Non-tuberculous mycobacteria (NTM) are ubiquitous in the environment and survive in various settings such as soil and water. The purpose of this project is to evaluate the ability of humoral responses produced from NTM pre-exposure on the prevention of Mycobacterium tuberculosis (M.tb) disease progression in mice. Additionally, we want to determine if antibody responses from immunization with bacille Calmette-Guerin (BCG) and/or candidate tuberculosis subunit vaccines contribute to protection against M.tb. To test this, we collected serum samples from mice that received various combinations of treatments before and after infection with M.tb. Groups either received NTM in the drinking water or normal drinking water, with or without BCG vaccine, and with or without a candidate vaccine or a combination of the treatments. M. avium ATCC 25291 NTM-lysate was used as an antigen for ELISAs and it was created by growing the mycobacteria and lysing the bacterial cells by multiple freeze-thaws. NTM-specific total IgG ELISAs were performed on serum samples following different vaccine strategies. Before M.tb infection, mice vaccinated with a candidate subunit vaccine regardless of NTM exposure or BCG vaccination had a significantly higher total IgG response. After M.tb infection, mice that received immunization with BCG followed by the candidate vaccine had significantly higher NTM lysate-specific total IgG responses compared to the saline group, however, this was observed only in NTM unexposed mice. NTM-exposed mice had equivalent levels of NTM-specific total IgG in the saline group when compared with the different vaccine regimens. This suggests that NTM exposure is able to produce potent NTM-lysate total IgG antibody responses in unimmunized ‘saline’ control mice, which suggests that the mice were likely boosted following M.tb infection. Given these findings, we plan to investigate whether the NTM-specific antibody response is reducing vaccine-mediated protective responses against M.tb.
 

Current technology to control 3D cell function takes advantage of bioorthogonal photochemistry to immobilize proteins into materials by using photocages—photoremovable molecular groups that block protein activity. However, diffusion limitations necessitate patterning times ranging from hours to weeks, far longer than the timescales of many biological processes. Protein activation aids signaling events within the extracellular matrix (ECM), leading to downstream changes in cell fate and physiological responses in our bodies. In order to probe and investigate biological systems, hydrogel materials provide an ideal synthetic platform, due to their polymeric, water-swollen characteristics that mimic the native ECM. In this project, I will use light to control the spatial and temporal presentation of biochemical cues through the photoactivation of proteins within hydrogels. We hypothesize that the kinetics of the protein activity between solution and biomaterial studies should correlate, given their dose-dependent response to light exposure—by varying the intensities of light and time intervals of exposure. By characterizing the photoactivatable protein system and controlling protein activity, we intend to use this platform to photoactivate biologically relevant proteins to control signaling that occurs on shorter time scales, applicable to biochemical processes.

Current trends indicate that while adolescents and young adults are starting to engage in sexual activity later in life and less frequently, there is a notable shift towards experiences characterized by low commitment and higher risk. Despite numerous studies on sex-related attitudes and practices among young people in the United States, broader insights are necessary to achieve a holistic understanding of the newly emerging landscape of youth sexuality. Against this background, our research examines sexual attitudes and reported behaviors in a sample of 1,200 undergraduate students at the University of Washington. Our study employs a comprehensive survey to identify patterns in sexual attitudes and behaviors among a representative sample. Administering online questionnaires is crucial in ensuring increased reliability and validity of responses on sensitive topics. These include but are not limited to, pornography viewership, kink and fetish interests, sexual self-concept, and more. Preliminary findings indicate a complex interplay between cultural, educational, and social factors in shaping sexual norms and practices among young adults. This study contributes to the broader discourse on sexual health education and highlights the need for nuanced approaches in addressing the dynamic and multifaceted nature of young people's sexual relationships. The results of this investigation aim to inform the foundation of modern sex education in order to address the evolving challenges of today's sexual landscape.

Bioconjugation, or the covalent linkage between a biomolecule and another chemical group, creates hybrid "conjugates" that exhibit the properties of both biomolecules and exogenous moieties. The N-termini of proteins often fall outside of their final fold, making the N-terminus an optimal site for conjugation while preserving a protein’s native folding and bioactivity. Consequently, N-terminal modification of proteins and peptides has been a long-standing goal in fields like drug delivery, biotherapeutics, and cellular imaging. However, the current techniques for N-terminal protein conjugation are limited by either the introduction of bulky protein assemblies at the conjugation site, the need for multiple costly and complicated steps, or low site selectivity. In this project, we aimed to develop an improved route for N-terminal bioconjugation. We created a generalizable platform for single-step purification and near-scarless N-terminal bioconjugation of proteins by leveraging the chemistry of the atypically split intein VidaL. To evaluate the effectiveness of our platform, we first examined the kinetics and reaction conditions of VidaL bioconjugation, confirming its ability to modify the N-termini of proteins successfully and selectively. Then, we used our platform to conjugate an alkyne, biotin, or FAM-biotin moiety to the N-termini of fluorescent proteins (EGFP and mCherry), a model enzyme (beta-lactamase), and a model growth factor (EGF). Through measuring fluorescence and conducting nitrocefin and proliferation assays, I found that, regardless of the moiety added, bioconjugation did not impact the native function or activity of these proteins. In the future, we expect that this platform's ability to easily N-terminally bioconjugate proteins with minimal impact on their functionality will find use across the growing fields of applied chemical biology.