The Postdoc Poster Session is an opportunity for postdocs to share their research in a relaxed atmosphere with fellow researchers on campus. Posters are encouraged to be accessible to a broad academic audience including graduate students, postdocs, and faculty from a wide spectrum of disciplines.
The theme this year is From Textbook to Lab: Postdocs Present to UK Undergraduate Aspiring Scientists which is why winners will be selected by a panel of undergraduate research ambassadors based on quality of content, display, and oral presentation. For more details see the scoring rubric here: Postdoc Poster Competition Scoring Rubric
Prizes: $200 for 1st place, $150 for 2nd place, $100 for 3rd place, $50 for 4th place
Poster #1
Background: Currently, mouse aortic aneurysm studies are conducted at room temperature (RT, 20-22 °C). However, thermoneutrality (TN), with temperature at ~30 °C, is the ambient temperature at which energy expenditure maintains basal metabolic rate in mice. This study aimed to determine the impact of TN relative to RT on angiotensin II (AngII)-induced aortic aneurysm development in hypercholesterolemic mice.
Methods: Three hypercholesterolemic mouse models were used to assess the effects of TN on aortic rupture and aortic aneurysm development: Apoe-/- mice fed a normal laboratory diet, Ldlr -/- mice fed a Western diet, and C57BL/6J mice infected with adeno-associated viruses (AAVs) containing a mouse PCSK9 gain-of-function mutation (AAV.mPCSK9D377Y) and fed a Western diet. Male mice of the 3 mouse strains at 8 to 10-week-old were implanted with mini osmotic pumps subcutaneously to infuse either saline or AngII (1,000 ng/kg/min) for 4 weeks. During the infusion period, mice were housed at either 20 °C (RT) or 30°C (TN). The primary endpoint was ex vivo aortic diameters of the suprarenal aortic region.
Results: Despite body weights not differing between the two groups across the three hypercholesterolemic mouse models, mice housed at TN exhibited whitening of brown adipose tissue and significantly downregulated mRNA abundance of Ucp1, Pgc1α, Prdm16, Dio2, and Cidea compared to those housed at RT, regardless of infusion with either saline or AngII. Plasma total cholesterol concentrations were similar between the two groups within each mouse model. The mortality rate, caused by either ascending or abdominal aortic rupture, was comparable between RT and TN groups within each mouse model. The maximum diameters of ascending aortas were not different between the RT and TN groups [Apoe -/- mice: 1.89± 0.08 mm vs 1.71 ± 0.04 mm, P = 0.07; Ldlr -/- mice: 1.70 ± 0.04 mm vs1.86 ± 0.04 mm, P = 0.19; mPCSK9(D377Y)-AAV injected mice: 1.39 ± 0.04 mm vs 1.46 ± 0.07 mm, P = 0.38]. Maximal diameters of abdominal aortas were also not different between the two groups [Apoe -/- mice: 1.05 (0.96-1.68) mm vs 0.98 (0.92-1.05) mm, P = 0.27; Ldlr -/- mice: 1.29 (0.84-2.65) mm vs 1.60 (1.60-1.86) mm, P = 0.75; mPCSK9(D377Y)-AAV injected mice: 1.37 (0.90-2.21) mm vs 1.58 (1.26-2.75) mm].
Conclusion: Thermoneutrality during 4 weeks of AngII infusion did not influence formation of aortic aneurysms in hypercholesterolemic mice.
Poster #2
Introduction: Disinfection byproducts (DBPs) are one of the most prevalent drinking water contaminants world wide, including in Kentucky. Rural communities are especially vulnerable to the formation of DBPs due to degraded infrastructure, financial constraints, and large distribution networks with low demand. Nitrogenous DBPs (HANs) are currently unregulated but have raised increasing concern due to their toxicity and tendency to form when source waters have elevated concentrations of nitrogen and bromide.
Methods: This study analyzed the spatial-temporal complexity (e.g., seasonality, climate variables, water age, source water quality) characterized by rural Appalachian drinking water distribution networks and its relationship to HAN and THM concentrations. Monthly tap water (nhomes=38; ntotal samples=639) was sampled over two years for DBPs while source waters (n=2) were analyzed for bi-monthly concentrations of DBP precursors (e.g., organic carbon, nitrate, chloride, bromide).
Results: The average concentration of total THMs was 0.074 ±0.036 mg/L with 76 exceedances (17 households) of the EPA maximum contaminant level (0.08 mg/L). Total THM varied seasonally, with greater concentrations from July to November, correlating with increased temperature, source water organic carbon, and source water bromide concentrations. The average total HANs concentration was 0.005 ± 0.001 mg/L. Total HANs were also correlated to temperature and source water organic carbon, bromide, and total nitrogen concentrations. Spatially, THMs increased with increasing water age while HANs did not have an observable spatial signature.
Discussion: Findings elucidate the spatial-temporal relationships between source water chemistry and DBPs which will be used to support predictive models to help small drinking water utilities reduce their formation.
Poster #3
Introduction: Cyanobacteria are a source of bioactive compounds and have been studied for over 50 years in the search for new antibiotics, antiviral, and anticancer drugs. In our current projects, we are investigating marine cyanobacterial compounds and their synthetic analogs for Ca2+ modulation via interactions with the sigma-2 receptor. We have identified several cyanobacterial natural products such as veraguamide which has sigma-2 receptor affinity and alter store operated calcium entry (SOCE). Recent research has shown SOCE and the sigma-2 receptor to be involved in neuropathic pain.
Methods: Cyanobacterial samples were collected from the Las Perlas Archipelago in Panama, extracted, and the crude extract was fractionated over silica gel using a stepwise gradient solvent system. The fractions were then evaluated in a psychoactive drug screening program, and those with sigma-2 affinity were further purified to isolate veraguamide compounds. The structural modification and optimization of these cyanobacterial compounds using medicinal chemistry tools are ongoing to develop antinociceptive agents and to better understand the interaction of sigma-2 and SOCE.
Results and Discussion: Veraguamides and their analogs have shown to alter SOCE in a sigma-2-dependent manner in both mouse and human-derived neurons, and this effect can be modulated through simple structural modifications. Furthermore, the reduction of SOCE caused by these analogs has demonstrated in vivo antinociceptive effects. The sigma-2/TMEM97 receptor provides a promising target for alteration of the perception of noxious stimuli and could potentially lead to a new option for neuropathic and chronic pain management in the future.
Poster #4
Introduction: When you bite into a burger, you might judge its doneness by the color inside but should you? The pink-to-brown shift in cooked beef is driven by myoglobin, a heat-sensitive protein whose behavior varies with its redox state. Burgers may appear fully cooked before reaching the safe internal temperature of 71 °C—a phenomenon known as premature browning—posing potential food safety risks. This study evaluated how endpoint cooking temperature and storage duration affect internal color and myoglobin thermal stability in ground beef patties.
Materials and methods: Eight coarse-ground beef chubs (80% lean) were formed into 120 g patties and stored at 2 °C for 0, 2, 4, or 7 days. Patties were cooked to 54 °C, 63 °C, or 72 °C, then analyzed for internal color (L*, a*, and R630/580) and myoglobin stability using differential scanning calorimetry (DSC) to determine denaturation temperature (Tm) and enthalpy (ΔH). Data were analyzed using two-way ANOVA with significance set at P < 0.05.
Results and discussion: Patties cooked to higher temperatures (63 °C and 72 °C) showed greater lightness and reduced redness compared to 54 °C. Storage time did not significantly influence color metrics. Color stability declined with increasing endpoint temperature. DSC analysis revealed that higher cooking temperatures increased myoglobin Tm values, indicating greater thermal stability, while ΔH remained unchanged across treatments.
Conclusions: The results demonstrate that the internal endpoint temperature influenced the color and the thermal stability of myoglobin in cooked ground beef patties. While increased endpoint temperatures decreased internal redness, it increased myoglobin thermal stability.
Poster #5
Introduction: The melanoma incidence/mortality continues to rise. First line immune checkpoint blockade is only effective for some patients. BRAF and NRAS mutations activate the MAPK pathway, yet therapies targeting this pathway (MAPKi; BRAFi, MEKi) are only effective for BRAF-mutant melanoma, and resistance and metastasis inevitably develops. Melanoma invasion/metastasis is characterized by a shift in EMT transcription factor expression (ZEB2/SNAI2-->ZEB1/TWIST1). ABL1/2 are key nodes during MAPKi resistance, and here, we examine the contribution of the EMT switch to ABL1/2-driven invasion and MAPKi resistance.
Methods: Invasion assays, clonogenicity studies, gene silencing/overexpression, pharmacological inhibition, western blots, RT-qPCR, cycloheximide assays, gain-/loss-of-function studies and rescue experiments and melanoma samples analyzed with sample-wise GeneSet Enrichment Analysis were used for the below studies.
Results: ABL1/2 promote invasion and metastasis during MAPKi resistance. Silencing/inhibiting (nilotinib) ABL1/2 reduces ZEB1 and N-cadherin (NCAD) protein but not mRNA expression and increases SNAI2 mRNA/protein, whereas expression of constitutively active ABL1/2 has opposite effects. ABL1/2 stabilize ZEB1/NCAD; MG132 (proteasome inhibitor) treatment prevents nilotinib-mediated ZEB1/NCAD degradation; and silencing NEDD4L, a putative ZEB1 E3 ligase, rescues the ability of nilotinib to reduce ZEB1/NCAD expression. ABL1/2 inhibition prevents invasion and clonogenicity during MAPKi treatment, and overexpression of ZEB1 rescues the effects of nilotinib on clonogenicity (but not invasion) in an ERK-dependent manner. Finally, our data are clinically relevant as ZEB1/NCAD, and ABL1/2 activities are correlated in patient samples.
Discussion: ABL1/2 kinases promote resistance by stabilizing ZEB1 and NCAD, likely via NEDD4L inhibition. We identify a new role for the EMT switch downstream of ABL1/2 during resistance.
Poster #6
Introduction: Non-invasive sampling is crucial for enhancing equine welfare and biosecurity during EHV-1 outbreaks, where subclinical shedding facilitates transmission.
Methods: This Randomized Controlled Field Trial (RCFT) evaluated nasal wipes against the gold standard sampling approach of nasal swabs for qPCR detection of EHV-1 in experimentally infected horses across a range of inoculum doses. Standard curve qPCR was performed to quantify the EHV-1 genome copies.
Results: Nasal wipes demonstrated an overall agreement of 85.4% (including negative samples) with nasal swabs, accurately capturing viral shedding kinetics. Crucially, wipes identified EHV-1 in 14 swab-negative samples (8.5%) during low-shedding phases (<500 copies/mL), achieving 62% sensitivity in this tier and highlighting higher sensitivity for detecting silent shedders as compared to nasal swabs. Quantitative differences at peak shedding (>10⁶ copies/mL; regression slope: 0.78) were consistent with expected method-dependent variability. Clinically, wipes offer significant welfare advantages, enhancing compliance during repeated sampling at a large scale. While agreement decreased at low viral loads (r = 0.09–0.66), mirroring limitations of possible individual sampling techniques and PCR-based diagnostics, nasal wipes represent a robust, welfare-centric tool for EHV-1 surveillance.
Discussion: Their enhanced sensitivity for subclinical shedders, responsible for a significant proportion of EHM cases, supports their use in outbreak control, particularly for refractory horses or large-scale screening where traditional swabbing is impractical. Further protocol optimization and integration with environmental sampling are recommended.
Poster #7
Introduction: Equid alphaherpesvirus 1 (EHV-1) remains a major concern for the equine industry, with recent outbreaks at international equestrian events highlighting the need for improved surveillance during competitions.
Study design: A cross-sectional field study.
Methods: Air (37) and surface samples (205) were collected from temporary stabling facilities across four events in Spain and 2 in the U.S. at different seasons over 10 months. Viral DNA was measured and quantified by quantitative and digital PCR. During the Spain events, we sampled the air twice, early evening vs nighttime.
Results: EHV-1 was frequently detected in the air in 20/28 samples in Spain, in all 3 samples from Florida, and in 3/6 samples from Kentucky, U.S. Surface samples were positive 15.6% in Spain, 26.3% in Florida, and 0.0% in Kentucky. EHV-4 was found in air samples at rates of 12/28 in Spain, 5/6 in Kentucky, and 0/3 in Florida, with surface detection of 1.6% in Spain and 7% in Kentucky. No significant differences were observed between daytime (high activity) and nighttime (low activity) airborne viral loads, suggesting that daytime horse movement and management practices had minimal impact on detection levels in our study. A positive correlation (Cohen’s K= 0.401) and moderate agreement between surface and air positivity for EHV-1 emphasized the potential of air sampling as a non-invasive, cohort-based surveillance tool.
Discussion: These findings underscore the utility of air sampling for early EHV-1 detection at mass gatherings, although further validation is needed to connect genomic detection to active shedding and viral infectivity.
Poster #8
Introduction: Aggressive periodontitis in children is rare, rapidly progressive, and may affect both primary and permanent dentition. It is often associated with familial susceptibility. Understanding long-term outcomes and genetic influences may guide early diagnosis and targeted interventions.
Methods: We present a case series of 10 children diagnosed with aggressive forms of periodontitis, involving either primary or permanent dentition. Clinical records, radiographs, and periodontal charts were reviewed, with follow-up extending up to 10 years in select cases. Family history was documented, and genetic predisposition was explored through reported disease patterns among first-degree relatives.
Results: In the primary dentition, early onset of severe bone loss and mobility often led to premature tooth loss. In the permanent dentition, rapid attachment loss predominantly affected incisors and first molars. Treatment included scaling and root planing, systemic antibiotics, and supportive periodontal therapy. Long-term follow-up showed that, with consistent care, patients preserved functional dentition; however, recurrence occurred in non-compliant cases. Familial clustering was evident, with siblings and parents also presenting periodontal breakdown, highlighting potential heritable risk factors.
Discussion: This series highlights the impact of aggressive periodontitis in both primary and permanent dentitions of children. Early diagnosis, comprehensive treatment, and long-term maintenance are essential to preserve dentition and function. The observed familial patterns emphasize the importance of genetic susceptibility, reinforcing the need for family-based screening and preventive strategies in pediatric populations.
Poster #9
Introduction: Arabinoxylan, the primary hemicellulose component in grass biomass, constitutes approximately 20–40% of the cell wall and serves as an energy source for ruminant microbes. Structurally, arabinoxylan consists of a 1,4-β-D-xylopyranose backbone substituted with various side chains, including monomeric arabinose, esterified ferulic acid, glucuronic acid, and acetyl groups. Each structural feature influences ruminal digestibility and may confer health benefits. However, little is known about the fate of arabinoxylans from cool-season pasture grasses during ruminal fermentation.
Methods: In this study, two cool-season forage grasses (timothy, Phleum pratense L., and perennial ryegrass, Lolium perenne L.) grown in central Kentucky were harvested, and their insoluble cell-wall materials were subjected to in vitro fermentation with pure cultures of ruminal fibrolytic bacteria. Structural changes in arabinoxylan were subsequently analyzed.
Results and Discussion: During fermentation, microorganisms selectively removed arabinose and xylose units from arabinoxylan polysaccharides and showed a preference for cellulose utilization. The bacterial enzymatic system fragmented arabinoxylan into oligosaccharides of varying lengths, which were released into the fermentation supernatant with minimal further utilization by the bacteria. Additionally, bacterial feruloyl esterases cleaved a significant portion of ester-linked ferulic acid from arabinoxylan, releasing it as free ferulic acid into the media. Extrapolating to in vivo conditions, the liberated arabinoxylan fragments and free ferulic acids from cool-season forages are likely available for metabolism or biotransformation by other rumen microbes. These findings enhance our understanding of how forage cell wall carbohydrates are utilized by ruminants and may inform strategies to optimize feed efficiency and nutritional outcomes.
Poster #10
Quorum sensing (QS) is a communication system employed by bacteria, allowing them to coordinate their behavior based on population density. Bacteria produce and release signaling molecules called autoinducers into the environment. As the bacterial population grows, the concentration of these autoinducers increases. Once a critical threshold is reached, the autoinducers will modulate various physiological activities, including motility, virulence or biofilm formation. In the present work, using a series of biochemicals, bioinformatics and genetic manipulation techniques, we propose that Streptococcus agalactiae [Group B Streptococcus [GBS)] may use a QS regulatory pathway to modulate the expression of virulence factors such as β-hemolysin/cytolysin (β-H/C) and other secreted proteins. In addition, we identified that overstimulation of this QS pathway induces a temperature-dependent lysis of GBS.
In this study we used a peptide-free chemically defined medium to investigate a possible QS pathway involved in the regulation of GBS virulence. Our findings revealed that the chemical nature of the autoinducer triggering the activation of this QS pathway is a small peptide that requires Opp system for its translocation into the cell. Once inside the cell, this small peptide interacts with and activates the transcriptional regulator RovS. After Rovs is activated, it leads to the regulation of its targeted genes, like the cyl operon, which encodes for the genes involved in the production of β-H/C. Additionally, we identified two other dysregulated genes: a putative secreted transglutaminase and another small peptide. In other organisms, similar genes have been reported to play a crucial role in biofilm formation. Therefore, here we present a novel mechanism in GBS that employs a QS pathway that regulates the expression of different virulence factors.
Poster #11
Introduction: Human Schwann cells (hSCs), the peripheral nervous system (PNS) glial cells, can be isolated and expanded in vitro for several passages. However, it is a challenge to obtain homogeneous hSC cultures due to donor variability and contamination with non-glial cells. Therefore, the aim of this research was to develop rigorous assays and quality control measures for establishing and managing hSC cultures from various adult nerve tissues.
Methods: To determine the phenotype and purity of hSC cultures, we combined the use of transcriptomics and image-based immunofluorescence labeling using cell type and maturation state specific antibodies. To determine the functional properties of hSC cultures, we performed bioassays for cell proliferation, differentiation, and senescence. hSC proliferation was determined by EdU labeling in the presence of SC-specific growth factors. Cell differentiation was determined by immunodetection of myelin-related markers, such as O4 and MPZ, in the absence and presence of cAMP, a strong inducer of myelination. Cellular senescence was determined by SA-B-Gal staining.
Results: The implementation of the abovementioned quality control protocols allowed us to improve the accuracy, reliability, and reproducibility of hSC cultures from various sources. Not only we developed simple methods to discriminate hSCs from nonglial cells (namely fibroblasts) but also determine the potency of individual hSC batches using a range of bioassays.
Discussion: The well-characterized hSC cultures we have generated can be used in assorted applications in regenerative medicine, e.g. to develop cell-based grafts for implantation, in vitro modeling of neurobiological processes, and drug testing for neurodegenerative diseases.
Poster #12
Introduction: Oropouche virus (OROV) is a neglected arbovirus that has caused outbreaks in Central and South America since the 1950s. This study investigates the ecological and demographic determinants of Oropouche fever in Brazil between 2014 and 2025.
Methods: In this epidemiological study, we combined epidemiological, genomic, and demographic data. We examined the ecological and demographic profile of Oropouche fever in Brazil.
Results: Between January 1, 2014, and January 25, 2025, 19,720 laboratory-confirmed Oropouche fever cases were reported in all 26 states and the Federal District. Age-sex structural analysis revealed that adults had a higher incidence than individuals aged 19 or younger. Male and Asian populations were 1.2 and 46.3 times more likely to be diagnosed with OROV-positive compared to females and other racial-ethnic groups. Acre, Amazonas, and Rondônia states in North Brazil had experienced sustained outbreaks from late 2023 to early 2024. After the OROV geographical expansion from the Amazon region toward East Coast Brazil, Espírito Santo experienced the most intense and prolonged transmission from March 2024 to January 2025. The cumulative incidence of Oropouche fever cases in rural municipalities was statistically significant and 7.2-fold higher than in urban municipalities between 2014 and 2025. We found OROV exhibited a median urban-to-rural case ratio of 0.6, while dengue, chikungunya, and Zika range from 2.5 to 2.9.
Discussion: Our data demonstrated that OROV transmission has predominantly occurred in rural municipalities before and after the re-emergence, indicating a pattern that differs significantly from current major arboviral diseases that primarily occur in urban settings.
Poster #13
Introduction: Rhodococcus equi is an emerging opportunistic pathogen that can cause severe pulmonary and systemic infections in humans, particularly in immuno-compromised individuals. It is commonly found in the soil of horse farms and is primarily transmitted to humans through the inhalation of contaminated dust or soil particles. In some cases, the infection may disseminate to internal organs such as the liver, spleen, and brain. In addition to its zoonotic potential, R. equi is implicated in the development of purulent bronchopneumonia in foals under six months of age. Current treatment protocols rely on antibiotic regimens, primarily involving rifampin in combination with macrolides. However, the emergence of antimicrobial resistance has underscored the urgent need for alternative therapeutic strategies. Our study aims to screen and identify novel small molecules (SMs) with potent antimicrobial activity against R. equi. Small molecules are low-molecular-weight compounds that can target specific bacterial cellular processes and exert antimicrobial effects ranging from narrow to broad-spectrum activities.
Methods: A high-throughput screen of ~2,500 SMs was conducted to evaluate their inhibitory effects on R. equi growth. Compounds showing 100% inhibition were tested for efficacy against biofilms and intracellular R. equi within murine macrophages. Cytotoxicity, spectrum activity assays and electron microscopy scanning were performed.
Results: Ten SMs exhibited strong antimicrobial activity against multidrug-resistant R. equi, with MIC and MBC values as low as 0.078 µM and 0.156 µM. Five SMs completely inhibited biofilm formation (0.078–10 µM), and all significantly reduced intracellular bacterial survival at 5 µM (p < 0.05).
Conclusion: These novel SMs could serve as potential alternatives to conventional antibiotics for the effective control of R. equi infections.
Poster #14
Spinal cord injury (SCI) disrupts supraspinal influence on autonomic function leading to alterations in cardiovascular parameters such as blood pressure and heart rate, as well as thermoregulation. Exploring these parameters in animal models is pertinent to develop deeper understanding of cardiac physiology after SCI. There is no study, as per our knowledge, that has meticulously established the acute effects of severe, high thoracic (T3) SCI on these various cardiovascular parameters. In the present study, we analyzed the effect of SCI on systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), heart rate, temperature, and activity in female rats. Adult female rats were implanted with in vivo telemetric probes in the descending aorta. Two weeks later, animals received T3 spinal contusion at 400 kdyn (5s dwell time). We collected baseline telemetric recordings for 48 hours prior to SCI and then continuous recordings for the first week after SCI. After SCI, rats were divided into two groups; one group received 2 ml Clodronate (7mg/ml- to deplete peripheral macrophages) on days 1, 3, and 6 post- injury (once a day), and the control group received vehicle (2 ml saline). After spinal cord injury, we observed hypertension for 2 days after injury with an increase in SBP, DBP, and MAP relative to baseline measures, followed by a decrease. We also observed that SCI disrupted heart rate dipping (circadian rhythm), increased core body temperature irregularities, and decreased in activity levels. Collectively, the current work provides unprecedented insight into acute cardiovascular changes after SCI.
Poster #15
After spinal cord injury (SCI), neutrophils and monocyte-derived macrophages infiltrate the lesion site. Macrophages contribute both to secondary injury and to repair; however, pro-inflammatory macrophages dominate, limiting recovery. Azithromycin (AZM) has immunomodulatory properties and improves motor outcomes when administered after SCI, but systemic delivery can cause cardiac side effects and disrupt the gut microbiome. To reduce off-target effects, we investigated the use of nonPEGylated liposomes, which are preferentially taken up by phagocytes. We hypothesized that encapsulating AZM in non-PEGylated liposomes (L-AZM) would shift macrophage phenotype while minimizing microbiome alterations. Mice received a T9 contusion injury and were treated for 7 days with PBS, free AZM (F-AZM, 40 mg/kg), or L-AZM (10 or 40 mg/kg). RNA sequencing of intraspinal macrophages revealed that L-AZM10 suppressed expression of M1 markers (Ccr7, Il12b, Il6, Cd80) and enhanced the M2 marker Retnla. Differential expression analysis indicated that L-AZM10 upregulated genes involved in extracellular matrix remodeling, a key reparative function. Although F-AZM and LAZM40 also altered gene expression, L-AZM10 exerted the most pronounced pro-reparative effect. Metagenomic analysis of fecal samples demonstrated that L-AZM10 preserved gut microbiome composition. Treated animals displayed a higher Firmicutes:Bacteroidota ratio and clustered closer to PBS controls in UniFrac analyses, whereas F-AZM and L-AZM40 induced distinct microbial shifts. Specieslevel changes further confirmed that L-AZM10 maintained profiles more similar to PBS. Together, these findings suggest that L-AZM10 optimally reprograms macrophages toward a reparative phenotype after SCI while limiting disruption of the gut microbiome, supporting its potential as a targeted therapeutic strategy.
Poster #16
Tris(1,3-dichloro-2-propyl) phosphate (TDCPP) is a widely used organophosphate flame retardant that has been increasingly linked to adverse metabolic effects. This study investigated the impact of TDCPP exposure on body composition and glucose metabolism in male and female C57BL/6J mice housed under thermoneutral (TNT) conditions. Mice were fed a TDCPP-containing diet at a dose of 1 mg/kg/day ad libitum for 8 weeks. Body weight and composition, including percent fat (%fat) and percent lean mass (%lean), were assessed using EchoMRI, while glucose tolerance and insulin sensitivity were evaluated using intraperitoneal glucose tolerance tests (IPGTT) and insulin tolerance tests (IPITT), respectively. In male mice, TDCPP exposure resulted in increased body weight relative to controls. There were no significant differences in %fat or fat-to-lean mass ratio between TDCPP-exposed and control males housed at either TNT or room temperature. However, %lean mass was significantly reduced in TDCPP-exposed males housed at room temperature compared to those housed at TNT. Additionally, TDCPP-exposed males exhibited impaired glucose tolerance, evidenced by elevated blood glucose levels and increased area under the curve (AUC) during IPGTT in TNT-housed mice. Insulin sensitivity, as assessed by IPITT, showed a modest but non-significant reduction in exposed males. In contrast, female mice showed no significant differences in body weight, %fat, or %lean mass between TDCPP-exposed and control groups. Glucose and insulin tolerance were not impaired in exposed females, and IPITT revealed no meaningful changes in insulin sensitivity. Overall, these findings suggest that TDCPP exposure induces sex-specific metabolic alterations, with male mice exhibiting increased body weight and impaired glucose regulation, while female mice appear relatively unaffected under similar exposure conditions.
Poster #17
Abstract: A a novel, noncontact, portable, time-resolved laser speckle contrast imaging (TR-LSCI) technique is presented for fast, high-resolution mapping of cerebral blood flow (CBF) at multiple depths. TR-LSCI employs picosecond-pulsed near-infrared illumination and a gated single-photon avalanche diode (SPAD) camera to detect long-pathlength photons, enhancing sensitivity to deep-brain hemodynamics. Reconstruction of CBF maps was accelerated using convolution functions and parallel computation. System performance was validated with tissue-mimicking phantoms and in vivo rodents, demonstrating noninvasive, depth-resolved CBF mapping with high temporal and spatial resolution.
Introduction: Cerebral blood flow (CBF) is essential for brain metabolism, and its disruption contributes to conditions such as stroke, traumatic injury, and ischemia. Current optical methods, including LSCI and OISI, provide high-resolution cortical imaging but suffer from limited penetration (<1 mm), often requiring invasive cranial windows. To overcome this limitation, TR-LSCI is developed for noninvasive, depth-sensitive monitoring of brain hemodynamics.
Methods: The TR-LSCI system combined pulsed near-infrared illumination with a gated SPAD camera to capture photons of varying pathlengths. Accelerated algorithms enabled efficient reconstruction of depth-resolved CBF maps. Validation was performed using phantoms and in vivo rodent models under CO₂ challenge and carotid ligation protocols.
Results: TR-LSCI achieved sampling rates up to 3 Hz. Phantom experiments confirmed depth sensitivity, while rodent studies demonstrated reliable detection of CBF dynamics in agreement with established modalities.
Discussion: TR-LSCI provides portable, fast, and noninvasive imaging of depth-resolved CBF. Future work with advanced SPAD cameras and multi-wavelength approaches may improve performance and expand applications in neuroscience and vascular disease research.
Poster #18
Differential glycosylation, sometimes referred as glycorandomization, encompasses chemoenzymatic strategies to enable differential glycosylation of a wide range of small molecule-based drugs and probes. This technique expands pharmacophore chemical diversity and can improve potency, solubility, and ADMET and, in some cases, even alter fundamental molecular mechanism. In the present study, we explored the substrate scope of a permissive engineered glycosyltransferase (OleD Loki) toward a diverse range of heterocycles and drugs/leads bearing aromatic-, primary-, secondary- and tertiary-hydroxyls. Newly identified representative heterocyclic OleD Loki substrates from this study include indoles, thiopyrimidines, thiopurines, benzoxazoles, pyridopyrimidines, and imidazopyridines. Loki regio-/stereoselectivity with corresponding substrates was determined by scaled enzymatic production and structure elucidation of the corresponding products. Importantly, this study highlights Loki’s ability to form novel S-, N-, and O-glycosides and to glycosylate a set of sterically constrained acceptor nucleophiles. As the first step toward predicting vivo stability of corresponding glycosides, -glycosidase-catalyzed degradation and plasma stability of representative glycosides were also evaluated. The representative glycosides were found to display greater stability than a corresponding control 4-Methylumbelliferone β-D-glucoside in both enzymatic and plasma stability tests. Furthermore, the cytotoxicity of the glycosides was evaluated utilizing cancer cell lines. Further directions will focus on subsequent Loki-catalyzed differential glycosylation with a goal of fine-tuning sugar-dependent in vivo stability and tissue targeting.
Poster #19
Introduction: The transplantation of peripheral nervous system (PNS) cells has been exploited in clinical trials to treat central nervous system (CNS) lesions resulting from trauma and neurodegenerative disease. The identification of grafted PNS cells in postmortem tissues remains a challenge at least in part because human cells cannot be labeled before their implantation in the human CNS. Therefore, we developed simple immuno-microscopy approaches to discriminate PNS components from those of CNS origin in formalin-fixed human tissue sections.
Methods: First, we conducted an in-silico analysis in the open source database Human Protein Atlas to identify candidate PNS markers predicted to be absent in the CNS. Next, we conducted immunostaining experiments followed by image analysis to test the immunoreactivity and PNS vs CNS selectivity of candidate antibodies using adult human PNS (spinal nerves and dorsal root ganglia) and CNS (brain and spinal cord) biospecimens.
Results: As predicted by our bioinformatics analysis, our immunostaining experiments confirmed that NGFR and MPZ antibodies reliably distinguished PNS-derived cells and myelin, respectively, from CNS proteins. Likewise, antibodies against ECM components, including collagen IV and laminin, reacted strongly with PNS rather than CNS antigens. Other antibodies, including: S100β, SMA, Glut1, CD34, and CSPG, recognized both PNS and CNS antigens. Despite, these antibodies were useful to identify PNS-derived cellular and noncellular elements based on structural differences with the CNS counterparts.
Discussion: We developed different antibody combinations with potential to reveal grafted or endogenous PNS cells, and study their survival, differentiation, integration, and recipient reaction within host CNS tissues.
Poster #20
Recovering critical metals from electronic waste can alleviate domestic supply chain constraints and mitigate environmental risks associated with waste disposal. Hydrometallurgical processes using solvent extraction are effective for recovering critical metals from leach solutions while selectively separating mixed metal ions. Understanding how the molecular features of extractant ligands influence the thermodynamics and kinetics of metal ion transfer from aqueous to organic phases is essential for advancing separation efficiency. In this work, we elucidate the role of metal solvation structure in governing the extraction process through a combination of experimental characterization and molecular dynamics simulations. Our results reveal that, at the oil–aqueous interface, water molecules in the metal solvation shell are replaced stepwise by oxygen atoms from the extractant’s phosphate group. FTIR and NMR spectroscopy confirm coordination complex formation between the metal ions and extractant ligands. Free energy profiles obtained from metadynamics simulations show that cobalt ions encounter a lower energy barrier for ligand exchange than nickel ions. This suggests that extraction selectivity arises from kinetic control, with the rate-determining step identified as the intermediate stage of ligand substitution. This study provides mechanistic insights into the thermodynamic and kinetic aspects of metal solvent extraction, offering guidance for the rational design of extractants with targeted selectivity.
Poster #21
Introduction: Alzheimer’s disease (AD) often coexists with cerebrovascular pathology, including cerebral amyloid angiopathy, exacerbating gait dysfunction. Emerging evidence suggests that astrocytic insulin signaling may play a key role in modulating vascular function, yet its impact on cerebrovascular integrity is not fully explored. This study aims to elucidate how astrocytic insulin receptor (IR) overexpression (OE) affects vascular dynamics and neurovascular coupling in a model of amyloidosis.
Methods: We overexpressed a constitutively active, truncated human IR β-subunit (IRβ) in sensorimotor cortex astrocytes of amyloidosis (5XFAD) and control mice. Longitudinal in vivo two-photon imaging assessed vascular morphology, remodeling, and astrocytic density, vasoreactivity. Gait performance was quantified by ambulation speed, stride time, length deviation, average stride length, and paw placement.
Results: Two-photon imaging showed Astrocytic IRβ OE resulted in vascular alterations, including changes in vascular morphology and reductions in both vessel and astrocytic density Vasoreactivity response rate (%) was decreased, particularly in 5XFAD mice. Gait analysis revealed significant reductions in locomotor speed (p=0.0002) without changes in stride length or balance. These findings suggest astrocytic IRβ activation modulates cerebrovascular structure, vasoreactivity, and locomotor behavior, supporting a role for astrocytic insulin signaling in neurovascular and motor regulation.
Discussion: Constitutive activation of astrocytic IRβ modifies cerebrovascular structure, impairs vasoreactivity, and alters locomotor behavior, supporting a role for astrocytic insulin signaling in regulating neurovascular integrity and motor performance in AD-related pathology. Ongoing work includes mechanistic analysis of IR signaling pathways (pAKT, AKT, IRS-1, IGF-1R) and morphological validation in Cxcl12-GFP mice to further define astrocytic contributions to cerebrovascular remodeling.
Poster #22
Introduction: After traumatic spinal cord injury (SCI), inflammation and other reactive processes exacerbate tissue damage and impair long-term motor recovery. Extracellular traps (ETs) are an immune effector function first described in neutrophils wherein chromatin is decondensed, decorated with cytotoxic granule enzymes, and expelled from the cell body. Recently, ETs have been linked to poor functional outcomes in SCI; however, translatable agents to prevent ET-mediated damage after SCI have yet to be explored.
Methods: We assessed recombinant human (rh) DNase as a potential therapeutic that could be repurposed to break down ETs after SCI in a murine model of contusive SCI. Acute ETosis was assessed via ELISA, flow cytometry, and immunofluorescence. Long term outcomes were assessed via Basso Mouse Scale (BMS), ladder rung walking test (LRWT) and eriochrome cyanine assessment of tissue sparing.
Results: ETs levels increased in the injured spinal cord by 4 hours post injury (hpi), peaking within 24 hpi. When rhDNase was administered at 1 hpi, DNase activity in the serum remained elevated for 24 hpi with a corresponding increase in circulating ET fragments. At 6 hpi, blood-spinal cord barrier permeability was attenuated in rhDNase-treated animals. Long-term functional hind limb recovery, as assessed via LRWT, was improved at 35 dpi with rhDNase treatment; no improvements were noted via BMS. RhDNase-treated animals also exhibited shorter SCI lesion lengths and improved white matter sparing at 35 dpi.
Discussion: These data demonstrate the potential of rhDNase as an anti-ET therapeutic to improve long-term SCI outcomes.
Poster #23
Introduction: PFOS is a widespread environmental pollutant found in 45% of U.S. drinking water. Its resistance to natural degradation allows accumulation in intestinal tissues, potentially disrupting normal function. Although PFOS may contribute to tumor development, its effects on intestinal tissues remain poorly understood. Investigating these impacts is essential to develop strategies to mitigate its harmful effects. Methods: Wild-type mice were exposed to PFOS in drinking water for 7 weeks. RNA-seq analysis was performed on intestinal tissues to assess gene expression changes. APCMin mouse and human organoids were used to study PFOS-induced alterations, employing RT-PCR, western blot, proliferation assay, and immunofluorescence. Results: PFOS upregulated pathways associated with cancer, lipid metabolism, and immune system in normal intestinal tissues. PFOS decreased HMGCS2 protein levels, an enzyme essential for producing β-hydroxybutyrate (βHB), which has anti-tumor properties. Moreover, PFOS increased the levels of proteins associated with colorectal cancer (CRC) initiation. HMGCS2 knockdown in normal colon cells resulted in increased cell proliferation, lipid accumulation, and higher levels of CRC-related proteins. Exposure to PFOS increased cell proliferation in APCMin mouse organoids, and treatment with βHB decreased cell proliferation. Discussion: Our data suggest that PFOS may induce harmful changes in intestinal tissues by elevating cancer-related proteins, potentially increasing CRC risk. PFOS downregulates HMGCS2, which may reduce βHB levels and its protective effects on tissue homeostasis. Additional studies are needed to determine if βHB supplementation can prevent these PFOS-induced changes. Dietary interventions, such as βHB supplementation, might improve the quality of life for individuals in PFOS-contaminated areas.
Poster #24
Introduction: Type-2 diabetes-related hyperglycemia increases the risk for cognitive decline. Amylin is a centrally-linked pancreatic hormone that induces satiation. The beneficial metabolic effects of amylin led to regulatory approval of the amylin analog drug, pramlintide, for weight loss. Different research teams (including ours) report amylin co-aggregates with brain parenchymal and vascular β-amyloid in persons with Alzheimer’s dementia. Using cancer cells, other teams reported that amylin inhibits glycolysis. The present study sought to determine changes of glucose utilization in brain tissues associated with suppressed vs. oversecreted pancreatic amylin.
Method: Because murine amylin is not amyloidogenic, we generated mice “humanized” for amylin expression (hAON mice). Amylin knock-out mice (hAOFF mice) and mice expressing wild-type mouse amylin (WT mice) served as controls. All mouse groups underwent four-month overnutrition to induce prediabetic amylin hypersecretion (in hAON and WT mice), followed by endpoint novel object recognition and mass spectrometry analyses of brain tissue glucose 6-phosphate (G6P) (the first intermediate of intracellular glucose metabolism) and glycolytic amino acids serine, glycine and alanine. We then assessed the ratios of serine, glycine and alanine to G6P (i.e. the glycolytic amino-acid flux).
Results: Compared to WT and hAOFF males, hAON males had longitudinal increases of the blood glucose levels and decreased recognition indices, at the endpoint. Compared to hAOFF and WT males, hAON males had higher brain tissue G6P levels and much lower brain tissue glycolytic amino-acid flux, whereas hAOFF males had the lowest average brain tissue G6P level and highest average brain tissue glycolytic amino-acid flux. Age-matched males from similar amylin genotype groups that were on a chow diet showed unaltered glucose homeostasis. Compared to mice in chow diet groups, metabolically stressed hAON and WT males had increased brain tissue amylin levels. Brain amylin accumulation was higher in hAON vs. WT mice consistent with amyloidogenicity of human amylin.
Conclusion: Prediabetic amylin hypersecretion increases brain amylin level and exacerbates amylin receptor signaling controlling glycolysis, leading to impairments of glycolytic flux and memory.
Poster #25
Introduction: Recently, nicotine has been shown to exert effects on neuroimmune signaling within the brain reward pathway, and specifically the nucleus accumbens, which can drive nicotine seeking following nicotine self-administration (SA) when evaluated in male rats. However, no studies have evaluated the impact of chronic nicotine SA on the accumbens neuroimmune milieu in females, despite clinically meaningful sex differences in nicotine use.
Methods: Ovary-intact female Long-Evans rats underwent nicotine SA for a minimum of 10 sessions. Ventral striatum, which encapsulates the accumbens, was then dissected, and serum trunk blood was collected and analyzed for cytokine analysis when females were in the estrus phase of the estrous cycle.
Results: We observed a general suppression of cytokine levels within the ventral striatum, with tumor necrosis factor alpha (TNFα) being the most changed (p’s <0.05). However, there was an increase in fractalkine, which is a “help me” signal released from neurons during apoptosis and is responsible for microglia recruitment. However, immunostaining of perfused brain slices following nicotine self-administration failed to detect significant apoptotic debris in the nucleus accumbens.
Conclusion: Together, these results indicate that chronic nicotine use in females disrupts neuronal health and neuroimmune signaling within the brain's reward pathway. Thus, targeting neuroimmune health may be a pharmacotherapeutic strategy to reverse the deleterious impacts of nicotine.
Poster #26
Introduction: Arboviruses pose a major threat to global public and animal health. Genomic characterization is essential to understanding arbovirus diversity and evolution, and improves timely outbreak response.
Methods: In this study, we performed high-throughput sequencing and computational analyses on 46 previously unsequenced or partially sequenced arbovirus isolates collected from 23 countries between 1954 and 1984.
Results: These viruses belong to 11 genera in eight viral families, including Sedoreoviridae (n=26), Peribunyaviridae (n=12), Rhabdoviridae (n=3), Spinareoviridae (n=1), Phenuiviridae (n=1), Togaviridae (n=1), Flaviviridae (n=1), and Nairoviridae (n=1). Based on genomic and evolutionary characterization, we propose 15 novel arbovirus species, including likely mosquito-borne, tick-borne, and vertebrate-associated viruses. We also found that Ossa virus (genus Orthobunyavirus) is a reassortant, with a unique M segment and S and L segments derived from Madrid virus (Orthobunyavirus madridense), while Brus Laguna virus (Orthobunyavirus gamboaense) exhibits an M segment from Alajuela virus (Orthobunyavirus gamboaense). Furthermore, our genome composition-based analysis indicates that Lanjan, Zingilamo, Tindholmur, and Bauline viruses have a high likelihood of producing human infection, indicating a risk of emergence and need for further experimental investigation.
Discussion: Collectively, our findings contribute to the understanding of arbovirus diversity and evolution, providing a valuable genomic resource to support future research on arbovirus biology and global virome surveillance efforts.
Poster #27
Introduction: Prenatal opioid exposure (POE) is associated with poor fetal neurodevelopment outcomes; however, the mechanisms underlying these changes have yet to be well characterized. POE may impair fetal brain development directly via opioid accumulation in placental tissues, which could lead to impaired microglia, critical regulators of CNS development. Thus, dysregulation in microglia development could have lifelong neurodevelopmental consequences.
Methods: To unravel the mechanisms underlying how POE affects fetal neurodevelopment, we utilized a model of microglia-like cells derived from umbilical cord blood mononuclear cells (UCBMC), which mimic the morphology and function of microglia in vivo. Differentiation of microglia-like cells from UCBMC was confirmed by morphology and phenotyping. We then assessed the functional landscape of microglia using multiplex Luminex, phagocytosis assays, and bulk RNA sequencing. Finally, we further stratified our findings by maternal HCV status at the time of delivery, given the high prevalence of HCV infection in subjects who inject drugs.
Results: Our results indicate that POE with and without maternal HCV infection blunts microglia maturation and activation, as shown by decreased production/expression of canonical microglia markers as well as inflammatory mediators and genes. However, these microglia-like cells are functionally defective, exhibiting reduced phagocytosis and attenuated responses to LPS stimulation. Altered functional potential was accompanied by transcriptional changes in genes associated with nervous system development, synaptic pruning, and innate immune responses.
Discussion: Overall, these findings suggest a significant impact of POE on microglia development and function, providing insight into the poor neurocognitive outcomes observed in newborns with POE.
Poster #28
Introduction: Salmonella is a leading cause of foodborne gastroenteritis in animals and humans and is usually transmitted through contaminated poultry products. The global spread of antibiotic-resistant Salmonella necessitates the exploration of non-antibiotic strategies to mitigate infections. Quorum sensing (QS), the bacterial communication system governing virulence and biofilm formation, represents an attractive target of anti-virulence therapy. Here, we aim to identify and develop QS inhibitors against Salmonella.
Methods: We screened ~2,500 small molecules (SMs) for their ability to inhibit QS AI-2 production in Salmonella Typhimurium using a Vibrio harveyi BB170 as bioluminescence indicator bacteria. Selected compounds were further evaluated for their toxicity and their effects on virulence-related genes and biofilm formation. The selected SMs were also tested for their effect on Salmonella colonization on human colorectal carcinoma (HT-29) cells.
Results: We identified ten SMs that inhibited AI-2 activity by more than 95%, without impairing bacterial growth. All SMs significantly suppressed biofilm formation (95–100% inhibition) and downregulated the expression of genes associated with quorum sensing, virulence, biofilm development, and motility. Furthermore, six of the molecules exhibited no cytotoxic activity on HCT116 cells at 20 µM, indicating their safety for further preclinical investigation. Two of these compounds exhibit complete clearance for Salmonella inside the HT-29 cells.
Discussion: Our results showed the promising potential to develop QS inhibitors as antibiotic alternatives for controlling infections in humans and animals. The best two identified SMs are strong preclinical candidates, with future studies aimed at validating their efficacy and elucidating the precise molecular targets.
Poster #29
Introduction: Previous research demonstrated that abomasal 5-HTP increased serum serotonin in beef cattle. But it is unclear how ruminal administration of 5-HTP influences serotonin metabolism and peripheral blood flow. The objective was to evaluate the effects of ruminal 5-hydroxytryptophan (5-HTP) supply on peripheral serotonin metabolism and caudal artery hemodynamics in growing beef steers.
Material and Methods: Five ruminally-cannulated Angus × Holstein steers (body weight = 265 kg) were assigned to a 5 × 5 Latin square design. Pulse-dose infusion treatments included: water (negative control), 0.5 mg 5-HTP/kg BW in abomasum (positive control), or ruminal infusions of 1, 2, or 3 mg 5-HTP/kg BW. Steers were fed alfalfa hay cubes once daily. Periods were 5 d in length, followed by a 5-d washout. Blood samples were collected from the jugular vein on days 1 and 5 at 0, 1, 2, 4, 8, 12, 16, 20, and 24 h post-feeding. Caudal artery hemodynamics at the 4th coccygeal vertebra were assessed 4 and 8 h after the morning feeding on days −1 (baseline), 1, and 5.
Results and Discussion: Steers receiving 0.5 mg 5-HTP/kg BW in the abomasum had greater (P < 0.05) serum serotonin concentrations compared with negative control. Ruminal infusion of 2 or 3 mg 5-HTP/kg increased (P < 0.05) serum serotonin compared with the negative and positive controls. On day 5, the caudal artery cross-sectional area increased (P < 0.004) by +31.9% relative to baseline in steers receiving 3 mg 5-HTP/kg BW ruminally. In conclusion, ruminal supplementation of 5-HTP at 3 mg 5-HTP/kg BW increases circulating serotonin and promotes peripheral vasodilation in growing beef steers.
Poster #30
Introduction: Virus infection rapidly induces interferons (IFNs), which amplify antiviral responses in infected and neighboring uninfected cells. IFN regulatory factor 7 (IRF7), the ‘master transcription factor,’ is essential in IFN induction, particularly in myeloid cells. Ubiquitination of IRF7 is essential for its transcriptional activation; however, the underlying molecular mechanisms remain poorly understood.
Method: We hypothesized that deubiquitinases (DUBs) act as endogenous regulators of IRF7 activity that led us to conduct a genetic screen using a human DUB-targeted siRNA library.
Result: This screen identified a positive and a negative regulator of IRF7 activity. OTUD5, an inhibitor DUB, inhibited K63-linked ubiquitination of IRF7, thereby suppressing its activation. Conversely, USP2 promoted IRF7 activity by removing K27-linked polyubiquitination and was identified as an inhibitor of IRF7 activation.
Conclusion: Collectively, our genetic screen, combined with the mechanistic studies uncovered USP2 and OTUD5 as novel modulators of IRF7 function, providing new insights into the regulation of antiviral immunity.
Poster #31
Introduction: Chronic low-grade systemic inflammation is characteristic of individuals who have obesity-associated prediabetes and type-2-diabetes (T2D). Detailing the relationship between inflammation and disease progression will provide opportunities for treating inflammation to prevent/delay disease progression.
Method: We investigate how preT2D affected regulatory T cell (Treg) mitochondrial function using seahorse the measure oxygen consumption rate (OCR). We also measure cytokine production using bioplex.
Results: We function that preT2D Tregs that were expanded have higher OCR and glycolysis indicated by extracellular acidification rate (ECAR) from people with obesity and normal glucose tolerance (Ob/NGT) and lean people (Lean/NGT). Despite higher OCR and ECAR measurements in preT2D Tregs, there are no differences in mitochondrial number between Ob/NGT and preT2D Tregs. However, there are significant morphological differences in the mitochondria of Ob/NGT and preT2D Tregs. PreT2D Tregs are significantly smaller than those from Ob/NGT by several measured metrics including volume and area. We also found that preT2D Tregs produce Th1, Th17 inflammatory cytokines. Simultaneously, in the prediabetes state, Tregs have significantly higher expression of CD36, which mediates the update of fatty acid needed to fuel oxygen consumption. Inhibition of CD36 expression in preT2D Tregs resolve Th17 driven inflammation in these cells.
Discussion: PreT2D changes Treg mitochondrial morphology and CD36 expression. The changes in mitochondrial morphology are linked to changes in mitochondrial function in our preT2D Tregs. Further studies are intended to link the changes in mitochondrial function to inflammatory changes in preT2D Tregs.
Poster #32
Rhabdomyosarcoma (RMS) is the most common pediatric soft-tissue cancer and often presents with aggressive behavior and poor outcomes. Zebrafish patient-derived xenograft (zPDX) models offer a rapid and cost-effective platform for preclinical drug evaluation. Here, we optimized an RD-cell xenograft model in zebrafish larvae to assess chemotherapy responses. RD cells were injected into the dorsal perivitelline space (PVS) at 48 hours post-fertilization (hpf), achieving 61% engraftment at 24 hours post-injection (hpi) with viable xenografts persisting up to 4 days post-injection (dpi). Chemotherapeutic agents tested included dactinomycin (DAC) and vincristine (VIN) at 34 °C. In vitro, DAC significantly reduced RD cell viability at 2, 5, 10, and 25 nM, while VIN showed significant effects at 10 and 20 nM. Toxicity assays in zebrafish larvae confirmed survival up to 72 h at concentrations ≤100 nM for both drugs. Imaging analysis of xenografts demonstrated tumor persistence for at least 4 dpi and, in control groups, tumors displayed the characteristic aggressiveness and invasive capacity of RMS cells. Using a standardized ImageJ-based readout (metrics: FC_IntDen, FC_Convex, MIG, ΔSolidity), a pilot cohort (DMSO n=4, DAC 15 nM n=4, VIN 15 nM n=3) revealed distinct drug effects: DAC preferentially promoted dispersal over growth, while VIN restricted both tumor growth and spread. These findings establish zebrafish RMS xenografts as a functional in vivo model. Future studies will extend this approach to patient-derived primary tumor cells, enhancing the clinical relevance of zebrafish PDX platforms for precision oncology.
Poster #33
Background: Postprandial triglyceride (TAG) levels are an independent predictor of cardiovascular risk in humans. Chylomicrons (CMs) are the primary carrier of dietary TAG and they travel to the circulation via intestinal lymph. The protein components of CM’s can impact their metabolism in the circulation, however, the composition of CM’s is difficult to study because these particles are rapidly metabolized upon entry to the circulation.
Objectives: 1) Determine the protein and lipid composition of mesenteric lymph during fasting and after a lipid bolus, and 2) identify proteins in lymph that may impact the metabolism of newly secreted chylomicrons prior to entry into the circulation.
Methods and Results: A conscious lymph fistula procedure was used to collect mesenteric lymph from mice before and after infusion of a mixed lipid bolus into the duodenum. Wild type mice were compared to Dennd5b-/- mice (n=5-8 mice/group), a model of impaired chylomicron secretion. This study revealed that, compared to wild type mice, Dennd5b-/- mice exhibit significantly reduced TAG content in the lymph (-94%, p<0.001). Electron micrographs revealed that the lymph of wild type mice had both greater number and larger sized lipid particles compared to Dennd5b-/- mice. In addition, lipidomics analysis of fasting and postprandial lymph showed significant increases in triglycerides and free fatty acids in wildtype, but not Dennd5b-/- mice. Shotgun proteomics revealed significant changes in the lymph proteome of wild type mice after lipid bolus (14 proteins increased and 18 decreased; p<0.01). Key protein changes were confirmed by western blotting. One notable finding was an increase in lymph Saa1 after the lipid bolus (+200%, p<0.001). Size-exclusion chromatography was used to determine the distribution of Saa1 across lipoprotein fractions in lymph and demonstrated that Saa1 was associated primarily with fractions that contain chylomicrons and to a lesser extent with HDL-containing fractions. To understand the functional role of Saa association with lipoproteins under postprandial conditions intralipid gavage studies were performed in wild type and Saa-/- mice and plasma triglycerides and plasma IL6 levels were measured over 6 hours. While Saa-/- mice had no significant difference in postprandial plasma triglyceride concentrations compared to wild type mice, Saa-/- mice have significantly higher postprandial plasma IL6 concentrations compared to wild type mice.
Conclusions: We identified several proteins in intestinal lymph that respond to dietary lipid ingestion. In wildtype mice, lipid feeding increases lymphatic Saa1, which appears to be predominantly associated with chylomicrons. This response was not observed in Dennd5b-/- mice, indicating that it is dependent on successful CM secretion from intestinal epithelium. Our studies reveal that Saa contributes to attenuation of postprandial inflammation, but may not significantly impact chylomicron lipid metabolism in the lymph or circulation.
This research was funded by NIH grant R01DK133184.
Poster #34
Introduction: Radiographic assessment of the temporomandibular joint (TMJ) in horses can be challenging due to over imposition of cranial structures. While magnetic resonance imaging of the TMJ remains the most reliable diagnostic option, requirements for general anesthesia or specialized hospitals are often a barrier. Readily accessible diagnostic approaches for TMJ conditions are needed. The aim of this study was to compare two recently proposed, clinically unproven radiographic projections for the evaluation of the equine TMJ.
Methods: Using recently proposed oblique and tangential projections, 95 horses aged 15+ years (higher prevalence of TMJ pathology) had both TMJs radiographed, followed by a complete oral examination. Images were evaluated by two blinded radiologists, with radiographic findings (e.g., sclerosis, remodeling, lysis) recorded and correlated oral examinations.
Results: The oblique projection identified nine different findings (e.g., irregular articular surface, subchondral sclerosis, periarticular remodeling) in 50.5% of the horses with varying agreement between radiologists (range 0.61 < κ < 1, overall κ=0.64). The tangential projection identified four findings in 2.1% of horses with no interobserver agreement. When combined, three or more findings from oblique projections were associated with TMJ pain (right TMJ, p=0.021; left TMJ p=0.048). Increased probability of radiographic findings was associated with the presence of periodontitis (p=0.0001, observer A; p=0.0107, observer B) and pre-molar hooks (p=0.0124 observer A; p= 0.0003 observer B).
Discussion: The oblique radiographic projection was effective, reproducible and the findings correlated with dental-related pathology. These findings support the use of oblique projections on the radiographic diagnosis of equine TMJ conditions
Poster #35
Introduction: Spinal cord injury (SCI) causes motor paralysis and severe secondary complications. Individuals with SCI are highly susceptible to sepsis, which worsens functional outcomes and increases mortality. This study developed a clinically relevant rodent model to investigate long-term complications of sepsis in SCI survivors.
Methodology: Rats were assigned to four groups: Naïve, Sepsis, SCI, and SCI+Sepsis. SCI, and SCI+Sepsis groups underwent T10-laminectomy and contusion (200 kDyn) using an IH impactor. Sepsis was induced via intraperitoneal cecal slurry injection (3 ml) 15 minutes after SCI. Fluid resuscitation and antibiotics were initiated 8 hours post-SCI/sepsis and continued twice daily for 5 days. Over 12 weeks, assessments included bacteremia, survival, body weight, cytokine levels (blood/spinal cord), locomotor performance (BBB, horizontal ladder), in vivo muscle strength, and quantitative spinal cord histology.
Results and Conclusion: SCI+Sepsis animals showed markedly higher bacteremia at 6 hours post-sepsis induction versus Sepsis or SCI alone, correlating with the lowest survival. Locomotor recovery was further impaired in SCI+Sepsis compared to SCI, while Naïve and Sepsis groups exhibited no deficits. Muscle strength was significantly reduced in SCI+Sepsis relative to SCI. Acute-phase findings included splenomegaly, reduced hindlimb muscle mass, and elevated cytokines in blood and spinal cord.
This is the first rodent model to mimic post-SCI sepsis complications, enabling mechanistic studies and therapeutic evaluation.
Acknowledgement: This project was supported by funding from the National Institutes of Health (NIH), including grant 1R21NS128749-01A1 (SP/HS) from the National Institute of Neurological Disorders and Stroke (NINDS) and grant P20 GM148326 from the National Institute of General Medical Sciences (NIGMS).
Poster #36
Introduction: Renal Glut2 deficiency, like SGLT2 inhibition, induces massive glycosuria in mice. Yet, their fasting blood glucose levels remain normal indicating a compensatory production of endogenous glucose.
Methods: To investigate this, we performed snRNA sequencing combined with ATAC sequencing on the mouse kidney samples and identified the genes that may contribute to molecular pathways involved in glucose production. We used 13C6- glucose to determine the changes in metabolites in the liver, kidney and skeletal muscle tissues using NMR coupled with ion-chromatography-MS.
Results: RNA sequencing data revealed an increased expression of genes involved in oxidative phosphorylation and inner mitochondrial membrane proteins whereas transcripts for ribosomal subunits and amino acid metabolism were found to be downregulated in kidneys of renal Glut2 KO mice. The transcription factors involved in cellular stress response such as the Activator Protein-1 family members (JUN, JUNB/D, FOS, FOSL1/2/D) and their mutual regulators CREB5, SMAD2/3, and BATF were enriched in multiple cell types of kidneys in the KO mice, indicating reprogramming of glucose metabolism through upregulation of glycolysis and enhanced gluconeogenesis. The 13C6-glucose tracer analysis revealed an increase in isotopologue concentration of metabolites of glycolysis (G-6P, F-6P, F-1,6BP, 1,3-BPG, pyruvate), lactate, TCA cycle (citrate, isocitrate, cis-aconitate, α-ketoglutarate) and mannose pathway (mannose-6P, GDP-mannose) in kidneys of the KO mice.
Conclusion: Altogether, our data shows that the transcription factors and metabolic pathways involved in cellular stress-response may contribute to the compensatory glucose production observed in the KO mice. Targeting these molecular pathways may help improve the efficiency of SGLT2 inhibitors to treat type 2 diabetes.
Poster #37
Introduction: Fluorescence-based imaging and diagnostics are integral to modern biomedical research, particularly in cell tracking, biomarker detection, and molecular imaging. In this study, we introduce a method to induce and control fluorescence in polystyrene, a widely used bio-compatible material, via focused electron beam irradiation under controlled gas environments.
Methods: Spin-coated polystyrene thin films, which do not exhibit luminescence in the visible spectral region, were irradiated using a 20 keV electron beam in an environmental scanning electron microscope, under different gas pressures and electron doses. The resulting photoluminescence was characterized using confocal microscopy, while chemical and structural changes were examined using transmission electron microscopy (TEM), energy-dispersive spectroscopy, and Fourier-transform infrared (FTIR) spectroscopy.
Results: We observed a significant enhancement in fluorescence intensity (up to 18 times) and tunable emission wavelengths (451–544 nm), with the highest photoluminescence yields obtained under inert gases like helium and argon on insulating substrates. The method preserved the structural integrity of polystyrene without oxidation, as confirmed by TEM and FTIR analysis, indicating potential for long-term stability in biological environments.
Discussion: This research provides a scalable, efficient approach for producing bio-compatible, tunable fluorescent markers without the need for chemical dyes or additives. The ability to spatially control fluorescence within polystyrene films opens up exciting possibilities for biomedical applications such as real-time imaging of living cells, molecular diagnostics, and development of miniaturized biosensors.
Poster #38
Background: Brain arteriolosclerosis (B-ASC), a subtype of small vessel pathology, is present in more than 50% of individuals over the age of 80 years and is associated with cognitive impairment, motor dysfunction, and sleep disturbance. Multiallelic variants are likely to be ignored in genome-wide association study (GWAS) because standard statistical analysis methods are designed for biallelic variants. In this study, we applied score-based testing within the generalized linear model framework and explored multiallelic variant associations with autopsy-confirmed B-ASC in autosomal chromosomes.
Methods: We used whole-genome sequencing (WGS) data from the Alzheimer’s Disease Sequencing Project (ADSP) and B-ASC phenotype data from the National Alzheimer's Coordinating Center (NACC) neuropathology (NP) dataset (September 2023 data freeze). We dichotomized the B-ASC data (NACCARTE) into 0 = no/mild (n= 1,192) and 1 = moderate/severe (n=948) (Table 1). The model included sex, age at death, and the top three principal components as covariates. We then computed global scores and the corresponding p-values.
Results: After variant filtering and quality control, 1,388,681multiallelic variants were retained for study. The genomic regions and genes listed in Table 2 (with their p-values of less than 1×10-5) indicate potential involvement of these loci in BASC. Further investigation of the identified genes (HULC, CCDC3, DCUN1D2) and associated intergenic regions may provide insights into the genetic basis of B-ASC.
Conclusion: We investigated possible associations between human multiallelic variants and B-ASC risk. These are underexplored areas — both the genetic phenomena of multiallelic variants, and B-ASC as a dementia-driving pathology. Identifying novel genetic variants putatively contributing to the pathogenesis of B-ASC will move the field forward although validation with independent datasets is required.
Poster #39
Introduction: Fescue toxicosis in beef cattle, caused by the consumption of ergot alkaloids from toxic endophyte-infected tall fescue (TE), decreases circulating serotonin. Previous research has demonstrated that increasing serotonin can result in vasorelaxation of isolated bovine blood vessels. The objective was to evaluate the effects of 5-hydroxytryptophan (5-HTP) supplementation, a precursor of serotonin synthesis, on peripheral hemodynamics in cattle consuming ergot alkaloids.
Methods: Twenty Angus Holstein steers were ruminally-dosed with non-toxic novel endophyte-infected tall fescue seed (NTE) or TE to provide 15 g ergovaline/kg of body weight. In addition, steers were ruminally-dosed with either 0 or 3 mg 5-HTP/kg of body weight. Steers were housed indoors in individual pens, exposed to cyclical temperatures mimicking summer conditions (21°C–32°C), and fed alfalfa cubes for 15 days. Color Doppler ultrasonography was performed at 1200 h to measure the cross-sectional area of the caudal artery, resistance index, pulsatility index, and heart rate at the base of the tail on days 1, 8, and 15.
Results and Conclusion: Consumption of TE decreased (P=0.01) heart rate compared with consumption of NTE. Pulsatility index was decreased (P< 0.01) for steers ruminally-dosed with 5-HTP, and greater (P=0.01) for TE versus NTE. Resistance index was greater (P=0.03) for TE versus NTE and tended to be greater (P=0.07) for steers ruminally-dosed with 5-HTP. Caudal artery cross-sectional area relative to baseline decreased 35.81% (P=0.04) with ruminal TE dosing. These results indicate that TE increased vasoconstriction in steers and that 5-HTP supplementation may also influence cardiovascular responses, warranting further investigation.
Poster #40
Low-pass (skim) sequencing through genotype imputation has gained prominence as a cost-effective approach to be used in breeding programs to enhance genomic selection in livestock and farm animals. Application of this strategy in Thoroughbred horses, provides significant potential to improve population genomics and diversity studies while reducing costs associated with high coverage sequencing and helps to estimate unobserved genotypes from low coverage sequencing data, aiming to find an optimal balance between the cost of sequencing and the accuracy of the results.
Building on precise pipeline tools for low pass sequencing was successfully applied in cattle and other species. A reference panel constructed from Thoroughbred genotypes from whole genome shotgun sequence data is crucial to get high imputation accuracy. We will apply different imputation algorithms, to impute genotypes from low-coverage data. Preliminary findings show that imputation can achieve 98% accuracy.
Further refinement, and augmentation of the reference panel with more animals will likely improve genotyping accuracy to greater than 99%. This will make this approach reliable for various genomic applications in horses.
This study will demonstrate the advantages of using inexpensive low pass sequencing and imputation as a cost-efficient genomic approach for genomic studies in Thoroughbred breeding programs and research. By increasing the accessibility of genomic data and improving the prediction of complex traits, we will provide a solid genomic foundation for herd management. Our results suggest that this approach could be broadly applied across various equine populations, making a significant impact on genomic selection strategies in horse breeding.
Poster #41
Introduction: Accumulating evidence suggests that lipoproteins contribute to thrombosis by modulating procoagulant and anticoagulant pathways, though the mechanisms remain unclear. Von Willebrand factor (vWF), a circulating mechanosensor, unfolds under pathological shear stress, exposing binding sites for low-density lipoprotein (LDL) and high-density lipoprotein (HDL). We found that unfolded vWF binds and inhibits the anticoagulant protein S (ProS), an interaction amplified by an unknown plasma cofactor. We hypothesize that unfolded vWF sequesters ProS onto lipoprotein particles, impairing its anticoagulant function.
Methods: We mimicked shear-induced vWF unfolding by vortexing human plasma, and subsequently separated lipoproteins using fast protein liquid chromatography. Proteins were measured by ELISA.
Results: Vortexing caused a 30% increase in VLDL-like cholesterol fractions, without changes in ApoA1 or ApoB distribution, suggesting LDL aggregation. This effect was absent in plasma from a Type III von Willebrand disease patient, supporting a vWF-dependent mechanism. After vortexing, ProS levels decreased by 16% in the HDL and free protein fractions, and increased by 15% and 50% in LDL and VLDL fractions, respectively. Protein C and total protein profiles remained unchanged, indicating specificity for ProS. This redistribution was associated with impaired anticoagulant activity in plasma thrombin generation assays, with VLDL-associated ProS exhibiting 19% anticoagulant activity compared to its HDL-associated counterpart.
Discussion: In conclusion, shear-induced vWF unfolding promotes ProS redistribution to VLDL-like particles, diminishing its anticoagulant function. We propose this mechanism as a direct causal link between abnormal shear stress, such as that found in stenotic or tortuous vessels, or venous valves, ApoA1-containing lipoparticles, and increased thrombotic risk.
Poster #42
Introduction: RANKL is a critical cytokine that drives macrophage differentiation into osteoclasts and is responsible for bone resorption. Recently, we demonstrated that 14-3-3ζ- knockout (YwhazKO) animals are susceptible to increased bone loss. However, the function of 14-3-3ζ in macrophage activation or bone homeostasis has not been studied. Hence, we examined the function of 14-3-3ζ in the RANKL signaling, osteoclast differentiation, and activity.
Methods: We utilized 14-3-3ζ-deficient primary bone marrow–derived macrophages obtained from wildtype and YwhazKO animals and RAW264.7 cells generated using CRISPR-Cas9.
Results and Discussion: Compared with wildtype, 14-3-3ζ-deficient primary macrophages differentiated into bigger osteoclasts with increased nuclei number. These osteoclasts were TRAP-positive multinucleated cells with higher bone resorption activity. Importantly, 14-3-3ζ suppressed RANKL-induced MAPK and AKT phosphorylation, transcription factors (NFATc1 and p65) nuclear translocation, and subsequently, gene induction (RANK, Acp5, and Ctsk). Mechanistically, 14-3-3ζ interacts and promotes TRAF6 ubiquitination and degradation via the proteasomal pathway, thus inhibiting the RANKL signaling. These groundbreaking results indicate that 14-3-3ζ inhibits RANKL signaling and plays a critical role in bone biology. Regulating the TRAF6 levels by 14-3-3ζ is a novel mechanism for regulating bone homeostasis in inflammatory diseases.
Poster #43
Background: Intrinsically disordered regions (IDRs) of proteins play critical roles in various cellular functions including post-translational modification and cell signalling in eukaryotes. However, their roles in bacteria are poorly understood.
Methods: In this study, a deep neural network method was used to predict extracytoplasmic IDRs in the proteome of group A streptococcus (GAS). Lectin affinity chromatography followed by proteomic analysis, mutagenesis, immunoblotting, and in vitro assays were utilized to identify the role of IDRs in GAS biology.
Results: We identify that serine/threonine-rich C-terminal IDRs of five membrane proteins are glycosylated with α-glucose, indicating the presence of O-glycosylation pathway in S. pyogenes. The identified glycoproteins participate in protein folding and peptidoglycan synthesis. Mutagenesis approach reveals that two glycosyltransferases, GtrBSpy and PgtC2Spy, are involved in protein glycosylation. Kinetic analysis demonstrates that GtrBSpy possesses a micromolar affinity for UDP-glucose and undecaprenyl phosphate (Und-P), suggesting that GtrBSpy catalyses the first step of protein glycosylation in the cytosol transferring UDP-glucose to the lipid career, Und-P. We suggest that PgtC2Spy transfer a glucose moiety from a glycolipid to the IDRs of membrane-proteins. Finally, we identify that GtrBSpy and PgtC2Spy are crucial for GAS protection from antimicrobial proteins and peptides, cathepsin G and LL-37.
Conclusion: These data suggest that O-linked IDR-glycosylation might contribute to GAS evasion of innate immune defences.
Poster #44
Marine cyanobacteria provide a plethora of bioactive compounds for development of pharmacological tools and therapeutics. The marine cyanobacterial product, barbamide was originally isolated in 1996 as a molluscicidal agent. 1 In 2023, it was shown to have affinity for CNS receptors and the ability to modulate Ca2+ in in vitro systems. 2 Barbamide shows affinity for the dopamine transporter, D3 receptor, kappa opioid receptor (KOR), and both sigma receptors. Its polypharmacological profile provides a unique basis for the development of therapeutics against several diseases including neuropathic pain. A total synthesis of barbamide was undertaken to generate quantities to explore its biological activity. The synthetic route is amenable to rapid generation of a library of analogs exploring alterations of thiazole, benzyl, and the modified lipid regions of the molecule. This library is being screened for affinity and activity at sigma-2/TMEM97 and KOR. The analogs will also be analyzed for their effects on metabolic stability and potential to cross biological membranes. Subsequent investigations will focus on the mechanisms through which these novel analogs impact calcium modulation and exhibit antinociceptive properties.