Multiple studies have demonstrated an association between a long-term exposure to air pollutants and health outcomes in area of cardiovascular and respiratory diseases. However, there is suggestive, but insufficient, evidence linking the association between long-term exposure to air pollution and mental health outcomes, such as depression. In South Korea, a nationwide population-based, cross-sectional study investigated the relationship between long-term exposure to ambient air pollutants and mental health status. The study of 124,205 participants found a positive associations between Particulate Matter (PM10 and PM2.5 ), Nitrogen Dioxide (NO2), and Carbon Monoxide (CO) exposures and mental health status. The goal of this study is to determine whether the air pollution-depression relationship holds true in the ACT (Adult Changes in Thought) cohort by conducting a similar study to the cross-sectional study conducted in South Korea. In this study, I will consider air pollutants (PM2.5 and PM10) as exposure and depression as a measure of health outcome. The central working hypothesis of this study is that higher levels of PM2.5 and PM10 will be associated with an increase in CES-D (Center for Epidemiologic Studies Depression) score. Participants from the ACT cohort (members of the Kaiser Permanente in Seattle, Washington) who had an ACT study visit between 11/1/2000-4/1/2003 will be the study population. The primary analysis aims to evaluate the association between PM (PM2.5 and PM10) and CES-D scores using linear regression model after adjusting for age, sex, smoking, income, education, cardiovascular disease, and cerebrovascular disease. The secondary analysis aims to evaluate the effect modification of age, alcohol history and early vs. late onset of depression has on this association. It is hoped that the results from this study will expand our understanding of the relationship between air pollution and depression.

The application of quantum dots (QDs) is increasing in optical manufacture (solar cells), biological and chemical devices (cancer research). Exposure to QDs can potentially lead to a variety of adverse health effects especially for children and pregnant women. The goal of this study is to determine the toxicity of CdSe/ZnS QDs on brain development of both sexes using in vitro techniques. To determine sex-specific response, we used human neural progenitor cells (hNPC) lines derived from male (NSC-H14) and female (hNP1). An in-vitro model was established using an adverse outcome pathway (AOP) framework. We plated cells in proliferation or differentiation media for treatment to assess the morphology and biochemistry response in cells. Two QDs, differing in their surface chemical properties: carboxyl functional group – ITK and polyethylene glycol (PEG) without reactive functional group (Qtracker) were assessed. Cadmium chloride (Cd) was also tested to determine whether the toxicity of QDs is associated with ion release. The NSC-H14 and hNP1 cells were exposed to QDs of concentration from 0, 2.5, 5, 10, 20 and 40 nM on days in vitro (DIV) 0. After 24 hours, we evaluated the number of cells remaining (cell viability) with an LDH assay at each dose treatment. The results showed that exposures to ITK QDs demonstrated significant dose-dependent decreases in viability of both cell lines during the proliferative stage. Male (NSC-H14) cells were more responsive than female (hNP1) cells to the cytotoxicity of ITK. We did not observe significant dose response for Qtracker and Cd at the same dose range as ITK, either in proliferation nor differentiation stages of both male and female cell lines. Our results indicate that the coating on quantum dots may play a significant role in the exposure and toxicity of the cells, suggesting potential strategies to reduce the adverse health effects of quantum dots.

Climate change has increased the prevalence and intensity of wildfire events in the Pacific Northwest. Exposure to wildfire smoke has been linked to several adverse health outcomes, most notably respiratory-related morbidity and mortality. Local public health response to reducing a community’s exposure has primarily focused on communicating risks to the public and provide suggestions to reduce exposure risk. Effectively communicating the health effects of wildfire smoke and the interventions possible to reducing exposure is imperative for reducing adverse health outcomes during smoke events. In order to discern the messages the public received, we conducted a content analysis of wildfire smoke risk information communicated by local and state government organizations and by the mainstream media during the August 2018, statewide wildfire smoke event in Washington state. The Extended Parallel Process efficacy and risk framework was used to assess the presence and message content of wildfire smoke risk and efficacy information. Summary statistics were calculated to identify common messages about efficacy recommendations, vulnerable populations, and trusted sources of public health information. With these results, we hope public health jurisdictions will better understand the extent of their wildfire smoke risk messages and improve their standing as trusted sources of information in the community. Through the use of the extended parallel process framework-- a “recipe” for future wildfire season communication can be created.

Short-term exposure to diesel exhaust particulate matter can cause headaches, dizziness, and irritation of the nose, throat, and eyes. Prolonged exposure has been shown to increase the risk of developing cardiovascular disease, respiratory disease and lung cancer. Because heavy machinery is often fueled by diesel, occupations such as coal miners, truck drivers, railroad workers, and construction workers are at high risk of exposure. Non-occupational exposures are also of concern, especially in locations impacted by high volumes of vehicle traffic. A potential way to determine diesel exhaust exposure is by measuring the amount of nitrated polycyclic aromatic hydrocarbons (NPAHs) in household dust. There is no current method for measuring NPAHs in dust. We conducted a literature review of methods for measuring related chemicals in household dust and measuring NPAHs in other matrices. We tested three different methods before establishing the optimized sample preparation and cleanup process using silica gel solid phase extraction followed by analysis using high performance liquid chromatohraphy with tandem mass spectrometry detection (HPLC/MS/MS). To evaluate method performance we analyzed replicates of spiked and unspiked household dust and silica gel. The method was used to analyze 42 household dust samples collected in two communities – one with expected high and one with expected low traffic-related air pollution. Future research should include comparisons between dust NPAH measurements and other measures of diesel exhaust exposure, including NP (nitropyrene) metabolites in urine, air filter NP, or a priori predicted exposure based on home location.

Domoic acid (DA), a common marine neurotoxin found in marine organisms such as shellfish and finfish, is gaining public attention with increasing outbreaks in the USA, Australia, China, and some countries in Europe. Most DA outbreaks are observed along coastal regions, increasing the risk of exposure by seafood consumption. However, little is known about the effects of low level prenatal exposure of DA. To address this growing public health issue, we administered behavioral and cognitive tests on 27 Macaca fascicularis infants prenatally exposed to 0, 0.075, and 0.15 mg/kg/day of DA. Their cognitive capacities were analyzed based on their speed of learning new tests using the Wisconsin General Testing Apparatus (WGTA). The WGTA requires testers to present stimuli to the infant and, depending on the response, reward the infant for a correct response. I was one of several testers that worked with the infants during testing sessions 5 days per week. I also worked with the investigators and data analysis team to analyze the results of the WGTA tests. Results from early cognitive tests of object and spatial discrimination did not indicate a statistically significant detrimental effect of DA on these basic learning tasks. This suggests that DA does not have an impact on basic discrimination learning in the early stages of life (< 1 year). However, due to the potential for delayed toxicity of DA, more complex learning and memory tests are being administered as the infants mature. The results of this study will provide insights into future research and influence policies being implemented regarding seafood consumption and food safety.

Maternal exposure to persistent environmental toxicants polybrominated diphenyl ethers (PBDEs) is associated with many human diseases, notably hepatotoxicity, thyroid disorders, and reproductive toxicity. PBDEs, previously seeded into manufactured products as flame retardants in the United States, were phased out during the 2000s. A 2017 study of women in California showed modest average annual percent increases in serum concentrations of PBDEs. Accordingly, the gut microbiome is increasingly recognized as the “second genome”, impacting crucial health-related factors like host xenobiotic biotransformation and energy homeostasis. Persistent dysbiosis may contribute to altered susceptibility of disease in adulthood. My goal for this study was to test the hypothesis that perinatal exposure to BDE-47, a common PBDE congener that can cross the placenta and enrich in breast milk, modifies the developmental trajectory of gut microbiota. Pregnant Wistar rats were orally exposed to BDE-47 (0.2mg/kg) from gestational day 8 to postnatal day (PND) 21. Fecal samples were collected from mothers at PND21 and male pups at PND65 and 120. I isolated Microbial DNA using 16S rDNA sequencing (n=5~9/group) and analyzed my data through QIIME. Perinatal BDE-47 exposure had minimal effect on the richness of gut microbiota in PND65 pups and mothers, but profoundly increased richness of gut microbiota in PND120 pups. In PND65 pups, only 4 taxa were persistently regulated by perinatal BDE-47 exposure. Interestingly, the effect of perinatal BDE-47 exposure was evident in PND120 pups, with an increase in 3 taxa in the Lactobacillales order and 11 taxa in the Clostridiales order. Targeted metabolomics has confirmed alterations of distinct microbial metabolites (short-chain fatty acids and secondary bile acids) corresponding to BDE-47-induced dysbiosis. In summary, the present study showed that perinatal BDE-47 exposure modified the developmental trajectory of the offspring’s microbiota, possibly producing delayed onset of diseases in adulthood.
 

Theiler's murine encephalomyelitis virus (TMEV) is a positive-sense RNA Picornavirus used to model epilepsy, poliomyelitis and multiple sclerosis. Previous research has demonstrated a dependence of some Picornaviruses on host glutathione (GSH) in order to produce viable progeny. GSH depletion in host cells has resulted in unstable viral capsids, reducing viral fitness due to a lack of oxidative homeostasis. The objective of this study was to investigate the dependence of TMEV on GSH and determine the consequences of oxidative stress in the host cell due to the absence of GSH. Incubation of TMEV at denaturing temperatures with increasing doses of reduced GSH demonstrated that GSH stabilizes viral particles and 54% viral infectivity was retained with doses as low as 25mM GSH. Using L-Buthionine-sulfoximine (BSO) to deplete GSH in host cells, a 3-fold decrease in viral production was observed when compared with untreated cells. Interestingly, when GSH was restored in BSO treated cells using cell permeable GSH ethyl ester, TMEV viral production was partially restored and viral plaque formation was comparable in yield to an untreated viral infection. Virus production and viability were quantified using plaque assays on host cell monolayers. Next steps include monitoring viral genome production via RT-qPCR and quantifying GSH levels using fluorescence plate assays. RT-qPCR will be further utilized to examine other host stress pathways. This project will use TMEV to look at a host-virus relationship as it relates to an important host antioxidant system interacting with the capsid, assembly, and genome replication of the virus life cycle.

Oral swab analysis (OSA) is a possible alternative sample type for tuberculosis diagnostics. It has been observed that tongue swabs contain greater amounts of Mycobacterium tuberculosis DNA than cheek swabs (p<0.0001) from tuberculosis patients. After determining that oral microbiota follows this same pattern, several factors including time-of-day swabbed and health status were analyzed to understand factors affecting the amount of bacteria on the tongue. This project aims to optimize the oral swab sampling methods in order to facilitate more sensitive diagnostic tests, using universal bacterial 16s rDNA as a proxy for Mtb DNA. Previously tested samples from South Africa were further analyzed to investigate amount of oral microbiota by day collected, HIV status, health status, and other demographic factors. To evaluate whether collecting multiple swabs per sample yielded more universal bacterial DNA, tongue swabs were taken from healthy volunteers in Seattle. Each subject provided a 1-swab sample and a 3-swab sample, which was then extracted and analyzed by a previously optimized universal bacterial PCR. Additionally, tongue scrapers are being assessed as an alternative to oral swabs. Swabs collected early in the morning had more bacterial DNA than swabs collected later (p<0.03). 3-swab samples yielded an average of 2-fold greater amounts of bacterial DNA than 1-swab samples. Bacterial biomass correlated with M. tuberculosis signal in most comparisons. Bacterial biomass may serve as a useful proxy when developing better oral swab sampling strategies for TB diagnosis.

Over the past several summers, Washington state has faced high levels of wildfire smoke exposure. Significant research has indicated that wildfire smoke is a public health hazard. Public health professionals require evidence-based communication and intervention methods for use in exposed communities. It is valuable for researchers to understand current practice-based risk communication and management challenges to inform their research strategy and priorities. In response, the University of Washington's School of Public Health faculty and students convened a wildfire smoke risk communication symposium on October 30th, 2018, to foster academic-practice partnerships among regional stakeholders, identify existing risk reduction and communication challenges, and identify research needs. We conducted pre and post symposium surveys to collect information on participants’ goals, demographic characteristics, and symposium satisfaction. Descriptive statistics were calculated and displayed on graphs and tables. The registration survey identified the most common participant goals for the symposium which were to learn about lessons learned from public health practitioners related to wildfire smoke and to identify collaboration opportunities. Participants had a variety of roles related to risk communication and research. The evaluation survey revealed that over half of participants reported knowledge increases on wildfire risk communication, the future of wildfires in the Pacific Northwest, and practice-based responses to wildfire smoke. Over three-quarters of participants reported developing new connections that could lead to new collaborations in the future. Our findings suggest that there is an interest in working collaboratively to improve risk communication during wildfire events to address knowledge gaps and challenges impacted communities may face. We recommend engaging stakeholders and participants to identify the best communication methods and work with multidisciplinary partnerships that will be crucial to prepare for future wildfire seasons.

Building upon our rat testicular o-culture system developed to improve evaluation of chemicals and identify their potential adverse health effects on humans, we have designed in vitro mouse system to expedite such assessments. Models of in vitro cell cultures are, however, limited in their approach typically using a monoculture of single cell types. Using a 3 dimensional organotypic approach can provide a framework for evaluating systematic interactions between cells with a goal to reduce the need for in vivo assessments. We utilized a novel organotypic, in vitro model of testicular development that mirrors the development shown by in vivo studies. We used isolated testicular tissue harvested from C57BL/6 male mice on post-natal day 9 and digested into a single cell suspension. Testicular co-cultures were maintained for up to 16 days in 24 well plates. Data collections occurred at days in vitro (DIV) 3, 7, and 16. These timepoints represent “windows of potential susceptibility” for testicular development. Plates were stained with antibodies providing markers corresponding to morphological features of specific cell type; six different primary antibodies were used as markers to visualize the change in cell populations; markers for Sertoli cells corresponded to Vimentin, Leydig cells with 3b-HSD, Germ cells to DAZL, SCP3 and c-kit, and proliferation with PCNA. We used an immunofluorescence dual staining technique with Scanning Laser Image Cytometery (iCys) to quantify the proportion of cell types and to determine changes in phenotypic distribution of the cell population. Our observations in vitro demonstrated concordance of changes in Sertoli cells, Leydig cells and Germ cells throughout testicular development with similar trends in vivo. Our observation of similarities in testicular development in our 3 D in vitro experiment and in vivo studies is important and supports the current trends in minimizing the need for in vivo studies for chemical toxicological.

Gut Microbiome is increasingly recognized as a pivotal player in toxicological responses, thus dysbiosis or microbial imbalance may worsen chemical-induced adverse outcomes such as inflammation, metabolic syndrome, and cancer. Early life exposure to environmental contaminants may produce long term toxicities in adulthood, and little is known to what extent early life exposure to environmental contaminants modulate the gut microbiome beyond adulthood. Therefore, this study tested the effects of perinatal exposure to 3 human health relevant environmental contaminants (BDE-47, TBBPA, and BPS), on the composition and functions of the gut microbiome of perinatally exposed adult male mice. CD-1 mouse dams were orally exposed to vehicle (corn-oil, 10ml/kg), BDE-47 (0.2mg/kg), TBBPA (0.2mg/kg), and BPS (0.2mg/kg) once daily from gestational day 8 to the end of lactation (postnatal day 21). Feces from male pups were collected at 12-weeks of age (n=14-23/group). Microbial DNA was isolated, subjected to 16rDNA sequencing, and analyzed using QIIME. Microbial biomarkers for each chemical exposure were predicted using LefSe. Microbial functions and key taxa that drive functional changes were predicted using PICRUSt and FishTaco, respectively. None of the 3 chemicals markedly altered the overall richness of the gut microbiome in adult male pups. However, principle coordinate analysis showed a distinct separation among different exposure groups, and especially between BPS and vehicle exposure groups. A total of 73 taxa were persistently altered by at least 1 chemical exposure, among which 12 taxa were commonly regulated by all 3 chemicals. The most representative microbial biomarkers for each exposure condition were Clostridiales for vehicle, S24-7 for BDE-47, Rikenellaceae for TBBPA, and Lactobacillus for BPS. Together, these observations suggest early life exposure to these human health relevant environmental contaminants produce persistent gut dysbiosis in adult male offspring, leading to functional shifts that may play important roles in regulating certain diseases of the host.

The Methow Valley is a community located in Okanogan County, WA that has experienced wildfires in 4 of the past 5 years. The Methow Valley Clean Air Project (MVCAP) is a local volunteer group that promotes air quality awareness through outreach and programming. MVCAP’s Purple Air Network was designed to provide access to spatial air quality information to help community members make decisions about protective actions, to identify relatively clean air spaces, and to serve as a public health invervention for wildfire smoke. In collaboration with MVCAP, we worked to install a network of 20 purple air monitors in the summer of 2018. The monitors were placed in homes of “Clean Air Ambassadors” who committed to maintain and promote the monitor. For calibration, each monitor was placed according to federal guidelines and two were collocated with nephelometers maintained by Washington Department of Ecology. During the summer of 2018, two wildfires burned nearby which allowed the monitors to be utilized in a wildfire smoke event. The data from the monitors was downloaded and compared to the nephelometer data using linear regression. I worked to establish a correction factor by analyzing the data and found that the Purple Air monitor over reported the PM2.5 concentration by a factor of 0.53 and even more at high concentrations. The network proved the importance of high spatial concentration monitoring by capturing the air quality variation. In some areas, the air quality was good while in others it was hazardous. Having the network is a tool for individuals to know what the air quality is near their homes and other places in their community. It allowed individuals to access local air quality data and make choices about poor air quality. The next step for MVCAP is designing an online interface that applies the correction factor directly to accurately communicate risk.

The PestiSeguro/PestiSafety app aims to give agricultural workers reliable information on pesticide safety by providing accurate Spanish translations. The health and safety information of the app is important because handling pesticides incorrectly may harm workers, crops, and the environment. The PestiSeguro/PestiSafety app was evaluated by the community itself, agricultural workers ranging from workers, supervisors, and farm owners. The app was also evaluated by state wide stakeholders. The evaluation consisted of 15min interviews where we gathered qualitative and quantitative data on demographics, translation quality, and recommendations. The interviews were conducted through phone on early mornings and late afternoons to accommodate agricultural workers schedules. During the Analysis of the qualitative data we started coding the content by topic and codes were generated from the data. This process was repeated 3 times adjusting codes and coding undil they were consistent. Then we identified themes and patterns that told us, what the community was benefiting from the most, errors made in the accessibility of translations, and improvements on user friendly experience, such as adding graphics. Among the testers 94% of them agreed that information in spanish helps to understand how to handle pesticides safely. Another 94% agreed that the translations were clear and understandable. Future work includes, releasing a demonstration app that can be downloaded directly from the app or play store and to continue to disseminate information to valuable stakeholders. 

Tuberculosis (TB) remains a major international health concern and one of the top 10 causes of death worldwide, according to the World Health Organization. Previous clinical work in our lab demonstrated that oral swab analysis (OSA) can successfully diagnose tuberculosis by detecting Mycobacterium tuberculosis DNA in the mouths of infected patients. In order to strengthen OSA against traditional but more invasive methods, such as sputum sampling, improvements to DNA extraction and swab type must be investigated. I am comparing different degrees of automation with MolBio’s Trueprep, the AudioLyse, and ThermoFisher’s KingFisher against our previously validated manual Qiagen extraction protocol—with the comparison lying in sensitivity and efficiency. In order to increase the versatility and sensitivity of OSA, I am also investigating boil preparations and dissolvable swabs. Boil preparation of swabs is a relatively simple extraction procedure, and early results have demonstrated its comparability against the Qiagen extraction. Meanwhile, dissolvable swabs have a hypothetical 100% yield of sample material. I have successfully dissolved calcium alginate swabs from Puritan in acidic sodium citrate solutions, and Luna swabs were dissolved in chaotropic agents. DNA yields are compared to non-dissolvable, previously validated swabs.

Clallam County has recently been experiencing concerning air quality conditions due to smoke drift from wildfire events in nearby areas. The objective of this study was to assess the wildfire smoke health-risk communication needs of organizations that serve the public in Clallam County. Communication needs were assessed by surveying 10 organizations that serve sensitive populations. Surveys were conducted in person or over the phone, and summary statistics were calculated. In addition, a low-cost air monitor was installed to track summer time air quality. The low-cost air monitor was installed adjacent to a nephelometer administered by the Olympic Region Clean Air Agency (ORCAA). This was done to determine the accuracy of the low-cost air monitor by comparing air quality values registered from both devices. The majority (60%) of organizations responding to the survey reported that they had not received information about the health risks of wildfire smoke. Nearly all organizations (90%) reported that they have the capacity and are willing to communicate the health-risks of wildfire smoke to the people they serve in Clallam County. Analysis of air quality data shows the low-cost monitor may be a useful device in determining air quality conditions. The correlation coefficient for the daily averages (from July 12 to August 24) between the ORCAA nephelometer and the low-cost air monitor was determined to be 0.98, but closer examination of data for hourly averages show a correlation coefficient as low as 0.82. Future wildfire smoke events in Clallam County require public health interventions to address health-risk communication needs of sensitive populations. Implementation of a low-cost air monitor network accessible by the public is a promising prospect to protect sensitive populations in Clallam County.