal injury induces both NAD+ and ATP loss, which raises a critical question: is loss of axonal ATP-production causal for degeneration? ATP production occurs primarily via glycolysis in the cytoplasm and oxidative phosphorylation (oxphos) in the mitochondria to meet axonal energy demand. We investigated how glycolysis and oxphos maintain axon survival, and whether perturbations to these pathways affect the rate of degeneration following injury.

Methods: Manipulations to these pathways were achieved by the application of various pharmacological inhibitors of enzymes involved in glycolysis or oxphos at various timepoints before and after injury. Post-injury ATP levels and rates of degeneration were measured by an ATP-sensitive luciferase assay and immunostaining, respectively.

Results: The application of pharmacological inhibitors against the two major axonal metabolic pathways will help elucidate their relative contributions towards basal axon energetics and survival.

Conclusions: Determining the contribution of glycolysis and oxphos to the rate of axonal degeneration will provide insight into the contribution of ATP production towards axon survival.

Easel #59 - Hunter Moran - (10-11am; 3-4pm)

Geography influences susceptibility to SARS-CoV-2 serological response in patients with inflammatory bowel disease: multinational analysis from the ICARUS-IBD Consortium

Serre-Yu Wong,¹ Judith Wellens,² Drew Helmus,¹ Luke Marlow,³ Stephanie Brann,³ Vicky Martinez Pazos,¹ Alan Weinberg,¹ Hunter R. Moran,¹ Colleen McGregor,³ Séverine Vermeire,² Kenji Watanabe,⁴ Koji Kamikozuru,⁴ Vineet Ahuja,⁵ Shubi Vermani,⁵ James Lindsay,⁶ Ashley Kingston,⁶ Usha Dutta,⁷ Harmandeep Kaur,⁷ Mark S. Silverberg,⁸ Raquel Milgrom,⁸ Siew Chien Ng,⁹ Joyce Wing Yan Mak,⁹ Ken Cadwell,¹⁰ Craig Thompson,¹¹ Jean-Frédéric Colombel,¹ Jack Satsangi³

¹The Henry D. Janowitz Division of Gastroenterology, Icahn School of Medicine at Mount Sinai, New York, USA

²Department of Gastroenterology and Hepatology, Leuven University Hospitals, Leuven, Belgium

³Translational Gastroenterology Unit, Nuffield Department of Medicine, University of Oxford, Oxford, UK

⁴Center for Inflammatory Bowel Disease, Division of Internal Medicine, Hyogo College of Medicine, Japan

⁵Department of Gastroenterology, All India Institute of Medical Sciences, New Delhi, India

⁶Center for Immunobiology, Blizard Institute, Queen Mary University of London - Barts Health NHS Trust, London, UK

⁷Postgraduate Institute of Medical Education and Research, Chandigarh, India

⁸Mount Sinai Hospital, Sinai Health System, University of Toronto, Toronto, Canada

⁹Division of Gastroenterology and Hepatology, The Chinese University of Hong Kong - Prince of Wales Hospital, Hong Kong

¹⁰ Division of Gastroenterology and Hepatology, Department of Medicine, Department of Systems Pharmacology and Translational Therapeutics, Department of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Pennsylvania, USA

¹¹Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Warwick, UK

Hypothesis/Statement of Problem: Beyond systematic reviews and meta-analyses, there have been no direct studies of serological response to COVID-19 in patients with inflammatory bowel disease (IBD) across continents. There has been limited data from Asia, with no data reported from India. The ICARUS-IBD consortium assessed serological response to SARS-CoV-2 in patients with IBD in North America, Europe, and Asia.

Methods: ICARUS-IBD is a multicenter observational cohort study spanning sites in seven countries. We report seroprevalence data from 2,303 patients with IBD before COVID-19 vaccination between May 2020 and November 2021. SARS-CoV-2 anti-spike (S) and anti-nucleocapsid (N) antibodies were analyzed.

Results: The highest and lowest SARS-CoV-2 anti-S seropositivity rates were found in Asia (81.2% in Chandigarh and 57.9% in Delhi, India, and 0% in Hong Kong). By multivariable analysis, country (India OR 18.01, [12.03-26.95], p<0.0001; United Kingdom OR 2.43, [1.58-3.72], p=<0.0001; United States OR 2.21, [1.27-3.85], p=0.005), male sex (OR 1.46, [1.07- 1.99], p=0.016), and diabetes (OR 2.37, [1.04-5.46], p=0.039) conferred higher seropositivity rates. Biological therapies associated with lower seroprevalence (OR 0.22, [0.15-0.33], p<0.0001). Multiple linear regression showed associations between anti-S and anti-N titers with medications (p<0.0001) but not with country (p=0.3841).

Conclusions: While the effects of medications on anti-SARS-CoV-2 antibody titres in patients with IBD were consistent across sites, geographical location conferred the highest risk of susceptibility to serologically detectable SARS-CoV-2 infection. Over half of IBD patients in India were seropositive prior to vaccination. These insights can help to inform shielding advice, therapeutic choices, and vaccine strategies in IBD patients for COVID-19 and future viral challenges.

Easel #60 - Marissa Mumford - (9-10am; 1-2pm)

Do Textured Shoe Insoles, a New Technology, Improve Balance and Gait in Cancer Patients with Sensory Impairments?

 Marissa Mumford,^1,2 and Dr. Lisa Ruppert, MD²

¹Department of Chemistry, Hunter College

²Memorial Sloan Kettering Cancer Center, New York, NY

Hypothesis/Statement of Problem: Cancer or its treatment may cause patients to have sensory impairments, which may compromise balance, and gait. Textured shoe insoles are a new technology that may improve symptoms of sensory impairment. These insoles were specifically created to activate small nerve proprioceptors on the soles of the feet. Use has not been studied in the cancer population. This case series aims to report initial findings on use in the cancer setting.

Methods: With IRB approval, a case series was completed for patients who received insoles through the Rehabilitation Medicine Service at Memorial Sloan Kettering Cancer Center (MSK). The series included 7 patients. Patient demographics, cancer diagnosis, treatment modalities, indications for insoles, use of other orthoses were recorded.  We collected and analyzed qualitative patient-reported outcome data regarding sensation and balance with insoles use, adherence, and adverse events.

Results: Among 7 cancer patients for whom insoles were recommended, 4 (57%) were female and 3 (43%) were male. Six (86%) were white and 1 (14%) was black. Median age was 61 (range: 41-70) years. Primary neoplasms varied among all individuals and included bone, colon, cervical and endometrial, lung, spinal, uterine, and vascular cancers. Patients underwent a variety of cancer treatments including radiation in 5 (71%), chemotherapy in 4 (57%), and surgery in 2 (29%). Four (57%) patients utilized more than one cancer treatment. Sources of sensory impairments included disease in 4 (57%), and chemotherapy in 3 (43%). Indications for insoles include hypesthesia in 7 (100%), gait dysfunction in 7 (100%), impaired proprioception in 6 (86%), impaired balance in 5 (71%), and paresthesia in 5 (71%). All 7 (100%) patients had more than one indication for insoles. Two (29%) patients used an additional orthosis with the insoles, including an ankle foot orthotic in 1 (14%) and a knee brace in 1(14%). Two (29%) participated in physical therapy, 1(14%) received acupuncture, 1 (14%) low-intensity focused ultrasound and 1 (14%) aquatic therapy. Reported improvements of symptoms with insole insertion included proprioception in 6/6 (100%), sensation in 6/7 (86%), gait in 6/7 (86%), balance in 4/5 (80%), and paresthesia in 2/5 (40%). Follow up data was available for 6/7 (86%). Of these, 6 (100%) were adherent. No adverse effects were noted.

Conclusions: Our case series suggests that textured insoles are well tolerated and may yield qualitative improvement in proprioception, sensation, gait, balance and paresthesia in cancer patients with sensory impairment. Prospective studies with quantitative measures and standardized questionnaires are needed to validate the benefits of textured insoles in patients with cancer.

Easel #61 - Selma Music - (9-10am; 3-4pm)

Teasing Apart the Role of Depression, Parenting, and Maternal Inflammation on the Intergenerational Transmission of Negative Affect
Kaelyn Kohlasch¹, Cassandra Hendrix¹, Integra Feliciano¹, Selma Music^1,5, Manpreet Kaur¹, Alejandra Lemus², Natalie Brito², & Moriah Thomason^1,3,4
1Department of Child and Adolescent Psychiatry, New York University Langone, New York, NY
²Department of Applied Psychology, New York University, New York, NY
³Department of Population Health, New York University Langone, New York, NY
⁴Department of Neuroscience, New York University Langone, New York, NY
⁵Macaulay Honors College, City University of New York (CUNY)

Hypothesis/Statement of Problem: Maternal depression can have wide-ranging impacts on child development. These associations are detectable early in life, with prior research demonstrating links between maternal depressive symptoms and greater infant negative affect within the first postnatal year. Altered parenting in the context of depression is one mechanism by which risk is transmitted, but biological mechanisms remain less understood. Increased production of pro-inflammatory cytokines across the perinatal period may enhance risk for depressive symptoms, and in turn be impacted by maternal depression.

Methods: ​​The proposed analyses will examine concurrent associations between maternal depressive symptoms and infant negative affectivity using hierarchical linear regression. We will also explore two mediational pathways that may link maternal depressive symptoms to infant negative affect: a social transmission pathway assessed via videotaped parent-child interactions, and a biological transmission pathway assessed via maternal blood samples. Assayed cytokine markers (TNFα, IL-1β, IL-6, IL-8, and IFNγ) will be combined into a latent factor to produce a single maternal inflammation score. Follow up sensitivity analyses will examine whether significant mediation findings are driven by specific inflammation markers.

Results: ​​Maternal depression did not predict maternal reported infant negative affectivity or observed infant negative affect during parent-child interactions. Maternal postnatal inflammation did not moderate the association between maternal depression and observed infant negative affect, but there was a trending interaction between maternal depression and postnatal inflammation to predict reported infant negative affectivity.

Conclusions: In contrast to work showing increased inflammation in the context of heightened depressive symptoms, maternal depression was not associated with concurrent levels of pro-inflammatory cytokines in our sample. While maternal depression did not predict observed dyadic reciprocity in parent-child interactions, less infant negative affect was reported and observed among mother-infant dyads with higher levels of reciprocity. However, among the group of mothers with the highest levels of 6-month postnatal inflammation, greater maternal depression was strongly associated with greater reported, but not observed, infant negative affectivity. Examining depression in collaboration with inflammation levels may enable us to parse out an inflammatory subtype of depression that might yield differential impacts on infant emotional development

Easel MN14 - Adrianne Nemchik - (11am-12pm; 3-4pm)

Analysis of Bone Regeneration in a Murine Calvarial Defect Model Using Immunofluorescent Staining

Adrianne Nemchik^1,2,3; Leya Groysman^1,2,3; Fernando D. Arias, MD¹; Jenn J. Park, MD¹; Alexandra Verzella, BS¹; Roberto L. Flores, MD¹; Piul S. Rabbani, PhD¹

¹Hansjorg Wyss Department of Plastic Surgery, NYU Grossman School of Medicine

²Macaulay Honors College, Hunter College

³Department of Biology, Hunter College

Hypothesis/Statement of Problem: Multipotent stromal cells (MSCs) have the ability to differentiate into specialized cells of the bone and other connective tissues making them capable of bone regeneration. However, little is known about the regenerative potential of the small extracellular vesicles (exosomes) secreted by pediatric bone MSCs (pMSCs). This project explores the effect of pMSC exosomes on different cell types involved in bone regeneration in a murine calvarial defect model.

Methods: Exosomes from pMSCs were applied to 3mm diameter defects made in the calvaria of mice, using a collagen sponge as a vehicle of delivery and stability. Immunofluorescent staining of cross-linked calvarial tissue sections and high-resolution inverted microscope imaging techniques were used to identify relevant cell types and analyze cellular morphology and tissue architecture during regeneration.

Results: At 12 weeks post creation and exosome implantation into the calvarial defect, vasculature patterns within the new bone of the defect were visualized using dual markers EMCN and CD31, which stain endothelial cells. Transcription factors OSX and RUNX2 characterize osteoblast differentiation at different stages of the osteogenic differentiation pathway. Nuclear protein Ki67 marks cell proliferation while POSTN, an extracellular matrix protein of the periosteum, marks osteoblast activity during osteogenesis. We found signals for RUNX2, OSX, and/or Ki67 at the edge of the new bone and fibrous bridge connecting the new growth to intact bone. POSTN was concentrated in the periosteum and surrounded the new bone. These biochemical markers characterized cell types and their spatial locations as they participate in bone regeneration. Our analysis revealed that bone tissue engineering using pMSC exosomes can regenerate bone and demonstrates similar patterns of cellular organization as seen in intact bone.

Conclusions: Understanding of the mechanisms involved in bone regeneration, and identifying key cells, can help optimize exosome treatment options for bone defects for future applications in bone tissue engineering procedures in pediatric cases, such as cleft lip and palate disorders.

Easel #62 - Robert Novo - (1-2pm; 3-4pm)

1-Hydroxylethylidene-1, 1-diphosphonic Acid-Modified Hydroxyapatite: Characterization via Q_max and K_l

Robert Novo¹, Angelina Chu¹, Spiro Alexandratos, PhD¹

¹Department of Chemistry, Hunter College

Statement of Problem: Metals including arsenic and lead contaminate numerous water bodies and pose threats to human health when ingested. Thus, development of substrates that remove these metals from water is critical. Hydroxyapatite (HAP) is a safe, inorganic substance that can be extracted from readily-available materials (e.g., fish bones) at virtually zero cost. HAP is capable of removing metal ions from water but does so with low affinity. 1-hydroxylethylidene-1, 1-diphosphonic acid (HEDP) has high affinity for metal ions. We investigated whether integration of HEDP into HAP would increase HAP's affinity for toxins in water.

Methods: HEDP-modified HAP (mHAP) was synthesized using previously described methods. Synthesis conditions (temperature, time, HEDP content) were varied. Fourier transform infrared spectroscopy (FTIR) was utilized to characterize structural changes in HAP. To study mHAP binding behavior, adsorption isotherms using Janus Green B (JGB) solutions were conducted. Q_max and K_l were determined using the Langmuir model.

Results: Q_max is positively associated with HEDP content. For mHAP synthesized for 24 hours at 80˚C, Q_max in mg/g were: 1330.80 (0.5 M HEDP); 551.89 (0.12 M HEDP); 262.78 (0.06 M HEDP); 156.41 (0.02 M HEDP). mHAP FTIR spectra revealed higher transmission at phosphate peaks (1100 cm^-1 and 990 cm^-1) with higher HEDP content. Temperature and synthesis time had less influence on mHAP FTIR spectra and Q_max.

Conclusion: Modifying HAP with HEDP dramatically increases its ability to remove JGB from water. Future studies should investigate mHAP's affinity to particular metals to further characterize its water remediation potential.

Easel #63 - Lewis Nunez - (12-1pm; 2-3pm)

Methamphetamine Induces Conditioned Place Preference in Sex and Strain Specific Manner in Adolescent Mice

Lewis Nunez,¹ Andre B. Toussaint,¹ and Nesha S. Burghardt^{1, 2} 

¹Department of Psychology, Hunter College

²Behavioral and Cognitive Neuroscience, The Graduate Center, City University of New York

Hypothesis/Statement of Problem: Substance abuse in humans often begins during adolescence, a developmental period characterized by novelty-seeking and high-risk behavior. In addition, there are sex differences in use and response to methamphetamine, with women initiating use earlier and transitioning to regular use faster than men. However, most studies investigating the cellular basis of addiction only test male rodents during adulthood.

Methods: We used a conditioned place preference (CPP) paradigm to test whether there are sex differences in the rewarding effects of methamphetamine (1mg/kg) in adolescent (postnatal day 41) male and female mice. We tested mice of two strains (C57Bl/6 and 129Sv/Ev) to evaluate strain differences in addiction-related phenotypes. To evaluate the neural basis of methamphetamine-induced CPP, brains were removed 90 minutes after the CPP test (drug-free) and behaviorally-induced expression of the neural activity marker c-Fos was quantified in the nucleus accumbens (NAc) with ImageJ software.

Results: In the C57Bl/6 strain, methamphetamine induced CPP and increased c-Fos+ cells in the NAc of females (n = 10) (drug females = 4664 cells vs.  saline females = 3839 cells; t(8) = 2.437, p < 0.05), but not males (n = 10) (drug males = 3201 cells vs. saline males = 2987 cells;  t(8) = 0.9561, p = 0.37). Conversely, in the 129Sv/Ev strain, methamphetamine induced CPP and upregulated c-Fos+ cells in the NAc of males only (n = 8/sex) (drug males = 4056 cells vs. saline males = 3069 cells; t(6) = 4.971, p < 0.01; drug females = 5664 cells vs. saline females = 4684 cells; t(6) = 1.856, p = 0.11).

Conclusions: Our findings demonstrate that there are sex and strain differences in the rewarding effects of methamphetamine. The relationship we find between CPP and increases in cell activity is consistent with literature indicating that the NAc drives the rewarding effects of drugs of abuse.

Easel #64 - Mashhura Nurilloeva - (1-2pm)

Rescuing the Function of Mutant pVHL in ccRCC

Mashhura Nurilloeva¹, Mariam Fouad², Christopher Parry², Ezra Shimabenga^2,3, John Karanicolas²

¹ City of New York Hunter College, New York, NY

² Program in Cancer Signaling & Microenvironment, Fox Chase Cancer Center, Philadelphia, PA

³ Cedarville University, Cedarville, OH

Statement of Problem: Clear cell renal cell carcinoma (ccRCC) is a subtype of kidney cancer which is highly metastatic. VHL mutations are the initiating event in the pathogenesis of renal cancer, and there is no established drug that targets its protein tumor suppressor (pVHL) for reactivation. Aim of the work: In this study, we aimed to characterize HIF-dependent and HIF-independent activities of small molecules that stabilize and re-activate mutant pVHL.

Methods: Small molecule CP4.29 was developed in the Karanicolas Lab to target pVHL. In this project, the on-target activity of CP4.29 was tested in various assays using two missense pVHL mutant cell lines: RCC-MF and 769-P, which have P86S and I180N mutant pVHL, respectively. CP4.29 is expected to reactivate mutant pVHL by binding to the folded protein conformation, thus stabilizing it and shifting the equilibrium away from misfolded or unfolded states. Thus, pVHL's oncogenic substrates, HIF2a and AURKA, would be ubiquitinated and degraded. To test the on-target effect of CP4.29 on pVHL, we used a direct inhibitor of pVHL (VH298).

Results: For RCC-MF cells treated with 5uM of CP4.29, there was an 11% reduction in HIF2a levels and a 25% reduction in AURKA levels compared to cells treated with 0.1% DMSO. By increasing the concentration of CP4.29 to 10uM, further reduction in HIF2a and AURKA levels was observed (39% and 55% relative to DMSO treated cells). Similar responses to CP4.29 were observed by carrying out these experiments in 769-P cells, which harbor a different VHL mutation. HIF2a level was 33% and 41% reduced in 769-P cells treated with 5uM and 10uM of CP4.29, respectively compared to cells treated with 0.1% DMSO. Combining CP4.29 with VH298 blocked the VHL-mediated degradation of both substrates. VH298 reduced the activity of CP4.29 on these substrates where the average expression of HIF2a and AURKA were about 90% to 120% to their levels in cells treated with 0.1% DMSO alone.

Conclusions: We confirm that indeed CP4.29 has an on-target effect on missense mutant pVHL, as exhibited through induction of HIF2a and AURKA degradation. As hypothesized, this effect was abrogated by co-treatment with VH298. These findings pave the way for developing new therapeutic drugs that can reactivate tumor suppressors to stop multiple oncogenic substrates with one a single VHL-reactivating compound, instead of inhibiting downstream effectors with multiple compounds.

Easel #65 - Shlomo Pallas - (10-11am; 2-3pm)

Comparative Therapeutic Inhibition of KRAS in Cancer

Shlomo S. Pallas^1,2,4 Leonard J. Ash^1,3,4, Dennis Lam^1,4 Andrew L. Wolfe^1,3,4

¹Hunter College, Department of Biological Sciences, City University of New York, New York, NY

²Research Initiative for Scientific Enhancement (RISE) Program, National Institute of General Medical Sciences, Hunter College, City University of New York, New York, NY

³Graduate Center, City University of New York, New York, NY

⁴Weill Cornell Medicine, Department of Pharmacology, New York, NY

Hypothesis/Statement of Problem: KRAS is a small GTPase mutated in 15-30% of all cancers, making it one of the most commonly altered oncogenes. Development of targeted KRAS inhibitors has been a major objective in cancer pharmacology. In 2013, the first direct inhibitor of the KRAS G12C mutant was developed, leading to the development of the first clinically approved drug for direct KRAS inhibition, Sotorasib. However, patients treated with Sotorasib develop resistance to KRAS inhibition after a median of 6 months, indicating a need to understand relevant mechanisms of resistance and study new ways to inhibit KRAS. This project has focused on characterizing resistance mechanisms to six structurally and mechanistically distinct KRAS G12C and G12D inhibitors.

Methods: We have conducted time and dose course experiments on ASPC1 (G12D) and H358 (G12C) lines and characterized cell viability and downstream cellular signaling using western blotting.

Results: We found that all tested inhibitors significantly reduced cell viability in both a dose and time dependent manner, and that inhibition was specific to the targeted KRAS mutant allele.

Conclusions: Our results indicate that the structurally distinct KRAS G12C inhibitor, Garsorasib, is more effective than currently approved KRAS G12C inhibitors against sensitive H358 cells. Future directions will be to conduct a drug anchored synergy screen on our six direct KRAS inhibitors using a cancer drug library of FDA approved antineoplastic agents.

Easel #66 - Cassey Pantzi - (2-3pm; 3-4pm)

Metabolic Regulation of Gene Expression in Oligodendrocyte Progenitor Cells

Cassey Pantazi^1,2,  Sami Sauma^3,4 Patrizia Casaccia^3,4

¹ Department of Chemistry, Hunter College, the City University of New York, New York, NY

² Velay Summer Undergraduate Research Fellow,  the City University of New York, New York, NY

³  Graduate Program in Biology, Graduate Center of the City University of New York, New York, NY

⁴ Neuroscience Initiative, Advanced Science Research Center, Graduate Center of the City University of New York, New York, NY

Hypothesis/Statement of Problem:  Oligodendrocyte progenitor cells (OPCs) are highly proliferative cells that can differentiate into oligodendrocytes, the myelinating cells in the central nervous system (CNS). Previous studies have shown that the timing of OPC differentiation is partly dependent on the levels of histone acetylation. Histone acetylation is an epigenetic modification that positively regulates gene expression. In OPCs, high levels of histone acetylation positively regulate the expression of growth and division-related genes, and histone deacetylation is required for OPCs to stop proliferation and begin differentiation. The substrate of histone acetylation is a high-energy metabolite called acetyl-Coenzyme A (acetyl-CoA). We hypothesized that OPCs sense the metabolic conditions in their environment as changes to histone acetylation based on the availability of acetyl-CoA. We therefore asked if changes in the levels of acetyl-coA have an effect on histone acetylation and the timing of differentiation.

Methods: To test our hypothesis we targeted the enzyme responsible for the synthesis of acetyl-CoA in the nucleus, ATP Citrate Lyase (ACLY), for the deletion in OPCs in the developing mouse brain. We used a tamoxifen-induced CRE line to selectively delete Acly in PDGFRa+ OPCs. Tissue samples were processed and tested for levels of histone acetylation and number of OPCs by immunohistochemistry (IHC).

Results: We detect a trend decrease in histone acetylation. We intend to repeat this model with new samples and further investigate the role of acetyl-coA metabolism in histone acetylation in OPCs and the timing of OPC differentiation.

Conclusions: We detect a decreasing trend in histone acetylation and decreased number of OPCs thereby supporting the hypothesis that Acly is necessary for glucose derived acetyl-coA driven cellular division. This mouse model is significant to the investigation of the regeneration of oligodendrocytes which can aid in remyelination, investigating inflammatory diseases such as multiple sclerosis where remyelination is incomplete.

Easel #67 - Suraj Patel - (2-3pm; 3-4pm)

Senescence as a Survival Mechanism in PTEN-deficient Tumor Cells in Response to Ionizing Radiation Treatment

Suraj S. Patel^1,2, Xinyi Fan², Meng Ouyang², Wen H. Shen²

¹ Macaulay Honors College at CUNY, Hunter College, New York, NY

² Department of Radiation Oncology, Weill Cornell Medicine, New York, NY

Hypothesis/Statement of Problem: Implemented for over a century, a common treatment modality for cancers has been radiotherapy, where high dose radiation is used to eliminate cancer cells by inducing DNA damage. The phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is one of the most frequently mutated tumor suppressor genes in human cancers. Canonically, PTEN hinders aberrant cellular proliferation as an antagonist of the PI3K/AKT cell survival and proliferation pathway, but new evidence has emerged supporting its critical role in maintaining genomic stability. The goal of our study was to investigate the molecular mechanisms behind the surprising ability of PTEN-deficient cancer cells to thrive after exposure to high dose ionizing radiation in vitro. We hypothesize that spontaneous escape of cell cycle arrest after therapy-induced senescence (TIS), is a cell-autonomous mechanism PTEN-deficient cancer cells primarily utilize to promote resistance to radiation therapy.

Methods: CRISPR-Cas9 genome editing was used to induce random mutations on the PTEN gene in a metastatic mouse mammary adenocarcinoma cell line, and PTEN deficiency was confirmed by western blot. Standard cell culture techniques were used to maintain cell lines. The Xstrahl small animal radiation research platform (SARRP) was used for various radiation treatments (0 - 10 Gy). Wild-type and PTEN-deficient cells were compared using cell growth curves, and cell death post radiation treatment was measured by propidium iodide labeling followed by flow cytometry analysis. The clonogenic phenotype was observed using colony formation assays that test the ability for single cells to develop into colonies. β Galactosidase was used to quantify senescence levels in untreated and radiation treated cells.

Results: PTEN-deficient cells demonstrate increased cellular senescence after high dosage ionizing radiation, an increase in non-clonal senescence-like cells post radiotherapy, decreased post radiotherapy cell death, and spontaneous recurrence of colony formation following radiotherapy-induced tumor dormancy.

Conclusions: This preliminary study indicates that PTEN may be an important regulator of maintaining therapy induced senescence, and that spontaneous escape of therapy-induced senescence may be an acquired mechanism of dormant tumor relapse post ionizing radiotherapy.

Easel MN15 - Swara Patel - (9-10am; 3-4pm)

Targeting  Glioblastoma with Chimeric Antigen Receptor-T Cells

Swara Patelٰ¹ ² ³, Touraj Aligholipour Farzani¹ ², and Khalid Shah¹ ²

¹Center for Stem Cell and Translational Immunotherapy, Brigham and Women's Hospital, Harvard Medical School

²Department of Neurosurgery, Brigham and Women's Hospital

³Department of Biological Sciences, Hunter College

Hypothesis/Statement of Problem: Glioblastoma (GBM) is a highly malignant primary brain tumor in adults with a 2 year survival rate of 8-12%. Chimeric Antigen Receptor (CAR) T cells are T lymphocytes that are genetically engineered to express a receptor selectively targeting a mutant or overexpressed antigen on cancer cells. CAR-T cell therapy has been successfully translated to the clinic as a therapeutic option for hematologic malignancies such as B-Cell Lymphoma and Acute Lymphoblastic Leukemia. Efforts are now underway to identify and target antigens on solid tumors with CAR-T cells. Interleukin 13 Receptor α2 (IL13Rα2) is a decoy receptor for the cytokine Interleukin 13 (IL13) expressed on about 60% of GBM cells while being relatively absent from healthy brain tissue making it an optimal target for engineered CAR-T cells.

Methods: We developed CAR-T cells expressing the IL13 Zetakine containing an E13Y mutein which increases their binding affinity to the mutant IL13Rα2 50 folds while decreasing their affinity to the wild type IL13Rα1 5 folds. We genetically engineered the CAR-T cells using third generation lentiviral vectors produced via transfecting 4 plasmids including a transgene plasmid to express IL13 Zetakine in Human Embryonic Kidney (HEK) 293T cells. We subsequently transduced T cells which were isolated from Peripheral Blood Mononuclear Cells through negative immunomagnetic selection. We evaluated the therapeutic efficacy of the CAR-T cells in-vitro through a cytotoxicity assay after co-culturing the cancer cell lines and CAR-T cells for 24 hours.

Results: The experimental GBM-IL13Rα2 cell line has been genetically engineered to stably express IL13Rα2 to demonstrate the specific interaction of the CAR-T cells with this mutant antigen compared to the control GBM cell line U87 fmc which does not uniformly express IL13. Here, our cytotoxicity assay demonstrates the specific targeting of the cells expressing IL13Rα2 by the IL13 Zetakine CAR-T cells. We conducted an Enzyme-Linked Immunosorbent Assay to quantify the production of interferon gamma produced by activated T cells and a Western Blot to detect the production of cleaved Caspase 3.

Conclusions: Our results demonstrate the targeted killing of cancer cells expressing IL13Rα2 by the IL13 Zetakine CAR-T cells compared to the control groups. Based on our in-vitro results, these CAR-T cells are a promising therapeutic option for GBM. Future directions include evaluating the therapeutic efficacy of these CAR-T cells in-vivo in immunodeficient NOD-scid mice.

Easel #68 - Sophia Pisarevskiy - (12-1pm; 2-3pm)

Adsorption of Copper (II) Ions by Tea Leaves

Sophia Pisarevskiy^1,2, Brandon Campos¹, Armin Osmanovic¹, & Abel E. Navarro¹

¹Science Department, Borough of Manhattan Community College, City University of New York

²Hunter College, City University of New York

Hypothesis/Statement of Problem: Due to the extensive use of copper water pipes and dumping of industrial wastewater into streams and rivers, copper makes its way through drinking water sources. The contamination of copper in drinking water then creates the risk of developing deteriorating health issues. Therefore, easily available and cost-effective adsorbents that could remove these copper contaminants in drinking water are highly necessary. Adsorption is an effective method in the removal of organic and inorganic compounds due to its surface and textural properties. The purpose of this study was to observe the adsorption kinetics of Copper (II) ions by PepsiCo tea leaves.

Methods: By measuring levels of uptake, we were able to determine the effectiveness of tea leaves at removing Copper (II) ions from a solution. The morphological and textural characteristics of the adsorbents were assessed by scanning electron microscopy (SEM) before and after adsorption. Batch experiments testing the pH, mass, particle size, salinity, heavy metal, crowding, dye, and time dependence effects were run to understand how each of the variables affected the uptake of copper.

Results: Maximum adsorption was reached with 100 ug of sample and a particle size of 75-106 micrometers at a pH of 6.5. As the particle size increased, the amount of copper removed from the solution increased. The time dependence test showed the speed of the reaction to be approximately 5 minutes long. Using SEM, the T and TB samples were found to have heterogeneous surfaces which increased the surface area of the absorbent, as well as its absorption capabilities. The TA and TO samples lost their rough exteriors but contained a greater amount of pores, demonstrating a physical change. Increased porosity in SO and SA decreased the surface area and amount of absorption sites available. The EDS analysis indicated an increase in copper after adsorption, confirming the ability of tea leaves to adsorb pollutants. TGA readings indicated that TB was the most heat resistant, had the highest weight percent, and the strongest structure than the other absorbents.

Conclusions: Overall, TB was found to be the most successful absorbent. This study shows high support towards tea leaves as a major contribution to a cost-effective solution for the removal of pollutants from wastewaters. This could help decontaminate the harmful health effects of pollutants such as Copper located in sources of drinking water.

Easel #69 - Amit Puthan - (1-2pm; 2-3pm)

Using Bioinformatic Tools to Determine Specific Cell Marker Genes for Pharyngeal Development in Zebrafish 

Amit Puthan, Kiyohito Taimatsu, Daniel A. Castranova , Madeleine Kenton Gennady Margolin, and Brant M. Weinstein

Division of Developmental Biology, NICHD, NIH, Bethesda, MD 20892 

¹ Department of Affiliation, National Institutes of Health 

² Eunice Kennedy Shriver National Institute of Child Health and Human Development

³ College Summer Opportunities to Advance Research (C-SOAR) Program, The National Institutes of Health 

Hypothesis/Statement of Problem: The organogenesis of the pharynx during human embryonic development is not yet fully understood due to surrounding bone, muscle, and cartilage tissue that make it hard to visualize the organ in a developing mammalian fetus. The zebrafish has proven to be a good model organism for studying embryonic organogenesis because they reproduce quickly, and their embryos are translucent. I aided Dr. Kiyohito Taimatsu in his study of pharyngeal organogenesis in zebrafish embryos by generating a list of specific cell-marker genes, genes that are specifically involved in the embryonic development of the pharynx. 

Methods: I compiled and analyzed data from two online databases, The Zebrafish Information Network and the UCSC cell browser. Dr. Taimatsu provided me with a preliminary list of potential cell marker genes. I then gathered and cross-referenced in-situ hybridization images from The Zebrafish Information Network (ZFIN.org), a database of genetic and genomic data for zebrafish, and scRNA-seq plots from the UCSC cell browser, a database for single-cell RNA sequencing data, to determine which genes were specifically involved in pharyngeal development. 

Results: At the end of my research, I compiled a list of 10 specific cell marker genes (Gng13a, trpm5, Desmb, Cav3, myl13, Krt1-19d, Colec-10, Col2a1a, Muc5.3, and Muc13a), each involved in the development of different cell types in the pharynx. 

Conclusions: Dr. Taimatsu might use these genes to generate transgenic lines for better imaging and visualization of pharyngeal cell migration and differentiation, or to perform knockout studies to better understand the roles of these genes in proper organ development.  

Easel MN16 - Afsana Rahman - (11-12pm; 1-2pm)

Argonaute-2-dependent gene regulation in the mammalian brain

Afsana Rahman,¹ Benjamin Kleaveland MD-PhD^,2

¹Yalow Scholar Program, Hunter College, The City University of New York

²Weill Medical College of Cornell University

Hypothesis/Statement of Problem: Humans with haploinsufficiency of Argonaute 2 (Ago2) exhibit neurological deficits and the molecular basis of these diseases are currently not known. Ago2 is the most abundant Ago protein in the mammalian brain, part of the RNA-induced silencing complex, and unlike other Ago proteins, it is also able to slice target RNAs. This slicing activity is essential for mouse development, red blood cell production, and oogenesis. However, its importance in the brain has not been thoroughly studied. The goals of my project are to determine the impact that Ago2 haploinsufficiency and Ago2 slicing have on neuronal gene expression in mice.

Methods: To circumvent the perinatal lethality caused by complete loss of Ago2 slicing activity, we used a Cre recombinase system on mice to generate genotypes. We crossed neuron-specific Actl6b-Cre mice heterozygous for a catalytically dead allele of Ago2 (Ago2^CD/+) with homozygous Ago2 floxed mice (Ago2^fl/fl) to generate the following four genotypes: Ago2^+/fl, Ago2^CD/fl, Actl6b-Cre; Ago2^+/fl, and Actl6b-Cre; Ago2^CD/fl. These genotypes allowed us to determine the effects of both Ago2 haploinsufficiency and Ago2 slicing activity. We isolated six central and peripheral nervous system tissues from these mice and sequenced their libraries. I then implemented a paired-end RNA-seq pipeline using bioinformatics software such as STAR alignment, featureCounts, and DESeq2 to perform differential pairwise expression analysis between genotypes of each brain region.

Results: From our results, we confirm that Ago2 slicing activity, and not haploinsufficiency, is a primary regulator of a gene known as Cdr1as, a previously only-predicted target of Ago2 slicing with a single near-perfect binding site for microRNA-671. Cdr1as was significantly differentially expressed in all tissues analyzed. All other dysregulated genes were brain-region specific and the effects from either Ago2's impaired slicing and haploinsufficiency were modest. We also conclude that retrotransposons are not affected by perturbations of Ago2 as no differentially expressed retrotransposons were detected in mice. Our next steps are to create additional RNA-seq pipelines identifying enriched circular RNAs and transcript isoforms as other types of transcripts may be regulated by Ago2 slicing in neurons.

Conclusions: This study provides insight into the molecular mechanisms underlying neurological diseases associated with Ago2 haploinsufficiency and highlights the importance of Ago2 slicing activity in neuronal gene expression regulation by confirming that Ago2 slicing activity, and not haploinsufficiency, is a primary regulator of Cdr1as.

Easel #70 - Sidorela Reci - (9-10am; 2-3pm)

Understanding the Effects of Disease Causing Mutations on Self Assembly of Collagen
Sidorela Reci,^1,2 Aisha Haroun,^1,2 Faizunnahar Dewan,^1,3 Yujia Xu¹

¹Department of Biochemistry, Hunter College

²Macaulay Honors College, Hunter College, City University of New York, New York, NY, USA

³PhD Program in Biochemistry, The Graduate Center, City University of New York

Hypothesis/Statement of Problem: Collagen is a structural protein that provides support in the extracellular matrix of connective tissues such as bones. Mutations in Type I collagen lead to genetic disorders including Osteogenesis Imperfecta (OI) that cause abnormalities in connective tissues. Glycine is required at every third residue of the amino acid sequence of collagen to ensure triple helix formation without distortions, and replacements of glycine lead to molecular distortions in the triple helix. The severity of OI varies based on the properties of the amino acid replacing glycine in a missense mutation and the location of the mutation. Generally, highly destabilizing and lethal mutations of collagen are found towards the C-terminal, while nonlethal mutations cluster on the N-terminal due to the C to N directionality of collagen folding. Past studies have shown that glycine substitutions decrease the stability of the triple helix but the effects on fibrillogenesis are largely unknown.

Methods: To further investigate the impact of mutants on fibrillogenesis, we engineered a collagen mimetic model with a Gly → Val missense mutation at the C-terminal region. Using an Escherichia coli expression system, the mutant was expressed and purified from transformed cells.

Results: The purified C-mutant is in the stages of characterization using Circular Dichroism (CD) to identify triple helix assembly and thermal stability, and Transmission Electron Microscopy (TEM) to validate fibrillogenesis.

Conclusions:  Gly → Val mutants have previously shown to be lethal in cases of OI and we hope to elucidate the differential effects of mutations at the C or N- terminals on fibrillogenesis.

Easel #71 - Lauren Renzoni - (9-10am; 1-2pm)

Reduced Acute Stress Susceptibility and Depression in Post-Menopausal Female Mice C57bl6: The Role of Dopamine Neuron Sensitivity and Plasticity

Lauren Renzoni ¹, Rania Darwish¹, Emine Ustundag ¹, Yuka Miura^,2, Allyson K. Friedman^1,2

¹ Department of Biological Sciences, Hunter College of the City University of New York, New York, NY, 10065, USA

² Graduate Center of the City University of New York, New York, NY, USA.

Hypothesis/Statement of Problem: Aging is a multifaceted process that affects the physiological, psychological, and cognitive aspects of an individual's life. Our study investigates the role of aging in reducing acute stress susceptibility and depression in post-menopausal C57bl6 female mice. The focus is on understanding the underlying mechanisms that involve reduced dopamine neuron sensitivity and plasticity. Previous research has shown that women tend to exhibit a decline in depression rates late in life, particularly after menopause. Here, we aimed to elucidate the biological basis for this phenomenon. We hypothesized that the reduction in acute stress susceptibility and depression observed in post-menopausal female mice is due to decreased dopamine neuron sensitivity and plasticity.

Methods: To test our hypothesis, we subjected young and aged post-menopausal female C57bl6 mice to acute stress tests and evaluated their behavioral responses and dopamine neuron plasticity.

Results: Our results revealed that aged female mice demonstrated reduced susceptibility to acute stress compared to their younger counterparts. Additionally, we observed a significant decline in depression-like symptoms in aged female mice, which correlated with the reduced sensitivity of dopamine neurons. Our findings support the idea that reduced dopamine neuron sensitivity and plasticity may be key factors contributing to the decline in depression rates observed in post-menopausal women.

Conclusions: In conclusion, our study provides valuable insights into the role of aging, dopamine neuron sensitivity and plasticity in reducing acute stress susceptibility. These findings may have significant implications for understanding the neurobiological basis of late-life depression and developing targeted interventions for mental health in aging populations.

Easel #72 - Stephen Rosario - (12-1pm; 2-3pm)

Highly Conductive PAN-based Hybrid Aqueous/Ionic Liquid Solid Polymer Electrolytes with Tunable Passivation for Li-ion Batteries
Stephen Rosario, Steven Greenbaum
Hunter College of the City University of New York, New York, USA

The global demand for lithium-ion batteries has created safety concerns due to the use of an organic solvent-based electrolyte, which is highly flammable. This study investigates a solution to these safety concerns using a newly developed electrolyte strategy called the hybrid aqueous/nonaqueous electrolyte (HANE) to adjust passivation at the anode in solid polymer electrolytes (SPEs). The HANE strategy, which incorporates ionic liquids as the nonaqueous component, was found to be effective in enhancing the performance of SPEs, while adequately addressing safety concerns. This research focuses on HAILSPEs (Hybrid Aqueous/Ionic Liquid Solid Polymer Electrolytes), which are designed to maintain ionic conductivity while forming a sturdy passivation layer. Two HAILSPE systems were synthesized and showcased remarkable room temperature ionic conductivities of up to 5.39 mS/cm, which is a significant improvement over previous generations of materials. Ionic diffusivity measurements performed at Hunter College, were conducted by pulsed field gradient nuclear magnetic resonance methods, and revealed Li+ ion transference numbers (fraction of the total current carried by the Li+ ions) of 0.6 or higher. The findings suggest that straightforward techniques, such as those used in this work, could serve as a viable solution to address the persistent challenge of cathodic degradation in aqueous SPEs. This research could have broad implications for the development of advanced and durable energy storage systems.

Affiliations for this experiment include: Kyle B. Ludwiga, Riordan Correll-Browna, Max Freidlina, Monesha Garagab, Sahana Bhattacharyyaa, Patricia M. Gonzalesa, Arthur V. Crescec, Steven Greenbaum, Chunsheng Wanga, Peter Kofinasa
(a) Dept. of Chemical & Biomolecular Engineering, University of Maryland, 4418 Stadium Dr., College Park, MD, 20740, USA
(b) Dept. of Physics & Astronomy, Hunter College of the City University of New York, 695 Park Ave., New York, NY, 10065, USA
(c) Combat Capabilities Development Command US Army Research Laboratory, 2800 Powder Mill Rd., Adelphi, MD, 20783, USA

Easel #73 - Elisa Sambataro - (9-10am; 12-1pm)

In Vitro Model to Study the Role of the Nasopharyngeal Microbiome in SARS-CoV-2 Infections

Elisa Sambataro^1,2, Alba Boix-Amorós², Tiffany Chan²

¹ Yalow Honors Scholars Program, Hunter College of the City University of New York. New York, NY 10065 

² Department of Genetics and Genomic Sciences, Precision Immunology Institute, Icahn School of Medicine at Mount Sinai. New York, NY 10029

Hypothesis/Statement of Problem: Published studies suggest that SARS-CoV-2 infections can cause changes in the microbiome's diversity and composition. Also, the nasopharyngeal microbiome, the first site of encounter between SARS-CoV-2 body cells, may protect against several viral infections. There is currently no evidence, however, whether a healthy nasopharyngeal microbiome or the presence of specific commensal bacteria could lower the risk of infection.  

Methods: Our group developed an in vitro model of SARS-CoV-2 infection that allowed us to study viral-cell interactions in the presence of bacteria. In this model, we expressed the human angiotensin-converting enzyme 2 (hACE2) receptor on the surface of a monolayer of HEK293T cells. We expressed the receptor-binding domain (RBD) of SARS-CoV-2 spike protein in the Expi293 cell line. The hACE2-expressing cells were co-incubated with SARS CoV-2 RBD-expressing cells in the presence of commensal bacteria commonly present in the nasopharyngeal microbiome. In this experiment, we used S. epidermidis, S. aureus, C.  pseudodiphteriticum, C. striatum, M. catarrhalis, V. dispar, and  P. bivia, as well as bacteria obtained from nasal swabs from three healthy donors to co-incubate alongside the hACE2 and RBD-expressing cell lines. To analyze the results, we used an integrated system for screening for RBD binding to the hACE2-HEK293T cells based on fluorescence microscopy. It allowed us to visualize the fluorescent signal of hACE2 and RBD, as the proteins were both tagged to a different fluorophore.

Results: We calculated correlation coefficients between the hACE2 and RBD fluorescence. We compared the values containing the commensal bacteria or bacteria from nasal swabs with a positive control using t-tests. Based on preliminary results, co-incubation with S. epidermidis (p=0.002) and S. aureus (p=0.002) resulted in significantly lower protein-receptor correlations than in the condition without bacteria, suggesting that the presence of these two bacteria could reduce the binding of the SARS-CoV-2 RBD protein to its receptor. Donor samples showed varying results. While one of the samples showed a reduction in protein-receptor binding (p=7.23e-05), another one led to an increase in binding (p= 0.016), suggesting that variations in microbiome composition across donors could have a role in SARS-CoV-2 infections.

Conclusions:  S.epidermidis and S. aureus showed limitations in the SARS-CoV-2 receptor-binding domain protein and hACE2 binding, and the nasopharyngeal microbiome plays a protective role in response to SARS-CoV-2 infections.

Easel #74 - Abir Sarker - (9-10am; 2-3pm)

The Effect of Iron-Oxide Nanocage Size on Brownian Motion and siRNA Delivery in Cancer Cells

Abir Sarker¹, Hiroshi Matsui^2,3

¹Macaulay Honors Scholar, Hunter College of The City University of New York

²Department of Chemistry and Biochemistry, Hunter College of the City University of New York

³Department of Biochemistry, Weill Cornell Medicine

Hypothesis/Statement of Problem: The potent magnetic behavior of iron oxide nanocages (IO-NCs), a class of nanoparticles, is of particular interest because of the biomedical techniques, such as drug delivery and hyperthermia, that can be developed from the combination of the nanomagnetism of the nanocages with external applied magnetic fields (AMF). Under AMF application, nanocages experience Brownian and Neel relaxation- two types of magnetic motions. As our interest is to apply Brownian motion to deliver siRNA more efficiently to the cytoplasm of cells, we investigated the effect of the size of IO-nanocages to Brownian motion in the magnetic fields and how such magnetic Brownian motion can enhance siRNA delivery in cancer cells. In this study, we hypothesized that the Brownian motion induced by AMF of IO-nanocages bigger than 17 nm can influence siRNA delivery with increased endosomal escape and in consequence, decrease expression of luciferase-expressing B16-F10 melanoma cells.

Methods: To test this hypothesis, we first developed a new synthesis protocol and discovered that varying the total volume of water used during synthesis, as well as the concentration of iron led to the successful synthesis of different sized iron-oxide nanocages without compromising its hollow, cubic shape. The 15 and 20 nm IO-nanocages obtained were then applied to B16-F10 melanoma cancer cells in vitro after culturing, passaging the cells, and incorporating the nanocages with siRNA. AMF was applied under the coil at 335 kHz and the bioluminescence assay was performed to quantify the degree to which luciferase expression was quenched. Transmission electron microscopy (TEM) was also performed to observe the state of the endosomes.

Results: It was found that with the increase of both water volume and iron concentration in the water injection synthesis procedure used, there was an increase in the size of nanocages to a certain extent, decreased hollowness of the nanocage, and increased polydispersity within the samples. It was also observed that siRNA delivery in cancer cells was the greatest when IO-nanocages of 20 nm were utilized. When firefly luciferase siRNAs were delivered by 20 nm IO-nanocages in the presence of AMF, 51% of luciferase expression was quenched. In contrast, when delivered by 15 nm nanocages, only 11% of expression was quenched. Brownian relaxation (induction of rotational motion) was noted to be dominant for IO-NCs in the size range of >17 nm while Néel relaxation (heat generation) became the dominant mechanism for IO-NCs < 17 nm. In TEM images, 15 nm IO-NCs also remained in endosomes as evident by the intact endosomal membranes which failed to rupture even after AMF.

Conclusions: This improved delivery efficiency can be attributed to maximum endosomal escape at this size allowing for increased diffusion of siRNA into the cytoplasm because of the dominating Brownian motion contribution. This, in consequence, enables the 20 nm IO-nanocages to serve as the most effective transfection agent inside cancer cells following endocytosis when compared to the other sized IO-nanocages tested. In conclusion, we believe that this development demonstrates size dependence of Brownian relaxation further supporting that the smallest IO-nanocage size used within the Brownian dominant region would result in the most quenching of luciferase expression indicating maximization of endosomal release. My results help us in better tailoring IO-nanocages for magnetic drug targeting allowing for more efficient delivery of therapeutic agents such as siRNA.

Easel #75 - Kaylin Sevilla Lopez - (1-2pm; 2-3pm)

Utilization of iLastik Pixel Classification Algorithms for Image Analysis in a Mouse Model of Repetitive Mild Traumatic Brain Injury and Anosmia

Kaylin J. Sevilla Lopez,¹ Anthony B. Crum, MS^1,2,3 Julian Meeks, PhD^1,2,3

¹NEUROCITY, City College of New York

²Meeks Lab, University of Rochester Medical Center

³Del Monte Institute for Neuroscience, University of Rochester

Hypothesis/Statement of Problem: Traumatic Brain Injury (TBI) is a form of dysfunction in the brain caused by outside forces such as concussions, direct impact injuries, hemorrhages, etc. Following a TBI, microglial cells and astrocytes are activated in response to neuroinflammation. Mild TBIs are one of the most underdiagnosed injuries, if diagnosed at all, and due to the presence of neuroinflammation may cause serious long-term side effects. One of the most common side effects is anosmia, or loss of smell. This condition has been shown to contribute to both anhedonia and depression. In order to better understand the effects of mTBI on anosmia, an accurate model must be developed.

Methods: Utilizing a modified weight-drop model, we endeavored to induce repetitive mild TBI (rmTBI) in a mouse model and determine if this was sufficient for causing an mTBI with induced anosmia. In order to establish the presence of neuroinflammation following the impacts, animals were sacrificed approximately 2-3 weeks post-TBI in conjunction with social behavior experiments. Immunohistochemical staining and analysis was then performed to determine whether or not neuronal markers of reactive astrocytes and activated microglia were present. Previously dissected and preserved brain samples were sectioned into 25 μm slices via cryostat. Immunofluorescent staining was conducted for GFAP and Iba1. Representative slides were imaged using an Olympus VS110 Slide Scanner with the resultant images analyzed via pixel classification algorithm included in the iLastik predictive software package.

Results: Clearly discernible GFAP and Iba1 staining was found following slide scanning. Additionally, an apparent increase in gliosis and astrocyte activity was shown in those samples which had undergone the rmTBI protocol. Furthermore, the pixel classification algorithm in iLastik was able to successfully predict cell types following user-supervised training.

Conclusions: Our preliminary data suggests the TBI weight-drop model has the capacity of inducing neuroinflammation, allowing the pixel classification algorithm iLastik to accurately label the focused cell types. This joint approach utilizing a modified weight-drop with supervised machine learning image analysis can be a useful and appropriate model moving forward. This model will allow us to further understand the long-term effects of rmTBI and anosmia by providing potential correlations between areas of neural inflammation and behavioral deficits in rmTBI cases.

Easel #76 - Safa Sheik - (1-2pm; 3-4pm)

Changes in Cognitive Processes Across Pregnancy

Safa E. Sheik,¹ Lauren G. Bailes,² Kathryn L. Humphreys²

¹Department of Biological Sciences, Hunter College, New York, NY

²Department of Psychology and Human Development, Vanderbilt University, Peabody College, Nashville, TN

Hypothesis/Statement of Problem: Pregnancy represents a critical period of rapid fetal development, as well as significant developmental changes for the pregnant individual.  Pregnancy brain is a phenomenon when pregnant people report declines in cognitive functions such as memory, processing speed, recall, and general attentiveness. This study aimed to examine the relationship between pregnancy and alterations in three cognitive processes, namely cognitive flexibility, processing speed, and episodic memory. Our driving research question is what is the effect of gestational age on pregnant people's cognitive flexibility, episodic memory, and processing speed? We hypothesized that as gestational age increases, pregnant people's performance in the three cognitive domains will decrease.

Methods:  A total of ten pregnant participants completed cognitive functioning assessments from the NIH Toolbox at multiple time points during their pregnancy. To test our research question, we studied three multi-level models which tested whether task performance differs across participants. This was followed by three models which tested whether gestational age was related to task performance.

Results: Our findings revealed that participants exhibited improvements in performance on episodic memory and processing speed tasks as the pregnancy progressed, while cognitive flexibility displayed a decline throughout the same period. A potential mechanism explaining the observed decrease in cognitive flexibility could be attributed to the well-documented reduction in grey matter that occurs during pregnancy.

Conclusions: These results highlight the dynamic nature of cognitive functioning during pregnancy, suggesting that specific cognitive domains may undergo divergent changes as gestation advances. Additionally, exploring the long-term implications of these cognitive changes could provide crucial information for designing interventions aimed at supporting pregnant individuals and facilitating positive outcomes for both the mother and the developing fetus.

Easel #77 - Natasha Sternberg - (12-1pm)

DNA-PKcs regulates the cGAS-STING signaling axis upon infection with Listeria monocytogenes, an intracellular bacterium

Natasha M. Sternberg¹, Rosalie Morales², Zuley A. Dominguez¹, Teague C. Dilgen¹, Ricardo A. Rodriguez³, Melissa Flores¹, Sara Rejman², & Abigail J. Morales²

¹Department of Human Biology, Hunter College, City University of New York

²Department of Medical Laboratory Sciences, Hunter College, City University of New York

³The City College of New York, City University of New York

Hypothesis/Statement of Problem: The cyclic guanosine monophosphate (GMP)-adenosine monophosphate (AMP) (cGAMP) synthase (cGAS) and STING, the ER localized stimulator of interferon genes, are essential components of innate immune signaling. This intracellular signaling pathway was initially characterized as a crucial host defense against viral infection. However, recent evidence suggests that the pathway is also activated during infection with some types of bacteria, though the differential impact of type I interferon on the development of distinct bacterial infections complicates its importance. In response to some species of bacteria, type I interferon production acts in a protective function, while it has a detrimental effect during infection with others. DNA-PKcs is a critical kinase in the repair of and response to DNA double-stranded breaks in mammalian cells. DNA-PKcs also regulates the cGAS-STING signaling axis in virally-infected cells as demonstrated in several recent reports, proposing that it may have previously unknown roles in regulating cytoplasmic signaling.

Methods: Whole bone marrow was isolated from wildtype (WT) and Scid mice between 4 and 6 weeks of age. Bone marrow-derived macrophages (BMDM) were generated by culturing the bone marrow for 6 days in media conning macrophage colony stimulating factor (M-CSF). Macrophages were activated with live Listeria monocytogenes at a multiplicity of infection of 5 (MOI5). At the indicated time points, cells and cell-free supernatants were isolated and cells were lysed to collect total RNA or total protein. Gene expression was analyzed using quantitative real-time PCR (qRT-PCR) and protein expression was analyzed via immunoblotting or ELISA. All were done in accordance with standard protocols and manufacturer instructions.

Results: During infection with L. monocytogenes in macrophages, DNA-PKcs is required for optimal transcriptional induction of IFN-β and various other well-characterized interferon-stimulated genes (ISGs). Furthermore, relative to wild type controls during infection with L. monocytogenes, the phosphorylation of kinase TBK1 (S727) and activation of IRF3 are substantially diminished in DNA-PKcs^-/- macrophages.

Conclusions: These findings may provide an additional explanation as to why Listeria infected Scid (DNA-PK^-/-) mice have a greater percentage of survival than their wildtype counterparts, as type I interferon is detrimental to effective Listeria clearance in murine infection models.

Easel #78 - Nabiha Subzwari - (2-3pm; 3-4pm)

The Effect of Exosomes on Oligodendrocyte Differentiation

Nabiha Subzwari,¹ RJ Bradley-Ortiz,² Hui Hui Jiang,³ and Carmen Melendez-Vasquez³

¹Macaulay Honors College, Biology Department, CUNY Hunter College, New York, NY

²Biology Department, Hunter College Masters Program, CUNY Hunter College, New York, NY

³Biology Department, CUNY Hunter College, New York, NY

Hypothesis/Statement of Problem: Exosomes are a subset of extracellular vesicles that carry information between cells. It has been shown that mature oligodendrocytes (OL) can use exosomes as a signal that inhibits myelination. We have found that ablation of the non-muscle myosin IIB (NMIIB) gene in OL potentiates their differentiation, myelin formation, and myelin repair. Since inhibition of NMIIB inhibits exosome production by OL, we asked if ablation of NMIIB changes how exosomes regulate these processes in OL.

Methods: Rat oligodendrocyte progenitors (OPC) cultures were given exosomes derived from wild-type and NMII cKO mice that were treated with demyelinating agents (lysolecithin) or inflammatory cytokines (IFN-γ). We will examine how these treatments affect OL differentiation by comparing MBP expression in cultures given exosomes before and after treatments.  

Results: We hypothesize that exosomes from NMIIB cKO mice might have a beneficial effect on OPC differentiation following demyelination.  

Conclusions: This study aims to further characterize how exosomes regulate oligodendrocyte differentiation. The overall goal is to use this information to treat, and even prevent, demyelinating diseases such as Multiple Sclerosis. 

Easel MN17 - Pooja Suganthan - (10-11am; 3-4pm)

Intermittent Theta Burst Stimulation in the Prelimbic Cortex Drives Brain-wide Circuit Reorganization and Rescues Stress-induced Spine Elimination

Pooja Suganthan^1,2, David J. Estrin², Shane B. Johnson², Thaira Ahmed², Christine Kuang², Kenneth Johnson², Conor Liston^2,3.

M¹Macaulay Honors College, Hunter College, City University of New York, New York, NY, USA.

²Feil Family Brain & Mind Research Institute, Weill Cornell Medicine, 413 East 69^th Street, New York, NY, USA.

³Department of Psychiatry, Weill Cornell Medicine, New York, NY, USA.

Hypothesis/Statement of Problem: Stress-induced spine elimination in the prefrontal cortex (PFC) is believed to be a primary driver of depressive symptoms. In parallel to PFC dendritic spine elimination, there is evidence that chronic stress leads PFC circuits to possess desynchronized activity. Transcranial magnetic stimulation (TMS) is a noninvasive stimulation of nerve activity that runs an alternating current through a conductive coil in a patient's scalp to induce an electric field. The objective of this study is to determine whether spine restoration is sustained due to chronic intermittent theta burst stimulation (iTBS), a mode of TMS. We hypothesize that iTBS of the PFC following chronic stress will rescue dendritic spine elimination in the PFC and subsequently drive changes in dendritic spine reorganization throughout the entire brain.

Methods: To test this, we utilized optogenetics to stimulate the PL neurons following 14 consecutive days of corticosterone exposure, a common endocrine model of stress in mice. Mice underwent behavioral tests or were used for Golgi staining, imaging, and spine quantification.

Results: After iTBS treatment, the treated brains demonstrated significantly higher spine density than the control brains. Individual branches of neurons found in Layer 1 treated brains exhibited significantly higher spine density than the branches found in the other layers of treated and control brains.

Conclusions: The rescue of stress-induced spine elimination indicates a potential mechanism for TMS. Next steps include quantifying spine density in downstream brain regions.

Easel #79 - Adelajda Turku - (12-1pm; 2-3pm)

Does ROCK2 inhibition promote remyelination in cuprizone mouse model?

Adelajda Turku,¹ Ace Alcantara,² and Carmen Melendez-Vasquez^,1

¹Department of Biology, Hunter College, The Graduate Center, City University of New York
²Ph.D. Program in Biology, The Graduate Center, City University of New York

Hypothesis/Statement of Problem: We have found that ROCK2 is a negative regulator of oligodendrocyte (OL) differentiation and myelination, and these effects are mediated via phosphorylation on non-muscle myosin II (NMII) (Wang et al. 2012). We have also found that in mice where NMII is ablated, myelin repair is accelerated (Rusielewicz, 2014). The goal of this study is to assess the efficacy of a new generation ROCK2 inhibitor on myelin repair in vivo.

Methods: The cuprizone model mimics some of the pathology found in patients with chronic multiple sclerosis. Mice were fed for 9 weeks with cuprizone chow to induce close-to-chronic demyelinated lesions. Chronic demyelinated (CDL) lesions are induced with 12 weeks of feeding cuprizone chow. The treatment is provided for 3 weeks. 

The distal region of the corpus callosum was stained with fluoromyelin for myelin content, DAPI for nuclei, Olig2 for oligodendrocytes, and CC1 for mature oligodendrocytes. Brain sections were also stained with glial inflammatory markers, the Iba1 for microglia, and GFAP for astrocytes.

Results: No significant difference for the Fluoromyelin content was detected between the chronic demyelinated groups (with and without the treatment). A small but significant decrease in the number of Olig2+ cells was detected in the CDL therapeutic group. No significant difference for CC1+cells was detected between the CDL groups (without or with treatment). No significant difference for Iba+ cells and GFAP+ cells was detected between the CDL groups (without or with treatment). 

Conclusions: Treatment with ROCK inhibitors does not show a significant acceleration of myelin repair, nor does it decrease glial cells activation.

Easel MN18 - Maisha Uddin - (11-12pm; 1-2pm)

Investigating Genetic Markers of Vocal Learning in Tursiops truncatus

Maisha Uddin,¹ Brigid Maloney,² Erich Jarvis ^{2, 3}

¹Department of Biological Sciences, Hunter College, New York, NY

²Lab of Neurogenetics of Language, The Rockefeller University, New York, NY

³Howard Hughes Medical Institute, Chevy Chase, MD

Hypothesis/Statement of Problem: This study asks if the genetic specializations seen in the human and songbird regions are also present in the brain of the bottlenose dolphin (Tursiops truncatus), a distantly related species with the trait, and if it is possible to detect such differential gene expression in sub-optimal tissue samples.

Methods: To determine if bottlenose dolphins exhibit molecular specialization of vocal learning-associated genes, we screened the motor cortex for genes found to be the strongest markers for vocal learning human and songbird brains. Fluorescence in situ hybridization (FISH) probes were generated for Parvalbumin and Slit1, which share convergent expression across the LMC, HVC, & RA. Dolphin cortical samples from stranded specimens were flash-frozen and cryosectioned into 20 micrometer coronal sections and stained for these markers.

Results: We have successfully generated RNA probes from stranded specimens and successfully imaged genes in a cetacean species. Preliminary results indicate that there is differential expression within the cetacean motor cortex.

Conclusions: This study established an RNA imaging protocol for large cetacean brains collected from stranding necropsies. By proving its feasibility, we can expand our work in the future to detect other markers of vocal learning, and use a comparative approach to study expression across all vocal learners. This is a major step in studying the evolution of spoken language, as dolphins share many more characteristics with humans than traditional models. The techniques we established allow for scientists to ask comparative questions across the animal kingdom to better understand evolution and the role of specific genes in the development of unique traits.

Easel #80 - Emine Ustundag - (2-3pm)

Increasing Potassium Channel Function In The Ventral Tegmental Area decreases Behavioral Susceptibility To Stress During Estrus

Emine Ustundag¹, Lauren Renzoni^1,2, Rania Darwish^1,2, Yuka Muira^1,3, Mary R. Shanley^1,3, Allyson K. Friedman^1,3

¹Department of Biological Sciences, Hunter College of the City University of New York;

²Macaulay Honors College, Hunter College, City University of New York;

³Graduate Center of the City University of New York.

Hypothesis/Statement of Problem: The estrous cycle plays a crucial role in modulating neuronal physiology and behavior. We hypothesized that increasing potassium channel activity in the VTA could reduce the effect of estrus on dopamine neuron excitability and the stress response. We assessed social interaction behavior following acute variable social stress.

Methods: We used whole-cell slice electrophysiology of VTA DA neurons in naturally cycling, adult female C57BL/6J mice to characterize the effects of the estrous cycle and the role of 17β-estradiol on intrinsic neuronal activity. Social interaction behavior was assessed following a series of acute variable social stressors performed during direct pharmacological manipulation of estrogen receptors in the VTA.

Results: We found that pharmacologically increasing potassium channel activity in the VTA before stress reverses stress susceptibility found during estrus as assessed by social interaction behavior. With the onset of puberty and fluctuating sex steroids, sex differences in the prevalence of DA-related disorders emerge.  Specifically, mood disorders like major depressive disorder (MDD) are diagnosed twice as frequently in women starting in adolescence. MDD is marked biologically by neuronal adaptations in the mesolimbic circuit, and behaviorally by disruptions in social behavior. Despite the sex difference in the prevalence of MDD, rodent models of depressive behaviors have historically used male subjects to determine the neuronal substrates underlying stress susceptibility and depression. These studies show that alterations in the firing patterns of DA neurons in the VTA occur after chronic stress through changes in intrinsic excitability and potassium (K+ ) channel activity.

Conclusions: These adaptations are crucial in regulating a healthy stress response and determining a behavioral response to stress.

Easel #81 - Angelica Vega - (2-3pm; 3-4pm)

Pointing Out Differences? Comparing Human-Given Pointing Cue Following in Pet and Guide Dogs

Angelica Cristina Vega¹, Angie Lee¹, Madison Murray¹, Liza Rothkoff¹, Gillian Schmitt¹, Sarah-Elizabeth Byosiere¹

¹Thinking Dog Center, Department of Psychology, Hunter College, City University of New York, United States

Hypothesis/Statement of Problem: Historically, dogs, compared to other non-human animals, have demonstrated a unique ability to follow human-given communicative cues. Like human infants, dogs have been found to attune to human-given ostensive cues, communication motivated by an informative intention, and ignore similar but non-ostensive cues. However, little is known about whether dogs interpret pointing as a social communicative gesture or as an associative cue, as well as whether prior training experience influences the ability to follow these cues.

Methods: Two populations of dogs were assessed, including 16 pet dogs at the Thinking Dog Center (NY, NY) and 23 guide dogs at Leader Dog for the Blind (Rochester, MI). Using a two-way object choice task, we assessed dogs' ability to follow non-ostensive and ostensive contralateral momentary pointing cues to find hidden food rewards.

Results: No significant differences between the ostensive and non-ostensive conditions were observed within pet dogs (N=16, p = 0.657) or guide dogs (N=23, p = 0.251). Overall, pet dog and guide dog performance was at chance (~50%) across both conditions, indicating no significant differences between the two populations.

Conclusions: These results highlight the importance of the type of pointing cue used and body language in human-dog interspecies communication and emphasize that pointing cue following may not be influenced by training experience.

Easel #82 - Anna Vera - (9-10am; 10-11am)

Molecular Basis of Recognition Between Yeast Spliceosomal U2-U6 snRNA Complex and Pre-mRNA-Splicing Factor Cwc2

Anna Vera¹, William Perea¹, Nathan Astrof¹, and Nancy L. Greenbaum¹

¹Department of Chemistry, Hunter College, The City University of New York, NY NY 10065

Hypothesis/Statement of Problem: The spliceosome is a large nucleolytic ribozyme (enzyme composed of RNA) whose role is to remove non-coding regions (introns) from precursor (pre)mRNA transcripts and ligate together coding regions (exons). This RNA-protein assembly uses five small nuclear RNA (snRNAs) that associate with spliceosomal proteins to perform pre-mRNA splicing. Of these snRNAs, the U2 and U6 snRNA are the most highly conserved, and that the only two that form a complex to comprise the catalytic center of the spliceosome. In yeast spliceosomes, the protein Cwc2 is proposed to have a role in stabilizing the catalytic center of the U2-U6 complex to enable it to adopt its catalytic conformation. We seek to discover the molecular basis of recognition between the Cwc2 protein and the yeast U2-U6 snRNA complex. We seek to determine whether the C-terminal region of Cwc2 is essential for its structural role. Towards this goal, we are making a mutant version of the Cwc2 protein in which the C-terminal region has been deleted.

Methods: We are conducting an assay to measure the binding affinity of the U2-U6 snRNA complex to a native Cwc2 protein, and plan to compare this to an assay of a Cwc2 protein whose C-terminus has been removed.

Results: To date, the RNA strands representing the yeast U2 and U6 snRNAs have been transcribed and purified, and the wild-type sequence of the yeast protein Cwc2 has been expressed and purified. There is some evidence of pairing of the two RNA strands, as well as evidence that the Cwc2 protein interacts with the U2 and U6 snRNA as determined by non-denaturing polyacrylamide gel electrophoresis.

Conclusions: This study aims to add to the existing body of literature about the molecular basis of recognition between the spliceosome and the C-terminal region of pre-mRNA-splicing factor Cwc2. This information will contribute to our understanding of how proteins help facilitate formation of the RNA catalytic core of the spliceosome.

Easel MN19 - Casey Walsh - (9-10am; 1-2pm)

Thermal and Concentration Effects on ¹H NMR Relaxation of Gd³⁺ -aqua using MD Simulations and Measurements

Casey Walsh¹, Thiago J. Pinheiro dos Santos²,  Arjun Valiya Parambathu², Carla C. Fraenza², Steve G. Greenbaum¹, Walter G. Chapman², Dilip Asthagiri^1,3,  and Philip M. Singer¹

¹ Department of Physics & Astronomy, Hunter College of the City University of New

York, New York, NY 10065, USA

² Department of Chemical and Biomolecular Engineering, Rice University, Houston,

TX 77005, USA. E-mail: ps41@rice.edu

³ Oak Ridge National Laboratory, Oak Ridge, TN 37830-6012, USA.

Hypothesis/Statement of Problem: MRI is a method of medical imaging that employs the use of a large, constant magnetic field. MRI contrast agents are used to create sharper images, by shortening T₁ and T₂  relaxation times of protons, thus creating better contrast in these images. Gadolinium is one of the more popular elements used in contrast agents, because of the highly paramagnetic nature of the Gd(3+) ion. However it is toxic and must be chelated in order to make it safe to inject into patients. Unfortunately, not much is understood about how these molecules behave once inside the body, and the quest for new and safer contrast agents requires a firmer understanding of NMR relaxation dynamics of Gd(3+) in water and various "bio-solvents".

Methods: We studied the molecular dynamics of GdCl3 in water in varying temperatures and concentrations, by means of molecular dynamics simulation as well as experimental measurements using NMR techniques, including fast field-cycling NMR relaxometry.

Results: It was found that the dynamics of the water closely surrounding gadolinium ions are highly sensitive to temperature changes, the experimental relaxivity results have validated the molecular dynamics simulations at human body temperature and concentrations used in clinical MRI.

Conclusion: From our results, we can better understand the mechanisms behind contrast agents. Further research is needed to simulate conditions more similar to the human body to improve our model. This can lead to the design of less-toxic contrast agents.

Easel #83 - Zixian Wang - (2-3pm; 3-4pm)

Intracellular localization of YAP/TAZ during oligodendrocyte development in vitro: implications for myelin formation and repair

Zixian Wang¹, Abdul Rahmani¹,Ace Alcantara^1,2, Aysha Tabassum¹, Carmen V. Melendez-Vasquez^1,2

1. Department of Biological Sciences, Hunter College, City University of New York, New York

2. Molecular Cell and Developmental Biology, Biochemistry and Behavioral and Cognitive Neuroscience, The Graduate Center, City University of New York

Research in mechanically-driven differentiation of mesenchymal stem cells (MSC)  have shown that the subcellular localization of YAP/TAZ changes in response to matrix stiffness. Thus, prolonged exposure of MSC to either soft or stiff matrices, leads to the acquisition of a mechanical memory. YAP/TAZ subcellular localization in MSC is correlated with this epigenetic memory and their final cell fate. Specifically in stiffer substrates YAP/TAZ is retained in the nucleus and cells become osteocytes, while YAP/TAZ is translocated out of the nucleus in softer substrates and MSCs become adipocytes.

To investigate if oligodendrocyte progenitor cells (OPC) acquire a mechanical memory in response to changes in ECM stiffness, we examined the subcellular localization of YAP/TAZ in primary cultures of rodent and iPSC-derived human OPC, in conditions that mimic healthy or chronically-demyelinated brains. Initial data obtained from myelinating co-cultures of rat OPC and DRG neurons indicate a negative correlation between MBP expression and YAP nuclear localization. At high stiffness conditions , there is less MBP expression and YAP is primarily  localized in the nucleus of Olig1+ cells. By contrast, Olig1+ cells grown in soft hydrogels, mimicking a healthy brain , exhibit higher MBP expression and cytoplasm localization of YAP. Collectively, our findings suggest that nuclear retention of YAP by OPC in stiffer matrices can contribute to remyelination failure in chronically demyelinated lesions.

Easel #84 - Andrew Wenger - (10-11am; 3-4pm)

Colon Cancer Mouse Models using Organoid Implantation 

Andrew Wenger¹, Sujen Rashid², Alison Juray¹, Maria Paz Zafra PhD³, Erika Hissong MD⁴, Marie Parsons PhD², Lukas Dow PhD^2,5, Despina Siolas MD, PhD^2,5 

¹ Hunter College, City University of New York, New York, NY, USA

² Department of Medicine, Division of Hematology and Oncology, Weill Cornell Medicine, New York, NY, USA

³ Universidad Granada, Granada, Spain

⁴ Department of Pathology, Weill Cornell Medicine, New York, NY, USA

⁵ Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA

Hypothesis/Statement of Problem: Colorectal cancer is the 2nd most common cause of cancer death in the United States. For unclear reasons, patient prognosis is notably affected by the anatomical site of tumor origin, with more aggressive tumors originating in the right side of the colon compared to the left side. KRAS is one of the most frequently involved genes in colon cancer and has been observed to be mutated more often in tumors originating in the right-side colon. To further understand the disparity between left-side and right-side localized colorectal cancer, we have developed a preclinical model to study tumorigenesis in specific parts of the colon. 

Methods: Mouse-derived Kras mutated colon cancer organoids (500,000 cells) were cultured in-vitro and injected into either the proximal colon or distal colon of a mouse. Tumors were dissected at four weeks post-surgical implantation, where they were subsequently weighed, fixated with formalin, preserved in ethanol, and paraffin embedded. Hematoxylin and eosin-stained tumor sections were examined histologically by a colon cancer pathologist. 

Results: Tumors were successfully formed in the sigmoid colon and cecum 4 weeks after implantation with high reproducibility. Immune cell infiltration may contribute to tumor development depending on the primary tumor's site of origin, so additional sections were stained via immunofluorescence for specific immunological components of the tumor microenvironment such as CD3+ T cells, CD8+ cytotoxic T cells and Foxp3+ T regulatory cells. Ongoing studies will examine overall survival for mice with tumors of different anatomical origin as well as a comprehensive analysis of potential immune cell interactions.

Conclusions: Here we have described a novel preclinical model for studying tumorigenesis originating in either the cecum or distal colon of a mouse. The results have demonstrated that tumor formation can be achieved with a quick turnaround rate and high reproducibility. This mouse model can be used to study multiple components of colon cancer such as the effects of tumor anatomical location, immunological interactions, mutation differences, drug reactions and much more.

Easel #85 - Yusef Wray - (2-3pm; 3-4pm)

Stability of Nipah Virus Attachment Glycoprotein and Glycan-Selective Binding by Receptors

Yusef Wray,^1,2 Mateusz Marianski^1,3,4,5

¹Department of Chemistry and Biochemistry, Hunter College, The City University of New York, 695 Park Ave, New York, NY 10065, USA

²Department of Chemistry and Biochemistry, Macaulay Honors College, The City University of New York, 35 W 67th St, New York, NY 10023, USA

³The Phd Program in Biochemistry, The Graduate Center of The City University of New York, 365 5^th Ave, New York, NY, 10016, USA

⁴The PhD Program in Chemistry, The Graduate Center of The City University of New York, 365 5^th Ave, New York, NY, 10016, USA

⁵Advanced Science Research Center of The City University of New York, 85 Nicolas Terrace, New York, NY, 10031, USA

Hypothesis/Statement of Problem: N-glycans are essential to enveloped viruses' capability to infect, reproduce and evade immuno-response. The unexpected emergence of viruses with potential for interspecies transmission - for instance, the Nipah and SARS CoV-2 viruses - has highlighted the need to develop broad-spectrum anti-virals. These anti-virals could take advantage of some universal viral mechanism, such as viruses' use of glycans to mediate infection.

Methods: Using GROMACS, several glycosylated models of the Nipah attachment glycoprotein were computationally prepared. Resultant interactions of the glycoprotein with synthetic carbohydrate receptors will be analyzed.

Results: Since synthetic carbohydrate receptors (SCRs) bind to the glycan portion of the virus, they could prevent interaction between the viral glycans and the host cell receptors. Viral infection is dependent upon the docking of the virus to cell receptors, meaning that the binding of SCRs to viral glycans could disrupt the first portion of the viral life cycle.

Conclusions: This study suggests that the viral N-glycans interactions with synthetic carbohydrate receptors can be used to exploit the viral glycan binding mechanism.