Pseudomonas aeruginosa (P. aeruginosa) pneumonia is characterized by alveolar- capillary inflammation and respiratory failure with high mortality. Current treatments remain supportive beyond antibiotics. Our earlier work showed pharmacological activation of large conductance Ca2+-activated K+ (BK) channels protects mice from P. aeruginosa pneumonia by decreasing inflammatory cytokine response in bronchoalveolar lavage (BAL) and plasma. This study investigates changes in BAL and plasma metabolites after treatment of P. aeruginosa-infected mice with BK channel activator NS1619, injected intratracheally once daily; BAL and plasma were collected after 72 hours. Metabolon’s Global Platform measured the effect of NS1619 on BAL and plasma metabolomes in infected mice compared to controls. We created a pipeline using R, Venny, and MetaboAnalyst, integrating metabolite profiling, network pathway enrichment analysis to identify pathways affected by NS1619. Using log2FC ≤ -1 or ≥ 1 (p < 0.05) on LC/MS and GC/MS data, 65 BALF and 2 plasma metabolites increased after infection and were reduced by NS1619. 10 BAL and 5 plasma metabolites decreased after infection and increased with NS1619. Network analysis revealed hub metabolites integrated into a multi-omics model via STITCH. The protective effects of BK channel activation against P. aeruginosa infection are linked to specific changes in plasma and BAL metabolite profiles; targeting these hub metabolites may offer a therapeutic strategy.

Air pollution is a widespread risk factor for pulmonary inflammation and cardiovascular disease. When inhaled, alveolar macrophages (AMs) are the first cells in the lungs to encounter ultrafine particles (UFPs), which are particles smaller than 0.1µm. AMs play a major role in regulating inflammatory responses by phagocytosing particulate matter (PM) and apoptotic cells. Macrophages can upregulate Heme oxygenase-1 (HO-1) expression to counteract PM-induced effects. While research has identified associations between larger PM (PM2.5) and pulmonary inflammation, the effects of UFPs are not well defined. This project investigates the impact of UFPs collected from 2 different methods on lung inflammation in myeloid-specific HO-1 knockout mice, while also assessing the role of HO-1 in regulating inflammatory cell recruitment. Mice were exposed to either filtered air (FA) or UFP at 300µg/m3 for 5 consecutive days. Bronchoalveolar lavage samples were harvested and prepared into slides via cytospin. Identification of specific immune cell populations in the lung was determined by Hema3 staining. We also characterized the pro-inflammatory profiles of alveolar macrophages with quantitative PCR (qPCR). Our results suggest that UFP exposure influences immune cell recruitment and alters pro-inflammatory gene expression in AMs while simultaneously implicating HO-1 as a potential modulator of these responses. These findings clarify how air pollution triggers health issues, helping to shape future prevention strategies.

Sequoia and Kings Canyon National Parks (SEKI) harbor a series of remote, mid-elevation populations of the northwestern pond turtle (Actinemys marmorata), a species of conservation concern. Although range delimitations and previous genetic studies indicate that the turtles in the park fall squarely in the range of pure A. marmorata, variation in inguinal scute morphology suggests co-occurrence or admixture with its sister species, Actinemys pallida. Recent anthropogenic disturbances, including a poaching event and the illegal introduction of suspected Actinemys hybrids, further emphasize the need to understand the genetic structure of this vulnerable, declining population. We generated genomic resequencing datasets for 80 individuals sampled from three river basins within SEKI to quantify genomic diversity at the individual, population, and landscape scales. Preliminary results reveal genetic substructuring consistent with isolation by distance, suggesting limited dispersal among riverine habitats. The detection of individuals with mixed ancestry also provides evidence of possible introgression from A. pallida into the population. Ongoing field and molecular work will expand these analyses to further resolve genetic structure and patterns of isolation. Ultimately, this research seeks to provide a genomic framework that will guide recovery and management actions for A. marmorata in Sequoia and Kings Canyon National Parks.

Tumor growth depends on vascular networks that deliver oxygen and nutrients to malignant tissue. In healthy tissue, vascular systems often follow principles of efficient transport, including power-law scaling between vessel radius and length and space-filling branching hierarchies. However, tumor vessels form under mechanical stress and may deviate from these optimal patterns. This study tests whether tumor vascular networks obey scaling laws and branching structures predicted by optimal network theory. Three-dimensional images were processed using a segmentation and graph extraction pipeline to reconstruct tree-like networks. Vessel radii, lengths, and Horton-Strahler orders were computed from these graphs. Scaling exponents were estimated using log-log regressions between vessel radius and length and compared to predictions from optimal transport models. Strahler analysis evaluated branching distributions and hierarchical depth. Preliminary results show linear trends on log-log plots consistent with power-law behavior but with scaling exponents that differ from classical predictions. Branching distributions are skewed toward lower-order vessels, suggesting localized sprouting rather than space-filling organization. These findings indicate that tumor vasculature partially follows optimal scaling but diverges in branching architecture. This provides a quantitative framework for understanding how physical constraints shape tumor vascular structure and may inform future studies about function and therapy response.

Microfluidics platforms offer the potential to miniaturize and automate conventional wet-lab assays while improving experimental consistency through precise control of discrete reaction volumes. Digital microfluidics based on electrowetting-on-dielectric (EWOD) enables programmable manipulation of individual droplets, supporting highly reproducible and scalable biochemical workflows. Here, we present a thin-film-transistor (TFT)-based EWOD microfluidics platform for performing polymerase chain reaction (PCR) at nanoliter-scale volumes. Using a custom-integrated heating system, we successfully amplified a 171 bp fragment of mouse GAPDH from cDNA derived from murine splenocytes across reaction volumes ranging from 10 µL to 0.2 µL. Amplification products were validated via gel electrophoresis, demonstrating consistent DNA yield across decreasing volumes when normalized per unit volume, despite reduced total product at smaller scales. Notably, volume-normalized signal remained comparable or enhanced relative to a 10 µL off-chip control, indicating preserved reaction efficiency at reduced scales. These results demonstrate that TFT-EWOD platforms can support robust enzymatic reactions with high efficiency and reproducibility, establishing a foundation for translating conventional molecular biology workflows into programmable, miniaturized lab-on-a-chip systems.