Characterizing E-Prostanoid Receptor G-Protein Binding and Downstream Signaling

Speaker: Daniel Moskoits

Co-Authors: Kelly Culhane, P

Abstract: Prostaglandin E2 (PGE2) is a paracrine/autocrine signaling molecule responsible for a broad range of crucial physiological functions including temperature regulation, bone remodeling, and immune support. PGE2 molecules bind and activate four distinct E-prostanoid receptors, which are all G-protein coupled receptors (GPCRs). Primary and secondary G-protein interactions determine the complex, context-dependent nature of the physiological role of these receptors. However, the characterization of the EP receptors and their secondary G-protein interactions remains incomplete. Therefore, we transfected recombinant EP receptors 1 and 2 with NanoBiT sensors into HEK293T cells and induced giant plasma membrane vesiculation to provide mechanistic insights into G-protein binding. To probe downstream signaling through the Gs pathway, we performed a luminescence-based cAMP assay. When HEK293T cells are initially stimulated with forskolin, the cAMP assay can be repurposed to measure a decrease in cAMP production in response to Gi pathway activation. An ELISA was used to elucidate levels of IP1, a key member of the Gq pathway. We find that EP1 interacts with and activates signaling downstream of Gs and Gq, while EP2 binds and signals only through Gs. Preliminary data indicate that EP3 may signal through the Gs pathway in addition to the Gi signaling observed in previous studies. A deeper mechanistic understanding of E-prostanoid receptor activation and downstream signaling could meaningfully contribute to the rational design of targeted therapeutics.

F-actin-dependent activation of NADPH oxidase 5 by the vacuolar-type ATPase

Speaker: Gwen Heywood

Co-Authors: Samantha Richter, Archi Sundar Paul, Elizabeth Sweeny

Abstract: NADPH oxidase 5 (NOX5) is a transmembrane oxidative signaling enzyme that produces superoxide in response to increases in intracellular calcium. NOX5 has been shown to play essential roles in processes such as specialized cell differentiation, proliferation, and cell contraction, while its overexpression and hyperactivation are associated with human diseases, including cardiovascular disorders, cancer, and diabetic kidney disease. Despite its importance in human health and disease, the regulatory factors and mechanistic signaling details of NOX5 remain incompletely resolved. We previously reported that chemical stabilization of F-actin by jasplakinolide induces a superoxide burst by NOX5 independent of changes in intracellular calcium. Here, we show that this F-actin-dependent activation of NOX5 superoxide production relies on the vacuolar-type ATPase (v-ATPase). In cells, jasplakinolide treatment stabilizes a complex comprising actin, actin-binding proteins, the v-ATPase, and NOX5. Chemical inhibition of the v-ATPase or v-ATPase subunit knockdown decreases NOX5 superoxide production in response to jasplakinolide while only minimally affecting NOX5 superoxide production in response to increases in intracellular calcium or NOX5 phosphorylation. Finally, in untreated cells, the v-ATPase and NOX5 localize to the cell leading edge and contribute to cell migration. Our findings represent a novel mechanism of NOX5-mediated superoxide production by the v-ATPase and provide a new foundation for developing strategies to disrupt protein interactions and activities that drive aberrant cell migration.

Discovery of an Enzyme Capable of Utilizing RNA as an Energy Carrier

Speaker: Zoe Wardle

Co-Authors: David N. Frick

Abstract: For generations, nucleic acids (DNA and RNA) have been thought to be simple information carriers that store and transmit the code of life. However, RNA World theory posits that early life relied on RNA to perform dual roles: storing genetic code and driving essential life reactions. Metabolism requires electron-carrying molecules, including metabolic coenzymes like nicotinamide adenine dinucleotide (NAD+), which can also act as initiating nucleotides during transcription, yielding coenzyme-capped RNAs. While NAD-capped RNAs exist across all domains of life, their functional roles beyond degradation remain largely unexplored, leaving open whether RNA can participate directly in redox reactions as a metabolic coenzyme.

We report the first identification of a dehydrogenase enzyme capable of utilizing intact NAD-capped RNA as a functional coenzyme. Screening candidate redox enzymes, AMP-transferases, and ADP-ribosyltransferases, we discovered that an alcohol dehydrogenase from a thermophilic archaeon reduces NAD-capped RNA to NADH-capped RNA in vitro. The wild-type enzyme showed robust activity with both free NAD+ and NAD-capped RNA, while a more promiscuous variant strongly preferred NAD-capped RNA. We used enzymatic characterization, including turnover rate, RNA sequence dependence, and transient and steady-state kinetics, to probe the catalytic mechanisms governing this reaction. To test if this capability extends beyond archaea, we expressed and purified E. coli homologs to assess conserved NAD-capped RNA utilization.

By demonstrating that an enzyme can utilize NAD-capped RNA to drive redox reactions, this work provides concrete evidence that RNA can function directly as an energy carrier, uniting information storage with metabolic activity in support of RNA World theory.

Conserved Active-Site Residues Redirect Early Redox Events in Cytochrome c Nitrite Reductase Under Weakly Reducing Conditions

Speaker: Stephen Nkwocha

Co-Authors: Bradley John Dimock, Oke Temidayo Joseph, Andy Pacheco, Jarett Wilcoxen

Abstract: Cytochrome c nitrite reductase (ccNiR) is a homodimeric, periplasmic enzyme with five c-type hemes per protomer. It catalyzes the six-electron reduction of nitrite to ammonium, a key step in the biological nitrogen cycle. Under strongly reducing conditions, catalysis proceeds to ammonium too quickly for intermediates to accumulate, so the steps beyond the two-electron-reduced {Fe_H1NO}⁷ species remain poorly defined. To address this, we reacted nitrite-loaded wild-type ccNiR and three active-site variants (R103Q, Y206F, and H257Q) with hexaammineruthenium(II) in the presence of indigo trisulfonate (I3S). The I3S oxidized-to-reduced ratio reports the solution potential, which we poised between approximately −55 and −92 mV vs. SHE. We monitored the reactions by stopped-flow UV–visible spectroscopy and resolved the spectral components by numerical fitting. Wild-type ccNiR showed three kinetic phases (k = 139, 31, and 0.72 s⁻¹) and substantial bis-His heme reduction, with features near 552 and 573 nm. Each variant followed a distinct early path. R103Q favored a six-coordinate {FeNO}⁷ intermediate. Y206F retained evidence of early bis-His heme reduction. H257Q showed little early heme reduction. Despite these differences, all three variants converged at longer times on the five-coordinate {FeNO}⁷ species. These results show that the conserved Arg103, Tyr206, and His257 residues control early electron- and proton-transfer events during nitrite reduction. Ongoing EPR studies aim to trap and characterize these short-lived intermediates.

Rigorous Absolute Binding Free Energy Calculations Elucidate Mutation–Driven Resistance Mechanisms of Non–Covalent Bruton’s Tyrosine Kinase (BTK) Inhibitors

Speaker: Justice J Mallen

Co-Authors: Shilpa Sharma, Md Nazmul Hasan, and Arjun Saha

Abstract: Drug resistance driven by acquired mutations in Bruton’s tyrosine kinase (BTK) threatens the long–term clinical efficacy of non–covalent BTK inhibitors (NCIs). While clinically observed variants such as A428D, C481S, and the double mutant C481S/T474I display distinct resistance profiles, their differential impact on inhibitor recognition remains poorly characterized at the atomic level. In this study, we employed rigorous alchemical absolute binding free energy calculations to quantify the thermodynamic consequences of these mutations across five chemically diverse NCIs. Benchmarking against wild–type experimental binding free energy values demonstrates that the Binding Free Energy Estimator 2 (BFEE2) framework yields highly accurate and robust predictions, significantly outperforming end–point methods like MM–PBSA. Subsequent structural and interaction fingerprint analyses reveal a distinct hierarchy of resistance mechanisms. The A428D mutation triggers widespread electrostatic and hydrophobic disruption near the hinge region, driving severe, broad–spectrum affinity loss. In contrast, the C481S substitution largely preserves the active site architecture, allowing NCIs to maintain full native potency. Finally, the C481S/T474I double mutant introduces localized steric clashes at the gatekeeper position, resulting in moderate, inhibitor–specific binding penalties. Together, these findings establish a robust thermodynamic framework for predicting mutation–induced resistance and offer actionable structural principles for the rational design of next–generation, mutation–resilient NCIs.

Trapping Nitric Oxide Intermediate of Variant Cytochrome c Nitrite Reductase

Speaker: Bradley Dimock

Co-Authors: A. Pacheco, B. Bennett, J. Wilcoxen, none are co-presenting.

Abstract: Cytochrome c nitrite reductase (ccNiR) is a multi-heme metalloenzyme responsible for the reduction of nitrite into ammonium as part of the nitrogen cycle. Three amino acids (R103, Y206, and H257) surround the active site heme where reduction takes place, all of which are conserved in ccNiR homologues from different bacteria, have been shown to be important to enzymatic function. Variant ccNiR was developed and tested, creating an unknown intermediate during reduction which required characterization.

Three different variant proteins were created for isolated analysis, each replacing one of the three known amino acids (R103Q, Y206F, and H257Q) which were then grown and purified. Changes in enzyme absorbance were monitored over the course of 90 minutes using time-resolved UV/Vis spectrophotometry when exposed to nitrite and the weak reductant, reduced indigo tetrasulfonate (I4Sred). Under extended anerobic reduction, electron paramagnetic resonance confirmed the existence of nitric oxide binding to the active site iron. This nitrosyl product resulted in a 5-coordinated iron-heme structure, confirming the removal of the covalently bound lysine that typically binds to the heme.

The creation of a 5-coordinated nitrosyl product on variant ccNiR and the loss of its covalently bound lysine confirms the need for the amino acids in the active site pocket to successfully orient the nitrite and its intermediates during reduction. This shows the impact of a protein’s primary and tertiary structures and their role in providing a suitable environment for enzymatic activity.

Development of Novel Selective 5-HT2B Receptor Antagonists

Speaker: Maya Fernando

Co-Authors: Shaun Harrington (attending), Paul Wiebe, Marley Makowiak (attending), Devin Esch (attending), Mubaraq Toriola (attending), Braden Barkley and Leggy A. Arnold (attending)

Abstract: A palladium catalyzed Buchwald-Hartwig synthesis was used to generate new amine substituted benzodiazepines for the development of novel serotonin receptor antagonists. An optimized method has been developed using 5% Pd2(dba)3 in conjunction with 10% Xphos as the catalytic system. This method proves to be successful with substituted anilines, aliphatic amines and heterocyclic amines with yields ranging from 23% to 89%. In collaboration with the Psychoactive Drug Screening Program, it was determined that some of these compounds are binding selectively the serotonin 2B receptor (5HT2B). The receptor is part of a larger serotonin receptor family containing multiple subtypes with almost identical structures, complicating the development of subtype selective ligands. Developing highly selective ligands is important because the serotonin receptor subtypes are expressed tissue specifically, mediating specific physiological processes. Our most promising compound has a Ki of 62 nM for 5HT2B with 15-fold selectivity over 5HT2C and does not bind 5HT2A. Functional cell-based assays identified this compound as antagonist enabling the development of new treatments for cardiopulmonary diseases, including pulmonary arterial hypertension (PAH) and valvular heart disease (VAD). We present the synthesis and preclinical characterization of a large number of related compounds with a clear structure-activity relationship. In addition, computational approaches aid our understanding of subtype selectivity. Overall, our findings confirm the adaptability of the benzodiazepine scaffold for developing new GPCR ligands without any affinity to the GABA(A) receptors.

Synthesis of Dinucleoside Polyphosphate Analogs Using Recombinant Lysyl-tRNA Synthetase (LysU)

Speaker: Mujidat Shittu

Co-Authors: David N. Frick

Abstract: Dinucleoside polyphosphates are a family of signaling molecules made of two nucleosides joined by a variable number of phosphate groups. The most common one is diadenosine tetraphosphate (Ap4A), which accumulates during cellular stress or oxidative damage. Aminoacyl-tRNA synthetases (AARSs), which typically charge tRNA for translation, synthesize Ap4A upon DNA damage, possibly due to insufficient tRNA levels. Past research has shown that Lysyl-tRNA Synthetase (LysU) is the AARS that contributes to most Ap4A synthesis in E. coli. The aim of this project is to determine whether LysU can be used to synthesize novel Ap4A analogs. We therefore cloned the E. coli LysU gene into the pET28 vector, fusing the protein to an N-terminal 6X-Histidine tag. We purified the protein using Ni-NTA and gel filtration chromatography, then concentrated it. We evaluated His-LysU’s ability to adenylate lysine by measuring pyrophosphate release. First, after nucleotide removal using activated charcoal, we used ATP sulfurylase, adenosine 5’-phosphosulfate (APS), and luciferase to quantify pyrophosphate. Second, pyrophosphatase was added, allowing the colorimetric quantification of liberated inorganic phosphate using malachite green and ammonium molybdate. Adding pyrophosphatase also enhanced LysU-catalyzed Ap4A or Ap3A synthesis, enabling potential synthesis of a variety of dinucleotide analogs (e.g., RXN 4). The identity and purity of the synthesized dinucleotides were verified using 31P NMR spectroscopy and mass spectrometry. They were also used in assays of enzymes that hydrolyze Ap4A, like the human NUDT2 enzyme. Ultimately, the dinucleoside polyphosphate analogs should be useful as mechanistic probes of Ap4A signaling pathways.

TBD

Speaker: Anna Bauer

Co-Authors:

Abstract:

TBD

Speaker: Alex Eng

Co-Authors:

Abstract:

TBD

Speaker: Juliannie Herrera

Co-Authors:

Abstract:

Phrenic nerve stimulation increases genioglossal output

Speaker: Felix Sanchez

Co-Authors: Ms. Elizabeth Rozycki, Dr. Dave D. Fuller, Dr. Kristi Streeter

Abstract: Obstructive sleep apnea (OSA) is characterized by repeated episodes of partial or complete obstruction of the upper airway during sleep, leading to periodic cessation of breathing. Poor adherence to current therapies, including continuous positive airway pressure, highlights the need for alternative treatment strategies. Recent evidence suggests that phrenic nerve stimulation may reduce upper airway collapse during drug-induced sleep endoscopy in individuals with OSA. Here, we tested the hypothesis that phrenic nerve stimulation increases genioglossus electromyography (EMG) activity. Experiments were performed in anesthetized, spontaneously breathing male (n = 8) and female (n = 5) rats. Bilateral genioglossus EMG activity was recorded while the left phrenic nerve was stimulated at increasing currents (15, 160, and 300 μA) during inspiration, pre-inspiration, and expiration for either one breath or five consecutive breaths. Stimulation at 160 and 300 μA delivered during inspiration or pre-inspiration for a single breath significantly increased ipsilateral genioglossus EMG amplitude compared with baseline. During five-breath stimulation, inspiratory stimulation similarly increased EMG amplitude relative to baseline and expiratory stimulation, whereas pre-inspiratory stimulation produced a significant increase only at 300 μA. While phrenic nerve stimulation is currently used in several patient populations to contract the diaphragm muscle and sustain breathing, these data suggest phrenic nerve stimulation has potential as a novel therapeutic option of maintaining upper airway patency during OSA.

Chemotherapy-induced Kupffer cell instability establishes a pro-metastatic liver microenvironment by initiating a pro-damage feedback loop

Speaker: Isabella Facchine

Co-Authors: M. Budithi, BS; O. Cortez-Toledo, MS; P. Jayakrishnan, PhD; A. Bauer, BS; K. Liptow, BA; N.K. Lytle, PhD

Abstract: Most patients with pancreatic cancer will succumb with significant metastasis. While early-stage patients receive neoadjuvant chemotherapy to debulk disease, improve surgical margins, and prevent futile surgeries, the majority will recur with distant metastasis. This invites the question: how does chemotherapy modulate the metastatic microenvironment?

Our studies have demonstrated that treatment with a standard-of-care combination chemotherapy, FOLFIRINOX, generates a pro-metastatic niche in murine models, evidenced by accelerated liver metastasis. Single-cell RNA sequencing analysis and immunophenotyping of livers following FOLFIRINOX revealed a profound depletion in Kupffer cells, a resident macrophage population critical for early metastatic control, complemented with an expansion of pro-tumorigenic Trem2+Spp1+ metastasis-associated macrophages. Lineage tracing studies demonstrated that FOLFIRINOX leads to either Kupffer cell death by ferroptosis or loss of Kupffer cell identity in remaining populations. Loss of Kupffer cell identity results in a pro-inflammatory neutrophil-like state enriched in S100A8/A9 expression that may perpetuate initial FOLFIRINOX-induced liver damage. Furthermore, resident Kupffer cells that persist following FOLFIRINOX treatment have diminished capacity to clear liver damage, evidenced by their loss of apoptotic clearance marker, Tim4. Finally, while total Kupffer cell number is restored to pre-injury levels following a four-week recovery period from FOLFIRINOX, Tim4 expression remains significantly depleted, indicative of enduring chemotherapy-induced Kupffer cell population shifts.

These data demonstrate that FOLFIRINOX leads to accelerated liver metastasis progression by driving Kupffer cell death and loss of Kupffer cell identity in remaining populations, with both contributing to the accumulation of damage that establishes a pro-metastatic microenvironment.

Safety and efficacy of transfusional iron overload screening and treatment in children and young adults following anti-neoplastic therapy or hematopoietic stem cell transplantation

Speaker: Rhea Singh

Co-Authors: Melissa Acquazzino, Michelle Brenner, Mahua Dasgupta, Rachel Phelan, Nathan Schloemer, Debra Schmidt, Ke Yan, Allison Remiker.

Abstract: Repeated blood transfusions can cause transfusional iron overload (TIO), in which excess iron accumulates in organs and can cause damage. Children and young adults receiving cancer treatment or hematopoietic cell transplantation (HCT) often need transfusions, but standardized approaches to identify and treat TIO in these groups are limited.

We retrospectively reviewed medical records for patients aged 0-25 years with cancer or HCT who developed TIO at a single pediatric center between 2012 and 2024. Patients were screened using a standardized institutional algorithm based on their diagnosis and timing of treatment completion or transfusion independence. We collected information on transfusion history, laboratory values, liver and cardiac magnetic resonance imaging (MRI), treatment, and outcomes.

The study included 157 patients with a mean age of 10 years; 44% were female. Diagnoses included blood cancers (54%), central nervous system malignancies (10%), solid tumors (30%), and bone marrow failure syndromes (6%). Of 63 treated patients, 59 received therapeutic phlebotomy, a procedure that removes blood to lower body iron stores. Patients completed a median of eight phlebotomy sessions over 0.6 years. Complications were uncommon (6.8%) and included lightheadedness, dizziness, and soreness.

Following treatment, liver iron concentration decreased by a median of 3.4 mg/g dry weight, and ferritin, a blood marker of iron stores, decreased by a median of 720 ng/mL. Greater transfusion exposure was associated with more phlebotomy sessions.

Standardized screening identified patients with TIO, and therapeutic phlebotomy was associated with meaningful iron reduction and few adverse events.

Pulmonary vagal sensory neurons as regulators of ventilator-induced lung injury

Speaker: Taylor Holmes

Co-Authors: Nicole Hennen, Luke Ehlert, Brooke Dir, Doriann Pina, Gary C. Mouradian

Abstract: Mechanical ventilation (MV) is a life-sustaining treatment that can also cause ventilator-induced lung injury (VILI). Despite global adoption of lung-protective ventilation strategies, VILI persists in nearly 25% of ventilated patients, suggesting that biological mechanisms beyond ventilator settings contribute to VILI susceptibility. Pulmonary vagal sensory neurons (PSNs) are positioned to detect the abnormal mechanical and inflammatory environment created during MV. We hypothesized that PSNs regulate VILI across physiological, structural, and inflammatory domains. To test this, we first established a VILI model in wild-type (WT) mice (n=14), characterized by physiological, structural, and inflammatory pulmonary injury, and subsequently used this model to determine whether broad PSN manipulation mitigates VILI development. Vglut2Cre+ mice (n=12) were intratracheally treated (postnatal day 7-10) with a Cre-dependent retrograde AAV expressing excitatory (Gq) or inhibitory (Gi) synthetic receptors. Mice (8–10weeks) were then exposed to VILI with simultaneous activation of Gq or Gi receptors via intraperitoneal injection of deschlorozlozapine (DCZ). PSN manipulation preserved inspiratory capacity (Gq: p=0.0410; Gi: p=0.0109) and dampened changes in tissue resistance (Gq: p=0.0092; Gi: p=0.0042) compared to WT controls. Despite similar structural disruption (p=0.8446), IL-6 in Gq (p=0.0285) and TNFα in Gi (p=0.0410) were elevated in lavage fluid compared to WT controls. These findings support our hypothesis that PSNs regulate VILI and reveal domain-specific effects, with PSN manipulation preserving pulmonary physiology while enhancing inflammatory signaling following injury-inducing MV. Future studies will define PSN subtype-specific mechanisms underlying these divergent responses and determine their contribution to VILI susceptibility and recovery.

Impact of a Single Day of Reduced Activity on the Systemic Metabolic and Proteomic Exercise Response

Speaker: Brandon Donahue

Co-Authors: Gries, K.; Grimm, J.; Elmhorst, A.; Parve, J.; Goetz-Sutinen, E. None of them will be co-presenting.

Abstract: Physical inactivity is linked to many cardiovascular risk factors and appears to minimize the benefits of aerobic exercise (AE). Our laboratory has recently shown that one day of reduced activity (<5,000 steps, IN) in young adults increased triglycerides, changed the metabolic demands of AE, and altered the skeletal muscle transcriptomics at rest and after exercise compared to normal activity (>8,500 steps, A). Using a similar approach, we compared the systemic metabolic (respiratory gas analysis, plasma lipidomics, and plasma untargeted metabolomics) and plasma proteomics response to an acute bout of exercise (65% VO2max on cycle ergometer for 60 min) after a day of RA (4574±378 steps) or NA (11274±2985 steps) in young adults (n=6, 3M/3F; 23±2y; BMI 27.7±3.7; VO2max=3.3±1.0 L O2/min) in a cross-over design study. Respiratory gases were analyzed at minutes 20, 40, and 60 of exercise to measure carbohydrate and fat oxidation, and venous blood draws were taken before and immediately after exercise. After centrifugation, plasma was analyzed for quantitative untargeted proteomics, untargeted metabolomics, and lipidomics. While fat oxidation increased and carbohydrate oxidation decreased throughout exercise (p<0.05), carbohydrate utilization trended higher at 20-min in IN vs. A (p=0.08). 14 metabolites (14 up), 8 lipids (1 up, 7 down), and 14 proteins (8 up, 6 down) differed pre vs. post AE. Inactivity resulted in two metabolites (phenol, up; trigonelline, down), two lipids (PC(16:0_18:3)+OH, up; PS(18:0_18:1), down), and no protein changes. Collectively, these data further illustrate that a single day of reduced activity causes modest alterations in the exercise response.

Growth Outcomes and Limb-Length Discrepancy After Knee Hemiepiphysiodesis: A Retrospective Cohort Study

Speaker: Christian J. Chang

Co-Authors: Noah Knezic, Jordan G. Wang, Adam Thiessen

Abstract: This study examined whether hemiepiphysiodesis for correction of knee valgus or varus is correlated with an increased risk of developing a clinically significant LLD, defined as greater than 20 mm. We conducted a single-institution retrospective cohort study of pediatric patients undergoing hemiepiphysiodesis from 1/1/2014 to 8/1/2026. Limb measurements (mm) including R/L long leg, R/L femur, R/L tibia and R/L mechanical axis of deviation derived from the Stevens mechanical-axis zone classification were obtained from radiographs preoperatively and at clinical dictation of deformity correction or end of growth (DCA/EG). 95 patients were included (mean age, 10.9 ± 2.4 years; mean BMI, 26.5 ± 7.7 kg/m²). Sixty-seven patients (70.5%) had valgus deformity and 28 (29.5%) had varus deformity. The mean final absolute LLD was 8.7 ± 8.8 mm. Absolute LLD did not change significantly between preoperative and DCA/EG (p = .074). Patients with varus deformity had greater final LLD than those with valgus deformity (median, 10.1 mm [IQR, 5.6–17.9] vs 4.8 mm [IQR, 2.5–8.6]; p < .001). Our findings suggest that hemiepiphysiodesis correction was not associated with a clinically significant LLD. Notably, varus patients demonstrated greater final LLD than valgus patients (median, 10.1 vs 4.8 mm; p < .001), suggesting deformity type may influence residual inequality. Future work should target multicenter populations and examine factors that might influence LLD between valgus and varus.

Validation of Spatial Frequency Analysis of Pediatric Lower Leg Ultrasound and Preliminary Comparison in Neuromuscular Patients

Speaker: Noah Knezic

Co-Authors: Dr. Scott Crawford, Dr. Rachel Lenhart

Abstract:

INTRODUCTION: Spatial frequency analysis (SFA) of ultrasound (US) images is a method of quantifying muscle organization through parameters peak spatial frequency radius (PSFR) and maximum frequency amplitude percentage (Mmax%), which reflect fascicular spacing and prominence of the dominant pattern normalized to echogenicity, respectively. SFA-US has not been applied to pediatrics.

METHODS: Typically developing (TD) and cerebral palsy (CP) children, aged 4-17 years, underwent US imaging of the Achilles, soleus, gastrocnemius, and the gastro-Achilles musculotendinous junction (MTJ), bilaterally. B-mode images were acquired in triplicate and analyzed using MATLAB-based SFA algorithms. Intrasession repeatability and intersession reliability used intraclass correlation coefficients (ICC) and coefficients of variation (CV). ICC values were interpreted as poor (&lt;0.50), moderate (0.50–0.75), good (0.75–0.90), or excellent (&gt;0.90). Comparison of TD and CP used Welch’s T-test.

RESULTS: TD (n=9, 5 males, age 10.1 +/- 3.1 years) children completed two visits, yielding 212 sites for intrasession repeatability and 86 matched sites for intersession reliability. Intrasession ICC ranged from 0.794-0.968 with CV% ranging from 0.13-6.68%. Intersession ICC ranged from 0.550-0.934 and CV% ranged from 0.520-12.5. Analysis of TD vs CP (n=3, 2 males, age 12.0 +/- 4.4 years) showed no statistical significance (p&gt;0.05).

DISCUSSION: Strong reliability and repeatability support SFA use as a reproducible method in the pediatric lower leg. Preliminary analysis of TD vs CP showed no statistical significance, but expected trends were observed with CP trending higher for PSFR and lower for Mmax% suggesting increased fascicle spacing and reduced pattern prominence. Continuing work looks to increase power.

Investigating How Liver Disease Risk Is Modified By Genetic Variation And Environmental Contaminants

Speaker: Samuel Hernandez

Co-Authors: Katie L. Holl, Wendy Demos, Monika Tutaj, John J. Reho, Lynn Malloy, Justin L. Grobe, Melinda R. Dwinell, Chien-Wei Lin, Anne E. Kwitek

Abstract: Per- and polyfluoroalkyl substances (PFAS) are present in >98% of the U.S. adult population. Exposure to the persistent PFAS, perfluorooctane sulfonic acid (PFOS), is associated with lipid dysregulation and increased risk of metabolic dysfunction-associated steatotic liver disease (MASLD). While various human genetic loci are associated with MASLD, it is currently unknown how the genetic risk for MASLD is impacted by PFAS. I hypothesize that genetic variation plays a key role in determining the magnitude and directionality of PFOS-induced hepatic lipid and transcriptomic disruption. Therefore, we performed a 12-week PFOS exposure study on seven inbred rat strains (ACI, BN, BUF, F344, M520, MR, and WKY) with high phenotypic and genetic variation, followed by circulating lipid, liver lipid, and liver mass measurements. We found PFOS induces MASLD in a genetic background dependent manner and focused on two of the most affected strains (BUF/MnaMcwi and WKY/NCrl). These strains showed liver hypertrophy. After PFOS exposure, in male rats, liver triglycerides were increased only in the BUF strain, while both strains showed decreased circulating triglyceride levels suggesting diverging strain-specific lipid disruption. Employing RNA-sequencing on BUF and WKY livers, we identified distinct transcriptomic profiles across strains, sexes, and exposures. Weighted correlation network analysis revealed gene networks in which the gene expression-phenotype correlation was significantly connected to PFOS-induced phenotypes (e.g. liver weight and triglycerides) and enriched for the biosynthesis of fatty acids. Thereby, our model suggests coordinated pathway-level disruption. This is the first study to assess genes and pathways dysregulated by PFOS exposure across genetically heterogeneous cohorts.

TBD

Speaker: Alexis Sawicki

Co-Authors:

Abstract:

TBD

Speaker: Brea Brennan

Co-Authors:

Abstract:

TBD

Speaker: Verushca Gasiorowski

Co-Authors:

Abstract:

TBD

Speaker: McKenzie Henkelman

Co-Authors:

Abstract:

Hidden Lives of the Grenada Frog: Using Video Surveillance to Study Behavior and Ecological Interactions

Speaker: Sam Smid

Co-Authors: Dr. Emily Latch, Billie Harrison.

Abstract: The Grenada Frog (Pristimantis euphronides) is an endemic species restricted to high-elevation habitats on the island of Grenada and is currently classified as critically endangered. Despite its conservation status, relatively little is known about the species’ ecology and behavior. Grenada frogs occupy cool, wet, montane habitats that are increasingly threatened by climate change and overlap with the introduced Johnstone’s Whistling Frog (Eleutherodactylus johnstonei). Documenting the behavior of this understudied species is critical for understanding its ecology and informing future conservation efforts. We used more than 12 hours of passive video recordings to characterize previously undocumented behaviors of Grenada frogs, with particular attention to video observations documented multiple instances of prey capture by Grenada Frogs, reproductive calling and interactions among individuals, coexistence with larger animals that were originally thought to be predators of the species. These observations are among the first detailed accounts of the natural behavior for this species and identify several areas for future hypothesis-driven research, including reproductive interactions and predator-prey relationships. This work highlights the value of extended passive video monitoring for characterizing the behavior of cryptic, understudied species.

Camouflage and aposematism do not simultaneously lower predation risk in Eastern Grey Treefrogs

Speaker: Angela Marie Edwards Mish

Co-Authors: Gerlinde Höbel

Abstract: Coloration is often used for predator avoidance, with common tactics being camouflage (blending into surrounding) and aposematic coloration (conspicuous warning colors). Some organisms, such as Eastern Gray Treefrogs, may use both strategies; they can change color to improve camouflage of their bodies and reveal hidden patches of aposematic coloration on their legs. We conducted a clay model study to determine if camouflage, aposematic coloration, or their interaction provided greatest protection from predation. Clay models were placed on transects and collected if found with marks from a predator. We also took photographs of each placed model and its surroundings, which allows us to compare model-background contrasts (estimated as Just Noticeable Difference, JND) between models and treatments. Using these photographs, we calculated vision models for different putative avian and mammalian predators. Overall, our results show that camouflage did not influence attack rate on frog models. Across all model types, lower contrast was not related to lower levels of attack. Although, gray models were more conspicuous (they had a significantly higher model-background contrast) than green models. Attacks were lower for gray models with aposematic leg color than those without, but there was no relationship between attacks and aposematic coloration for green models. Perhaps this means that aposematic coloration is primarily protective for models that already stand out.

TFPI and Neuroplacentology

Speaker: Alyssa Braeger

Co-Authors: Alan E. Mast, Susan A. Maroney

Abstract:

Background: Tissue factor pathway inhibitor (TFPI) is a Kunitz-type anticoagulant protein involved in vasculature development.1 Mice with TFPI’s first Kunitz domain knocked out (K1KO) are embryonic lethal, dying at mid and late-gestation, prior and post E11.5.2 This differs from total TFPI KO lethality, occurring at mid-gestation only. Preliminary data shows K1KO cerebrovascular abnormalities with mostly normal placental histology. Placental and cerebral vascular development uniquely occur primarily through angiogenesis (blood vessels form from existing vessels). Previous studies of other mice have found placental defects in early lethality embryos (E9.5 – E14.5) that correlate with brain, heart, and vasculature defects and developmental delay.3 Placental and cerebral vascular development were examined in embryos lacking the entire TFPI protein.

Hypothesis: Placental development is a biological checkpoint for brain development. TFPI total KO mice will have abnormal placental and cerebral vascular development.

Methods: Timed matings were performed; E9.5-11.5 embryos were harvested, genotyped, and histologically processed. Brains were stained for laminin, an endothelial basement membrane protein. Placental tissue was examined for antibodies to SyncytinA, MCT4, Endomucin, GCM1, CD34, and Ki67 to analyze syncytiotrophoblast layers and developing fetal endothelium.

Results: Preliminary findings indicate total TFPI KO embryos had decreased to no vasculature development in placental labyrinth, and smaller total placentas.

Conclusions: K1KO mice contain TFPI’s K2/K3C domains, while total KOs have no TFPI. Although these both lead to 100% embryonic lethality, K1KO lethality’s predominant target appears to be the brain, while total KOs’ is the placenta. Future goals are to investigate TFPI’s different domains’ effects on angiogenesis.

The Role of RBM26 and RBM27 in Neuronal Development

Speaker: Sagnik Das

Co-Authors: Linh Thi Nhat Nguyen, Sri Kavya Durga Devineni and An Phu Tran Nguyen

Abstract: The RBM27 gene encodes an RNA-binding protein and is recognized as an autism candidate gene by the Simons Foundation Autism Research Initiative (SFARI). In cultured human cells, RBM27 and its paralog, RBM26, function as components of the poly(A) exosome targeting (PAXT) connection, an adaptor complex for the nuclear RNA exosome. In C. elegans, loss of RBM-26, the ortholog of mammalian RBM26/27, causes defects in axon development and mitochondrial function, suggesting a protective role for RBM-26 in preventing axon degeneration during larval development. The roles of RBM26 and RBM27 in the development of mammalian neurons, particularly their contributions to axon development and degeneration, remain poorly understood. In this study, we aim to determine the impact of RBM26 and RBM27 depletion on neurite development in mammalian neurons. We generated lentiviral vectors expressing short hairpin RNA (shRNA) constructs to knock down RBM26 and RBM27 in rat cells. Lentiviral-mediated expression of RBM26 shRNA and RBM27 shRNA in rat dopaminergic N27A cells and rat adrenal medulla pheochromocytoma PC12 cells resulted in robust reductions in the respective mRNA and protein expression levels compared with cells expressing the control shRNA. Interestingly, PC12 cells expressing RBM26 or RBM27 shRNA exhibited reduced neurite outgrowth at both 72 and 120 hours following stimulation with rat nerve growth factor (NGF), suggesting that RBM26 and RBM27 may play important roles in neurite development. Future experiments will investigate the molecular mechanisms underlying the impaired neurite outgrowth phenotype observed in PC12 cells following RBM26 and RBM27 depletion. We also sought to determine whether RBM26 and RBM27 depletion affects neurite outgrowth in human neurons. To this end, we generated lentiviral vectors harboring single-guide RNAs (sgRNAs) targeting human RBM26 and RBM27. To achieve RBM26 and RBM27 depletion, we delivered the sgRNAs into human induced pluripotent stem cells (i3-iPSCs) that stably express the transcriptional repressor dCas9-BFP-KRAB. Our preliminary data demonstrate that delivery of an sgRNA targeting RBM26 into these cells resulted in robust depletion of RBM26 at both the mRNA and protein levels. We are currently developing RBM27-depleted i3-iPSCs using a similar approach. Future studies will examine the effects of RBM26 and RBM27 depletion on neurite outgrowth in human neurons by further differentiating the i3-iPSCs into glutamatergic neurons. Together, these studies will help clarify the roles of RBM26 and RBM27 in neurite development in mammalian neurons.

The Role of PPM1H in LRRK2-Mediated Neurotoxicity and Neurodegeneration in Parkinson’s Disease

Speaker: Thi Nhat Linh NGUYEN

Co-Authors: Sagnik Das, Julia Hickey, Sri Kavya Durga Devineni, and An Phu Tran Nguyen

Abstract: Missense mutations that lead to LRRK2 kinase hyperactivation contribute to neuronal cell death in Parkinson’s disease (PD). LRRK2 phosphorylates a subset of Rab GTPases within their Switch-II motifs, thereby regulating their interactions with effector proteins. Recently, PPM1H was identified as a highly selective phosphatase that counteracts LRRK2 activity in cells. PPM1H belongs to the PPM family of Ser/Thr phosphatases, which are characterized by their dependence on Mn²⁺ or Mg²⁺ ions. In cellular models, PPM1H selectively dephosphorylates Rab GTPases at LRRK2-targeted phosphorylation sites. However, the contribution of PPM1H to LRRK2-mediated neuronal cell death remains incompletely understood, and its neuronal function is still poorly defined. In this study, we first sought to determine whether PPM1H protects against neuronal toxicity and cell death. Lentiviral-mediated transduction of an shRNA targeting rat PPM1H resulted in a robust reduction in PPM1H protein expression and increased phosphorylation of Rab8A at T72 and Rab10 at T73 in rat dopaminergic N27A cells. Intriguingly, rat adrenal medulla pheochromocytoma PC12 cells expressing PPM1H shRNA exhibited reduced neurite outgrowth at both 72 and 120 hours following stimulation with rat NGF. Future studies will assess whether pathogenic LRRK2 mutations further exacerbate the impaired neurite outgrowth phenotype caused by PPM1H depletion in PC12 cells. We next sought to determine whether enhancing PPM1H catalytic activity through overexpression can mitigate the neurotoxic effects induced by pathogenic LRRK2 mutations. To this end, we generated lentiviral vectors encoding V5-tagged PPM1H wild-type (PPM1H WT) and a substrate-trapping, catalytically inactive mutant carrying the D288A mutation (PPM1H D288A). We found that overexpression of PPM1H WT, but not the PPM1H D288A mutant, markedly reduced R1441C LRRK2-induced Rab10 phosphorylation at T73 in HEK-293 cells, demonstrating that PPM1H catalytic activity effectively counteracts LRRK2-driven Rab phosphorylation. Future studies will assess whether PPM1H overexpression can rescue the neurotoxic effects induced by pathogenic LRRK2 expression in rat cortical neurons. Together, these findings will help establish whether PPM1H acts as a key modulator of LRRK2-mediated neurotoxicity and evaluate its therapeutic potential to counteract LRRK2 kinase hyperactivity in PD.

Selective expression of a mutant AT1R with prolonged Gq signaling in AT1R-expressing neurons of the Subfornical Organ enhances Na preference and consumption in mice

Speaker: Joshua House

Co-Authors:

Abstract:

Sex Differences in the Effects of Chronic Stress, Chronic Corticosterone and Prefrontal Cortical REDD1 Overexpression on Pavlovian Autoshaping Behavior in Rats

Speaker: Beliz Kurtoglu

Co-Authors: Minhaj Abedin, Samuel Lawton, Daniel T. Brozoski, Justin L. Grobe, Pablo Nakagawa, Curt D. Sigmund

Abstract:

Background: Hypertension contributes substantially to cardiovascular disease and is linked to overactivation of the brain renin–angiotensin system via angiotensin II type 1 receptors (AT1Rs). The subfornical organ (SFO) regulates hydromineral balance and blood pressure through AT1R signaling. We hypothesized that prolonged AT1R G-protein signaling in SFO AT1R-expressing neurons increases sodium intake and alters fluid regulation.

Methods: BAC-AT1R-Cre;Ai14 mice received SFO-targeted injections of Cre-dependent AAVs encoding either wild-type AT1R (AT1R-WT) or a Gq-biased AT1R mutant with impaired β-arrestin recruitment and receptor internalization (AT1R-TSTS). Baseline drinking was measured using a ramped two-bottle choice paradigm comparing deionized water with 0.03–0.45 M NaCl. Intake was also assessed after water restriction or sodium depletion using deionized water and 0.15 M NaCl. Time-domain nuclear magnetic resonance assessed body composition, total-body water, and hydration status.

Results: Mice expressing AT1R-TSTS in SFO AT1R-expressing neurons consumed and preferred more NaCl than AT1R-WT controls, especially at lower NaCl concentrations. This pattern persisted after water restriction and sodium depletion, with AT1R-TSTS mice consuming more total sodium. Groups did not differ in fat mass, total-body water, or hydration status.

Conclusions: Prolonged Gq-biased AT1R signaling in SFO AT1R-expressing neurons promotes sodium preference and intake at baseline and during angiotensin II-associated homeostatic challenges

TBD

Speaker: Hossein Asgari

Co-Authors:

Abstract:

TBD

Speaker: Madison Rittinger

Co-Authors:

Abstract:

Step off: territorial calling behavior in the tropical Grenada Frog (Pristimantis euphronides)

Speaker: Leila Goodrum

Co-Authors: Billie Harrison, Emily Latch & Gerlinde Hoebel – I believe all will be in attendance at the conference, but none will be co-presenting with me.

Abstract: In anurans, sound is the primary mode of communication, and species display a variety of acoustic signals. Generally only males call, and most frogs have at least two call types: an advertisement signal, meant to alert the female to the male’s presence and attract her, and an antagonistic signal, meant to defend calling territory against other males. In the Grenada frog or Pristimantis euphronides, an endangered montane frog endemic to the island of Grenada, the advertisement signal is well documented and consists of several clicks, starting with a burst and gradually slowing. However, because Grenada frogs do not aggregate in water to chorus, little has been known about the territorial dynamics between males or between the Grenada frog and its relatively new acoustic competitor, the invasive Johnstone’s frog. The Johnstone’s frog, or Eleutherodactylus johnstonei, has a call which is much louder and in an overlapping frequency range to that of the Grenada frog. Further, because the Grenada frog requires a cool, minimally disturbed montane habitat, climate change and human development are driving it into a smaller and smaller area, so acoustic competition with the sympatric Johnstone’s frog could have high stakes. Using four treatments of loud and quiet Johnstone’s and Grenada frog calls, we conducted playback trials in the field to assess male Grenada frogs’ responses to the acoustic presence of conspecific and invasive males at simulated near and far proximities. Here we discuss preliminary results and describe a newly recorded Grenada frog signal, presumed to be territorial/antagonistic.

Counting Incorrect Fubini Rankings

Speaker: Kayla A Picasso

Co-Authors: Joakim Jakovleski, Pamela E Harris

Abstract: A Fubini ranking $\alpha=(a_1, a_2,\ldots, a_n)$ is a tuple of length $n$ whose entries are in $\N={1,2,3,\ldots}$. Fubini rankings encode the final outcome of a competition with $n$ competitors where ties are allowed. For example, $(3,1,1)$ is the Fubini ranking for a race in which competitor $1$ places third, and competitors $2$ and $3$ tie for first. We consider $\alpha^\uparrow=(a’_1, \ldots, a’n)$ to be the weakly increasing rearrangement of $\alpha$ – it is known that $\alpha$ is a Fubini ranking if and only if for any $a_i’ \in \alpha^\uparrow$, $a_i’=a{i-1}$ or $a_i’=i$. Given a weakly increasing Fubini ranking $\alpha$, we say that $\beta$ is an incorrect Fubini ranking if, for at least one $i \in [n] = {1,2,\ldots,n}$, $b_i \neq a_i$. We define $b_i$ to be a \emph{happy} competitor if $b_i > a_i$ and a \emph{sad} competitor if $b_i < a_i$; we denote their counts using $h$ and $s$ respectively. Given a specific $\alpha$ and values for $h$ and $s$, the set of possible incorrect Fubini rankings is denoted $\IFR{\alpha}{h}{s}$. We give formulas for the cardinality of $\IFR{\alpha}{1}{0}$, $\IFR{\alpha}{0}{1}$, and $\IFR{\alpha}{2}{0}$ for any given $\alpha$. We also give a formula for the cardinality of $\IFR{\alpha}{h}{0}$ for any $h \in \N$ where $\alpha=(1,1, \ldots, 1)$.

Parking Necklaces

Speaker: Cyrus Young

Co-Authors: Pamela E. Harris, Jennifer Elder, Gregor Grote

Abstract: If you try to make a necklace out of a certain number of colored beads, there are only a certain number of distinct necklaces you can come up with. Counting the number of distinct possible necklaces is a classic problem in combinatorics, which was first investigated by Moreau in 1872. From determining this count, we can derive one of the cleanest known proofs of Fermat’s little theorem. We are going to count necklaces with are made out of combinatorial objects called “parking functions”; hence the name “parking necklaces”. In doing so, we will obtain new analogues of Fermat’s little theorem, and new methods for counting necklaces with any specified pattern.

Lean Coloring Hypergraphs

Speaker: Harrison Piehowski

Co-Authors:

Abstract: Inspired by the notion of tricolorability of knots, we introduce the concept of lean coloring for hypergraphs and the associated lean number of a hypergraph.

Lean coloring often involves very few colors, yet still requires the methods of usual graph coloring, forcing the overall complexity to be NP-Hard. We provide two alternative formulations of the lean coloring problem that involve a type of coloring on abstract simplicial complexes and a partial coloring on bipartite graphs.

We then provide bounds for the lean numbers of hypergraphs that are k-uniform, k-partite, wide-path connected, or r-complete.

Python-like script is included to allow the implementation and study of a lean coloring algorithm. We conclude with some directions for future work and present the lean numbers of 130 knots and links.

Pullback Parking Functions

Speaker: Ilana Lavene

Co-Authors: Jennifer Elder, Pamela E. Harris, Lybitina Koene, Lucy Martinez, Molly Oldham

Abstract: We introduce a generalization of parking functions in which cars are limited in their movement backwards and forwards by two nonnegative integer parameters k and ℓ, respectively. In this setting, there are n spots on a one-way street and m cars attempting to park in those spots, and 1≤m≤n. We let α= (a_1,a_2,…,a_m)∈[n]^m denote the parking preferences for the cars, which enter the street sequentially. Car i drives to their preference a_i and parks there if the spot is available. Otherwise, car i checks up to k spots behind their preference, parking in the first available spot it encounters if any. If no spots are available, or the car reaches the start of the street, then the car returns to its preference and attempts to park in the first spot it encounters among spots a_i+1,a_i+2,…,a_i+ℓ. If car i fails to park, then parking ceases. If all cars are able to park given the preferences in α, then α is called a (k,ℓ)-pullback (m,n)-parking function. Our main result establishes counts for these parking functions via a recursive formula.

Classical and Topographical Combinatorial Statistics on Words in Natural Languages

Speaker: Joakim Jakovleski

Co-Authors: Pamela E Harris, Matt McClinton, Harrison Piehowski (all are attending, none are co-presenting)

Abstract: Combinatorial statistics are integer-valued functions defined on a family of combinatorial objects. Most commonly, words studied via combinatorial statistics are permutations or multiset permutations. Since words in a natural language are akin to elements of a multiset permutation, Elder, Harris, and Simpson consider and give the distributions of the flattened and the run statistic on words in the languages Danish, Dutch, English, French, German, Italian, Norwegian, and Spanish. Inspired by their work, we consider the same languages and determine distributions for classical statistics (ascent, descent, major index, inversion, excedance) and topographical statistics (tie, peak, valley, mesa, canyon, altitude). Provided are visualizations of our findings in the form of data tables, line graph interpolations, and heatmaps. We conclude with many directions for future study.

Structural Equation Modeling: Predictors of Collaborative Problem Solving

Speaker: Nishchitha Girana Pathirannehelage

Co-Authors: Sabreet Kaur Dhatt

Abstract: This study investigated the structural relationships among collaborative dispositions, science-related motivational beliefs, and collaborative problem-solving (CPS) performance using data from the Programme for International Student Assessment 2015 assessment. Drawing on Expectancy-Value Theory and Social Cognitive Theory, the study examined whether Science Interest, Science Self-Efficacy, and Achievement Motivation mediated the relationship between Collaborative Disposition and CPS performance. Data were obtained from 10,505 students from Singapore (N = 5639) and the United States (N = 4866). Structural equation modeling with robust maximum likelihood estimation was employed due to violations of multivariate normality. The measurement and structural models demonstrated acceptable fit and strong factorial validity. Science Interest emerged as the strongest positive predictor of CPS performance, whereas Collaborative Disposition and Science Self-Efficacy exhibited significant negative direct effects after controlling for other variables. Achievement Motivation did not significantly predict CPS performance. Mediation analyses indicated that Collaborative Disposition indirectly enhanced CPS performance through increased Science Interest, while a small negative indirect effect was observed through Science Self-Efficacy. Multi-group analyses supported configural, metric, scalar, and structural invariance across both countries, suggesting that the constructs and structural relationships were comparable between groups. The findings highlight the importance of domain-specific interest in collaborative problem-solving contexts and suggest that collaborative orientations may not necessarily translate into higher CPS performance without the support of strong cognitive and motivational engagement.

Extension of McDougall’s Circle Theorem

Speaker: Olivia Bouthot

Abstract: Ptolemy’s Theorem states that for a cyclic quadrilateral, the sum of the products of opposite side lengths is equal to the product of the diagonal lengths. A ratio form of this property extends to McDougall’s Circle Theorem, which shows that a similar property is true for all cyclic n-gons where n is even. McDougall’s Circle Theorem extends further. Arbitrary point(s) are added within the circle, and their squared distances from the vertices of the cyclic polygon are used as weights. This project uses geometric and analytic methods to prove this further extension of McDougall’s Circle Theorem. We find a set of points in the complex plane that fall in a balanced alignment. Additionally, we describe an alternate elementary proof of McDougall’s Circle Theorem from the balanced alignment.

Rank Based Bayesian Variable Selection in High-Dimensional Data

Speaker: Sakib Salam

Co-Authors: Anjishnu Banerjee, PhD

Abstract: High-dimensional biomedical data pose challenges for statistical analysis, including multicollinearity, small sample sizes, and unstable variable selection across analyses. As a result, feature selection is a key part of modern statistical learning, especially in translational research, where interpretability, reproducibility, and robustness matter as much as predictive accuracy. Many feature-selection methods have been developed, including penalized regression, Bayesian shrinkage, and spike-and-slab models. However, these methods are often sensitive to tuning, depend on data scale, or lack clear ways to prioritize features when competing goals arise.

The objective of this study is to develop and evaluate a rank-based feature selection framework that addresses the major challenges of existing methodologies in high-dimensional data contexts. The proposed approach is built around a measure-utility function that maps feature-specific evidence into a unified ranking, allowing variables to be prioritized by stability, strength of association, and value across multiple modeling settings rather than by binary inclusion decisions alone. This framework is designed to complement, rather than replace, established Bayesian variable-selection paradigms.

The new framework is compared with common Bayesian methods, such as stochastic search variable selection (SSVS) and spike-and-slab priors, which employ hidden indicators and mixture priors to model sparsity. These methods offer clear probabilistic interpretations, but their results can depend on prior choices and may be unstable with small sample sizes. In contrast, the rank-based approach focuses on combining evidence across features, yielding more stable rankings when data or models change. The framework is demonstrated using a high-dimensional biomedical example that includes complex clinical and molecular features without revealing sensitive details.

Comparisons show that the rank-based framework yields more stable feature rankings than traditional selection methods, especially when predictors are correlated or sample sizes are small. Rather than selecting a single subset of features, this method produces a clear order of candidate features, which supports later validation, sensitivity checks, and expert review.

This work presents a flexible and intuitive alternative to standard feature-selection methods by treating variable selection as a ranking and prioritization problem guided by clear utility goals. This framework can be applied to many high-dimensional biomedical studies and provides a solid basis for combining Bayesian variable-selection methods with approaches that emphasize stability in real-world settings.

TBD

Speaker: Vanshika Agarwal

Co-Authors:

Abstract:

TBD

Speaker: Emma Tillison

Co-Authors:

Abstract:

TBD

Speaker: Justin Benoit

Co-Authors:

Abstract:

You BETcha: Modulating Macrophage Inflammation through BET Bromodomains

Speaker: Paul Sidlowski

Co-Authors: Savannah J. Makowski, Joshua A. Nord, and Brian C. Smith

Abstract: Dysregulated inflammation is a critical driver in the initiation and progression of many inflammation-related diseases, accounting for >60% of global deaths. Prolonged macrophage activation to a proinflammatory state is a primary contributor to dysregulated inflammation, particularly through the secretion of proinflammatory mediators. Here, we examine the roles of the bromodomain and extraterminal domain (BET) protein family in regulating macrophage inflammatory activation and transcription of proinflammatory mediators. BET proteins (BRD2/3/4/T) are epigenetic readers that contain two tandem bromodomains (BD1 and BD2) capable of binding acetylated lysine residues on nuclear proteins. Pan-BET inhibitors, which block both bromodomains of the entire BET family, strongly attenuate inflammatory responses across multiple cell and tissue types. BET bromodomain inhibitors attenuate macrophage-mediated inflammation by reducing the expression of NF-κB target genes. We previously demonstrated that selective inhibition of BET BD1 suppresses a subset of NF-κB-dependent genes in β-cells by disrupting an interaction of BRD4-BD1 with the p65 subunit of NF-κB. At the same time, robust suppression of the pro-inflammatory response in macrophages requires inhibition of both BET bromodomains, with BRD2 playing a substantial role. Furthermore, recent sequencing data suggest novel transcription factor interactions that drive both BET-dependent and BET-independent inflammatory responses in macrophages. Taken together, our findings suggest that BET proteins hold distinct cell- and context-specific roles in macrophage inflammatory activation. These distinctions will inform the future development of therapeutics that selectively target macrophage-driven inflammation while sparing other BET transcriptional roles.

mTORC1 Activity Coordinates Translational Programs to Organize Epithelial Morphogenesis in a Human Model of Epiblast Formation

Speaker: Lauren Elberfeld

Co-Authors: Anusha Rengarajan, and Kenichiro Taniguchi

Abstract: Immediately after the human embryo, known at this stage as the blastocyst, initiates implantation into the uterine wall, a seemingly unpolarized group of pluripotent epiblast cells begin to radially organize and deposit a central lumen. This morphogenic event provides a foundation for essential subsequent processes such as amniogenesis and gastrulation. Recently, it has been shown that mammalian target of rapamycin (mTOR) signaling is a driver of developmental progression in pre-implantation blastocysts. However, it is currently unclear whether mTOR signaling continues to support developmental processes during implantation. Interestingly, our previous study showed that LAMTOR1, a component of the pentameric Ragulator complex associated with mTOR complex 1 (mTORC1) activation, is enriched in the apical territory of a human pluripotent stem cell-based model of epiblast formation (hPSC-epiblast model), raising the possibility that mTORC1 signaling drives human epiblast lumen formation. Using an hPSC-epiblast model allowing for inducible depletion of the LAMTOR1 protein, we found that LAMTOR1 depletion leads to decreased mTORC1 signaling, accompanied by reduced protein synthesis, while overall hPSC-epiblast size is unchanged. Moreover, the LAMTOR1-depleted hPSC-epiblasts show defective apical membrane morphogenesis. Importantly, such defects are also seen when control hPSC-epiblasts are treated with rapamycin, an mTOR kinase inhibitor, or with small molecules that suppress mTORC1-dependent translational effectors. Together, this study uncovers a previously unrecognized notion that LAMTOR1-mTORC1 signaling drives human epiblast formation by maintaining the translation of proteins that contribute to apical membrane formation. Currently, we are exploring transcripts translated in LAMTOR1-mTORC1-dependent manners by performing ribosome profiling in control and LAMTOR1-depleted backgrounds.

Defining Pseudo-Haplotype Analysis Reveals Multi-Gene Genetic Pattern Across BAF Chromatin Remodeling Complexes

Speaker: Xiaowei Dong

Co-Authors: Neshatul Haque, Jessica Wagenknecht, Michael T. Zimmermann

Abstract: BRG1-associated factor (BAF) is a crucial chromatin remodeling complex. Variants in genes encoding BAF complex components cause human diseases, including cancers and developmental disorders. However, the genetic diversity and variant co-occurrence patterns within BAF genes remain incompletely understood. It is feasible, though largely untested, that rare patterns of common variations could alter function similarly to rare deleterious variants. Further, there is no modern census of how often individual people simultaneously carry multiple rare and common variations, nor means for genomics practitioners to assess their combined effects. Approaches are needed to characterize complete sequences from individual samples.

In this study, we introduce a pseudo-haplotype analysis (PHA) framework, combining multiple protein-coding sequence variants, observed concurrently within individual samples, into discrete BAF patterns. In this cohort, 78.44% of pseudo-haplotype (PH) copies carry at least one BAF coding variation. Among these, 56.18% contain at least two distinct variants, and 32.39% contain three or more, indicating a substantial burden of multi-variant configurations across individuals. Notably, 25.30% of unique PHs are observed only once, highlighting a considerable proportion of people who are affected by rare or private combinations of genetic variations. We identify multiple significant (FDR &lt; 0.05) co-occurrence combinations across global populations. These findings underscore the importance of considering population-specific genetic structures, and complete individual variant configurations when investigating disease associations and genetic mechanisms. Our approach provides a generalizable framework for characterizing multi-variant architectures within chromatin remodeling genes at a population scale, with potential applications in elucidating disease etiology and advancing precision medicine.

Retinoid Supplementation Reveals a Role for STRA6 in the RPE Beyond Retinoid Transport

Speaker: Allison Hall

Abstract: The retina has enormous metabolic demands to support vision. The retinal pigment epithelium (RPE) and photoreceptors are physically and metabolically interconnected, exchanging metabolites and relying on each other for mutual support. Therefore, understanding pathways that maintain metabolic homeostasis in the RPE is critical for determining how RPE dysfunction contributes to retinal disease.

STRA6 is a transmembrane protein in the RPE that facilitates retinol uptake for delivery to photoreceptors for phototransduction. In mouse and zebrafish knockout models, stra6 loss causes photoreceptor disruption and vision loss. In these animals, oral treatment with pharmacological doses of vitamin A morphologically and functionally rescues photoreceptors. However, in stra6-/- zebrafish treated with supplemental dietary retinoid esters, RPE abnormalities persist. STRA6’s contribution to retinal health outside of retinoid transport remains unclear. Our primary evidence suggests that STRA6 has additional roles in the RPE beyond supplying retinoid chromophores to photoreceptors.

To test this hypothesis, we used a zebrafish stra6 knockout model and performed transcriptomic analysis on isolated RPE from wildtype and mutant animals with or without dietary retinyl palmitate supplementation. Gene set enrichment analysis revealed depletion of several mitochondrial pathways, indicating coordinated downregulation of mitochondrial pathways in stra6-/- RPE. Consistent with persistent RPE dysfunction, confocal imaging of RPE in rpe65a:LifeAct-mGreenLantern transgenic zebrafish revealed alterations in cytoskeletal organization of stra6-/- RPE that were not rescued by retinyl palmitate supplementation.

Together, these results support a role for STRA6 in maintaining RPE physiology beyond retinoid transport and identify mitochondrial dysfunction as a potential therapeutic target for preserving retinal health and function.

The Adaptor Protein Complex 1 Regulates Human Amnion Development

Speaker: Bailey Schultz

Co-Authors: Carleton A, Wang S, Duncan M, Taniguchi K

Abstract: Amnion is a critical component of the fetal membranes, providing essential immune, mechanical, and endocrine protection to the fetus. The formation of the amnion depends on a transcriptional cascade initiated by BMP signaling, which promotes the transition of cells out of their pluripotent state and toward an amnion specific differentiation. Disruption of amnion formation, which can occur in conditions such as infection or inflammatory stress, can compromise fetal membrane integrity and increase the risk of preterm birth, congenital abnormalities, and fetal death. AP-1 is a membrane trafficking protein that mediates clathrin-dependent vesicular trafficking and polarized sorting of proteins to specific cell membrane compartments. While impaired AP-1 function has been associated with developmental disorders, its role in early embryonic development remains largely uncharacterized. Previous studies from the lab have identified that the loss of the AP-1-y1 subunit, which plays a critical role in maintaining the AP-1 complex structure, leads to defects associated with lumen formation. To further characterize the role of AP-1-γ1 in early development, we have begun to utilize a Gel-3D hPSC-amnion cyst model, which relies on a soft mechanical cue to initiate amniogenesis. Interestingly, we have seen in the absence of AP-1-γ1 the exit from pluripotency associated with the initiation of amnion formation is halted and the amnion lineage progression is not initiated. Surprisingly, we have identified that with the loss of AP-1-γ1, SMAD proteins regulating the BMP signaling pathway are unable to become phosphorylated, and lumen formation defects are present. Overall, suggesting the importance of AP-1-γ1 mediated trafficking involved in lumenal formation, maintenance of pluripotency, and initiation of amniogenesis.

Higher apoB-100 in Rh-negative blood group and comparable apo(a) across ABO and Rh blood groups in a healthy blood donor cohort

Speaker: Maya Vergara

Co-Authors: Maya Vergara-1,2,3, Hayley Lund-1,2,3, Nina T. Yang-4, Duo Yu-2, Ziyu Zhang-1,2,3, Ze Zheng-1,2,3,5

1-Versiti Blood Research Institute, Milwaukee, WI, United States

2-Department of Medicine, Medical College of Wisconsin, Milwaukee, WI, United States

3-Cardiovascular Research Center, Medical College of Wisconsin, Milwaukee, WI, United States

4-Summer Program for Undergraduate Research, Medical College of Wisconsin, Milwaukee, WI, United States

5-Medical College of Wisconsin, Department of Physiology, Milwaukee, WI, United States

Abstract: A, B, and AB blood types refer to erythrocytes that contain A, B, or both antigens, whereas O blood type contains neither A nor B antigens. Likewise, Rh factor is characterized by the presence or absence of the Rh antigen on erythrocytes. Previous literature shows that Rh- males have elevated total cholesterol levels, while other studies found no correlation between Rh status and cardiovascular disease risk. Likewise, several studies demonstrated that non-O blood types (A, B, and AB) are associated with higher incidence of coronary heart disease and higher levels of low-density lipoprotein (LDL) cholesterol. Apolipoprotein-B (apoB-100), the scaffolding protein of LDL, and apolipoprotein (a) (apo(a)), which binds to apoB-100 to assemble lipoprotein (a) (Lp(a)), are drivers of cardiovascular disease. However, the levels of apoB-100 and apo(a) in ABO blood types and by Rh status have not been reported. Through the Versiti Blood Center of Wisconsin blood bank, we collected de-identified whole blood samples from donors between ages 18-40, equally distributed by sex, and following the same distribution of ABO blood type and Rh status in the US (blood type A with 39%, B 12%, AB 4%, O 45%, Rh+ 85%, Rh- 15%). Whole blood samples were centrifuged at 1500 g for 15 minutes to isolate plasma. ELISAs were used to determine plasma apoB-100 and apo(a) concentrations. Wilcoxon rank-sum tests were employed for comparisons between O vs non-O, A vs non-A, and Rh+ vs Rh-. In this cohort, Rh- donors had 10.1% higher apoB-100 than Rh+ donors (mean, Rh- = 77.14 mg/dL, Rh+ = 69.33 mg/dL, p = 0.036). We observed no difference in apo(a) concentrations (median, Rh- = 7.55 mg/dL, Rh+ = 5.58 mg/dL, p = 0.323). Among ABO blood types, we observed comparable apoB-100 concentrations (mean, A = 75.19 mg/dL, B = 70.25 mg/dL, AB = 69.24 mg/dL, O = 71.66 mg/dL, p = 0.697), as well as between O and non-O blood types (mean, non-O = 73.6 mg/dL, O = 71.66 mg/dL, p = 0.658). There was no difference in apo(a) levels between blood types (median, A = 6.05 mg/dL, B = 5.59 mg/dL, AB = 5.97 mg/dL, O = 5.74 mg/dL, p = 0.759) or between O and non-O blood types (median, non-O = 6.00 mg/dL, O = 5.74 mg/dL, p = 0.464). However, we found that non-A blood type had a 5.6% significantly lower total cholesterol than A blood type (mean, non-A = 103.45 mg/dL, A = 109.64 mg/dL, p = 0.017). In this study, we found that Rh- donors had elevated apoB-100 levels, while non-A donors demonstrated decreased total cholesterol levels. Future prospective investigations should delineate the underlying mechanism of ABO blood type and Rh status in relation to cardiovascular disease.

Fli1 Regulates B-Cell Pathogenicity in Chronic Graft-Versus-Host Disease

Speaker: Rajdip basnet

Co-Authors: Yongxia Wu, MD, PhD; Xue-Zhong Yu, MBA, MS, MD

Abstract: Allogeneic hematopoietic cell transplantation (allo-HCT) can cure hematologic malignancies, but chronic graft-versus-host disease (cGVHD) remains a major long-term complication. Abnormal donor B-cell activation, germinal center (GC) responses, and class-switched autoantibodies contribute to cGVHD, yet the B-cell-intrinsic mechanisms driving these responses remain poorly understood. We investigated Fli1, an ETS-family transcription factor highly expressed in B cells.

Using B cell-specific Fli1 conditional knockout (Fli1 BcKO) mice, we examined B-cell development, B-cell receptor (BCR) signaling, antigen-induced GC responses, and cGVHD after murine allo-HCT. Purified follicular B cells were stimulated with anti-IgM and analyzed by flow cytometry, phospho-flow, and immunoblotting. NP-KLH immunization, splenic immunofluorescence, and single-cell RNA sequencing were used to characterize GC and transcriptional responses.

Fli1 deficiency preserved early bone marrow B-cell development but altered mature splenic B-cell homeostasis. Follicular and marginal-zone B cells were reduced, whereas CD21loCD23lo hyporesponsive B cells increased 5.4-fold. Fli1-deficient B cells showed impaired activation, proliferation, and plasmablast differentiation. Proximal BCR signaling was largely preserved, whereas downstream AKT-mTORC1-S6 signaling was reduced, with approximately sixfold lower p-S6 at 120 minutes. Following NP-KLH immunization, GC B-cell frequency decreased 6.14-fold, with reduced proliferation and fewer and smaller PNA-positive GC foci. Single-cell RNA sequencing further revealed altered B-cell differentiation and GC-associated transcriptional programs. In allo-HCT, Fli1-deficient B cells showed reduced pathogenic humoral responses and attenuated cGVHD while preserving IgM.

These findings identify Fli1 as a B-cell-intrinsic regulator linking BCR signaling, B-cell differentiation, GC formation, and pathogenic humoral immunity.

Investigating Genetic Determinants of Retinal Ganglion Cell Genesis

Speaker: Rocky Rausch

Co-Authors: Joel B. Miesfeld

Abstract: Retinal ganglion cells (RGCs) and their axons form the optic nerve, the sole retinal neuron that bridges communication from the retina to the brain. RGC axons are the primary targets of the neurodegenerative disease, glaucoma, a leading cause of vision loss with no cure. Critical to the development and survival of RGCs is the evolutionarily conserved transcription factor ATOH7, which is lost in patients with optic nerve aplasia and is essential to RGC development in all species studied. To determine if additional genes can rescue the loss of RGCs in atoh7 mutants, we created transgenic zebrafish lines to ectopically express RGC genes of interest in the atoh7 lineage. By utilizing immunohistochemistry with the pan RGC marker Rbpms2, we determined our transgenic overexpression paradigm can successfully rescue the loss of RGCs in atoh7 mutant animals through atoh7 re-expression. RGC rescue by known and newly identified genes – pou4f2, elavl4, klf7, isl1, eya2, and fstl4 – was either non-existent or limited in atoh7 wild type and mutant retinas. Surprisingly, ectopic isl1 expression was insufficient to generate RGCs in atoh7 mutant fish, while pou4f2 over expression generated RGCs throughout the retina, although, these RGCs did not form a proper ganglion cell layer. Future analysis will focus on the co-expression of these atoh7 regulated genes, as understanding the genetic components contributing to RGC genesis and survival downstream of ATOH7 will provide potential targets to aid regenerative and degeneration-slowing treatments.

MD-PHD Panel

Speaker:

Co-Authors:

Abstract:

TBD

Speaker: Tony Breitbach, PhD (FAC)

Co-Authors:

Abstract:

TBD

Speaker: Christopher Buehl, PhD (FAC)

Co-Authors:

Abstract:

TBD

Speaker: Mikelene (Mikke) Ray (FAC)

Co-Authors:

Abstract:

An Evolutionarily Conserved Ypk1/SGK1 Kinase Pathway Regulates the Unfolded Protein Response by Modulating the Ire1 Protein Abundance

Speaker: Saswata Chakrabarty

Co-Authors:

Abstract: The unfolded protein response (UPR) is a cellular stress response that helps maintain protein balance when proteins accumulate improperly in the endoplasmic reticulum (ER). A central regulator of the UPR is Ire1, an enzyme that detects ER stress and activates the unconventional splicing of HAC1 mRNA in yeast and XBP1 mRNA in human cells. These spliced transcripts produce transcription factors that activate genes required to restore protein homeostasis.

Our previous work showed that the MAP kinase Slt2 promotes UPR signaling by increasing IRE1 expression through the transcription factor Rlm1. Here, we investigated whether other protein kinases also regulate this pathway. We found that loss of several kinases, including Cdc28, Pkc1, Rio2, Tor2, Pkh1, and Ypk1, reduced HAC1 expression, suggesting that multiple signaling pathways contribute to UPR regulation. We focused on Ypk1, a yeast kinase related to human serum/glucocorticoid-regulated kinase 1 (SGK1). Genetic and biochemical analyses indicate that Ypk1 acts through the Pkc1 pathway to maintain Ire1 protein abundance. Importantly, this regulatory relationship is conserved in human cells, where SGK1 influences IRE1 protein levels and XBP1 mRNA splicing.

Together, our findings identify a conserved signaling mechanism linking cellular kinase signaling to the abundance of Ire1 and the activation of the UPR. This work provides new insight into how cells coordinate signaling pathways with ER stress responses to maintain protein homeostasis.

Function of key ciliary protein Dzip1 may be modulated by the Rab8 signaling pathway

Speaker: Signe Kopp

Abstract: Cell signaling pathways are crucial for normal development of an organism. To facilitate proper differentiation and growth, individual cells must be capable of responding various small changes in the surrounding environment. One of the major cellular organelles that facilitates signal transduction is the primary cilium. Primary cilia are highly specialized microtubule-based protrusions in the cell membrane, hosting and modulating essential components of multiple signaling cascades active throughout development, especially Hedgehog signaling. Thus, the proper formation of primary cilia, or ciliogenesis, is essential in development. Ciliogenesis is a highly complex and coordinated process involving translocation of the basal body to the cell membrane, membrane docking of ciliary vesicles, axoneme elongation, and recruitment of functional protein machinery such as Interflagellar Transport complexes. Previous studies in mice and zebrafish have identified Dzip1 as an important regulator of ciliogenesis. Zebrafish lacking Dzip1 (dzip1 -/-) fail to form cilia and present with a host of developmental issues such as ocular coloboma, body curling, neural tube defects, and eventual lethality around five days post fertilization. In vitro studies have identified interactions between Dzip1 and Rab8 pathway components as essential for ciliogenesis, but no further studies have been conducted in vivo. In this research, we use a zebrafish model to investigate the role of Rab8 pathway components in Dzip1-dependant ciliogenesis. Preliminary results demonstrate that supplying dzip1 -/- fish with constitutively active Rab8a may have a positive effect on mutant phenotypes, confirming this interaction as an important functional component of ciliogenesis.

Coupling the MAPK Slt2/ERK1 Pathway and IRE1-driven UPR Through Transcription Factor Rlm1/MEF2

Speaker: Anish Chakraborty

Co-Authors: Saswata Chakrabarty, Jagadeesh Kumar Uppala, Kimberly Ann Mayer1, Anna J Evans, Jasmine George, Chandrima Ghosh, Ritisha Dey, Franca Ohikhuare, Sham Mirza, An Phu Tran Nguyen, Nadege Gouignard, Pradeep Chaluvally-Raghavan, Madhusudan Dey.

Abstract: Unfolded protein response (UPR) is a conserved cellular strategy that enhances the protein folding capacity of cells under stress conditions. In Saccharomyces cerevisiae, the dual kinase RNase IRE1 initiates the UPR by catalyzing the cytosolic splicing of HAC1 mRNA, a process conserved in humans where IRE1 splices XBP1 mRNA. The spliced HAC1/XBP1 mRNA yields a transcription factor that upregulates the expression of protein-folding enzymes and chaperones, thereby boosting the cell’s ability to cope with unfolded proteins. Our study demonstrates that the UPR involves two distinct phases. The early phase operates predominantly through the canonical IRE1 signalling pathway, while the later phase involves additional regulation by the MAP kinase Slt2 or its human ortholog ERK1/ERK2/ERK5 and the downstream target the MADS-box transcription factor Rlm1 (an ortholog of human MEF2C) (Published in Communications Biology, April 2026.).

We further show that Slt2 promotes IRE1 expression through Rlm1. Together, these findings reveal a previously unrecognized crosstalk between the MAPK and IRE1-mediated arm of the UPR.

Efficient Pd-Catalyzed Amination of Benzodiazepines for GPCR Ligand Development

Speaker: Shaun Harrington

Co-Authors: Maya Fernando, Mubaraq Toriola, Kayode Medubi, Michelle Meyer, Leggy Arnold

Abstract: Palladium catalyzed reactions have become prominent reactions for creating C-N bonds and are widely used in the synthesis of pharmaceuticals. In this study, a palladium catalyzed Buchwald-Hartwig synthesis was used to introduce new amine substituents to a brominated benzodiazepine for the development of compounds with novel activity. We established an optimized protocol utilizing 5% Pd2(dba)3 in conjunction with 10% XPhos as the catalytic system. Conversions with substituted anilines, aliphatic amines and heterocyclic amines were observed with yields ranging from 23% to 92%. All products were analyzed and characterized using liquid chromatography-mass spectrometry and NMR spectroscopy. We identified that some of these compounds are having promising affinities for the Alpha 2B adrenergic receptor (AR𝛼2B) and the 5-Hydroxytryptamine 2B receptor (5-HT2B). We are currently synthesising new molecules to identify ligands with high specificity and better affinity enabling us to evaluate their pharmacological effects in vivo. ​

Redox-Driven Selectivity of a Photoinducible Phenyl Selenide: A Dual-Lock Mechanism for Targeting Triple-Negative Breast Cancer

Speaker: Regina Oddiri

Co-Authors: Thilini Nimasha Fernando

Abstract: Triple-negative breast cancer (TNBC) remains a significant clinical challenge due to its aggressive nature and the lack of selective therapeutic options. Photoinducible DNA cross-linking agents offer a promising strategy for achieving selectivity through spatiotemporal control. In this work, we synthesized and investigated a binaphthalene phenyl selenide (Compound 1) as a photoactivatable DNA interstrand cross-linking (ICL) agent. Photochemical studies showed that Compound 1 induces DNA cross-links upon 350 nm irradiation but remains inactive in the absence of light. Mechanistic and alkylation-site analyses revealed that cross-linking proceeds via radical and carbocation intermediates with preferential reactivity at guanine residues. Cytotoxicity studies demonstrated strong selectivity toward TNBC cells (MDA-MB-468, IC₅₀ = 1.7 μM) compared to normal epithelial cells (MCF-10A, IC₅₀ = 170 μM) under irradiation, while an in vivo safety study confirmed minimal toxicity. Despite these promising results, the molecular basis for this selectivity remained unclear. To decode the chemical logic of this selectivity, we modeled the cellular redox environment using in vitro DNA cross-linking assays to investigate how glutathione (GSH) and hydrogen peroxide (H₂O₂) govern the drug’s reactivity. We found that elevated GSH concentrations, characteristic of normal cells, suppressed DNA cross-linking, with complete inhibition at 24 mM. Conversely, elevated H₂O₂ concentrations characteristic of cancer cells increased ICL yields by 25% upon irradiation, likely through the oxidation of the phenyl selenide to a more reactive selenoxide intermediate. Notably, no cross-links formed in the absence of light, demonstrating a synergistic activation mechanism. This study reveals a sophisticated dual-lock mechanism: high basal GSH in normal cells acts as a protective “off-switch” by quenching reactive carbocation intermediates, while elevated H₂O₂ in cancer cells acts as an “on-switch” that sensitizes the compound to synergistic activation. This redox-driven framework provides a powerful new blueprint for the precision treatment of TNBC by exploiting the inherent chemical vulnerabilities of the tumor microenvironment.

Development of Nebulized Pegylated GABAA Receptor Ligands as Novel Therapeutics for Asthma Treatment

Speaker: Ethan Kowalczyk

Co-Authors: Maya R.T. Fernando, Michelle J. Meyer, Kayode M. Medubi, Mubaraq A. Toriola, Gene T. Yocum, Daniel A. Webb, Charles W. Emala, Leggy A. Arnold.

Abstract: The GABAA receptor (GABAAR) is a ligand-gated chloride ion channel that’s located primarily on neurons but is also expressed on airway smooth muscle and inflammatory cells. Allosteric GABAAR ligands that don’t cross the blood brain barrier (BBB) have been shown to elicit smooth muscle relaxation and anti-inflammatory properties. The rationale of this research is to develop a new class of GABAAR ligands that don’t cross the BBB. Analogs of PI320, a previously introduced GABAAR ligand, were synthesized with varying lengths of polyethylene glycol (PEG) chains. Analogs with increased PEG chain length had increased solubility. The apparent cell membrane permeability was tested using a parallel artificial membrane permeability assay (PAMPA). All analogs exhibited a high permeability, comparable to verapamil. Compounds were screened for toxicity against human embryonic kidney cells and showed low cytotoxicity. No behavioral or cognitive changes were observed in the mice measured by rotarod and open field testing for nebulized compounds. Phase I metabolism was observed for all analogs reducing systemic exposure when inhaled. This important safety feature resulted in low systemic toxicity. Novel compounds bind to a1, a2, and a3 GABAAR subtypes. Analogs with a shorter PEG chain exhibited better binding. The compounds were able to relax constricted mice trachea rings using ex-vivo tissue and we observed a reduction of airway hyperresponsiveness (AHR) for AJ mice when challenged with methacholine. All analogs showed reduction of AHR. Overall, we show that nebulized pegylated imidazodiazepines are effective to reverse bronchoconstriction and represent excellent new drug candidates for asthma.

TBD

Speaker: Nikole Tamayo

Co-Authors: Jon-Marc Rodriguez

Abstract: The American Chemical Society (ACS) Committee on Professional Training (CPT) publishes guidelines for bachelor’s degrees in chemistry to make recommendations related to training undergraduate students. For this research project, we analyzed documents provided by CPT that were released by ACS from 1939 to 2026 to understand how the requirements for institutions have changed over time. With our analysis, the documents were grouped chronologically and then categorized based on the length of the guidelines, similarities in table of contents, and large changes in content. In depth review of each initial group further separated the guidelines which resulted in six groups or “eras” based on the titles of the guidelines, how the purpose of the guidelines was introduced, and then the education level described in the contents. Following this, we tracked how the following requirements changed across the eras: facilities, faculty, coursework, laboratory skills, student skills, and chemical safety. Additionally, to provide context for the changes in the guidelines, we also analyzed published committee reports, comments, and editorials. As an implication of our work, we intend to offer suggestions to further the development of the next-generation of chemists.

Elucidating Parkin’s Mechanism Of Activation Via Integrative Computational Tools

Speaker: Carlos Herrada

Co-Authors: Dr. Kesaban Sankar Roychoudhuri. Dr. Vanessa Leone

Abstract: Parkin, a Ring-Between-Ring (RBR) E3 ubiquitin ligase encoded by the PRKN gene, is the most common genetic cause of autosomal recessive Parkinson’s disease. Parkin is a key player in mitophagy, whose role is regulated by a set of complex conformational changes. Understanding these structural states and transitions is critical for elucidating disease mechanisms and identifying novel therapeutic strategies.

To investigate Parkin conformational dynamics, we combined AlphaFold3 structure prediction with hydrogen-deuterium exchange mass spectrometry (HDX-MS) data. First, we benchmarked AlphaFold3 against experimentally characterized Parkin states and found that it accurately distinguished between post-translationally modified conformations associated with activation. We then generated structural models of an uncharacterized Parkin:E2~ubiquitin complex, which reproduced interaction features observed in homologous RBR E3 ligases.

To assess the biological relevance of predicted conformations, AlphaFold3 structural ensembles were reweighted using an experimental HDX-MS dataset. Preliminary results reveal a single preferred conformation of Parkin’s active structure that receives the highest weight from the experimental data used. This would suggest that Parkin’s RING2 domain adopts a well-defined orientation in its active state rather than having flexible behavior.

Ongoing work will further validate this preferred conformation by comparing it to complementary experimental data sets before moving onto molecular dynamics. Together, this integrated computational-experimental framework will provide new insight into Parkin’s conformational dynamics and establish a foundation for future structure-based therapeutic design.

JRM-28, a Novel HDAC2 Inhibitor Enhances Cognitive Function and Memory Gene Expression in the Hippocampus of AD Mouse Models.

Speaker: Omolola Adedeji

Co-Authors: A.F.M Towheed Rahman, Avik Roy and Mahmun Hossain

Abstract: Neurodegenerative diseases such as Alzheimer’s disease (AD) are characterized by progressive cognitive decline and synaptic dysfunction, processes increasingly linked to dysregulated epigenetic regulation. Histone deacetylase 2 (HDAC2) has emerged as a key transcriptional repressor of genes involved in learning, memory, and neuronal plasticity, positioning it as a promising therapeutic target. Guided by the pharmacophore of the FDA-approved pan-HDAC inhibitor romidepsin, we designed and synthesized MJM-1 and subsequently developed JRM-28, an HDAC2 inhibitor. Compound identity and purity were confirmed by NMR spectroscopy and mass spectrometry.

In vitro studies using induced pluripotent stem cell (iPSC)–derived cortical neurons from AD patients demonstrated that JRM-28 treatment was associated with enhanced CREB pathway activation, increased expression of synaptic plasticity markers (MAP2, NR2A, and GLUR1), and partial recovery of dendritic spine density. To evaluate functional relevance in vivo, healthy mice were administered JRM-28 and assessed using the Barnes maze and Open field tests. Treated animals exhibited improved spatial learning performance, reduced escape latency, and increased exploratory behavior relative to controls. This was then translated into Alzheimer’s disease mouse models to determine the therapeutic effects of JRM-28.

5xFAD mouse models were administered orally with JRM-28 and subjected to behavioral tests and immunohistochemistry analysis was carried out. Immunofluorescence analysis of hippocampal tissue revealed reduced expression of HDAC2, within the CA1 region, alongside upregulation of CD10-positive interneurons implicated in amyloid beta degradation. Collectively, these findings demonstrate that HDAC2 inhibition by JRM-28 modulates neuroplasticity-associated signaling pathways, supports cognitive function in vivo and may reduce amyloid plaques in the hippocampus region of the brain. Future work includes pharmacokinetics study and synthesis of analogs.

TBD

Speaker: Annabelle Presly

Co-Authors:

Abstract:

TBD

Speaker: Isabella Wyatt

Co-Authors:

Abstract:

Caring for the Carers: A Mixed-Methods Study of the Mental Health Needs of Anti-Trafficking Professionals

Speaker: Margaret McManus

Co-Authors: Krishna Priya Arunkumar, Jade Ann Wierzchowski, Kamiya Arya, Carisa Cota Wright; Addie Mumey, MSc* Dr. Srishti Meera Sardana, MSc, PhD* (senior authors*)

Abstract: Anti-trafficking staff are routinely exposed to survivors’ trauma, but little research has examined their wellbeing. This mixed-methods study aims to describe the mental health needs of anti-trafficking professionals, identify what drives vicarious trauma and burnout, and generate practical recommendations for staff care. A global online survey will measure stress, burnout and secondary traumatic stress, analyzed descriptively and compared by role (frontline vs. leadership). Virtual focus group discussions (upto 6 participants each) will explore impacts, coping, support gaps and recommendations, using team-based thematic analysis. The two strands will then be integrated. We expect high levels of secondary traumatic stress, different stressors by role, and gaps between the supports staff want and what agencies provide. The results will inform evidence-based staff-care strategies that support workforce sustainability and quality of services for survivors.

Reimagining the Therapy Room: A Roadmap Toward Personalized and Scalable Nature-Integrated Mental Health Care for Underserved Communities Globally

Speaker: Krishna Arun

Co-Authors:

Abstract: Srishti Meera Sardana

Objectives: This study protocol proposes a three-phase model aimed at conceptualizing the impact of therapy environments on the effectiveness of evidence-based psychotherapy for historically underserved communities. Methods: Phase 1 will conduct a pilot randomized trial comparing eight sessions of manualized Cognitive Behavioral Therapy (CBT) delivered in designated campus greenspaces (Nature-Integrated CBT; NI-CBT) versus a traditional clinic setting among university students with mild-to-moderate depression and/or anxiety. Psychophysiological indicators at both, the patient- and provider-levels including clinical symptoms and heart rate variability, as well as treatment outcomes data (example, engagement, therapeutic alliance, nature relatedness, cognitive flexibility, and participant and therapist perceptions) will be assessed. Phase 2 will investigate for whom and under what conditions NI-CBT is most beneficial with the goal of developing a person–environment–intervention matching framework. Phase 3 will translate these findings into a formal clinical decision framework, therapist training resources, and implementation guidance to support scalable adaptation across diverse communities and settings. Anticipated Results: Our phasic approach will help establish proof of concept and identify preliminary mechanisms indicative of improved mental health for both, the patients and the providers. Conclusions: Altogether, this program aims to create guidelines for cost effective psychotherapy that is personalized, evidence-informed, and supportive for historically underserved communities globally.

Subjective Cognition: The Decline And Gains Older Adults Notice Most

Speaker: Patricia Iwok

Co-Authors: Cara Charles, Dr. Cait Bowman

Abstract: Many cognitive abilities are known to change with age, but much of the existing research has documented these changes using laboratory-based tasks that focus on differences in memory, attention, and processing speed. While this work has been important, it does not always reflect how cognitive change is actually experienced in everyday life. Understanding subjective cognition is important because how older adults perceive changes in their thinking can shape well-being, daily functioning, and even objective performance. This study examined subjective cognition in adults aged 60 years and older in the Milwaukee area using a self-report survey informed by the Everyday Cognition framework. Participants compared their current cognitive functioning with how they remembered functioning in their 20s and rated perceived change across multiple domains, including memory, attention, processing speed, problem-solving, language, learning, spatial skills, and emotional regulation. They also rated how important it was to maintain or improve each ability in daily life, and open-ended responses provided additional context. The findings suggest that older adults’ subjective experiences of cognitive aging do not fully mirror the patterns most often emphasized in laboratory-based research. Instead, participants described both perceived declines and perceived gains across domains. These results highlight the importance of understanding cognitive aging not only through objective measures, but also through the lived experiences and priorities of older adults, and they may help inform future research and community-based approaches to healthy aging.

Decoding the Regulatory Circuit Underlying Biosynthesis of the Novel Antimicrobial Compound RejuAgro A in Pseudomonas soli 0617-T307

Speaker: Ton Nu Bao Vy Huyen

Co-Authors: Ching-Hong Yang

Abstract: Antibiotic resistance is not only a major challenge in human medicine but also an emerging threat to sustainable crop production. Fire blight, caused by Erwinia amylovora, is one of the most destructive bacterial diseases affecting apple and pear production. For decades, growers have relied on antibiotics such as streptomycin (STR) for disease management; however, the widespread use of these compounds has contributed to the emergence of STR-resistant E. amylovora populations and increased regulatory restrictions on antibiotic use, particularly in organic production systems. Therefore, alternative antimicrobial strategies are urgently needed.

We isolated Pseudomonas soli 0617-T307 from Wisconsin soil and discovered that it produces RejuAgro A (RAA), a novel antimicrobial metabolite with strong activity against E. amylovora, including STR-resistant strains. Multi-state field trials demonstrated that RAA provides fire blight control comparable to streptomycin, highlighting its potential as an alternative tool for sustainable disease management. Although the biosynthetic gene cluster responsible for RAA production has recently been identified, the regulatory mechanisms controlling RAA biosynthesis remain unknown. Here, we integrated bioinformatic analysis, genetic mutagenesis, and green fluorescent protein reporter assays to identify RasR, a transcriptional regulator required for activation of the RAA biosynthetic pathway. Furthermore, we revealed an autoregulatory feedback mechanism that modulates RAA production. These findings provide new insights into the regulation of antimicrobial metabolite biosynthesis and establish a foundation for engineering P. soli 0617-T307 as a microbial cell factory for enhanced RAA production.

AI-Enabled Hyperspectral Microscopy for Detection and Quantification of Salmonella Typhimurium in Mixed Cultures

Speaker: Meili Papa

Co-Authors: Margaret Broderick, Bosoon Park, Jiyoon Yi.

Abstract:

Introduction: Image-based pathogen detection enables rapid screening, but accurate detection and quantification of target pathogens in mixed cultures are limited by background microbial interference, motivating the application of artificial intelligence (AI). 

Purpose: This study aims to develop and validate an AI-enabled hyperspectral microscope imaging workflow for detection and quantification of Salmonella Typhimurium in mixed cultures, leveraging synthetic training data to enable scalable applications. 

Methods: Five bacterial species (Salmonella Typhimurium, Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, and Pseudomonas fluorescens) were cultured for 16 h in tryptic soy broth with yeast extract at 37°C prior to imaging. In mixed cultures, non-target species were maintained at 10^5 CFU/mL, while Salmonella concentrations were varied from 10^3–10^6 CFU/mL to evaluate detection and quantification performance. Samples were prepared by placing 2 µL of culture onto glass slides, air-drying, and rehydrating with 1 µL of sterilized deionized water, followed by darkfield hyperspectral microscopy (100× objective) to acquire hyperspectral data (n=500). These pure culture data were normalized and reduced by principal component analysis to generate composite images used to train deep learning-based object detection models augmented with synthetic mixed culture data. Model performance was evaluated using accuracy, and image-based quantification and limit of quantification were established by relating detected instance counts to inoculum concentration and validated against selective plate counts. 

Results: Detection accuracy for Salmonella Typhimurium in mixed cultures improved from approximately 61% using pure culture only training to 76% with synthetic mixed culture augmentation on held-out test data (n=100 images). Image-based enumeration in mixed cultures showed a clear proportional relationship between detected instance counts and initial inoculum concentration (10^3–10^6 CFU/mL), consistent with selective plate counts. 

Significance: These results demonstrate a scalable AI approach for pathogen detection and quantification that reduces reliance on extensive mixed culture training datasets, supporting adoption of image-based methods in food safety applications. 

Antibacterial Efficacy of Alcohol Functionalities and Copper Metal-Organic Frameworks on Modified Cotton Fabrics

Speaker: Faith Rothkopf

Abstract: Antibacterial resistance has been on the rise from extended overuse of antibiotic medications. Previous research has indicated that Copper Metal-Organic-Frameworks (MOFs) possess the ability to inhibit bacterial growth through the regulated release and rationing of copper ions. A MOF is a porous molecular structure composed of a metal ion and an organic linker. This research project uses copper as the metal ion for its inherent antibacterial properties and 1,3,5-benzene tricarboxylic acid (BTC) as the organic linker. However, various alcohols have been used as disinfectants in healthcare and laboratory settings for many decades utilizing a process called denaturation, which is currently impermeable to adaptive changes the organisms could attempt. Previous research utilized the foundational knowledge of alcohol as a disinfectant to synthesize a reusable antimicrobial bandage. This research project aims to compare the antibacterial ability of copper MOFs and alcohol functionalities bound to cotton fabrics as a bandage. After the modification of the cotton, synthesis involved either esterification to covalently connect alcohol moieties or MOF addition. The exploration of the antibacterial abilities began with E. coli controls and Kirby Bauer type experimentation. Preliminary results indicate the presence of effective antibacterial properties on both bandage types.

Isolation of Carbapenem-Resistant Non-fermentative Gram-negative Bacteria in Fresh Microgreens and Sprouts

Speaker: Clovis Cormier-Tardif

Co-Authors: Xinhui Li

Abstract: Antimicrobial resistance (AMR) is a growing public health issue. Simply put, bacteria can be resistant to the antibiotics used to treat them through different mechanisms. Fresh vegetables may serve as a vehicle for spreading antibiotic-resistant bacteria, as they’re minimally processed (not cooked) prior to consumption. Carbapenem-resistant non-fermentative Gram-negative bacteria, such as Stenotrophomonas, Acinetobacter, and Pseudomonas, are among the organisms that can cause nosocomial infections and are very difficult to treat. This study investigated carbapenem-resistant non-fermentative Gram-negative bacteria in fresh microgreens and sprouts. Commercial products of microgreens and sprouts were purchased from grocery stores in Wisconsin, Minnesota, and Illinois. Samples were enriched in Dijkshoorn medium, and CHROMagar™ Acinetobacter with the MDR supplement was used to isolate resistant bacteria. Disk-diffusion assay was used to examine carbapenem resistance. Selective isolates were then identified through 16S rRNA gene sequencing. Carbapenem-resistant bacteria were isolated from different sprout and microgreen samples from different farms. These bacteria include Acinetobacter baumannii, Stenotrophomonas pavanii, S. maltophilia, S. hibiscicola, Pseudomonas plecoglossicida, and Empedobacter brevis. Some of these isolates showed intermediate resistance to carbapenem antibiotics. Some of the bacteria, such as Acinetobacter and Stenotrophomonas, are clinically significant. In addition, resistance-conferring plasmids have been found to contribute to the dissemination of AMR in Acinetobacter. Fresh microgreens and sprouts could serve as reservoirs for AMR, underscoring the importance of controlling AMR in these food products. Additional characterization of these isolates, such as whole-genome sequencing, could provide more insight into their roles in AMR.

Beyond Antibiotics: RejuAgro A as a natural antimicrobial metabolite for citrus disease management

Speaker: Shreyashi Mitra

Co-Authors: Jian Huang, Dr. Ching-Hong Yang

Abstract: Citrus production is increasingly threatened by Huanglongbing (HLB) and citrus canker, which have caused substantial yield and economic losses, including severe impacts on the Florida citrus industry. In citrus-producing regions where these diseases remain absent or under regulatory control, their introduction also poses a major threat to production and market access. The 2024 U.S. EPA ban on streptomycin use in citrus has further intensified the need for effective non-antibiotic alternatives. We evaluated RejuAgro A (RAA), a natural antimicrobial metabolite produced by Pseudomonas soli 0617-T307, for the management of HLB and citrus canker. In a two-year field trial initiated with Candidatus Liberibacter asiaticus (CLas)-free young sweet orange trees, foliar applications of RAA at 10, 20, and 30 mg/L delayed HLB symptom development and slowed disease progression in a dose-dependent manner, accompanied by reductions in cumulative fruit drop. Mechanistic studies indicate that RAA acts against HLB primarily through a novel host-directed mechanism that mitigates CLas-induced oxidative stress, reduces excessive ROS/H₂O₂ accumulation and associated physiological and phloem dysfunction, and improves tree health and resilience. In parallel, foliar applications of RAA at 20–30 µg/mL significantly suppressed citrus canker caused by Xanthomonas citri subsp. citri, with efficacy comparable to streptomycin applied at 1,200 µg/mL. Together, these findings demonstrate the potential of RAA as a low-dose, natural alternative.

Careers Panel

Speaker:

Co-Authors:

Abstract:

TBD

Speaker: Prof. Kavi Mehta (FAC)

Co-Authors:

Abstract:

TBD

Speaker: Nathaly Cormier, MS, PhD (FAC)

Co-Authors:

Abstract:

Design, Synthesis and Preclinical Evaluation of Boron-Based Niclosamide Prodrugs for Nonalcoholic Steatohepatitis

Presenter: Megan Honn

Co-Authors:

School:

Abstract:

Evaluation of Pediatric Cervical Spine Response in Far-Side Oblique Impacts Using a 3-Year-Old Human Body Model

Presenter: Kushal Devkota

Co-Authors: Ganesh Shete, Karthik Somasundaram

School: Medical College of Wisconsin

Abstract: Far-side oblique impacts can produce substantial lateral head excursion and complex cervical spine loading in pediatric occupants. Previous sled testing with the Q3s 3-year-old anthropomorphic test device (ATD) demonstrated that increased top-tether restraint reduced lateral head excursion but was accompanied by elevated upper-neck axial tension, suggesting a potential trade-off between occupant containment and cervical loading. However, the simplified mechanical neck of the ATD cannot reproduce the multi-segmental motion of the pediatric cervical spine. This study evaluates this response using the biofidelic THUMS 3-year-old human body model (HBM). Explicit LS-DYNA simulations replicate previously published far-side oblique sled conditions consisting of a 20° oblique FMVSS 213 pulse (ΔV = 31.3 km/h; peak acceleration = 21.0 g), a forward-facing child restraint system, and an adjustable-head-restraint seat configuration. The Q3s ATD is replaced with the THUMS 3-year-old HBM while maintaining comparable boundary and restraint conditions. Model response will be evaluated using three-dimensional head excursion, head rotational kinematics, BrIC, and upper-neck axial force. In addition, cervical spine kinematics will be examined at upper, middle, and lower cervical regions to characterize segmental contributions to overall neck flexion and head motion. Simulations and quantitative analyses are currently underway. Comparison of HBM responses with the previously reported Q3s response will help determine whether anatomical cervical flexibility alters head excursion, rotational response, and neck load transfer during far-side loading. This work demonstrates the potential value of pediatric HBMs for investigating neck injury mechanisms that cannot be fully characterized using conventional ATDs.

Enhancing Robot-Assisted Lower-Limb Rehabilitation with MPRL-Driven Dynamic Difficulty Adjustment in Serious Games

Presenter: Steven Zhang

Co-Authors: Md Enamul Haque (co-presenting), Mohammad H. Rahman

Bio-Robotics lab, University of Wisconsin-Milwaukee

School: University of Wisconsin-Milwaukee

Abstract: Traditional lower-limb rehabilitation often relies on therapist-assisted, repetitive exercises that can become monotonous and lead to reduced adherence. We developed a suite of three serious games designed to improve engagement for robot-assisted rehabilitation: a 2D flappy bird-style vertical runner for motion priming, a skiing endless runner for physiotherapeutic exercise, and a task-based soccer game for simultaneous cognitive and motor planning. Each game utilizes a Multiple-periodic reinforcement learning(MPRL) difficulty adjustment framework coupled with the patient’s lower-limb joint angle measurements. Unlike traditional heuristics-based or single-period difficulty adjustment methods, the MPRL system decouples short-term performance targets from long-term mastery trends to maintain the patient’s “flow state” without sudden challenge disruptions. To verify the game’s control system, the game was interfaced with a Webots simulation of the SmartB2R three degree-of-freedom lower limb robot. This demonstration proved the capability of the control system to accurately map and animate the patient’s joint movements. The closed-loop Webots simulation successfully demonstrates real-time telemetry ingestion, smooth kinematic control of the virtual avatar, and automated difficulty scaling based on dynamic performance metrics. For safety and additional individualization to the patient, a complimentary calibration system is included in each game, ensuring that the game movements stay within the joint range of each patient. Future work will involve deploying the game onto physical robot hardware and validating the long-term effectiveness of the MPRL framework through comprehensive clinical trials.

Microgravity-Induced Mechanical Memory in Mesenchymal Stem Cells and Modulation by Low-Dose Radiation

Presenter: Grace Noerenberg

Co-Authors: Joy Zhao, Priyatha Premnath, Laura Bowman, Anthony Lau, Qingsu Cheng.

School: University of Wisconsin-Milwaukee

Abstract: Mechanical cues such as substrate stiffness are known to induce lasting “cellular memory” in mesenchymal stem cells (MSCs), influencing their structure and differentiation potential. However, it remains unclear whether microgravity can induce similar mechanical memory and how space-relevant stressors, such as low-dose radiation, modulate this effect. This study investigates the presence, persistence, and functional consequences of microgravity-induced cellular memory in bone marrow–derived MSCs (BMMSCs), with implications for spaceflight-associated musculoskeletal degeneration and terrestrial disuse conditions.

Developing a Cost-Effective and Portable Visible Spectrophotometer for Academic Application

Presenter: Layla Ogango

School: University of Wisconsin-Oshkosh

Abstract: The purpose of this research project is to design a cost-effective and portable visible spectrophotometer, a helpful research tool that can be used for STEM-related courses in higher education. Spectroscopy studies how light interacts with different elements and different forms of electromagnetic radiation on the spectrum. Spectrophotometers are research devices used for this particular area of study that measures light once it comes in contact with a sample, that absorbs and measures this light within a spectrum that consists of a range of wavelengths. Visible spectrophotometers measure light in a visible spectrum whose wavelengths range from 400nm-700nm, roughly speaking. The primary issue found within spectroscopy techniques, whether that be molecular spectroscopy, elemental spectroscopy, or mass spectrometry, is both ensuring portability and stable costs for spectrometric research devices. What we want to control and personally modify in constructing this analytical device is both the nature of the design coupled with its estimated cost by creating a system that can perform quantitative and rapid analyses and deliver accurate results while having the necessary but affordable components that allow for this type of function. We are hoping to improve and expand the quality of research and learning in higher education by building a system that can be used to complete research tasks both in a laboratory and in a field. The final prototype was tested using different light sources and was shown to produce readable but less defined graphs with high resolution for each experiment conducted.

Mechanistic Modeling of Mitochondrial Calcium Regulation of Bioenergetics

Presenter: Yisak Dereje Asefa

Co-Authors:

School:

Abstract:

Progressive Deterioration of Social Recognition Memory Linked to Entorhinal-CA2 Network Dysfunction Across Aging

Presenter: Alexis Becerra Casillas

Co-Authors: Said H Audi, Allen W Cowley Jr, Ranjan K Dash

School: Medical College of Wisconsin

Abstract: Mitochondrial Ca2+ uptake is mediated by the mitochondrial calcium uniporter (MCU) complex, in which MICU1/2 act as cytosolic Ca2+-sensing gatekeepers that impose a Ca2+-dependent activation threshold. Coordinated control of MCU activity is critical because mitochondrial Ca2+ entry couples cytosolic Ca2+ signals to metabolic activation and must be regulated to avoid Ca2+ overload and bioenergetic dysfunction. We developed a biophysical MCU model that incorporates explicit MICU1/2-dependent gating together with a thermodynamically constrained Ca2+ transport formulation that accounts for Mg2+ inhibition and membrane potential dependence. Subsequently, mitochondrial Ca2+ dynamics were simulated by integrating the MCU model within a mitochondrial cation-handling model under multiple Ca2+ stimulation protocols. Because matrix-side Ca2+ regulation of MCU remains controversial, we additionally evaluated a putative matrix-side regulatory mechanism by comparing simulations performed with and without an added matrix-side regulatory module, rather than assuming such regulation as a required feature of the base model. The model reproduces key experimental behaviors across genotypes. In wild-type mitochondria, MCU flux is negligible below a cytosolic Ca2+ threshold (~0.2 µM), whereas MICU1-knockout mitochondria show constitutive uptake and MICU2-knockout mitochondria exhibit an intermediate, lowered threshold. In comparison simulations, inclusion of matrix-side regulation transiently attenuated MCU flux over an intermediate matrix [Ca2+] range, producing higher transient cytosolic Ca2+ and lower transient matrix Ca2+, while both cases approached similar steady states. In addition, cytosolic Mg2+ acts as a graded inhibitor of MCU-mediated Ca2+ uptake, limiting matrix Ca2+ loading. These results provide a quantitative framework for MCU-mediated mitochondrial Ca2+ uptake and dynamics across physiological and perturbed conditions.

The neurobiological link between itch and anxiety

Presenter: Avleen Benipal

Co-Authors: Nicole Kooij, Laney Waydick, Tayler Sheahan

School: Marquette University

Abstract: Itch is a debilitating sensory disorder that is associated with stress, anxiety, and other mood disorders. Despite this, the neuronal mechanisms linking itch and anxiety remain poorly understood. To evaluate this, we performed immunohistochemistry on brain sections from mice treated with the acute itch caused by compound 48/80 (C48/80) and vehicle-treated controls. Expression of cFOS, a marker of neuronal activity, was measured within brain regions that are associated with itch and anxiety. We found that mice treated with C48/80 exhibited a greater number of cFOS-positive neurons in the parabrachial nucleus and central amygdala. Following this, we wanted to investigate if chronic itch induced by squaric acid dibutyl ester (SADBE), an ACD model, influences anxiety-like behaviors in mice. Our recent studies have shown that SADBE-treated mice have increased spontaneous itch behavior compared to vehicle-treated controls. Furthermore, other studies have demonstrated an association between chronic allergic contact dermatitis and anxiety-like behaviors. We therefore hypothesized that SADBE-treated mice would exhibit increased anxiety-like behaviors compared to vehicle-treated controls. To measure anxiety-like behaviors, we performed a series of validated behavioral assays (open field test, elevated plus maze, and marble burying test) and examined whether anxiety-like behaviors were increased in SADBE-treated mice compared to vehicle-treated controls. Future analyses will examine neuronal activation in brain regions implicated in anxiety to better understand how chronic itch drives anxiety. A clearer understanding of the neurons involved could help guide future studies supporting the development of more effective therapeutic strategies for patients experiencing both chronic itch and the associated psychological burden.

Sex- and time-dependent cortico-striatal sub-circuit regulation of opioid taking

Presenter: Cassidy Kraemer

Co-Authors: Shalana Atwell, Jason Cruz, Salma Elsherbiny, Ayah Sbeih, Nathan Kehl, Alex Cain, Matthew Hearing.  

School: Marquette University

Abstract: The importance of the rodent prelimbic cortex (PrL) in drug-related behaviors has been highlighted in part by in vivo electrophysiological recordings in behaving animals. While insightful, these seminal studies have lacked selectivity for projection targets, contributing to incongruent findings during similar behaviors. The PrL projections to the nucleus accumbens core (PrL-NAc) have frequently been implicated in most forms of drug-seeking; however, work to date has assessed this exclusively under reinstatement or relapse conditions. Thus, how the PrL as a whole and the PrL-NAc circuits contribute to volitional opioid taking compared to non-drug reward taking is lacking, with no data regarding whether this contribution shifts with increased opioid exposure. Here, we used excitatory or inhibitory chemogenetic approaches to manipulate the PrL globally and PrL-NAc pathway to test their necessity in drug (remifentanil) and non-drug (Ensure) taking. Animals received intracranial viral surgeries and catheters to self-administer remifentanil for up to 30 days with clozapine-n-oxide (CNO) administered on approximately days 14 and 30. Inhibiting the PrL globally significantly reduced drug taking behaviors in both males and females at 14 days, but only in males at 30 days. Interestingly, inhibiting the PrL-NAc pathway significantly reduced drug taking in males and females at 14 and 30 days of self-administration while excitation of this pathway had no effect. Furthermore, we found no effect on non-drug taking behavior when inhibiting this pathway. Additionally, these data identify sex differences in global PrL regulation of drug taking but overlapping roles within the PrL-NAc and the necessity of subcircuit specificity in regulation of drug intake. This research highlights a potential therapeutic target during early phases of drug taking that could attenuate opioid use progression.

Entorhinal Representational Geometries Underlying Social Memory and Generalization

Presenter: Davis Lesperance

Co-Authors: Jose Martinez-Cabral, Lorenzo Fusi, Lara Boyle, Stefano Fusi, Jeffrey Lopez-Rojas

School: University of Wisconsin-Milwaukee

Abstract: Episodic social memory requires abstraction across encounters with the same individual while preserving episode-specific details. How these competing demands are represented in cortical circuits projecting to hippocampus remains poorly understood. Here, we used projection-specific calcium imaging of lateral and medial entorhinal cortex (LEC and MEC) inputs to dorsal CA2 (dCA2) while mice explored familiar littermates and novel conspecifics whose identities and positions were swapped across trials. We analyzed the representational geometry of population activity across all combinations of familiarity, identity, position, and trial. These analyses revealed distinct population codes in LEC and MEC during social exploration. In LEC, population activity formed a high-dimensional representation in which familiar and novel partners occupied distinct regions of representational space. High-dimensional population codes typically support separability of multiple task variables but can limit generalization across changes in context. Nevertheless, LEC exhibited strong cross-condition generalization performance for social familiarity across individual identities and spatial positions. In contrast, MEC activity was dominated by spatial position, with a lower-dimensional, position-aligned geometry and minimal evidence for abstract social coding. Dimensionality analyses further showed that LEC representations were distributed across many axes, whereas MEC representations were more compact and spatially anchored. Together, these results indicate that LEC→dCA2 population activity contains information about multiple social and episodic variables while retaining an abstract familiarity-related signal, whereas MEC→dCA2 population activity provides a more spatially anchored code during social exploration.

Sterol Carrier Protein 2 (SCP-2) Expression and Lipid Uptake Dynamics: A Pathway Toward Novel Anxiety Targets

Presenter: Greta Vickery

School: University of Wisconsin-Milwaukee

Abstract: Anxiety affects over 350 million people worldwide, and with only one in four achieving remission, it is essential to explore the therapeutic potential of alternative signaling systems such as the endocannabinoid system (ECS). The ECS influences threat-processing networks in the central nervous system by controlling the balance of inhibitory GABA and excitatory glutamate release via retrograde signaling. Endocannabinoids (eCBs) are transported within the aqueous cellular environment by amphipathic proteins, one of which is sterol carrier protein-2 (SCP-2). It has been shown in mice that inhibition of SCP-2 reduces anxiety, possibly due to a higher concentration of eCBs in the synapse. Our overarching hypothesis is that SCP-2 localizes near the cell membrane to bind eCBs and remove them from the synapse. Our current research focuses on where SCP-2 localizes and moves in the cell, as well as how its expression differs based on brain region and lipid cargo. Confocal microscopy allows us to assess where SCP-2 moves within HEK cells following binding to various lipids. We predict that SCP-2 will initially be localized at the membrane and move into the cell upon binding to a lipid. Immunohistochemistry (IHC) provides a way to study brain region and cell-specific localization of SCP-2 in mice. We are interested in where SCP-2 resides within specific brain regions to allow for the development of mechanistic hypotheses. Insight into the location and movement of SCP-2 will support the development of a novel therapy for anxiety.

Comparison of Escherichia coli Quantification Analysis for Winnebago County

Presenter: Ivan Abramovich

Co-Authors: Jackie Behrens, Brittanee Dupree, Brianna Elizondo, Greg Kleinheinz

School: University of Wisconson-Oshkosh

Abstract: Coliform bacteria, specifically Escherichia coli, represents bacteria that can cause negative changes of human health. Escherichia coli is frequently found in surface water that represents beach water and water surrounding boat launches, often increasing when heavy rainfall or avian fecal material is present. Due to weather, animal and human impacts, it is difficult to predict Escherichia coli concentrations. Measuring Escherichia coli is a reactive, dynamic effort that can be completed with multiple methods; however, emphasis is placed upon efficient and valid quantifications. Other methods are being considered that involve aspects of automation to reduce significant delays in reporting the level of Escherichia coli and determining if the location is safe to perform recreational activities. Three recreational beaches across Winnebago County were selected and monitored during the summer of 2026 to compare the concentration of Escherichia coli to observe for patterns between two quantification methods. The samples were prepared with their respective methods in order to use the IDEXX Quanti-Tray and the IDEXX TECTA to determine the amount of Escherichia coli in the beach water. The IDEXX Quanti-Tray-Colilert-18 and the IDEXX TECTA was evaluated via graphical analysis to produce similar Escherichia coli concentration values. Winneconne beach and Fresh Air Park produced more consistent concentration values than Menominee beach.

670 nm Photobiomodulation (PBM) Protects Vascular Integrity in Diabetic Retinopathy by Regulating Muller Cells and Balancing Angiogenic Factors

Presenter: Jehiel Smith

Co-Authors: James Baffoe, Quin McCarthy

School: University of Wisconsin-Milwaukee

Abstract: Diabetes mellitus is a chronic disease characterized by high blood sugar and insulin deficiency. Diabetes affects the pancreas, disrupting insulin production and causing insulin resistance, resulting in hyperglycemia. A major complication of diabetes is diabetic retinopathy (DR) Symptoms of DR typically include distorted vision, faded colors, floaters, and blurry vision. Disease progression could lead to total blindness. Molecularly, in DR, persistent hyperglycemia damages blood vessels in the retina via inflammation and excess angiogenesis. These changes are a result of hyperglycemia induced oxidative stress and mitochondrial disfunction. While treatments exist for DR, they are invasive and function to halt blood vessel growth. Earlier treatments that mitigate oxidative stress and mitochondrial dysfunction could prevent development and progression of DR. One such treatment that shows promise is the use of red light at 670 nm which functions as an oxidative stress reliever and mitochondrial activity promoter. Therefore, we hypothesize that 670nm red-light treatment mitigates early oxidative stress events leading to the decrease angiogenic protein expression in hyperglycemic retinal dysfunction. We will use an in vitro model of DR, Rat Müller cells exposed to high glucose to examine changes in angiogenic genes via RT-qPCR and angiogenic protein via Western blot We expect our findings to further define the mechanism by which 670nm is beneficial in preliminary treatment of DR.

 Population Density of Escherichia coli in the Kinnickinnic River – Milwaukee, WI

Presenter: Jorden Budny

Co-Authors: Rilee Koplinski, Cristina Contreras, Megan Krueger

School: Alverno

Abstract: This study evaluated coliform population densities in the Kinnickinnic River and Lake Michigan during summer 2026. Water samples were collected from three Kinnickinnic River sites and analyzed using the Most Probable Number (MPN) method, with bacterial identification supported by lactose broth, EMB agar, nutrient agar, and Gram staining. Lake Michigan samples were analyzed using two water-testing methods. Coliform densities varied by location and sampling date. Zablocki Park recorded the highest density at 160 MPN on June 23, while Wilson Park showed the most consistent results (16–35 MPN) and Jackson Park the lowest (2–12 MPN). Lake Michigan samples also detected coliform populations, with the 15-tube method producing higher MPN values. These results demonstrate spatial and temporal variation in coliform densities and support the need for additional sampling to better characterize bacterial populations in local waterways.

Perforant-Path Projections from Entorhinal Cortex to Dorsal CA2 in a Schizophrenic Mouse Model

Presenter: Liam Eickhoff

Co-Authors: Michael Mendez, Davis Lesperance, Jenni Andjelic, Jeffrey Lopez-Rojas

School: University of Wisconsin-Milwaukee

Abstract: Social recognition memory is a vital subtype of declarative memory that enables discrimination of familiar and novel individuals required for navigation of complex social settings. The Df(16)A+/- mouse line replicates the 22q11.2 microdeletion associated with significantly elevated schizophrenia risk. The hippocampal CA2 region is critical for social memory and exhibits structural, physiological, and social-encoding abnormalities in Df(16)A+/- mice. Direct input from lateral entorhinal cortex (LEC) to dorsal CA2 (dCA2) conveys multimodal social information required for normal social memory function. However, whether entorhinal perforant-path projections to dCA2 are altered in Df(16)A+/- mice remains unclear.

Unlocking Sterol Transport in Brettanomyces and Beyond Using a  S. cerevisiae Chassis

Presenter: Loria Dao

School: University of Wisconsin-Oshkosh

Abstract: One major advantage of yeast fermentation in foods, beverages, and industry is the ability to grow where oxygen is depleted. Two species of budding yeasts commonly used in the beer environment are Saccharomyces cerevisiae and Brettanomyces spp., which are facultative anaerobes capable of growing in the presence and absence of oxygen. To thrive in anoxic environments, S. cerevisiae must uptake exogenous sterols by expressing genes encoding sterol transporters and cell wall mannoproteins. As Brettanomyces thrives in anaerobic environments like S. cerevisiae, Brettanomyces should have its own putative sterol transporter and cell wall mannoprotein genes, which have been identified via sequence homology but never through functional analysis. To test the functionality of these Brettanomyces genes, we will begin with a S. cerevisiae strain that is missing both genes encoding sterol uptake proteins (aus1Δ and pdr11Δ), making it sterol-uptake-deficient and unable to grow without oxygen. We will delete the URA3 gene from this strain by selecting on plates containing 5’-FOA. Then, we will clone putative sterol uptake genes from different yeast species into a plasmid bearing the URA3 gene and transform them into the sterol-uptake-deficient S. cerevisiae strain. Lastly, we will test for anaerobic growth in anoxic conditions, where growth will confirm the functionality of Brettanomyces sterol transporters.

GATA2 and GATA3 Regulate Early Transcriptional Programs and Fate Determination in Human Embryonic Stem Cell Derived Amnion Model

Presenter: Maliha Kabir

Co-Authors: Lauren Juga, Nikola Sekulovski, PhD; Amber Carleton and Kenichiro Taniguchi, PhD

School: Medical College of Wisconsin

Abstract: GATA transcription factors play central roles in early embryonic development, yet their precise function during human amnion specification remains incompletely defined. During human embryonic development, amniogenesis is triggered in a collection of pluripotent epiblast cells as the human embryo implants. Utilizing human pluripotent stem cell (hPSC)-based models of amniogenesis (called Gel-3D and Glass-3D+BMP), our previous studies showed that amniogenesis requires the activation of BMP signaling to trigger the amniogenic transcriptional cascade, and 3-dimensional cysts with a central lumen uniformly surrounded by squamous amnion cells are seen by day 4 in these systems (Gel-3D, Shao et al., Nature Materials 2017; Glass-3D+BMP, Sekulovski et al., eLife 2023). GATA2 and GATA3 are zinc-finger transcription factors that act downstream of BMP signaling and are among the earliest transcriptional responders during amnion differentiation. Analysis of published in vitro stem cell models together with the Gel-3D and Glass-3D+BMP systems reveals that direct BMP targets, including GATA2 and GATA3, are rapidly activated at the onset of amnion specification. Consistent with prior studies, GATA2/3 expression is highly enriched in the amnion of human and non-human primate embryos relative to epiblast and primitive streak populations. Functional knockout experiments demonstrate that loss of GATA3 disrupts amnion differentiation and abolishes activation of downstream lineage markers such as TFAP2A, ISL1, and HAND1. Using a GATA2/3 double-knockout hPSC line, we generated cysts in both Gel-3D and Glass-3D+BMP models, and performed time-course bulk RNA sequencing. Interestingly, loss of GATA2/3 prevents amnion fate acquisition and instead leads to persistent pluripotent cyst identity in both systems. This differentiation failure is evident as early as 6 to 12 hours post BMP induction and persists at later stages, accompanied by sustained loss of core amnion genes including ISL1, HAND1, and CDX2. Together, these findings establish GATA2 and GATA3 as early, essential mediators of BMP-driven amnion specification and define a narrow temporal window during which GATA activity is required to initiate the amnion transcriptional program.

The Role of ACE2 in Intestinal Epithelial Recovery Following Radiation Injury

Presenter: Mobin Omdeh Ghiasi

School: University of Wisconsin-Milwaukee

Abstract: Radiation therapy is an important treatment for cancer, but it can also damage healthy intestinal tissue and impair its ability to regenerate. Angiotensin-converting enzyme 2 (ACE2) is highly expressed in the intestine and has been linked to intestinal homeostasis and recovery following radiation injury. Our project investigates whether ACE2 plays a direct role in intestinal regeneration after radiation exposure.

We established intestinal organoids from wild-type (WT) and ACE2 knockout (KO) mice to compare intestinal epithelial responses in the presence or absence of ACE2. Intestinal crypts were isolated, cultured in Matrigel, and allowed to develop into budding organoids. RNA was isolated from organoids, converted to cDNA, and analyzed using TaqMan RT-qPCR to evaluate stem cell and differentiation markers. Organoid survival, morphology, and crypt budding will also be monitored using Incucyte live-cell imaging following radiation exposure.

Preliminary RT-qPCR results before radiation confirmed reduced ACE2 expression in knockout organoids. We also observed increased MUC2 expression in male knockout organoids compared with female knockout organoids, suggesting a possible sex-dependent difference that is currently being investigated. Overall, this study will help determine how ACE2 affects intestinal recovery following radiation-induced injury and may provide insight into strategies to reduce gastrointestinal toxicity during cancer treatment.

The cancer-associated RHOA-R5W mutant is activated by RhoGEF SmgGDS and drives diffuse gastric cancer through activation of PKN effector signaling networks

Presenter: Nasya Miller

School: Medical College of Wisconsin

Abstract: The understudied diffuse gastric cancer (DGC) is the most lethal gastric cancer subtype, lacking clinically effective therapies. Contrary to other subtypes, DGC is increasing incidence in younger patients and is responsible for 30-45% of the >1.1M new gastric cancer cases annually worldwide. Approximately 25% of DGC patients harbor somatic missense mutations in the small GTPase RHOA. The most common RHOA mutations are R5W, Y42C, and L57V. These mutations are unique to RHO GTPases, making extrapolations from the key cancer driver RAS limited in value. Thus, the cancer-driving mechanisms underlying RHOA-mutant DGC are poorly understood, restricting the development of targeted therapies. We hypothesize that RHOA-R5W, -Y42C, and -L57V are activated by distinct RhoGEFs which induces mutation-specific effector signaling driving variant-dependent tumorigenic potencies. Previously, we identified RHOA-Y42C and -L57V as activating oncogenic mutations that stay longer in their active state and drive DGC through mutation-specific mechanisms. The mechanisms by which RHOA-R5W promotes DGC are unknown. We found that RHOA-R5W increases F-actin stress fiber formation, a classical readout for RHOA function. The R5W mutation impairs RHOA activation by canonical Dbl-family RhoGEFs but retains interaction with the non-canonical RhoGEF SmgGDS. Quantitative immunofluorescence microscopy studies suggest that SmgGDS is required for R5W-induced F-actin stress fiber formation. Currently, we are investigating the role of SmgGDS for RHOA effector signaling (e.g., PKN) in patient-derived DGC cell models. In conclusion, RHOA-R5W is, as Y42C and L57V, an oncogenic mutation in DGC that induces cellular RHOA functions through mutation-specific interactions with typical and atypical RhoGEFs.

Stimulation of Npy2r-expressing vagal sensory neurons inhibits breathing in health and chronic lung disease

Presenter: Nicole Hennen

Co-Authors: Luke Ehlert, Taylor Holmes, Brooke Dir, Doriann Pina, Gary Mouradian

School: Medical College of Wisconsin

Abstract: The respiratory system is not fully developed in infants born extremely prematurely, so they often need high levels of supplemental oxygen (hyperoxia; HX) for survival. However, sustained use of HX impedes ongoing lung development and leads to chronic lung disease. Pulmonary vagal sensory neurons (PSNs) marked by the expression of Neuropeptide Y receptor 2 (Npy2r) innervate the alveoli, which are damaged after HX. Therefore, the objective of this study was to define the influence of Npy2r+ PSNs in health and HX. To study this, mice were exposed to 90% O2 to induce lung damage or room air as control (normoxia; NX). For optogenetic studies, mice which express the light-sensitive protein ChR2 within Npy2r PSNs were used. For chemogenetic studies, Npy2r-Cre driver mice were intratracheally treated with a Cre-dependent retrograde AAV to drive expression of the excitatory DREADD, hM3Dq (Gq). Mean linear intercept of HX lung tissue increased 128% compared to NX lung tissue, confirming alveolar simplification. Optogenetic stimulation of Npy2r+ PSNs decreased frequency (21%), tidal volume (70%), and minute ventilation (18%) in NX and HX Npy2rCre+Ai32 mice (percent change of baseline; n>7 per group). Without anesthetic influence, chemogenetic stimulation revealed that the 60% reduction in minute ventilation was driven by frequency in NX mice but tidal volume in HX mice (n>5 per group). These data suggest an inhibitory breathing role of Npy2r+ PSNs, and that HX alters the influence PSNs have on ventilatory control. Current studies are testing the impact of Npy2r+ PSN inhibition in NX and HX mice.

Investigating the Contribution of Platelet Activation to Embryonic Survival in TFPI–Deficient Mice

Presenter: Obinna Anyamele

School: University of Wisconsin-Madison

Abstract:

Background: Tissue Factor Pathway Inhibitor (TFPI) is a major regulator of tissue factor–initiated coagulation. Mice completely lacking TFPI do not survive embryonic development. Because platelets contribute to coagulation, vascular integrity, and embryonic vascular development, we hypothesized that excessive platelet activation contributes to this lethal phenotype and that in utero platelet inhibition may improve TFPI-null embryo survival. Understanding this relationship may clarify how coagulation and platelet activity influence embryonic development with broader relevance to pregnancy complications and vascular diseases involving abnormal clotting.

Methods: TFPI heterozygous mice were intercrossed to generate wild-type, heterozygous, and TFPI-null offspring. Pregnant dams received aspirin and ticagrelor during gestation to inhibit cyclooxygenase-1- and P2Y12-dependent platelet activation. Embryos were collected at E15.5 and genotyped to assess survival and genotype distribution. Untreated crosses served as baseline controls. Blood collected from the maternal inferior vena cava was assessed by whole-blood aggregometry using ADP stimulation to confirm platelet inhibition.

Results: Whole-blood aggregation demonstrated reduced ADP-induced platelet responsiveness following treatment. Untreated pregnancies produced approximately 33% wild-type, 66% heterozygous, and 0% TFPI-null embryos. Following treatment, no viable TFPI-null embryos were detected at E15.5. However, surviving embryos shifted to 16.7% wild-type (6/36) and 83.3% heterozygous (30/36), with fewer viable progeny per litter.

Conclusions: Aspirin and ticagrelor did not rescue TFPI-null embryonic lethality. However, the altered genotype distribution and reduced progeny following treatment suggests genotype-dependent effects of the treatment on embryonic survival that warrant further investigation.

The Role of TAL1 in the Development and Function of NK Cells

Presenter: Oluwabusola Ajagbe

Co-Authors:

School:

Abstract:

Establishing a Histological Approach to Investigate the Blood-Retinal Barrier in the 13-Lined Ground Squirrel

Presenter: Tyra Tye

Co-Authors:

School:

Abstract:

Serotonergic psychedelics regulate neuroplasticity in the rat striatum

Presenter: Alyssa Ottum, BS

Co-Authors:

School:

Abstract:

From Slaughterhouse to Lab: Extraction of Primary Fibroblasts from Bovine Lung

Presenter: Brooke Johnson

Co-Authors:

School:

Abstract:

Effects of doxorubicin-treated BT549 cell conditioned media on DAMI megakaryocytes

Presenter: Carver Nelson

Co-Authors:

School:

Abstract:

Modulating stress susceptibility via ERβ- and circuit-specific mechanisms

Presenter: Emily Walters

Co-Authors:

School:

Abstract:

Metabolic Versatility and Biofilm Dynamics of Novel Deinococcus fluvensis Isolates from the Fox-Wolf Watershed.

Presenter: Grant Johnson

Co-Authors:

School:

Abstract:

Complementary Social and Spatial Representations Through Parallel Entorhinal-CA2 Projections

Presenter: Jose A. Martinez-Cabral

Co-Authors:

School:

Abstract:

Psychedelic Psilocybin and DOI’s Effect on Nucleus Accumbens Core Structural and Functional Synaptic Plasticity.

Presenter: Kaitlyn Rhyner

Co-Authors:

School:

Abstract:

Establishing an Astrocyte Model to Investigate SOD1 Function in ALS Through CRISPR-Cas9 Gene Editing

Presenter: Keith M. Gardner

Co-Authors:

School:

Abstract:

Upper Limb Myoelectric Prostheses With Arduino in Transradial Amputees: Development, Functionality, and Cortical Neuroplasticity

Presenter: Letícia Rodrigues

Co-Authors:

School:

Abstract:

TBD

Presenter: Najeebah junaid

Co-Authors:

School:

Abstract:

Estradiol in Health and Disease: Cell-type-specific regulation of Dorsal Hippocampal memory and Alzheimer’s-like pathology

Presenter: Rutuja Borawke

Co-Authors:

School:

Abstract:

Industry and Occupation Trends in West African Chronic Kidney Disease Research

Presenter: Sarah Hulett

Co-Authors:

School:

Abstract:

Investigating the role of pH in siderophore-iron binding in the yeast Kluyveromyces lactis.

Presenter: Vianna Dao

Co-Authors:

School:

Abstract:

Establishment of GFP/Luciferase Uveal Melanoma Xenograft Models

Presenter: Warren Sauer

Co-Authors:

School:

Abstract:

TBD

Presenter: Adeline Carrell

Co-Authors:

School:

Abstract:

Investigating New Corral Trap Designs to Improve Trapping of Single Nanoparticles

Presenter: Alexander Kuiken

Co-Authors:

School:

Abstract:

Synthesis and Screening of Novel insulin sensatizers

Presenter: Austin Bunkelman

Co-Authors:

School:

Abstract:

The Effects of Nicotine on PTH and PTH1R: How Smoking Can Lead to Osteoporosis

Presenter: Caroline Litsey

Co-Authors:

School:

Abstract:

Benchmarking DFT Methods for the Fe–NO Geometry in a Heme Nitrosyl Model

Presenter: Devin Sage

Co-Authors:

School:

Abstract:

DNAzymes for Protein Lysine Acylation

Presenter: Elek Anderson

Co-Authors:

School:

Abstract:

Total Iron detected in Milwaukee River Water

Presenter: Fatima Yasir

Co-Authors:

School:

Abstract:

The Replication Stress Response and Mutagenic Consequences to DNA Strand-Specific Stalling

Presenter: Gopishankar Thirumoorthy

Co-Authors:

School:

Abstract:

Synthesis of Luminescing Stable Organic Radical: ADPA∙

Presenter: HyungDong Seo

Co-Authors:

School:

Abstract:

Mutational Analysis of a Burkholderia Toxin and Substrate

Presenter: Lucky Paolone

Co-Authors:

School:

Abstract:

A Green Chemistry Approach to the Knoevenagel Condensation of Naturally Occurring Benzaldehydes

Presenter: Mark Mamalakis

Co-Authors:

School:

Abstract:

Impact of the Second Coordination Sphere of the Molybdopterin Cofactor on the Mechanism of Formate Dehydrogenase from Cupriavidus necator

Presenter: Matt Kapelis

Co-Authors:

School:

Abstract:

MnM-W-MMGBSA: Improving Computation of Relative Binding Free Energies of Protein–Protein Interaction Systems

Presenter: Md Nazmul Hasan

Co-Authors:

School:

Abstract:

TBD

Presenter: Mike LeClaire

Co-Authors:

School:

Abstract:

Complex Substrate and Inhibitor Effects on the Reactivity of an Altered Form of Cupriavidus necator Formate Dehydrogenase

Presenter: Mohammad Touhidul Islam

Co-Authors:

School:

Abstract:

Mitigation of chlorine-induced lung injury by nebulized GABA(A) receptor modulator, MIDD0301

Presenter: Mubaraq A Toriola

Co-Authors:

School:

Abstract:

Exploration of New Compounds to Serve as Chemical Inducers of Proximity

Presenter: Ryan Burmesch

Co-Authors:

School:

Abstract:

GAMA : A Robust and Automated Fragment Based Quantum Chemistry protocol for Biomolecular systems

Presenter: Sujan Kundu

Co-Authors:

School:

Abstract:

Determination of Orthophosphate, Hydrolyzable, and Total Phorphorous Concentration by Semi-Automated Colorimetry

Presenter: Ximena Garcia Soto

Co-Authors:

School:

Abstract:

Synthesis, Characterization, and Autoxidative Kinetics of Biomimetic Dicopper(II) Complexes as Catechol Oxidase Models

Presenter: Allen Maly

Co-Authors:

School:

Abstract:

Gut microbial effects on trophallaxis in the honey bee, Apis mellifera.

Presenter: Anushka Patil

Co-Authors:

School:

Abstract:

Synthesis and analysis of octahedral transition metal complexes: Exploring the effects of ligand and metal identity through spectroscopy, magnetism, and crystal field theory

Presenter: Armita Aghamiri

Co-Authors:

School:

Abstract:

Synthesizing benzodiazepines to target the 5-HT2B receptor via Buchwald-Hartwig amination ​

Presenter: Devin Esch

Co-Authors:

School:

Abstract:

Elucidation of the molecular mechanism and physiological consequences of the cross-talks between iron and sphingolipid metabolism to promote cell death in glioblastoma

Presenter: Franca Ohikhuare

Co-Authors:

School:

Abstract:

Optimizing Bovine Serum Albumin–Polyethylene Glycol Conditions as a Model System for Investigating Macromolecular Crowding by Atomic Force Microscopy

Presenter: Griffin Clark

Co-Authors:

School:

Abstract:

Corral trapping of small particles and single molecules in solution

Presenter: Jhoanna Molina

Co-Authors:

School:

Abstract:

Evaluation of metabolism, protein binding, and pharmacokinetics of novel pegylated imidazodiazepines as novel asthma drug candidates

Presenter: Kayode Medubi

Co-Authors:

School:

Abstract:

In Vitro Characterization of PbrRohR: Coupled Kinetic Assay and Crystallographic Studies

Presenter: Kimberly Osborn

Co-Authors:

School:

Abstract:

Molecular Docking Studies to Identify Preferentially Selective HDAC2 Inhibitors

Presenter: Madeline Rickert

Co-Authors:

School:

Abstract:

Synthesis of Novel Benzodiazepines via the Buchwald-Hartwig Amination for the Development of GPCR Ligands

Presenter: Marley Makowiak

Co-Authors:

School:

Abstract:

Hydrogen Peroxide Responsive Theranostics for Cancer-Selective Activation of DNA Alkylators and Real-Time Fluorescence Monitoring

Presenter: Mayurika Mahendran

Co-Authors:

School:

Abstract:

Synthesis of peptide – small molecule conjugates

Presenter: Natalie Chartrand

Co-Authors:

School:

Abstract:

Synthesis of Polyethylene Terephthalate (PET) Oligomer chains.

Presenter: Nathan Gorgius

Co-Authors:

School:

Abstract:

Preliminary Characterization of Biodegradable Detergent Candidates for Biological Applications

Presenter: Nethyanji S. Premananda

Co-Authors:

School:

Abstract:

Trapping putative intermediates in the catalytic reduction of nitrite to ammonia, catalyzed by S. Oneidensis Cytochrome c Nitrite Reductase (ccNiR)

Presenter: Oke Temidayo Joseph

Co-Authors:

School:

Abstract:

Effects of doxorubicin-treated BT549 cell conditioned media on DAMI megakaryocytes

Presenter: Samantha Edge

Co-Authors:

School:

Abstract:

Understanding the Roles of Tyr33 and Gln58 in Modulating the Rate of Met Truncated Hemoglobin N Reduction

Presenter: Sharmin Khandker Shampa

Co-Authors:

School:

Abstract:

TBD

Presenter: Shirisha Jonnalagadda, PhD

Co-Authors:

School:

Abstract:

Exploiting Redox Vulnerabilities for Selective Cancer Therapy via Ascorbate-Driven Prodrug Activation

Presenter: Thilini Nimasha Fernando Ponnamperumage

Co-Authors:

School:

Abstract:

Using seed priming to improve alfalfa feed quality and value

Presenter: Thomas Suthers

Co-Authors:

School:

Abstract:

Sieving For Stability: A Grain Size Analysis On Spit Systems in Lake Michigan

Presenter: Bryanna Ninham

Co-Authors:

School:

Abstract:

A Facies Analysis of the Mesoproterozoic (1.09 Ga) Copper Harbor Conglomerate Formation in the Keweenawan Midcontinent Rift System

Presenter: Kasserena Cvikel

Co-Authors:

School:

Abstract:

Magnetic Mineral Composition and Grain Size in the San Rafael Volcanic Field, Utah

Presenter: Noah Melcer

Co-Authors:

School:

Abstract:

Lava Under the Lense: Crystal Cargoes and the Chemistry of Capping Lavas at Four Tuyas in Northeast Iceland

Presenter: Valerie Brost

Co-Authors:

School:

Abstract:

LIPI: Linguistic Internal Planning for Inference

Presenter: Brian Ouyang

Co-Authors:

School:

Abstract:

Wildfire Smoke Impact on Emergency Room Visits – A Literature Review

Presenter: Cameron Lee

Co-Authors:

School:

Abstract:

Wildfire Smoke Impact on Emergency Room Visits – A Literature Review

Presenter: Kali Quade

Co-Authors:

School:

Abstract:

Robust Item Selection for Adaptive Testing of Slow Starters

Presenter: Matheesha Lakshitha

Co-Authors:

School:

Abstract:

Evil Parking Functions

Presenter: Sadie Baker

Co-Authors:

School:

Abstract:

Modeling Population-Level Dynamics of C. difficile Transmission Between the Hospital and Community

Presenter: Grace Mueller

Co-Authors:

School:

Abstract:

TBD

Presenter: Anand Anilkumar

Co-Authors:

School:

Abstract:

Friction Discrepancies of Monolayer Molybdenum Disulfide

Presenter: Ian Bigelow

Co-Authors:

School:

Abstract:

Extracting Continuous Conformational Heterogeneity from Cryo-EM Data

Presenter: Umeshika Dissanayaka M

Co-Authors:

School:

Abstract:

Land use and river residence time alter Milwaukee Estuary microbial community abundances

Presenter: Isaiah Chang

Co-Authors:

School:

Abstract:

Myxococcus xanthus globins exhibit heme binding, providing insight into proteins that enhance Gluconobacter oxydans growth

Presenter: Emily Hauke

Co-Authors:

School:

Abstract:

Characterization of HbN, a globin protein,  from the obligate aerobe Myxococcus xanthus

Presenter: Kate Fechner

Co-Authors:

School:

Abstract:

Characterization of HbN, a globin protein, from the obligate aerobe Myxococcus xanthus

Presenter: Olivia Baus

Co-Authors:

School:

Abstract:

Glen Hills Students’ NASA Journey

Presenter: Lalitha Murali

Co-Authors:

School:

Abstract:

Investigating Perineuronal Nets Across the Estrous Cycle in Mice

Presenter: Ella Cerkoney

Co-Authors:

School:

Abstract:

Melanopsin Signaling Modulates Light-Evoked Changes in Breathing

Presenter: Ayah Sbeih

Co-Authors:

School:

Abstract:

The Role of Estrogen Receptor Beta in Stress-Induced Sleep Impairments and Maladaptive Behavioral Responses in Mice

Presenter: Gargee Pandya

Co-Authors:

School:

Abstract:

Activity-Dependent Tagging of Socially Activated Neurons in the Lateral Entorhinal Cortex of Mice

Presenter: Isabella Souvannarath

Co-Authors:

School:

Abstract:

Sex-specific effects of HDAC inhibition on adolescent stress susceptibility and development of depression-related phenotypes

Presenter: Sydney Zimmerman

Co-Authors:

School:

Abstract:

Salinity and Total Iron detected in Kinnikinnick River Water

Presenter: Airam Velasco

Co-Authors:

School:

Abstract:

TBD

Presenter: Angela R Paredes Montero

Co-Authors:

School:

Abstract:

Idioms, Metaphors, and Explanatory Models of Suffering Across Cultures: A Systematic Review Seeding a Global Atlas of Human Suffering

Presenter: Nathan Chue Thao

Co-Authors:

School:

Abstract:

Beyond Novelty: Remembered Associations Guide Visual Attention

Presenter: Eunji Back

Co-Authors:

School:

Abstract:

TBD

Presenter: Natalie Nassar

Co-Authors:

School:

Abstract:

Does the MISO Planning Resource Auction Price Capacity Consistently with Seasonal Reliability Risk?

Presenter: Waleed Nassar

Co-Authors:

School:

Abstract: