Human Rhinovirus (HRV) is one of the primary causes of mild upper respiratory infections and is the most common infectious agent which affects billions of humans globally. To most healthy individuals, this illness causes mild symptoms. However, in populations who have immunosuppression, comorbidities, or predisposition health issues, this virus can cause severe symptoms which can lead to possible hospitalizations and even an increased mortality rate. Given the lack of approved therapeutics for this disease, our project aims to prepare a target Phenylpropenoid, which is an organic molecule that has been previously isolated from the plant Bupleurum fruticosum and has reported antiviral qualities against HRV. Our synthetic approach toward the target compound involves a three-stage process: synthesizing a phenylpropenol fragment, preparing a bis-enoate fragment, and combining the two through esterification to access the target phenylpropenoid. We have successfully prepared the phenylpropenol fragment and are working toward accessing the bis-enoate fragment for examination in the final esterification.  Our findings will enable preparation of derivatives to assess in bioactivity studies that may provide valuable insights for future target design.

Wildfire smoke is a significant environmental health risk for Latino agricultural workers, particularly in Washington State counties with high concentrations of both agricultural activity and smoke exposure, such as Yakima, Chelan, and Douglas. Young workers face unique and poorly understood risks due to potential biological susceptibility and a lack of safety communication tailored to their demographic.

This project explores how social media interventions can reach at-risk populations during smoke emergencies. A short Spanish-language video was developed to deliver protective guidance for agricultural workers in a clear, culturally relevant format. As the first in a planned series of short-form videos, this content promotes protective mask use and introduces key health messages in an engaging, accessible way. Designed for platforms such as TikTok, Instagram Reels, and YouTube Shorts, the video serves as an entry point to a broader educational campaign launching during the 2025 wildfire season.

This pilot will test the reach and engagement of short-form content among younger audiences, who may be less responsive to traditional public health channels. Engagement metrics—including views, likes, shares, and viewer retention—will be collected and analyzed to assess effectiveness and guide refinements in message design and delivery.

Insights from this evaluation will inform a summer campaign focused on improving wildfire smoke safety through practical, culturally grounded messaging. The campaign aims to promote protective behaviors and reinforce employer responsibilities under occupational health regulations. By integrating community-informed design and real-time platform analytics, this work contributes to the development of scalable tools for occupational risk communication and supports ongoing efforts to protect vulnerable workers during wildfire smoke events.

Chronic kidney disease is projected to be the fifth leading cause of death worldwide by 2040. Chronic kidney disease of unknown etiology (CKDu) makes up 70% of CKD cases in places such as India, Mexico, and Sri Lanka, largely through environmental factors. Ochratoxin A (OTA) accumulates in the kidney and is a nephrotoxic mycotoxin that contaminates grain products such as wheat, rice, beer, and most plant-based foods. Chronic OTA exposure has been linked to CKDu in rural agricultural areas, such as Sri Lanka. A prominent family of cell membrane transporters, Organic Anion Transporters (OATs), are one of the main drug transporter families in the kidney. Previous work in our lab elucidated that OAT1/3 and 4 are major OTA transporters. Certain antioxidants, found in common plant-based food products like green tea, coffee, and certain vegetables have been studied to reduce OTA-mediated nephrotoxicity. However, since our preliminary data indicate OAT transporter-dependent uptake into the kidney, we aim to test the competitive inhibition effect of OAT-substrate antioxidants in preventing kidney accumulation of OTA. Potential inhibitors include epicatechin gallate, miquelianan, caffeic acid, luteolin, and myricetin. Competitive inhibition in individuals consuming these products along with OTA exposure could lead to decreased uptake of OTA into the kidney, mitigating toxicity. Our preliminary uptake experiment with those inhibitors indicates that miquelianan reduces OAT3 mediated uptake of OTA by 48%. We will next assess the inhibition potential of miquelianan on OTA with an IC50 curve via mass spectrometry analysis. This study will provide evidence for a potential new mechanism of antioxidant amelioration of kidney toxicity to OTA exposure.

Ochratoxin-A (OTA) is a ubiquitous food contaminant linked to nephrotoxicity and carcinogenicity. Yet, its exposure risk and metabolic pathway in humans remain poorly understood. This research aims to investigate the intrinsic clearance of OTA in the human liver and to identify cytochrome P450 (CYP450) isozyme(s) responsible for its biotransformation. I employed a substrate depletion assay on OTA-treated human liver microsomes and used ultraviolet–visible spectroscopy to determine the kinetic parameters of clearance rates. To identify specific CYP450 isozyme(s) involved in metabolism, a parallel substrate depletion assay was conducted with recombinant CYP450 supersomes at defined intervals. Findings from this study reveal human susceptibility to OTA-induced toxicity and offers insight to our understanding to the hepatic metabolism of this widespread dietary toxin. Future research will explore human proximal-tubule specific OTA bioactivation, ultimately guiding regulatory decisions and public health interventions to reduce OTA-associated health risks.

In vitro models (cells in a dish) are a powerful tool in toxicology, allowing for advanced research in biological mechanisms while decreasing our reliance on in vivo animal models. Reproductive development is a critical endpoint in toxicology and requires a large number of animals, making reproductive studies a priority for in vitro alternatives. The current in vitro testis models are insufficient to recapitulate human reproductive development as they still rely on cells from laboratory rodents due to low human testis tissue availability and the need to capture dynamic developmental stages. To address this, I am developing an in vitro model that recapitulates human spermatogonia development to generate human primordial germ cell-like cells (hPGCLCs) using two induced pluripotent stem cell (iPSC) lines. This approach relies on spontaneous differentiation of the iPSCs using an extracellular matrix overlay. My pilot experiments did not robustly differentiate; therefore, I adapted the protocol to first induce incipient mesoderm-like cells (iMeLCs), which are primed for differentiation to hPGCLCs. I observed distinct cell morphological differences in the iMeLCs relative to control iPSCs using phase-contrast microscopy and found increased expression of Vimentin in the iMeLCs using immunocytochemistry. I am completing additional experiments to visualize expression of the mesoderm marker Brachyury, proliferative marker ki67, and primordial germ cell markers ki67 and SOX17. Using these iMeLCs I will follow the overlay protocol to derive hPGCLCs. I will assess the hPGCLC phenotype using flow cytometry for TFAP2C, a marker of PGCs. The hPGCLCs will then be cocultured with primary testis tissue to drive development towards spermatogonia-like cells (SpLCs), determined by expression of DDX4. The primary tissue will include our labs standard rodent model, as well as human tissue from collaborators at the UW Male Fertility Lab. Developing a fully human in vitro model system will be a powerful tool to study infertility.

Influenza viruses are a causative agent of seasonal flu outbreaks, which are mitigated through routine vaccination. Due to antigenic drift, many illness-causing strains evolve slower and are therefore, well-characterized. However, new strains occasionally emerge from animal reservoirs through antigenic shift, which can evade pre-existing immunity and cause lethal pandemics. Currently, H5N1 strains are of global health concern. Influenza viruses have two major antigenic surface glycoproteins: hemagglutinin (HA) and neuraminidase (NA), which have opposing functions and depend on a host cellular receptor, sialic acid. HA binds sialic acid for virus entry while NA cleaves sialic acid for viral release. NA is a dimer of dimers with several distinct domains, and two of particular interest: a head domain with sialidase activity and a flexible, hypervariable stalk domain. It is suggested that stalk length alters the range of accepted substrate-enzyme geometries of the NA head. As such, it is hypothesized that stalk length influences NA expression levels, sialic acid cleavage, and head tilting. Recent literature also demonstrates that shorter NA stalks result in reduced viral fitness in human hosts. Characterizing the structural effects of different NA stalk truncation constructs will provide valuable insight into influenza host-virus interactions. HDX-MS is an excellent tool for determining the structural dynamics of NA head regions by measuring local backbone amide solvent accessibility. MS data provides a detailed profile of deuterium uptake kinetics, effectively identifying differences in NA head flexibility across constructs. Additionally, we will use negative stain electron microscopy to observe differences in NA quarternary configuration and head tilting. We plan to correlate structural changes across constructs to changes in NA native function using a variety of NA activity assays in further experiments. This ongoing study aims to inform about how NA stalk length affects the influenza replication cycle, pathogenicity, and broader implications on host immunity.

6PPD-Quinone (6PPD-Q) is a toxic transformation product of the tire rubber additive, 6PPD, that has been identified as the primary cause of Coho Salmon (Oncorhynchus kisutch) mortality in watersheds impacted by roadway runoff. Recent studies have focused on quantifying the lethal concentration of 6PPD-Q, identifying the major sources, and predicting the environmental release from rubber products. Organic chemical release from solids is typically evaluated with solvent extraction where organic solvent and solid are contacted, releasing the leachable chemicals for measurement. However, different solvents and methods introduce inconsistencies in leaching data from different laboratories. This study evaluates the impact of solvent choice on 6PPD-Q extraction from crumb rubber. I will quantify 6PPD-Q concentrations in methanol, ethyl acetate, or acetone during storage after rubber extractions. Determining the best solvent for 6PPD-Q that promotes the most recovery and stability is essential for data quality. After this study, desorption and resorption rates of 6PPD-Q onto various crumb rubbers will be measured.  These studies aim to improve study design for leaching assessments and enhance our understanding of the persistence and mobility of 6PPD-Q in the environment.

Liposomes are synthetic vesicles composed of phospholipids that are used as both a model biological membrane and drug-delivery system. Doxil® is a widely used liposome-based chemotherapy drug used to treat ovarian cancer, multiple myeloma, and Kaposi’s sarcoma. Liposome stability affects drug-delivery efficacy. Cholesterol is a key component of membranes that has been shown to regulate membrane fluidity, permeability, and overall structure. Electrostatic interactions between phospholipid headgroups also can impact liposome stability and are impacted by buffer conditions. While it is known that inclusion of cholesterol and electrostatic interactions can impact liposome stability, how these changes influence membrane structure and stability is poorly understood. Cryo-electron tomography (CryoET) is an electron microscopy technique that produces high resolution 3-dimensional images of macromolecular structures, allowing detailed visualization of lipid bilayers and membranes. Cryo-ET can be used to preserve native hydration of membranes in order to maintain lipid organization. Using Cryo-ET, we plan to study how inclusion of different cholesterol concentrations and phospholipid compositions can influence membrane architecture and stability. We hypothesize that we will be able to directly visualize and analyze structural changes in membrane leaflets and membrane fine structure, which will enhance our understanding of lipid membrane architecture. An in-depth understanding of how cholesterol concentrations in liposomes under various buffer conditions influences membrane architecture will provide insight into how these factors directly impact membrane architecture and thus liposome stability. This knowledge is crucial for optimizing liposomes as drug delivery systems, improving their stability and efficiency, and enhancing their use as model membranes for studying biological processes.

Organs-on-a-chip (OOAC) are biomimetic systems that replicate the physiological environments of human organs at a micro-scale. They are gaining industry acceptance due to their ability to control critical parameters including shear stress, concentration gradients, and cell-biofluid interactions. By mimicking the behavior of human organs, OOACs are transforming how pharmacokinetics, physiological, and toxicological studies are performed, offering a more relevant model than animal-based studies. Our studies focus on how drugs and toxins affect the human kidney, a crucial organ for processing medications and filtering out harmful compounds. A key component of kidney OOACs is a hydrogel, which provides a structural scaffold and a biological substrate for cells. The hydrogel consists of rat tail Collagen I (Col-I) and specialized cell culture media (PTEC and 199 (10x)). The media mimics the extracellular fluids that surround kidney cells in the body, providing a more realistic environment for cell growth/interaction. Collagen IV (Col-IV) is the most abundant protein in kidney tissue but lacks structural rigidity. A combination of these materials is crucial for achieving a more accurate representation of kidney structure and function. While adding more matrix to the hydrogel improves the model’s ability to replicate the native environment, it is challenging to maintain structural stability, hence the need for a stabilizing agent. The aim of this project is to determine the proximal tubule epithelial cell (PTEC) viability of a mixed collagen I and IV matrix. At this stage, we have shifted from determining optimal collagen ratios to evaluating cell viability. By refining these models with optimized kidney extracellular matrices, the Kelly-Yeung lab aims to develop OOAC systems that better predict how drugs, toxins, and diseases impact human kidneys. This progress will lead to more effective and personalized treatments, as well as a reduction in reliance on animal testing.