NIEHS SRP 2020 Annual Meeting December 14–15, 2020 Program Book

Virtual Meeting hosted by the Texas A&M Superfund Research Center

Contents NIEHS SRP 2020 Annual Meeting December 14–15, 2020 Program Book Contents...... 3

Main Program Agenda...... 4

Administrators Program Agenda...... 6

Welcome from NIEHS SRP • Bill Suk...... 8

Welcome from Texas A&M SRP Center • Ivan Rusyn...... 9

Keynote I • t.e.j.a.s. & Texas A&M University...... 10

Keynote II • US Enivironmental Protection Agency...... 11

Wetterhahn & Donnelly Awards...... 12

Karen Wetterhahn Memorial Award...... 13

KC Donnelly Externship Awards...... 14

Scientific Presentations I: Community-Informed Superfund Research...... 15

Scientific Presentations II: Data Science Solutions for Superfund Challenges...... 21

Poster Session I...... 26

Poster Session II...... 96

Poster Session III...... 162

Poster Session IV...... 232

Attendees...... 298

NIEHS SRP 2020 Virtual Annual Meeting Program • 3 ContentsMain Program Agenda

ALL TIMES LISTED ARE EASTERN US TIME ZONE Monday, December 14, 2020 Theme: Addressing Hazardous Substance Exposures at the Community Level – Solutions by the Superfund Research Program

Opening Remarks by the NIEHS Superfund Research Program Director (SRP): 10:00 - 10:05 a.m. Bill Suk (NIEHS) Opening Remarks by the Texas A&M University SRP Center Director: 10:05 - 10:10 a.m. Ivan Rusyn (Texas A&M University) Opening Remarks by the Texas A&M University Vice President for Research: 10:10 - 10:25 a.m. Mark Barteau (Texas A&M University) Opening Remarks by the NIEHS Director: 10:25 - 11:00 a.m. Rick Woychik (NIEHS)

Keynote I: (introduced by Weihsueh Chiu, Texas A&M University) Building Resilient Communities to Protect Against Exposure to Hazardous Substances: Multidisciplinary Engagement and Partnerships with the 11:00 - 11:45 a.m. Superfund Researchers • Juan Parras & Nalleli Hidalgo (Texas Environmental Justice Advocacy Services [t.e.j.a.s.]) • Galen Newman & Garett Sansom (Texas A&M University)

Karen Wetterhahn Award Announcement & Lecture 11:45 - 12:30 p.m. (moderated by Bill Suk, NIEHS)

12:30 - 1:00 p.m. Lunch Break KC Donnelly Externship Award Presentations: (moderated by Stephen Safe, Texas A&M University) • The Legacy of KC Donnelly’s Contribution to the Superfund Research Program 1:00 - 1:30 p.m. Stephen Safe (Texas A&M University) • 2019 KC Donnelly Award winners • Jill Riddell (West Virginia University, Northeastern University Superfund Research Center) • Nabil Shaikh (University of New Mexico Superfund Research Center) Scientific Presentations I: Community-Informed Superfund Research (moderated by Weihsueh Chiu, Texas A&M University) • Harrison Schmitt (University of Arizona Superfund Research Center) “Chronic environmental contamination: Psychosocial health consequences, risk factors, and pathways to community resilience” • Rui Zhu (Texas A&M University Superfund Research Center) 1:30 - 2:30 p.m. (introduced by Galen Newman, Texas A&M University) “Resilience through Regeneration: An engagement and performance-based approach to repurposing vacant community lots with green infrastructure” • Tyler Gripshover (University of Louisville Superfund Research Center) (introduced by Matthew Cave, University of Louisville) “Volatile organic compound exposures are positively associated with liver apoptosis in a residential cohort”

4 • NIEHS SRP 2020 Virtual Annual Meeting Program Main Program Agenda

• Clare Pace (University of California-Berkeley Superfund Research Center) 1:30 - 2:30 p.m. (introduced by Rachel Morello-Frosh, University of California-Berkeley) “Advancing California’s Human Right to Water Law: Characterizing inequities (continued) in drinking water quality among domestic well communities and community water systems” 2:30 - 3:00 p.m. Break (Poster Preview) 3:00 - 4:00 p.m. Poster Session I 4:00 - 5:00 p.m. Poster Session II 5:30 - 7:00 p.m. Center PI meeting (separate log-in to be provided)

Tuesday, December 15, 2020 Theme: Meeting the Challenges of Environmental Protection with 21st Century Science

Keynote II: (introduced by Ivan Rusyn, Texas A&M University): US EPA’s Path Towards More Rapid, Efficient, 11:00 - 11:45 a.m. and Protective Chemical Testing with Fewer Animals • Russell Thomas (Center for Computational Toxicology and Exposure, US EPA) 11:45 - 12:00 p.m. Break (Poster Preview) 12:00 - 1:00 p.m. Poster Session III 1:00 - 2:00 p.m. Poster Session IV 2:00 - 2:30 p.m. Lunch Break Scientific Presentations II: Data Science Solutions for Superfund Challenges (moderated by Ivan Rusyn, Texas A&M University) • Rance Nault (Michigan State University Superfund Research Program) (introduced by Tim Zacharewski, Michigan State University) “Development of minimum requirements and data management framework for animal toxicology experiments” • Dillon Lloyd (NC State University/Texas A&M Superfund Research Center) (introduced by Fred Wright, North Carolina State University) “An R Shiny tool for spatial and correlation analysis of biological and chemical 2:30 - 4:00 p.m. measurements” • Madison Hattaway (University of California-Davis Superfund Research Program) (introduced by Thomas Young, University of California-Davis) “A nontarget workflow for identifying transformation products of anthropogenic chemicals in biological treatment systems” • James Gibson (Dartmouth College Superfund Research Center) (introduced by Tracy Punshon, Dartmouth College) “The biological elemental imaging database: Development of a FAIR data sharing platform for the environmental and life sciences” Announcement of Trainee Awards and Concluding Remarks: 4:00 - 4:30 p.m. Bill Suk (NIEHS)

NIEHS SRP 2020 Virtual Annual Meeting Program • 5 Administrators Program Agenda

ALL TIMES LISTED ARE EASTERN US TIME ZONE

Monday, December 14, 2020 Theme: Addressing Hazardous Substance Exposures at the Community Level – Solutions by the Superfund Research Program

Time Administrators Program 10:00 - 1:30 p.m. See Main Program Administrator Session I 1:30 - 2:45 p.m. • 1:30 - 2:30 p.m.: Grants Management, including Q&A with Jenny Greer • 2:30 - 2:45 p.m.: General Review, including Q&A with Laura Thomas 2:45 - 5:00 p.m. See Main Program

Tuesday, December 15, 2020 Theme: Meeting the Challenges of Environmental Protection with 21st Century Science

Time Administrators Program Administrator Session II: Peer-to-Peer Discussions • 11:00 - 11:15 a.m.: “It Takes a Village…”—Tips & Tricks to Managing Center Grants 11:00 - 12:00 p.m. • 11:15 - 11:30 a.m.: Management of RETCC • 11:30 - 11:45 a.m.: Supplements • 11:45 - 12:00 p.m.: Open Discussion 12:00 - 1:00 p.m. See Main Program

Administrators Session III 1:00 - 4:00 p.m. • 1:00 – 4:00 p.m.: One-on-One Grants Management with Jenny Greer Sign-up required: https://form.jotform.com/203135228737050

6 • NIEHS SRP 2020 Virtual Annual Meeting Program

Welcome from NIEHS SRP • Bill Suk

First and foremost, I hope everyone is staying safe and healthy during these difficult and unprecedented times. The COVID-19 pandemic has an immense global impact and has claimed many lives. Our condolences go out to the people who have lost loved ones and have suffered due to this pandemic.

I also want to thank you all for what you have done to support your communities this year. We have heard about a lot of the important work by SRP grantees to support public health during the COVID-19 pandemic. I am tremendously proud of you.

As you can imagine, the Covid-19 pandemic has affected scientific meeting planning everywhere, and the SRP annual meeting is no exception. To protect people’s health, we had to cancel the in-person meeting in College Station, Texas and pivot to a virtual platform. We are very grateful for all the work that has been done by the Texas A&M team in organizing this year’s virtual conference and ensuring its success.

We were delighted with the number of registrations and poster submissions for the virtual meeting and we hope that it provides you with an opportunity to engage with colleagues. In addition to some excellent talks, we are looking forward to an exciting new interactive, multimedia poster format with more than 250 posters on display. Even though we would all prefer to be in person, I am pleased that we can still come together to share scientific discoveries and we hope that you all make the most of these virtual poster sessions.

Because our hosts this year are at Texas A&M, I also wanted to recognize the influence that K.C. Donnelly has had on this program. Before he passed away in 2009, Donnelly was a longtime SRP grantee at Texas A&M who worked tirelessly to improve our understanding of environmental exposure and genotoxicity of complex chemical mixtures. He was a dedicated mentor to his students and postdoctoral researchers, instilling in them the importance of applying their knowledge and findings to improve the health of communities exposed to environmental contaminants. You’ll see from the meeting agenda that we continue his legacy by featuring the next generation of environmental health professionals with trainee presentations throughout the agenda as well as the trainee poster competition.

Advancing environmental health has never been more important and I am looking forward to a safe and productive meeting. We are glad that each and everyone one of you can be a part of this meeting and help shape the future of the program.

With appreciation, Bill Suk

8 • NIEHS SRP 2020 Virtual Annual Meeting Program Welcome from Texas A&M SRPWelcome Center • IvanDr. Bill Rusyn Suk

I would like to join in Dr. Suk’s wishes of good health and well-being to all of you and your friends, family and colleagues. It is heart-warming to read the e-Notes and Superfund Briefs and learn about the vibrance and diversity of research, community outreach and translation to the Superfund stakeholders. All of us have been able to find the new normal and adjust without sacrificing the excellence and quality of the work from entire Centers, and individual scientists and staff. A special thank you to the Superfund Program staff for being concerned, reaching out to all of us, and providing guidance in these challenging times.

As much as we are saddened that we will not get the chance this year to host all of you in sunny and warm College Station, we are very pleased by the turnout this meeting will have. Texas A&M University is one of the largest academic and research centers in the United States, with total student enrollment on College Station campus and branches in Galveston and Qatar eclipsing 70,000 this year. Enrollment growth at Texas A&M University and other Universities of the A&M System has created new challenges for in-person instruction this year, but the campus is vibrant, mask-wearing and testing “early and often.” We are all looking forward to the end of the pandemic, but everyone pulled together to heed the recommendations from the federal and State health authorities and to not bend in the face of the challenges.

The science at this annual meeting will be truly exciting. We are focusing on two over-arching themes: (1) Addressing Hazardous Substance Exposures at the Community Level – Solutions by the Superfund Research Program, and (2) Meeting the Challenges of Environmental Protection with 21st Century Science. The first theme is timely because community-engaged research and work to address disparities in susceptibility and effects of environmental exposures, including those from the biological threats present and future, is even more important today as it was ever. The Superfund Centers, along with other NIEHS grantees and intramural scientists are well-equipped to be at the forefront and lead the way. The second theme is also of acute importance as not only the science of hazard, exposure and remediation is evolving more rapidly because of technology advances, but also because the regulatory agencies are starting to take notice and make plans for adjusting to the realities of new data streams replacing traditional methods and models.

While it was not difficult to acknowledge that these themes synergize well with the work by many in the Superfund program, it was very difficult to select 8 presentations to represent the breadth and depth of the work. It is also difficult to get the full immersive experience of a poster session in an online format. But I would like to extend the special thanks to Arlean Rohde, the manager of Texas A&M Superfund Center, and Brittany Trottier, Health Specialist in the NIEHS Superfund Research Program for spending countless hours researching online meeting options and interviewing the vendors to select the option for this meeting. Posters will be available from December 1, 2020 to May 31, 2021 and we hope that this will provide ample opportunity for review and electronic interactions with presenters.

In conclusion, I hope you participate in the entire program, enjoy the science and the interactions, and establish new collaborations. Stay positive and test negative! With best regards, Ivan Rusyn

NIEHS SRP 2020 Virtual Annual Meeting Program • 9 Keynote I • t.e.j.a.s. & Texas A&M University

Monday, December 14, 2020 | 11:00–11:45 a.m. Introduced by Weihsueh Chiu | Texas A&M University

Building Resilient Communities to Protect Against Exposure to Hazardous Chemicals: Multidisciplinary Engagement and Partnerships with the Superfund Researchers Juan Parras Founder & Exec. Dir. | t.e.j.a.s Recent research has demonstrated many benefits of engaging community members and interest groups in the conduct of research, development of interventions, and educational outreach programs within sensitive environmental regions. Further, recent research has also revealed the importance of establishing these partnerships prior to major hazards to improve rapid research needs and promote better preparedness. In order to accurately and swiftly understand the environmental conditions and public health needs within fence line communities, the Texas A&M Superfund Center has partnered with the Nalleli Hildalgo Texas Environmental Justice Advocacy Services (t.e.j.a.s.) to collect and Community Outreach & Education Liaison | t.e.j.a.s. synthesize data as well as co-learn with neighborhoods about the risks associated with hazardous events. Established in the mid-1990s, t.e.j.a.s is dedicated to providing community members with the tools necessary to create sustainable, environmentally healthy communities by educating individuals on the health concerns and implications arising from environmental pollution. t.e.j.a.s. also actively advocates for empowering individuals with an understanding of applicable environmental laws and regulations and promoting their enforcement, and offering community building skills and resources for effective community action and greater public participation. This presentation will discuss the engagement Galen Newman process and some of the pertinent results/findings from the relationship Associate Professor, between t.e.j.a.s and Texas A&M Superfund Center. Further, an PI Community Engagement Core Texas A&M Superfund emphasis will be placed on the best-practices and identifying some of Research Center the bi-directional communication pathways and pitfalls that have been identified when undertaking community/academic partnerships

This presentation will last approximately 30 minutes and will begin with a brief video created by t.e.j.a.s. showcasing many of the community’s environmental concerns. This will allow for ample opportunity for questions and comments at the end. There are three main goals of the presentation (1) discuss what t.e.j.a.s and the Texas A&M Superfund Center have accomplished over the last few years through utilizing Garett Sansom local interest groups and high school citizen scientists, (2) examples of Research Assistant Professor, successes and failures from this unique partnership, and (3) lessons Co-I Community Engagement Core learned on how other Centers can promote long-term deep community Texas A&M Superfund engagement efforts. Research Center

10 • NIEHS SRP 2020 Virtual Annual Meeting Program Keynote II • US Environmental Protection Agency (EPA)

Tuesday, December 15, 2020 | 11:00–11:45 a.m. Introduced by Ivan Rusyn | Texas A&M University

US EPA’s Path Towards More Rapid, Efficient, and Protective Chemical Testing with Fewer Animals

Abstract Tens of thousands of chemicals are used in commerce and are present Russell S. Thomas in the environment. Hundreds more chemicals are introduced each Director, year. In most cases, the chemicals have limited data that can inform Center for Computational the diverse range of decisions that state and federal regulators may Toxicology & Exposure, be required to make regarding potential human health and ecological U.S. Environmental Protection Agency risks. Advances in computational and technological approaches have begun to enable more rapid and cost-effective evaluation of potential toxicological and exposure-related properties. The new methods are largely animal free, but the endpoints and uncertainties associated with these new technologies are qualitatively and quantitatively different than the traditional approaches leading to challenges in integrating the advances into regulatory decision making. In 2019, EPA released a memo stating its intent to eliminate animal testing by 2035 while remaining fully protective of human health and the environment. A work plan was developed that integrates short- and long- term regulatory, scientific, and communication strategies that are necessary to achieve these goals. Scientific advances by external organizations such as those being developed in the Superfund Basic Research Program are an important component of the strategy and will be important in the longer-term success of the initiative. The talk will provide an overview of the various challenges faced by the Agency, the multi-disciplinary research being performed, and the short-and long-term strategies to realize the future vision of chemical safety testing. This abstract does not necessarily reflect U.S. EPA policy.

Biosketch Russell S. Thomas, PhD is the director of the Center for Computational Toxicology and Exposure at the U.S. Environmental Protection Agency (EPA). The Center is performing solutions-driven research to rapidly evaluate the potential human health and environmental risks due to exposures to environmental stressors and ensure the integrity of the freshwater environment and its capacity to support human well-being. Dr. Thomas has a broad, multidisciplinary background and experience. Dr. Thomas’ formal academic training includes a B.A. in chemistry from Tabor College, an M.S. in radiation ecology and health physics from Colorado State University, and a Ph.D. in toxicology also at Colorado State. Following his doctoral studies, Dr. Thomas performed postdoctoral research in molecular biology and genomics at the McArdle Cancer Research Laboratory at the University of Wisconsin. Following his academic training, Dr. Thomas gained experience in bioinformatics, genomics, and high-throughput in vitro screening in the biotechnology and biopharma industry. Dr. Thomas worked as an investigator and senior manager at a non-profit research institute. Prior to coming to the U.S. EPA, Dr. Thomas worked as an investigator and senior manager at a non-profit research institute.

NIEHS SRP 2020 Virtual Annual Meeting Program • 11 Wetterhahn & Donnelly Awards

Monday, December 14, 2020 | 11:45 a.m.–12:30 p.m. Moderated by Bill Suk| NIEHS Superfund Research Program Karen Wetterhahn Memorial Award About Dr. Wetterhahn Karen E. Wetterhahn, Ph.D., Professor of Chemistry and the Albert Bradley Third Century Professor in the Sciences at Dartmouth College, died June 8, 1997 at age 48. Her death was the result of dimethylmercury poisoning caused by the accidental spill of a few drops of the chemical on her latex glove-covered hand. Dr. Wetterhahn was an established authority on the effects of heavy metals on biological systems as well as a dedicated teacher and mentor. She played an integral role in the administration of the sciences at Dartmouth and co-founded Dartmouth’s Women in Science Project (WISP), which is aimed at increasing the number of women majoring and taking courses in the sciences, including mathematics and engineering. She served as the Program Director of the Dartmouth SRP from 1995 to 1997, and she was a PI who studied the effects of toxic heavy metals on cellular processes. She was an acknowledged international expert in chromium carcinogenicity. Her untimely death has given us an opportunity to reflect on the value of understanding how heavy metals can interfere with important cellular processes that are vital to life. As a way of honoring the life and scientific accomplishments of Dr. Wetterhahn, the SRP has established an annual award to recognize an outstanding graduate student or post-doctoral researcher that best demonstrates the qualities of scientific excellence exhibited by Dr. Wetterhahn.

Monday, December 14, 2020 | 1:00–1:30 p.m. KC Donnelly Externship Awards The Legacy of KC Donnelly’s Contribution to the Superfund Research Program Moderated by Stephen Safe | Texas A&M University

About Dr. Donnelly K.C. Donnelly, Ph.D., passed away on July 1, 2009. Donnelly was an excellent teacher, and for more than 30 years, his research impacted the world. His research—conducted in Eastern Europe, China, and locations across the United States—changed the way environmental health is perceived. He was also a dedicated mentor to his students and postdoctoral researchers, instilling in them the importance of applying their knowledge and findings to improve the health of communities exposed to environmental contaminants. Donnelly joined the SRP at Texas A&M University in 1995. His research projects used cell culture, animal and human studies to investigate the toxic interactions of polycyclic aromatic hydrocarbon (PAH) mixtures. He also headed Community Outreach and Education efforts. He established and maintained relationships with Environmental Protection Agency (EPA) scientists and remedial project managers that not only increased the relevance of his research but supported EPA’s efforts. He initiated and championed the SRP Bioassay Network, bringing together researchers from six SRP Centers and scientists from EPA Regions and Headquarters. In honor of Donnelly, the SRP has established this supplemental award to provide current SRP-funded graduate students and postdoctoral researchers with translational/transdisciplinary opportunities and experiences within other SRP-funded centers, government laboratories (EPA, ATSDR, NIEHS), or other agencies (state, local, Tribal).

12 • NIEHS SRP 2020 Virtual Annual Meeting Program Karen Wetterhahn Memorial Award

Monday, December 14, 2020 | 11:45–12:30 p.m. Karen Wetterhahn Memorial Award Announcement & Lecture The Superfund Research Program has named Jennifer Kay, Ph.D., from the Massachusetts Institute of Technology (MIT), as this year’s Karen Wetterhahn Memorial Award winner. Kay’s research evaluates how genetic factors affect susceptibility to mutations and cancer following exposure to N-nitrosodimethylamine (NDMA), a contaminant found near a Superfund site in Wilmington, MA. Kay was a postdoctoral fellow in the Bevin P. Engelward, Sc.D., Lab at MIT. She also directed the MIT SRP Center Research Translation Core.

Analysis of Genetic Susceptibility Factors for N-nitrosamine-Induced Toxicity, Genomic Instability, and Cancer

Jennifer E. Kay1,2,3, Joshua J. Corrigan1,2,3, Amanda L. Armijo1,2,3,4, Ilana S. Nazari1,2,3, Dorothea Torous5, Svetlana Avlasevich5, Sebastian E. Carrasco4, Dushan N. Wadduwage6, Stephen Dertinger5, Leona D. Samson1,2,3,7, John M. Essigmann1,2,3,8, and Bevin P. Engelward1,2,3

1Department of Biological Engineering, 2Superfund Research Program, 3Center for Environmental Health Sciences, 4Division of Comparative Medicine, 7Department of Biology, 8Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA; 5Litron Laboratories, Rochester, NY; 6Center for Advanced Imaging, Harvard University, Cambridge, MA

Abstract N-nitrosodimethylamine (NDMA) is a genotoxic IARC Group 2A probable human carcinogen. NDMA contamination from the Olin Chemical Superfund site led to widespread exposure in Wilmington, MA, and recently several pharmaceuticals have been found to contain high levels of NDMA. One major DNA lesion caused by NDMA is the replication-blocking 3-methyladenine (3MeA), which is excised by the alkyladenine DNA glycosylase (AAG) to initiate Base Excision Repair (BER). Notably, since 3MeA blocks replicative polymerases and BER intermediates consist of single strand breaks (SSBs), modification of AAG activity enables analysis of blocking lesions and strand breaks, two major classes of DNA damage with fundamentally different biological implications. Since DNA repair capacity varies among individuals, AAG activity is anticipated to substantially impact susceptibility to NDMA-induced disease. To uncover the differential impacts of low and high Aag expression, Aag-/- mice (which have persistent blocking lesions) and Aag-overexpressing (AagTg) mice (which have high levels of SSBs) were created, each carrying the gpt-delta and RaDR transgenes for detecting point mutations and genetic sequence rearrangements, respectively. In the days immediately following NDMA exposure, significant DNA damage was observed that was particularly acute in AagTg mice. This high level of DNA damage was correlated with high levels of toxicity, with elevated hepatocellular necrosis and apoptosis within 24 hours of exposure and animal lethality within two weeks. However, this acute toxicity in AagTg mice ultimately suppressed the development of mutations and cancer relative to WT. On the other hand, Aag-/- mice had elevated cell death over WT (but less severe than AagTg) one day after exposure and significantly higher induction of mutations and cancer later in life. Analysis of the molecular events immediately after exposure and mutations and cancer weeks and months later enables an integrated perspective of disease progression, wherein strand breaks tip the balance toward toxicity and replication blocking lesions promote mutations and cancer.

NIEHS SRP 2020 Virtual Annual Meeting Program • 13 KC Donnelly Externship Awards

Monday, December 14, 2020 | 1:00–1:30 p.m. 2019 KC Donnelly Externship Awardee Presentations

Jill Riddell | 2019 KC Donnelly Award Winner (West Virginia University, Northeastern University Superfund Research Center) Jill Riddell, a Ph.D. candidate at West Virginia University working under Dorothy Vesper, Ph.D., is a trainee with the Northeastern University Superfund Research Program (SRP) Center. For her four-week externship, Riddell will travel to Tucson, Arizona, where she will work at the University of Arizona SRP Center under the mentorship of Jon Chorover, Ph.D., a world-recognized soil chemist. With a Northeastern SRP Center project led by Ingrid Padilla, Ph.D., Riddell’s research looks at how organic contaminants, chemicals that are based on carbon such as solvents and pesticides, are transported and stored in karst aquifers. Karst aquifers are formed by water-soluble rock like limestone and dolomite. Due to the water-soluble nature of karst aquifers, they are susceptible to contamination. At the University of Arizona SRP Center, Riddell will build upon her research to describe how a new tracing technique interacts with karst sediments. Traditional tracing approaches use dyes and salts to mimic how contaminants move through groundwater. However, these tracers fail to mimic the movement of organic contaminants. Riddell will use advanced analytical techniques to better understand polymer microspheres, which are small, spherical particles with molecules attached, and how they can be used as a tracer to model the behavior of organic contaminants. “As a geologist, the opportunity to continue to diversify my skills and knowledge will give me an advantage in solving complex geologic problems throughout my career,” said Riddell. “Through this externship, I hope to build new relationships with scientific professionals. Having an extensive network of colleagues and friends in the scientific community is an invaluable resource.”

Nabil Shaikh | 2019 KC Donnelly Award Winner (University of New Mexico Superfund Research Center) Nabil Shaikh is a Ph.D. candidate at the University of New Mexico (UNM) SRP Center under the direction of Jose Cerrato, Ph.D. During his two-month externship, Shaikh will be traveling to the University of Iowa SRP Center to work with Keri Hornbuckle, Ph.D., and Andres Martinez, Ph.D. Shaikh’s work at the UNM SRP is focused on the cleanup of inorganic contaminants, like uranium, in water. During his externship, he will be learning an advanced technique called electrospinning that creates electrospun nanofiber mats (ENM) with specific chemistry for binding pollutants. He plans to test the ENMs for their ability to capture and remove uranium from contaminated water. The externship will provide Shaikh with new knowledge of nanomaterials, which can be applied to removal strategies at uranium-contaminated sites in New Mexico. The goal is for the ENMs to reduce risks for communities living near contaminated waste sites. “The externship will benefit my professional goals of learning new cutting-edge research techniques to become a successful independent researcher,” said Shaikh. “It will also help me contribute to developing cleanup strategies that will better protect human health, particularly in rural Native American communities.”

14 • NIEHS SRP 2020 Virtual Annual Meeting Program NIEHS SRP 2020 Annual Meeting

Scientific Presentations I: Community-Informed Superfund Research (moderated by Weihsueh Chiu, Texas A&M University) • Harrison Schmitt (University of Arizona Superfund Research Center) “Chronic environmental contamination: Psychosocial health consequences, risk factors, and pathways to community resilience”

• Rui Zhu (Texas A&M University Superfund Research Center) (introduced by Galen Newman, Texas A&M University) “Resilience through Regeneration: An engagement and performance-based approach to repurposing vacant community lots with green infrastructure”

• Tyler Gripshover (University of Louisville Superfund Research Center) (introduced by Matthew Cave, University of Louisville) “Volatile organic compound exposures are positively associated with liver apoptosis in a residential cohort”

• Clare Pace (University of California-Berkeley Superfund Research Center) (introduced by Rachel Morello-Frosh, University of California-Berkeley) “Advancing California’s Human Right to Water Law: Characterizing inequities in drinking water quality among domestic well communities and community water systems” Scientific Presentations I

Chronic environmental contamination: Psychosocial health consequences, risk factors, and pathways to community resilience

Harrison J. Schmitt, University of Arizona, Tucson, Arizona Daniel Sullivan, University of Arizona, Tucson, Arizona Eric Calloway, Gretchen Swanson Center for Nutrition, Omaha, Nebraska Whitney Clausen, Gretchen Swanson Center for Nutrition, Omaha, Nebraska Jamie Rayman, ATSDR Pamela Tucker, ATSDR Ben Gerhardstein, ATSDR

Superfund Mandates: None

Superfund Center: University of Arizona Superfund Research Center

A body of psychological and social scientific evidence suggests that the experience of technological disaster or long-term exposure to environmental contamination can be psychologically stressful. We undertook a mixed-methods narrative and systematic review to assess the current research, and provide a platform for future research, on the psychosocial impact of chronic environmental contamination. In addition, we sought to identify strategies for strengthening community resilience to this hazard. In particular, our objectives were to (1) develop a theoretical framework for understanding the unique psychosocial impact of chronic environmental contamination (in contrast to natural and technological disasters, and background pollution); and (2) assess the impact of chronic contamination on psychological health effects. We conducted a meta-analysis of available findings ( k = 57, N = 22,786) from 1995-2019 in which we observed small-to-medium effects of experiencing contamination on general stress, anxiety, depression, and PTSD. However, there was also evident risk of bias in the body of data. Relevant quantitative and qualitative studies were examined to derive a model identifying likely factors increasing risk for distress in chronic contamination experience, as well as actions that may be taken by public health professionals and local leaders to enhance community resilience. This review suggests that psychological stress in the context of chronic contamination is an important potential public health burden and a key area for additional research.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

16 • NIEHS SRP 2020 Virtual Annual Meeting Program Scientific Presentations I

Resilience through Regeneration: An Engagement and Performance-based Approach to Repurposing Vacant Community Lots with Green Infrastructure

Rui Zhu, Texas A&M University, College Station, Texas Galen Newman, Texas A&M University, College Station, Texas Dongying Li, Texas A&M University, College Station, Texas Dingding Ren, Texas A&M University, College Station, Texas Garett Sansom, Texas A&M University, College Station, Texas Jennifer Horney, University of Delaware, Wilmington, Delaware Xueqi Song, Texas A&M University, College Station, Texas

Superfund Mandate: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

Many urban areas affected by flood disasters are also becoming increasingly ecologically and socially fragmented due to the accumulation of vacant properties. While redevelopment is often viewed as the primary objective in regenerating vacant properties, they can also potentially provide ecological and hydrological land uses. Rather than chasing development-based incentives for regenerating vacant lots in high flood-risk and high-contamination exposure communities, a balance should be sought between new developmental land uses and green infrastructure to help counteract stormwater runoff, flood, and hazardous exposure effects, or “Resilience through Regeneration.” This research uses built environment performance models to evaluate the economic and hydrologic performance of green infrastructure regeneration projects for three marginalized neighborhoods in Houston, Texas, USA. Each project site is characterized by excessive vacant lots, flood issues, and contamination effects due to proximity to industrial land uses. Purposeful efforts were made to 1) utilize the same definition and inventory method for vacant properties, 2) utilize similar design processes and equivalent engagement procedures, and 3) utilize the same performance assessment methodology to increase the generalizability of conclusions. The master plan for each project was created following a participatory design approach which utilized feedback loops between university personnel. Local organizations, and community members to assist in design decision making. Results suggest that, when using green infrastructure to regenerate vacant properties, 1) flood risk continually decreases, 2) upfront economic costs increase in the short term (when compared to conventional development), and 3) the long-term economic return on investment is much higher. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 17 Scientific Presentations I

Volatile Organic Compound Exposures Are Positively Associated with Liver Apoptosis in a Residential Cohort

T. C. Gripshover, University of Louisville, Louisville, Kentucky B. Wahlang, University of Louisville, Louisville, Kentucky D.W. Riggs, University of Louisville, Louisville, Kentucky S.E. Smith, University of Louisville, Louisville, Kentucky T.V. Krivokhizhina, University of Louisville, Louisville, Kentucky R.J. Keith, University of Louisville, Louisville, Kentucky C.J. McClain, University of Louisville, Louisville, Kentucky S. Srivastava, University of Louisville, Louisville, Kentucky M.C. Cave, University of Louisville, Louisville, Kentucky

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Background. The Health, Environment and Action in Louisville (HEAL) cohort was assembled by the NIEHS- funded University of Louisville Superfund Research Center to allow determination of associations between residential VOC exposures and cardiometabolic syndrome (inclusive of liver disease) in the Louisville community. The objective of this interim cross-sectional analysis of 679 HEAL subjects collected in 2018 and 2019 is to determine associations between urinary VOC metabolites and the circulating hepatocyte apoptosis biomarker, caspase-cleaved keratin 18 (CK18 M30). Methods. Serum CK18 M30 (normal cutoff <200 U/L) was determined by ELISA. 15 creatinine-adjusted urinary metabolites of 10 parent VOCs were measured by UPLC-MS2. Generalized linear models were used to test for associations between disease and exposure biomarkers. Final models were adjusted for race, BMI, alcohol use, cotinine-confirmed smoking status, and education level. Results are presented as % change (95% CI) per interquartile range of VOC metabolite. Results. The following results pertain to 391 samples collected in the first round of HEAL recruitment (2018) while analysis on the remaining samples are pending. Mean age was 50.3±12.0 (SD). Mean BMI was 29.9±6.1 kg/m2. The cohort was 57.8% female, 78.5% white and 22.0% diabetic. Elevated CK18 M30 was present in 12.3% of participants. CK18 M30 was positively associated with 4 urinary VOC metabolites corresponding to acrolein (3HPMA); acrylamide (AAMA); and 1,3-butadiene (DHBMA and HPMMA) exposures. These associations were modified by sex and race for three of the four significant VOC metabolites and were more positive in males and whites. Conclusions. This interim analysis demonstrated positive associations between specific VOC exposures and hepatocyte apoptosis in the residential HEAL cohort. The role of VOC exposures in metabolic liver diseases warrants further investigation.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

18 • NIEHS SRP 2020 Virtual Annual Meeting Program Scientific Presentations I

Advancing California’s Human Right to Water Law: Characterizing inequities in drinking water quality among domestic well communities and community water systems

Clare E. Pace, University of California Berkeley, Berkeley, CA Lara Cushing, University of California, Los Angeles, CA Carolina Balazs, Office of Environmental Health Hazard Assessment, CA EPA Rachel Morello-Frosch, University of California, Berkeley, CA

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of California-Berkeley Superfund Research Center

California legally recognizes the Human Right to Water, however, roughly 10% of California’s community water systems (CWS) are currently out of compliance with drinking water quality standards. A disproportionate number of water quality violations occur in low income communities of color, raising environmental justice concerns. Additionally, significant data gaps exist regarding the location and water quality of unregulated water sources, such as private domestic wells. The UC Berkeley Water Equity Science Shop sought to characterize demographic differences in water quality for communities reliant upon domestic wells and CWS. We integrated data from the Department of Water Resources; Tracking California’s Drinking Water Systems Geographic Reporting Tool; and the US Census to locate domestic well communities statewide. We integrated residential parcel data and building footprint locations using dasymetric mapping techniques to generate high resolution population estimates for domestic well communities and CWS. We then used water quality data from CalEnviroscreen 3.0 and demographic data from the American Community Survey to estimate sociodemographic differences in water quality using a multivariate logistical regression modeling approach that accounts for spatial autocorrelation. We estimate that over 1.3 million Californians rely on domestic wells for drinking water. The likelihood of elevated arsenic increases by a factor of 1.08 (CI: 1.01-1.15) for every 10% increase in renters, and the likelihood of elevated nitrate increases by a factor of 1.10 (CI 1.04-1.16) for every 10% increase in Hispanic population among domestic well communities statewide. This work highlights the importance of including domestic well communities in policy, intervention and tracking efforts related to achieving environmental justice and the Human Right to Water.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 19

NIEHS SRP 2020 Annual Meeting

Scientific Presentations II: Data Science Solutions for Superfund Challenges (moderated by Ivan Rusyn, Texas A&M University)

• Rance Nault (Michigan State University Superfund Research Program) (introduced by Tim Zacharewski, Michigan State University) “Development of minimum requirements and data management framework for animal toxicology experiments”

• Dillon Lloyd (NC State University/Texas A&M Superfund Research Center) (introduced by Fred Wright, North Carolina State University) “An R Shiny tool for spatial and correlation analysis of biological and chemical measurements”

• Madison Hattaway (University of California-Davis Superfund Research Program) (introduced by Thomas Young, University of California-Davis) “A nontarget workflow for identifying transformation products of anthropogenic chemicals in biological treatment systems”

• James Gibson (Dartmouth College Superfund Research Center) (introduced by Tracy Punshon, Dartmouth College) “The biological elemental imaging database: Development of a FAIR data sharing platform for the environmental and life sciences” Scientific Presentations II

Development of minimum requirements and data management framework for animal toxicology experiments

Rance Nault, Michigan State University Hunter Moseley, University of Kentucky Kelly Pennell, University of Kentucky Matthew Cave, University of Louisville Gabriele Ludewig, University of Iowa

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

Complete and transparent data sharing is a laudable, but challenging goal in toxicology research. Recently, a working group consisting of stakeholders from academia, industry, government, and scientific publishing described a set of guiding principles to make data findable, accessible, interoperable, and reusable (FAIR). These principles are now a cornerstone of data management policies of the NIEHS Superfund Research Program (SRP). In order to promote the FAIRness of in vivo toxicology experiments, a working group from several Superfund Research Centers developed the Minimum Information about an Animal Toxicology Experiment (MIATE) requirements and surrounding data management framework which leverages the Investigation, Study, Assay (ISA) data model and related tools. A consensus of essential metadata was established covering animal details (e.g. species), housing conditions, diet, treatment, and study termination conditions. Recommended ontologies for specific fields are used to ensure interoperability. Metadata requirements were defined in relation to the ISA data model to leverage the mature data management ecosystem including the ISA software suite for the collection and validation of (meta)data. MIATE requirements were also used to develop publicly available data collection templates for ISA tools. We demonstrate the effective implementation of MIATE concomitantly with existing microarray, proteomic, and metabolomic datasets that also adhere to Minimum Information about Sequencing Experiments (MINSEQE), Minimum Information about Proteomics Experiments (MIAPE), and Metabolomics Standard Initiative (MSI) standards. Through use of minimum requirement standards such as MIATE, we hope that data from basic toxicology studies can be standardized to enhance their interoperability and reuse, and consequently improve data capture that is truly FAIR. Funded by Superfund Research Program data interoperability and reuse supplements to MSU (P42ES004911), UK (P42ES007380), UofL (P42ES023716), and UI (P42ES013661).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

22 • NIEHS SRP 2020 Virtual Annual Meeting Program Scientific Presentations II

An R Shiny tool for spatial and correlation analysis of biological and chemical measurements

Dillon Lloyd, North Carolina State University, Raleigh North Carolina Zunwei Chen, Texas A&M University, College Station, Texas Ivan Rusyn, Texas A&M University, College Station, Texas Fred Wright, North Carolina State University, Raleigh, North Carolina Yi-Hui Zhou, North Carolina State University, Raleigh, North Carolina

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

The visualization and processing of geospatial data often form an integral part of an analysis pipeline for toxicological datasets, which often involve location-based sampling. Measurements of the concentration of multiple chemicals and/or biologically relevant features provide the opportunity to test for spatial correlation and for correlation between measurements, but also present multiple testing challenges. Standard analysis workflows require a background in scripted computing languages, or the use of commercial specialized software that is general-purpose and not well-suited to toxicological datasets. Accordingly, we have developed an R Shiny tool for (i) informed visualization of location-sampling data, (ii) spatial correlation testing for one or multiple measurements, and (iii) cross-platform correlation testing, e.g. as encountered when both chemical and biological measurements have been gathered on the same samples. Correlations for multiple hypothesis testing using false discovery rates is built into the tool. The use of an R Shiny framework allows the translation of standard R code into a hosted web application, so that the underlying code is transparently available for trained data scientists to understand and modify, while the graphical user interface enables ease of use for any practitioner. We illustrate the tool using data on chemical and biological assays from samples in the Manchester Neighborhood in Houston, Texas, with a focus on polycyclic aromatic hydrocarbons and bioassays using induced pluripotent stem cells. We describe the steps to input the data, compute various distance measures, perform spatial correlation and feature testing, and overall data visualization. Overall, our tool makes a targeted suite of geospatial analyses easily available to a large user base, without requiring specialization data science training. This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 23 Scientific Presentations II

A Nontarget Workflow for Identifying Transformation Products of Anthropogenic Chemicals in Biological Treatment Systems

Madison Hattaway, University of California, Davis Olivia Wright, University of California, Davis Luann Wong, University of California, Davis Aniela Burant, California Department of Pesticide Regulation Jennifer Teerlink, California Department of Pesticide Regulation Heather Bischel, University of California, Davis Thomas M. Young, University of California, Davis

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of California, Davis Superfund Research Center

Nontarget analysis promises the ability to identify a broader range of contaminants, beyond the limitations of target or suspect screening, to find the “unknown unknowns”. However, this approach also can generate daunting lists of thousands of “features”, or aligned spectral peaks presumed to be chemicals. Beyond being computationally intensive, attempting to identify all features is unnecessary, as some may be spurious artifacts of the alignment algorithm, while others may be unthreatening biomolecules. Furthermore, increased attention is being given to transformation products of contaminants of concern, as disappearance of parent compounds across a treatment system does not always indicate a reduction in toxicity. Therefore, the goal of this research is to develop a workflow to process nontarget data aimed specifically at identifying breakdown products of anthropogenic chemicals. Tools such as the EAWAG Envipath pathway prediction model, feature based molecular networking, and machine-learning algorithms will prioritize nontarget features for further investigation to determine formula and structure. These methods are being developed to easily interface with output from MS-DIAL, an open source alignment software package for LC and GC-MS data. Applications to reactors that treat pesticide-containing water, including municipal wastewater treatment systems and woodchip-based bioreactors will be discussed.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

24 • NIEHS SRP 2020 Virtual Annual Meeting Program Scientific Presentations II

The Biological Elemental Imaging Database: Development of a FAIR Data Sharing Platform for the Environmental and Life Sciences

James Gibson, Dartmouth College, Hanover, New Hampshire Benjamin C. Bostick, Columbia University, Palisades, New York Sam Webb, Stanford Synchrotron Radiation Lightsource, Menlo Park, California Mary Lou Guerinot, Dartmouth College, Hanover, New Hampshire Tracy Punshon, Dartmouth College, Hanover, New Hampshire

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Dartmouth College Superfund Research Center

Synchrotron-based spectroscopy is an important technique for identifying the concentration, speciation, and distribution of contaminants in a range of biological media. However, datasets from synchrotrons come in a wide variety of formats, which make it difficult to easily share and exchange data. The elemental imaging community needs a database that enables reuse and reanalysis of their data, while subscribing to the data sharing policies of their funding organizations. This paper introduces the Biological Elemental Imaging Database (BEID). The BEID is a MySQL-PHP website that represents a first step in creating a Findable, Accessible, Interoperable, and Reusable (FAIR) data resource for the elemental imaging community. Currently, the BEID is a searchable database of multi-elemental imaging data collected primarily from the model plant species Arabidopsis, over a 10-year period. Registered users can browse the BEID, preview images of data, and download raw data files from experiments conducted at different synchrotron facilities around the US. If a user wishes to download data from the website for reuse, they can request a data reuse license for publication. Collaborators can also upload and contribute their own synchrotron data to help the database grow. Code from Sam’s Microprobe Analysis Toolkit (SMAK), will be used to periodically pre-process newly uploaded data files into logically scaled preview images. Future work will focus on further integrating SMAK with the BEID. Code from SMAK will be directly incorporated into the BEID to enable users to perform statistical analysis on data files without having to download SMAK or other synchrotron software separately. The BEID is a crucial first step to improve data sharing, standardization, and collaborations between different synchrotron centers.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 25 Poster Session I: December 14, 2020 | 3:00 – 4:00 p.m.

Last Name First Name Poster Session ID Abuawad Ahlam I-1 Adhihetty Prasadanie I-2 Ankrum James I-3 Balogun Fatai I-4 Beene Daniel I-5 Begay Jessica I-6 Bittle Maeve I-7 Black Gabrielle I-8 Bowers Christopher I-9 Camargo Krisa I-10 Cathey Amber I-11 Chen Zunwei I-12 Cholico Giovan I-13 Cuthbertson Amy I-14 NIEHS Dalaijamts Chimeddulam I-15 Fontana Catherine I-16 SRP Fung Samantha I-17 Ganesh Hari I-18 2020 Gawel James I-19 Gomes Aldrin I-20 Annual Meeting Halas Naomi I-21 Hamm Joseph I-22 Hearon Sara I-23 Huizenga Juliana I-24 Jang Suji I-25 Jobe Timothy I-26 Kaeli David I-27 Kastleman Catherine I-28 Khan D M Isha Olive I-29 Koshko Lisa I-30 Larson-Casey Jennifer I-31 Li Wenting I-32 Li YIng I-33 Macaluso Francesca I-34

26 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: December 14, 2020 | 3:00 – 4:00 p.m.

Last Name First Name Poster Session ID McFall Samantha I-35 McGraw Katlyn I-36 McReynolds Cindy I-37 MEZA MARIA I-38 Moorthy Bhagavatula I-39 Morales Juan I-40 Morales-McDevitt Maya I-41 Paranjape Neha I-42 Pettibone Kristi I-43 Phelps Drake I-44 Ranville James I-45 Rice Brittany I-46 Rigutto Gabrielle I-47 NIEHS Roman Carolyn I-48 Schilz Jodi I-49 SRP Shah Rishabh I-50 Shakya Aryatara I-51 2020 Sherr David I-52 Shukla Vidushi I-53 Annual Meeting Signes-Pastor Antonio I-54 Spaur Maya I-55 Sun Yao-Hui I-56 Taylor Breandon I-57 Tummala Chandra Mouli I-58 Vandiver Kathleen I-59 Varner Paige I-60 Wang Weicang I-61 White Forest I-62 Wilson Lindsay I-63 Wirth Charlotte I-64 Wu Jingyi I-65 Xenakis James I-66 Zhao Yuwei I-67

NIEHS SRP 2020 Virtual Annual Meeting Program • 27 Poster Session I: Poster #I-1

Methods for accounting for urine dilution in arsenic analyses: comparisons to blood arsenic

Ahlam Abuawad, Columbia University, New York, NY Faruque Parvez, Columbia University, New York, NY Tariqul Islam, Columbia University, New York, NY Joe Graziano, Columbia University, New York, NY Ana Navas-Acien, Columbia University, New York, NY Mary V. Gamble, Columbia University, New York, NY

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Columbia University Superfund Research Center

Exposure to arsenic (As) affects >140 million individuals worldwide. An established biomarker of As exposure is urinary As (uAs). Creatinine is excreted at a constant rate via urine and is commonly used to correct uAs for urine dilution. However, urinary creatine (uCr) levels are influenced by meat intake, muscle mass, and other factors that may also be related to As elimination. An alternative method, specific gravity (SG), has limitations in individuals with kidney damage and diabetes. We compared uAs corrected for uCr or SG vs. blood As (bAs), an increasingly used biomarker of As exposure that is unaffected by hydration status. Participants from the Folic Acid and Creatine Trial were examined (N=539) to analyze the relationship between log-transformed total bAs and log-transformed uAs with adjustments for age, sex, and BMI, with further adjustments for uCr or SG in separate models. Median (IQR) uAs and bAs were 112.8 (133.6) and 8.41 (5.88) µg/L, respectively. Urinary creatinine and SG were highly correlated with each other (ρ=0.86, p<0.001). Regression models revealed that a log 10-unit increase in total bAs concentrations was related to 0.69, 0.72, and 0.57 increases in log-10 uAs when unadjusted for dilution factors, adjusted for uCr, and adjusted for SG, respectively (p<0.001). We compared uAs to bAs and found that uCr outperformed SG in predicting bAs in this Bangladeshi population exposed to high water As levels. Further research is required for populations with different sociodemographic characteristics, and As exposure levels to evaluate the consistency of these findings and optimize urine dilution adjustment methods in As-related research.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

28 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-2

An Alkali Metal-Based Au MPCs Chemiresistor Sensor Array for Sensing Aromatic VOCs

Prasadanie K. Adhihetty, University of Louisville, Louisville, Kentucky Sujoy Halder, University of Louisville, Louisville, Kentucky Xiao-An Fu, University of Louisville, Louisville, Kentucky Michael H. Nantz, University of Louisville, Louisville, Kentucky

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Louisville Superfund Research Center

We aim to develop chemiresistive sensors to detect trace levels of aromatic volatile organic compounds (VOCs) in outdoor and indoor air. Many aromatic VOCs, such as benzene, toluene, ethylbenzene and xylene (BTEX), have harmful effects on human health. Consequently, methods have been developed to detect BTEX in ppb to ppt levels. However, the detection and analysis of these aromatic VOCs without analyte preconcentration has not yet been reported. Our interest is to design a sensor comprised of metal ion-functionalized thiol ligands coated on gold monolayer protected clusters (Au MPCs) as a chemiresistor for detecting BTEX VOCs at room temperature with high sensitivity and selectivity by accommodating microelectromechanical system (MEMS) technology. Aromatic compounds contain conjugated π systems that interact with electron deficient elements through cation-π interactions. In some circumstances, this specific noncovalent interaction can be stronger than hydrogen bonding. In this study, we have synthesized nanoparticles that feature alkali metal ions bound to the surface of thiol ligand-coated Au MPCs. The Au MPCs are deposited within interdigitated electrodes fabricated by MEMS technology to create a chemiresistor that senses trace levels of benzene toluene, ethylbenzene, and xylene, presumably through magnified cation-π interactions at the surface of the Au MPCs. Herein, we report our initial studies on harnessing cation-π interactions for sensing aromatic VOCs. Details on syntheses of the chelating thiol ligands and Au MPCs will be presented as well as the methods of alkali metal surface functionalization, particle characterization, and chemiresistor responses. N-Boc protected short alkane chain (6C) and long alkane chain (11C) aminooxy thiols were synthesized and used to prepare thiol ligand- coated Au MPCs with ~2 nm particle diameter.

Email: [email protected], [email protected] and [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 29 Poster Session I: Poster #I-3

Development of an adipose organoid system to screen PCB congeners and doses that lead to metabolic dysfunction

James A. Ankrum, University of Iowa, Iowa City, Iowa Jesse Liszewski, University of Iowa, Iowa City, Iowa Riley Behan, University of Iowa, Iowa City, Iowa Francoise Gourronc, University of Iowa, Iowa City, Iowa Aloysius Klingelhutz, University of Iowa, Iowa City, Iowa

Superfund Mandate: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Iowa Superfund Research Center

Polychlorinated biphenyls (PCBs) persist in the built and natural environment and pose a significant health risk to the public. Human exposure to PCBs have been linked to cancer, obesity, and metabolic disease. However, with over 200 distinct PCBs in the environment, research is needed to determine which are the most harmful to human health. Since PCBs are associated with obesity and known to accumulate in adipose tissue, we sought to develop a screening platform to determine which PCBs directly impact human adipose function. In order to properly screen these toxins, a robust adipose model system is needed. Previous adipose models have been incredibly limited due to the lack of available cell lines, difficulty culturing, and inability to correlate 2D models to 3D human adipose tissue. We’ve created a novel 3D adipose organoid without the need for an exogenous scaffold. Here we show how this technique can be applied to the creation of an adipose model system that can be used to determine the effects of PCBs on adipocytes. To achieve the highest level of test sensitivity, an optimal adipose model should have high levels of adiponectin secretion, a marker of differentiation, and low levels of IL-8 secretion, a marker of inflammation. By examining the effects of different seeding cell densities, differentiation medias, and PCB exposure on the secretions of adipocyte spheroids, we determined an intermediate size of 5,000 cells per spheroid provides a robust model system for toxin screening. Future work will include screening for the effects of different PCB congeners and metabolites on adipocyte function as well as integration of the model into a biomimetic chip to better understand the systemic effects of PCBs.

Email: [email protected] Presenter Status: Researcher

30 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-4

Elucidating the effect of dissolved organic carbon complexity in the fate of hexavalent chromium in multicomponent systems

Fatai Balogun, University of Oregon, Eugene, Oregon Miranda Aiken, University of California-Riverside, Riverside, California Lucas Silva, University of Oregon, Eugene, Oregon Owen Duckworth, North Carolina State University, Raleigh, North Carolina Matthew Polizzotto, University of Oregon, Eugene Oregon

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Other: University of Oregon

Chromium (Cr) contamination of soil and water is of significant environmental and health concern. The specific risks of Cr are dependent on its oxidation state, with Cr(III) being relatively non-toxic and Cr(VI) being very toxic and a known carcinogen . Manganese (Mn) oxides are ubiquitous minerals in many environmental systems and facilitate the oxidation of Cr(III). Conversely, suites of dissolved organic carbon (DOC) may directly impact Cr availability. Dissolved organic carbon limits Cr availability either by redox processes or limiting Mn-oxide-induced oxidation. These competing factors makes it challenging to assess the specific risks of Cr contamination in environmental systems. The objective of this research is to elucidate the mechanisms of Cr(III) oxidation as function of Mn-oxide induced oxidation, DOC concentration and functional group complexity. To address this objective, mixed batch experiments using synthesized Cr hydroxide, Mn-oxide (50 nm), citric and gallic acid as DOC sources are being conducted. Dissolved Cr(VI), total Cr and Mn are being quantified to develop a quantitative model for Cr(VI) production across DOC concentration gradients of 0.5 – 10 mM. Preliminary results have shown that at a starting concentration of 0.5 mM, citric acid caused rapid Cr(VI) production, which peaked at 5.2 µm within 48 hours. However, within the same period, 10 mM citric acid produced 7.5 times less Cr(VI), resulting in concentrations less than the 1.92 µm threshold for total Cr in drinking water set by the EPA. Ongoing work is seeking to quantify Cr(VI) production at pH 5 using gallic acid as DOC source. These results will elucidate the antagonistic effect of organic carbon during Cr oxidation and help better understand the fate of Cr(VI) in multicomponent systems.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 31 Poster Session I: Poster #I-5

Geospatial modeling to map environmental exposure to abandoned uranium mine waste on the Navajo Nation, USA

Daniel Beene, University of New Mexico, Alburquerque, New Mexico Yan Lin, University of New Mexico, Alburquerque, New Mexico Joseph Hoover, Montana State University, Billings, Montana Esther Erdei, University of New Mexico, Alburquerque, New Mexico Zhuoming Liu, University of New Mexico, Alburquerque, New Mexico

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of New Mexico Superfund Research Center

This research is focused on developing a geovisualization product that characterizes potential exposure to environmental metal contamination from abandoned uranium mines (AUMs), of which there are 523 on the Navajo Nation (NN), a sovereign indigenous Tribal nation in the Southwestern US. Previous health studies have articulated the many human health hazards associated with AUMs, and the environmental mechanisms and pathways (e.g., air, water, and soil) for contaminant transport. Despite this evidence, the limited extant modeling of AUM contamination relies solely on proximity to mines and considers single rather than combined pathways from which the contamination is a product. This can hinder implementation of Tribal public health policies and action plans mitigating health effects associated with AUMs. To address these limitations, the model adopts the following methods to develop a more sophisticated geovisualization illustrating the complex spatial distribution of AUM contamination: GIS- based multi-criteria decision analysis (GIS-MCDA), fuzzy logic, and analytic hierarchy process (AHP). Eight criteria layers were selected for the model: proximity to AUMs, roadways, and drainages; and topographic landforms, wind index, topographic wind exposure, vegetation index, and groundwater contamination. Model sensitivity was evaluated using the one-at-a-time method, and statistical validation was conducted using two separate environmental datasets. The sensitivity analysis indicated consistency and reliability of the model. Model results were strongly associated with environmental uranium concentrations. The model classifies 20.2% of the NN as high potential for AUM contamination while 65.7% and 14.1% of the region are at medium and low exposure potential, respectively. This study is a novel application for AUMs and crucial first step toward informing future studies and ongoing remediation efforts to reduce mobility of AUM wastes and to minimize human exposure.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

32 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-6

Regional Wildfire Pollution on Native Lands: Exposure Assessment and Toxicological Characterization Using a Mobile Inhalation Exposure Laboratory

Jessica Begay, University of New Mexico, Alburquerque, New Mexico Rita Sarracino, University of New Mexico, Alburquerque, New Mexico Selita Lucas, University of New Mexico, Alburquerque, New Mexico David Scieszka, University of New Mexico, Alburquerque, New Mexico Russell Hunter, Marsha Bitsui, Kyle Swimmer, Guy Herbert, Barry Bleske, Melissa Gonzales, Adrian Brearley, Joe Galewsky, Yan Lin, Savannah LaRosa-LoPresti, Jack Harkema, James Wagner, Masako Morishita, Matthew Campen, University of New Mexico; Michigan State University

Superfund Mandate: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

Over the late summer-fall of 2020, unprecedented wildfire activity struck the west coast of the United States, decimating millions of acres of forest as well as inhabited regions. The effluent from these fires travelled hundreds of miles and negatively impacted air quality for millions of people. The UNM METALS Superfund team has been extensively characterizing particulate matter levels and meteorological conditions in the Laguna Pueblo for the past year, as they relate to concerns for windblown toxic metals from a uranium mine site. Additionally, a tangential research project had been conducting inhalation toxicology experiments in the village of Paguate, NM (within the Laguna Pueblo). Specifically, sub-chronic (21d) exposures in male C57BL6/J mice, along with exposures in pregnant FVB/J mice (gestational day 3- 15) were conducted daily for 4h/d. While highly variable due to roaming jet stream patterns, mouse exposures ranged from daily averages of 100 µg/m3 to over 900 µg/m3 in the particle concentrator chambers. Comprehensive evaluations of respiratory inflammation, cardiac function, neuroinflammation, placental responses and uterine artery resistance were carried out. Daily PM2.5 measures often exceeded the annual and 24h average EPA Standards in the Village of Laguna, where air pollution is routinely low due to the rural, low population density setting. These data collectively highlight how detrimental wildfire emissions can be for populations that live hundreds of miles away from the fires.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 33 Poster Session I: Poster #I-7

Investigating Indoor and Outdoor PCB Sources at Homes Near a PCB- Contaminated Waterway

Maeve Bittle, University of Iowa, Iowa City, Iowa Rachel F. Marek, University of Iowa, Iowa City, Iowa Peter S. Thorne, University of Iowa, Iowa City, Iowa Andres Martinez, University of Iowa, Iowa City, Iowa Keri C. Hornbuckle, University of Iowa, Iowa City, Iowa

Superfund Mandate: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Iowa Superfund Research Center

This project determined airborne concentrations of persistent, toxic polychlorinated biphenyls (PCBs) inside and outside homes in urban East Chicago, Indiana. Possible sources of PCBs to home air include modern pigments present in house paint and cabinet varnish, as well as historical Aroclor mixtures of PCBs such as what is found in some old window caulking and light ballasts. PCB concentrations in outdoor air of these homes may be impacted by the dredging of the Indiana Harbor and Ship Canal (IHSC). The IHSC is a PCB-contaminated waterway that is currently being dredged by the Army Corps of Engineers. Indoor and outdoor air samples were collected by field staff using polyurethane foam passive air samplers (PUF- PAS) before and after dredging started. We established the following hypotheses for this study: 1. Aroclor and non-Aroclor PCBs and their possible sources can be identified inside East Chicago homes. 2. The effect of IHSC dredging can be detected and measured at East Chicago homes using PUF-PAS. Samples collected before dredging started (n=73) and after dredging started (n=62) were analyzed for all 209 PCBs. Indoor concentrations ranged from 0.4 to 22.1 ng/m3 (median = 2.1 ng/m3), and outdoor concentrations ranged from 0.1 to 3.0 ng/m3 (median = 0.2 ng/m3). Extraction efficiency, reproducibility, and accuracy were assessed using surrogate standards and replicates of method blanks. We expect the results of this study to inform our stakeholders, including our study participants, the East Chicago community, the Army Corps of Engineers, and the EPA, of the effect of this dredging and of common housing construction materials on airborne PCBs.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

34 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-8

Effects-Directed Analysis: Agonism Isn’t Everything

Gabrielle Black, University of California, Davis Guochun He, University of California, Davis Michael S. Denison, University of California, Davis Thomas M. Young, University of California, Davis

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California, Davis Superfund Research Center

The beneficial reuse of sewage sludge via land application holds immense potential for nutrient recycling, yet the safety of doing so is heavily debated. Consumer product chemicals are found profusely in wastewater treatment plant discharges, and the safety of discharging such chemicals into the environment as part of complex mixtures is not currently well understood. Non-targeted analysis of High- Resolution Mass Spectrometry (HRMS) was combined with in vitro bioassay analysis to identify estrogen active chemicals in 14 sewage sludge samples. Compounds correlated to estrogen agonistic behavior measured on VM7Luc4E2 bioassays were primarily investigated. Chromatographic fractionation and subsequent agonism assay analysis examined four highly estrogen agonistic samples, where 60% of total activity was isolated to a single fraction in 3 of the 4 of samples. However, in one of the four fractionated samples, a sizeable discrepancy between whole extract activity and the sum of the activity of its corresponding fractions highlighted the presence of antagonistic compounds. As a result, VM7Luc4E2 antagonistic bioassays were run on the same 14 extracts and a strong, negative correlation between antagonistic and agonistic activity was observed (linear regression R2=0.7825). The results emphasize the importance of evaluating both agonistic and antagonistic behavior when performing effects-directed analysis to achieve a more comprehensive understanding of biologically relevant chemicals, and thus evaluate risk associated with sewage sludge reuse.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 35 Poster Session I: Poster #I-9

Field-Scale Feasibility: A Priori Modeling of Shear-Thinning Fluids in Remediation

Christopher Bowers, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Cass T. Miller, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

Shear-thinning fluids are found in many industrial, biological, and geophysical processes, and have recently been used in remediation of subsurface pollution. Despite increasing need, modeling of these fluids at field-scales remains ad-hoc, requiring data that is unobtainable for real geological systems. Improving remediation through engineering design requires a new model that eliminates the need to test every possible fluid of interest for every possible geological application. Thermodynamically constrained averaging theory (TCAT) – which derives field-scale models by directly averaging from the pore-scale – was used to determine how the physics of shear-thinning flow are observed at the field-scale. Findings included that (1) the resistance to flow at the field-scale is dependent on the fluid-solid interfacial viscosity, and (2) the correlation between the two may be quantified using parameters of the geological formation that are typically neglected in traditional models. With these insights, a new model is proposed for shear thinning flow that does not require experimental or simulation data. Potential computational challenges are discussed that may warrant further investigation.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

36 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-10

Historical spatial and temporal distribution of legacy contaminants in Galveston Bay and the Houston Ship Channel (GB/HSC) – a systematic evidence map

Krisa Camargo, Texas A&M University, College Station, Texas Margaret Foster, Texas A&M University, College Station, Texas Brian Buckingham, Texas A&M University, College Station, Texas Thomas J. McDonald, Texas A&M University, College Station, Texas Weihsueh A. Chiu, Texas A&M University, College Station, Texas

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Texas A&M University Superfund Research Center

Background: To our knowledge, there have been no other systematic maps interested in characterizing the historical spatial and temporal distribution of legacy contaminants in Galveston Bay (GB) and the Houston Ship Channel (HSC) sediments. We apply a systematic evidence map to evaluate whether a baseline dataset exists for chemical concentrations with consideration given to identifying relevant exposure risks for Houston residents. Objective: The objective of this systematic evidence map is to determine whether there is a baseline reference dataset for GB/HSC that can characterize the historical spatial and temporal distribution of legacy contaminants in the region. Methods: Our inclusion/exclusion criteria utilize a Condition, Context, Population (CoCoPop) statement that addresses five areas: chemicals of interest (condition), geographic region and environment descriptor (context), sediments (population), and study design. In total, our search identified 487 studies with 24 studies analyzed after full text data extraction. Results: Most of the available studies reported on dioxins/furans or mercury; data for other organics and heavy metals was sparse and limited to certain geographic regions of GB/HSC. For example, most metal data were more frequently reported in Upper and Lower Galveston Bay, while the organic data were reported in the Houston Ship Channel. Conclusions: The resulting data sparsity within certain regions of GB/HSC make it difficult to determine baseline levels. Future efforts will need to broaden the spatial, temporal, and contaminant coverage as well as improve key word indexing and data reporting so as to facilitate study identification by future systematic evidence maps and reviews. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 37 Poster Session I: Poster #I-11

Mediation by hormone concentrations on the associations between exposure to phthalate mixtures and adverse birth outcomes

Amber Cathey, University of Michigan Max Aung, University of Michigan Deborah Watkins, University of Michigan Zaira Rosario, University of Puerto Rico Carmen Vélez Vega, University of Puerto Rico Akram Alshawabkeh, Northeastern University José Cordero, University of Georgia Bhramar Mukherjee, University of Michigan John Meeker, University of Michigan

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Northeastern University Superfund Research Center

Phthalates are used in the manufacturing of a myriad of consumer products, resulting in ubiquitous human exposure to a mixture of phthalate compounds. Previous work has suggested that phthalates display endocrine disrupting capabilities, and associations with adverse birth outcomes including preterm birth. Given the importance of hormone regulation during pregnancy, we hypothesized that phthalates may affect pregnancy outcomes via disruption of hormone concentrations. This work therefore aimed to assess the mediating effects of hormone concentrations on the associations between phthalate mixtures and adverse birth outcomes. Repeated urinary phthalate metabolite (N=13) and serum hormone (N=9) measurements were taken at 16-20 and 24-28 weeks gestation among 1011 women in the PROTECT (Puerto Rico Testsite for Exploring Contamination Threats) longitudinal birth cohort. We utilized adaptive elastic net to create phthalate environmental risk scores (ERS) at each study visit, which represent a weighted sum of each individual’s exposure to the mixture of metabolites. Causal mediation analyses were then conducted on a subset of 705 women for whom hormone data was available. All analyses were conducted separately by fetal sex. Among mothers carrying a male, a standard deviation increase in phthalate ERS was associated with 1.061 (95% CI: 1.008, 1.103) times greater odds of delivering preterm, and 28.5% of that association was mediated by progesterone. Incomplete mediation on the association between phthalate ERS and spontaneous preterm birth was observed with CRH (31.5%), estriol (34%), progesterone (40.2%), and testosterone (23.9%), possibly due to strong relationships between the exposure and mediators. There were no significant mediating results observed for mothers carrying a female fetus. These results provide introductory evidence of hormone disruption on the causal pathway between phthalate exposure and preterm birth.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

38 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-12

Risk Characterization of Environmental Samples Using in vitro Bioactivity and Polycyclic Aromatic Hydrocarbon (PAH) Concentrations Data

Zunwei Chen, Texas A&M University, College Station, Texas Dillon Lloyd, North Carolina State University, Raleigh, North Carolina Yi-Hui Zhou, North Carolina State University, Raleigh, North Carolina Weihsueh Chiu, Texas A&M University, College Station Texas Fred Wright, North Carolina State University, Raleigh, North Carolina Ivan Rusyn, Texas A&M University, College Station Texas

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

Methods to assess environmental exposure to hazardous chemicals have primarily focused on quantification of individual chemicals, although chemicals often occur in mixtures, presenting challenges to the traditional risk characterization framework. Sampling sites in a defined geographic region provide an opportunity to characterize chemical contaminants, with spatial interpolation to provide estimates for non-sampled sites. At the same time, the use of in vitro bioactivity measurements is informative for rapid risk-based decisions. In this study, we measured in vitro bioactivity in 39 surface soil samples collected immediately after flooding associated with Hurricane Harvey in Texas in a residential area known to be inundated with polycyclic aromatic hydrocarbon (PAH) contaminants. Bioactivity data were from functional and toxicity assays in human induced pluripotent stem cell-derived hepatocytes, cardiomyocytes, neurons and endothelial cells, as well as human umbilical vein endothelial cells. Concentrations of PAH in these samples was also available, offering a unique opportunity to assess the joint spatial variation. We found significant evidence of spatial correlation of a subset of PAHs and of cell- based phenotypes. In addition, we show that the cell-based bioactivity data can be used to predict several PAH concentrations, as well as overall PAH summaries and cancer risk. This study demonstrates that cell- based profiling can be used as a rapid hazard screening tool for environmental samples by anchoring the bioassays to PAH concentrations. This work sets the stage for identification of the areas of concern and direct quantitative risk characterization based on bioactivity data, thereby providing an important supplement to traditional individual chemical analyses by shedding light on constituents that may be missed from targeted chemical monitoring. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 39 Poster Session I: Poster #I-13

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-elicited substrate overload and thioesterase induction promotes a futile cycle that inhibits hepatic β-oxidation

Giovan N. Cholico, Michigan State University, East Lansing, Michigan Russ R. Fling, Michigan State University, East Lansing, Michigan Nicholas A. Zacharewski, Trinity College, Hartford, Connecticutt Kelly A. Fader, Michigan State University, East Lansing, Michigan Rance Nault, Michigan State University, East Lansing, Michigan Tim Zacharewski, Michigan State University, East Lansing, Michigan

Superfund Mandate: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Michigan State University Superfund Research Center

Non-alcoholic fatty liver disease (NAFLD) includes a spectrum of disorders ranging from simple steatosis, characterized by excessive lipid accumulation, to nonalcoholic steatohepatitis (NASH) with fibrosis. Diverse pharmaceuticals and xenobiotics induce hepatic fat accumulation including 2,3,7,8- tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. Lipidomic analysis of liver extracts from TCDD-treated mice identified marked increases in unsaturated fatty acids (FAs), triacylglycerols, and cholesterol esters. In the present study, we examined the β-oxidation of straight-chain FAs in mice treated with 0.01, 0.03, 0.1, 0.3, 1, 3, 10, or 30 μg/kg body weight TCDD every 4 days for 28 days to test the hypothesis that TCDD represses gene expression associated with β-oxidation. Untargeted metabolomic analysis revealed 30 µg/kg TCDD decreased hepatic decanoyl-, hexanoyl-, butyryl-, and acetyl-CoA levels 126.6-, 34.9-, 11.8-, and 6.3-fold, respectively, while inducing octenoyl-CoA 138.9-fold. Bulk RNA- sequencing suggested TCDD repressed the hepatic gene expression associated with lipolysis, binding proteins (i.e., Fabp1, Dbi, and Scp2), FA activation, and thiolysis while inducing thioesterases. Protein levels for the binding protein DBI, the acyl chain activators ACSM3 and ACSL1, and the acyl-CoA dehydrogenase ACOX1, all exhibited dose-dependent repression. During β-oxidation, acyl-CoAs are subjected to oxidation, hydration, a second oxidation and finally, coenzyme A (CoASH)-dependent thiolytic cleavage to produce acetyl-CoA and an acyl-CoA that is two carbons shorter. Decreased acyl-CoA levels and the accumulation of medium-chain enoyl-CoA species suggests incomplete oxidation. Integrating metabolomics and RNA-seq data suggests TCDD elicited a futile cycle involving the sequestration of CoASH within acyl-CoA species that was released by thioesterases resulting in the repression of β-oxidation. GNC and RRF are supported by the NIEHS Training in Environmental Toxicology Program (T32ES007255). This project is funded by the Superfund Research Program (P42ES004911).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

40 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-14

Identification of New Byproducts from the Oxidation of PAHs with Thermally Activated Persulfate using Liquid Chromatography High Resolution Mass Spectrometry

Amy Cuthbertson, University of California-Berkeley David L. Sedlak, University of California-Berkeley

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of California-Berkeley Superfund Research Center

In situ chemical oxidation (ISCO) is a technique used in the treatment of chemical contaminants, especially for compounds that have been difficult to treat by established methods such as hydrocarbons, organic solvents, polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons (PAHs). ISCO involves the addition of a relatively high concentration of an oxidant into contaminated groundwater or surface soils. Activated persulfate is growing in popularity as a treatment option because of potentially low soil oxidant demands, stability in the subsurface in conjunction with the steady generation of hydroxyl radicals and sulfate radicals. Ideally, oxidation would result in the complete mineralization of organic contaminants to carbon dioxide and water, but recent research shows this may not always be the case. Partial oxidation of organic contaminants can result in the formation of potentially toxic byproducts. Some of these byproducts have increased mobility and hydrophilic character in comparison to parent compounds, which poses a risk to groundwater resources. To understand these processes, we identified new oxidation byproducts from the oxidation of 16 PAHs in heat activated persulfate batch reactions using liquid chromatography high resolution mass spectrometry (LC-HRMS). To prioritize these potentially toxic oxidation byproducts, we tested their reactivity with specific biomolecules (N-acetyl-L-cysteine, N-acetyl- L-lysine, glutathione, and histidine). Results show that oxidation mixtures have higher reactivity with some biomolecules in the aqueous phase in comparison to controls, providing evidence of potentially toxic byproduct formation.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 41 Poster Session I: Poster #I-15

Physiologically-based Pharmacokinetic (PBPK) Modeling of Variability in Perchloroethylene Metabolism in the Collaborative Cross Mouse Population

Chimeddulam Dalaijamts, Texas A&M University, College Station, Texas Joseph A Cichocki, Texas A&M University, College Station, Texas Yu-Syuan Luo, Texas A&M University, College Station, Texas Ivan Rusyn, Texas A&M University, College Station, Texas Weihsueh A. Chiu, Texas A&M University, College Station, Texas

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

Quantitative analysis of inter-individual variability in hazardous substance-induced toxicity is a critical gap in risk assessment. Our previous studies indicated that quantitative estimates of toxicodynamic and toxicokinetic variability can be derived through the use of a population-based approach that is based on studies in multiple inbred mouse strains. For example, Bayesian physiologically-based pharmacokinetic (PBPK) modeling has provided evidence for inter-strain variability in metabolism of perchloroethylene (perc) based on data from three mouse inbred strains. These data suggested the need to better characterize toxicokinetic uncertainty and variability using a large population. In this study, the population PBPK model for perc was advanced to include toxicokinetic data for blood and tissues from male mice from 45 Collaborative Cross (CC) mouse strains. Hierarchical Bayesian population PBPK model calibration was conducted using Markov chain Monte Carlo simulation, with the most influential parameters selected using global sensitivity analysis. The resultant PBPK model provides accurate estimates in organ-specific inter-strain variability and uncertainty in toxicokinetics of perc and its metabolites. Using inter-strain variability as a surrogate for human inter-individual variability, this model was used to calculate toxicokinetic variability factors as a replacement for the default uncertainty factor of 3.16. We found greatest variability, exceeding the default uncertainty factor, in glutathione conjugation metabolites of perc, whereas parent compound and oxidative metabolites showed lower variability and were within the default uncertainty. Overall, these results show that the use of CC mouse population, coupled with PBPK modeling, enabled rigorous empirical data-driven estimates of toxicokinetic variability and that such studies are a sensible in vivo-in silico approach for replacing the default assumptions with chemical- and tissue-specific values for human health risk assessment. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

42 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-16

Assessment of Soil Development using Experimental Revegetation Treatments at an Arizona Copper Mine

Catherine G. Fontana, University of Arizona, Tucson, Arizona Sierra Lauman, University of Arizona, Tucson, Arizona Jessica Ledesma, University of Arizona, Tucson, Arizona Albert Kline, University of Arizona, Tucson, Arizona Raina M. Maier, University of Arizona, Tucson, Arizona Elise S. Gornish, University of Arizona, Tucson, Arizona Julia W. Neilson, University of Arizona, Tucson, Arizona

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Arizona Superfund Research Center

Reclamation of degraded, arid soils disturbed by mining in the Southwest is a critical step in mine closure. In addition to establishing a productive ecosystem, revegetation of degraded soil can significantly reduce dust transport and erosion, which impact surrounding communities. However, arid soils face a number of revegetation challenges, such as low nutrient levels and long periods without precipitation. Potentially, these challenges can be mitigated through seed application amendments, seeding method, and timed seeding with annual rains. This research aims to evaluate which of these mitigation options most positively impacts soil development – an indicator of potential revegetation success. At a copper mine in Arizona, existing vegetation and contaminated topsoil were removed from 0.93 km2 approximately 400 m north of its decommissioned smelter to reduce soil arsenic concentrations below the daily exposure limit. Eighteen randomized experimental plots were thereafter established on the residual unvegetated subsoil to investigate the outcome of hydroseeding season (summer monsoon or winter rains), amendment with arbuscular mycorrhizal fungi (AM120), and seedball (1” balls comprised of organic matter, clay, and seeds) application on soil development over time. Sampling and analysis of plots was conducted prior to seeding and will continue for four consecutive years after initial summer hydroseeding. Soil development parameters, including particle distribution, pH, electrical conductivity, soil biomass, and glomalin concentration (an indicator of soil aggregate formation), will be measured. We hypothesize that greatest soil development (eg, the highest soil biomass and glomalin concentrations) will occur within plots seeded with the AM120 amendment during winter rains. Assessment of temporal differences in these soil development parameters across treatments will illuminate the impact on long-term revegetation potential of these management strategies.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 43 Poster Session I: Poster #I-17

Impacts of stratification and mixing patterns on arsenic mobilization and health risks in urban lakes

Smantha Fung, University of Washington Erin A. Hull, University of Washington, Tacoma Ken Burkart, University of Washington, Tacoma Marco Barajas, University of Washington, Tacoma Alex Horner-Devine, University of Washington, Seattle James E. Gawel, University of Washington, Tacoma Rebecca Neumann, University of Washington, Seattle

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Washington Superfund Research Center

Legacy arsenic in lake sediments can make its way into the food chain and pose health risks for people consuming organisms from contaminated lakes. We found that oxic water arsenic concentration, a known predictor of bioaccumulation in primary producers (Barrett et al. 2018), is controlled by physical lake mixing characteristics such as stability of summer stratification and mixing frequency. This work examines the physical and biogeochemical mechanisms behind arsenic movement from the sediment to oxic waters. We analyzed seasonal patterns of lake mixing, water column arsenic concentrations and bioaccumulation in multiple lakes in the South-Central Puget Sound region, WA. In our shallow study system (max depth of 4 m) we found that, during the period of highest arsenic mobilization, two distinct mixing patterns exist and control the degree of arsenic in oxic waters. Specifically, during early summer, periods of stratification cause arsenic build-up at the sediment water interface and episodic mixing transfers arsenic into the lake epilimnion. During late summer, convection-driven diel overturning distributes arsenic homogeneously throughout the water column. We observed that our deep study system (max depth of 15 m) stays stably stratified throughout the period of highest arsenic mobilization. Subsequently, arsenic is not mixed up into oxic waters. Despite both systems having similar sediment concentrations, organisms collected from the shallow lake exhibit much higher levels of arsenic in their tissues than those collected from the deep system. This work shows that physical mixing processes control movement of arsenic into the aquatic food web. Understanding the physical mechanisms controlling pathways of arsenic transport is essential for identifying contaminated systems that pose health risks to lake biota and people consuming these organisms.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

44 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-18

Estrogen receptor activity prediction of environmental toxicants using image analysis and data-driven modeling

Hari Ganesh, Texas A&M University, College Station, Texas Burcu Beykal, Texas A&M University, College Station, Texas Adam T. Szafran, Baylor College of Medicine, Houston, Texas Fabio Stossi, Baylor College of Medicine, Houston, Texas Lan Zhou, Texas A&M University, College Station, Texas Michael Mancini, Baylor College of Medicine, Houston, Texas Efstratios N. Pisktikopoulos, Texas A&M University, College Station, Texas

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

During environmental emergency-related catastrophic events, many unknown chemicals are mobilized, which increases the risk of exposure and long-term adverse biological effects. Predicting the endocrine disruptive potential of unknown chemicals released in the environment, especially after such events, is critical for human health-focused mitigation strategies. In this work, we develop a classification model to predict the estrogenic activity, an important hormonal pathway in human pathophysiology, of environmental toxicants as estrogen receptor-α (ERα) agonists or antagonists by high throughput microscopy-based high content analysis (HCA) employing big-data analytics and machine learning algorithms. The single cell-level response of 60 benchmark compounds with known in vivo effects upon ERα pathway activities were measured using a novel HCA pipeline. This input dataset captures 77 size, shape, and intensity features per cell which describes ERα expression and subcellular/subnuclear distribution in an engineered cell model that facilitates analysis of multiple mechanistic steps of ERα functions. To reduce the dimensionality of the cell-level data, an appropriate probability distribution function (non-central gamma or normal) was fitted onto the data for each sample as a pre-processing strategy, reducing approximately 2000 cell-level measurements for each sample into a non-central gamma distribution with 3 parameters, or a normal distribution with 2 parameters. The resulting parameters of the probability distribution, which represent the mean and the shape of the distribution, were considered as the features for classification analysis using a Random Forest supervised learning algorithm. The resulting model showed a prediction accuracy and specificity of approximately 90% for classifying the estrogenic potentials of benchmark chemicals as ER agonist or antagonist. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 45 Poster Session I: Poster #I-19

Human health risk from consumption of aquatic species in arsenic-contaminated shallow urban lakes

James Gawel, University of Washington Erin Hull, University of Washington, Tacoma Marco Barajas, University of Washington, Tacoma Kenneth Burkart, University of Washington, Tacoma Ricky Pendergrass, University of Washington, Tacoma Samantha Fung, University of Washington Brian Jackson, Dartmouth College Pamela Barrett, University of Washington Rebecca Neumann, University of Washington Julian Olden, University of Washington

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Washington Superfund Research Center

Arsenic contamination of lakes in the US has occurred as a result of mining, smelting, and its use as an aquatic herbicide. This carcinogen is mobilized from lake sediments during stratification and hypolimnetic anoxia in the summer, but in deeper lakes this often results in a spatial separation between dissolved arsenic and oxygen-requiring organisms, reducing the potential for biotic uptake. However, during periodic summer mixing events typical in shallow lakes, bioavailability of arsenic is enhanced when arsenic mobilized in near-bottom waters is mixed into overlying oxygenated waters where biota reside. Our previous work measured sediment arsenic concentrations of over 200 µg/g in Puget Sound lakes affected by the former ASARCO smelter, and significant bioaccumulation of arsenic in phytoplankton (up to 970 µg/g) and zooplankton (up to 80 µg/g) in contaminated shallow lakes. We have now quantified arsenic concentrations and associated health risks in human-consumed tissues of sunfish, crayfish, and snails from lakes representing a gradient of arsenic contamination and differing mixing regimes. We found that consuming aquatic species from the arsenic-contaminated shallow lakes consistently resulted in greater health risks compared to the deeper lake with similar sediment arsenic concentrations at maximum depth in the lakes. Arsenic concentrations in littoral sediments were more indicative of arsenic uptake in the investigated aquatic species than profundal (deep) sediments. Consumption of crayfish and snails from arsenic-contaminated shallow lakes results in an increased cancer risk for recreational fishers, and subsistence fishers may be at increased risk from consumption of fish from the most contaminated shallow lakes. Further work is needed to identify at risk populations at our study lakes and the potential geographic extent of similar contaminated waterbodies.

Email: [email protected] Presenter Status: Researcher

46 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-20

Insecticides increases oxidative stress in cardiac cells

Aldrin V. Gomes, University of California-Davis, Davis, California Rasheed Sule, University of California-Davis, Davis, California Naoki Matsumoto, University of California-Davis, Davis, California Christophe Morisseau, University of California-Davis, Davis, California Bruce D. Hammock, University of California-Davis, Davis, California Nipavan Chiamvimonvat, University of California-Davis, Davis, California

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

In a recent study by Bao et al. in the JMAA internal medicine journal (doi:10.1001/jamainternmed.2019.6019) they found that prolonged exposure to a widely used household insecticide (pyrethrin) may increase the risk for cardiovascular disease and early death. The World Health Organization (WHO) states that “insecticides tend to be more toxic to humans than herbicides.” We investigated the insecticides esfenvalerate, fipronil, and the insecticide product methamidophos (formed from biotransformation of the organophosphate insecticide, acephate). H9C2 cardiac cells were exposed to 10µM of each pesticide and then reactive oxygen species (ROS) levels were detected. Although ROS are byproducts of normal cellular metabolic processes, high levels of ROS is associated with several diseases including cardiovascular disease. All three insecticides significantly increased ROS levels relative to negative control. Methamidophos has one of the highest ratios of handler poisonings per 1,000 applications in field workers. To further investigate the source of the ROS produced by these insecticides, H9c2 cells were exposed to methamidophos (10µM) and the mitochondrial specific ROS detected. Methamidophos was found to significantly increased mitochondrial superoxide ions. These results suggest that insecticides are potent inducers of oxidative stress by inducing higher levels of intracellular ROS and a significant proportion of the ROS induced originates from the mitochondria.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 47 Poster Session I: Poster #I-21

Extending PAH Identification by integrating Spectroscopies with Machine Learning Strategies

Naomi J. Halas, Baylor College of Medicine, Houston, Texas Aneel Damaraju, Rice University, Houston, Texas Mary Bajomo, Rice University, Houston, Texas Oara Neumann, Rice University, Houston, Texas David Renard, Rice University, Houston, Texas Peter Nordlander, Rice University, Houston, Texas Ankit Patel, Rice University, Baylor College of Medicine, Houston Texas

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Baylor College of Medicine Superfund Research Center

Polycyclic Aromatic Hydrocarbons (PAHs) are dangerous chemical contaminants found throughout our environment, and human exposure to PAHs is known to have deleterious health consequences. The detection of PAHs, particularly in complex fluids or materials such as placental fluid or soil, is a significant challenge. Conventional detection methods typically include extraction, followed by separation procedures such as chromatography, which are integrated with the analysis of all chemical components in the extracted fluid by mass spectrometry. While this approach is the gold standard, it is slow, expensive and time-consuming, requiring large laboratory facilities. This limits our ability to rapidly identify the many PAH exposure sources that may be impacting humans, particularly in well-populated urban areas block- by-block variations in PAH exposure can have impactful health consequences. In this project we are developing a new approach to PAH detection. We are utilizing our expertise in surface-enhanced spectroscopies (Raman (SERS) and Infrared (SEIRA)) for compact chemical detection and identification, combined with Machine Learning (ML) to rapidly identify PAHs in complex mixtures, eliminating or minimizing separation steps. Our first goals in this project are to collect a library of standard spectra of PAH molecules for training of the neural network (NN) that will be created and developed specifically for analysis. One of the initial challenges for NN training is to identify whether a Raman library of PAH spectra can be used directly for PAH identification, or whether a specialized, substrate-dependent SERS library is required. We will describe the initial steps in our chemical analysis and in our NN training we have taken to accomplish our PAH identification goals.

Email: [email protected] Presenter Status: Researcher

48 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-22

Building Trust in the Michigan Department of Health and Human Services

Joseph Hamm, Michigan State University Alaina Bur, Michigan State University Jenna VanFossen, Michigan State University Jennifer Carrera, Michigan State University Adam Zwickle, Michigan State University James Dearing, Michigan State University

Superfund Mandates: None

Superfund Center: Michigan State University Superfund Research Center

Trust in environmental health governance (EHG) agencies is an important element of community resilience to disturbances in the environmental health context. Given the geographic focus of our Superfund Research Center, we identified the Michigan Department of Health and Human Services as an important, public-facing EHG agency and three Michigan communities who were facing dioxin contamination: St. Clair Shores (Ten-Mile Drain Superfund Site), Saginaw (Saginaw River and Bay Area of Concern), and Allegan (Allied Paper INC/Portage Creek/Kalamazoo River NPL Site). This project took an important first step in a larger effort to build trust between the MDHHS and these communities by eliciting information regarding the current state of this relationship from the perspective of the public. Qualitative data were collected in two phases. During the first, we worked with community convenors to identify leaders in each community. These leaders were then invited to participate in the first round of virtual focus groups (one per community). We then recruited a second set of participants through these leaders’ network, this time focusing on residents more generally (i.e., individuals not in leadership roles). This second phase recruitment was supplemented by reaching out to closed Facebook groups with clear connections to one of the three communities. The first phase of data collection is currently complete and suggests several major themes in community leaders’ thoughts about their relationship with and trust in the MDHHS. Specifically, participants identified several specific roles that they expected the MDHHS to play in protecting their communities’ environmental health and almost universally highlighted geographic and social separation from the MDHHS as a key limiter of the level of trust they felt in them.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 49 Poster Session I: Poster #I-23

Reduction of the bioavailability of PFAS (per- and polyfluoralkyl substances) from soil and their translocation to plants in the presence of parent and amended montmorillonite clays

Sara E. Hearon, Texas A&M University, College Station, Texas Meichen Wang, Texas A&M University, College Station, Texas Asuka A Orr, Texas A&M University, College Station, Texas Haley Moyer, Texas A&M University, College Station, Texas Phanourios Tamamis, Texas A&M University, College Station, Texas Timothy D. Phillips, Texas A&M University, College Station, Texas

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

Consumption of food and water contaminated with per- and polyfluoralkyl substances (PFAS) presents a significant risk for human exposure. There is limited data on high affinity sorbents that can be used to reduce the bioavailability of PFAS from soil and translocation to plants and garden produce. A field- practical strategy was used to address this need, where montmorillonite clay was amended with the nutrients carnitine and choline to increase the hydrophobicity of the sorbent and the interlayer spacing. In this study, the binding of parent and amended clays to PFOA (perfluorooctanoate) and PFOS (perfluorooctanesulfonate) was characterized. Isothermal analyses were conducted at pH 7 and ambient temperature (to simulate environmentally relevant conditions). The data for all tested sorbents fit the Langmuir model indicating saturable binding sites with high capacities and affinities under neutral conditions. Amended montmorillonite clays had increased capacities for PFOA (0.51 and 0.61 mol kg-1) compared to the parent clay (0.37 mol kg-1). Molecular dynamics (MD) simulations suggested that hydrophobic and electrostatic interactions at the terminal fluorinated carbon chains of PFAS compounds were major modes of surface interaction. The safety and efficacy of the clays were confirmed in a living organism (Hydra vulgaris), where clays (at 0.02% inclusion) significantly reduced PFOA and PFOS toxicity (p ≤ 0.01). Importantly, soil studies showed that 2% sorbent inclusion could significantly reduce PFAS bioavailability from soil (up to 60%). Studies in plants demonstrated that inclusion of 2% sorbent significantly reduced PFAS residues in cucumber plants (p ≤ 0.05). These results suggest that nutrient- amended clays could be included in soil to decrease PFAS bioavailability and the translocation of PFAS to plants. Supported by NIEHS SRP P42 ES027704.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

50 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-24

Influence of growth substrate on aromatic hydrocarbon and chlorinated alkene cometabolism by Rhodococcus Rhodochrous 21198 and Mycobacterium Strain ELW1

Juliana Huizenga, Oregon State University, Corvallis, Oregon Lewis Semprini, Oregon State University, Corvallis, Oregon

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Oregon State University Superfund Research Center

Mixtures of hydrocarbon contaminants, such as those present in oil spills, present unique opportunities and challenges for microbial degradation. Many microorganisms can grow on small hydrocarbon compounds present in these mixtures, but larger hydrocarbons, particularly aromatic hydrocarbons, are not as easily degraded by microorganisms, and can remain in the environment long after smaller hydrocarbons have been consumed. However, some microorganisms can degrade these aromatic hydrocarbons via cometabolism, a process in which a compound is transformed but no carbon or energy is gained. Therefore, a system in which microorganisms grow on smaller hydrocarbons while also cometabolizing aromatic hydrocarbons would be advantageous for bioremedial strategies. In this study, resting cell rate tests were performed with pure cultures of Rhodococcus Rhodochrous 21198 and Mycobacterium Strain ELW1 grown on either their primary gaseous hydrocarbon growth substrate or an alternative growth substrate. Cometabolic activity was first confirmed with the degradation of cis-1,2- dichloroethylene and formation of the epoxide intermediate, and then assessed in batch tests with toluene as the model aromatic hydrocarbon. GC/ECD and GC/FID were used to measure chlorinated and volatile aromatic compounds, respectively. HPLC/UV and GC/MS were used to identify non-volatile aromatic products. Preliminary results indicate that the cometabolic capability of the pure cultures depends on both the growth substrate and the contaminants. Alternative growth substrates can decrease or eliminate the cometabolic activity of the microorganisms, but changes in enzyme expression due to growth substrate may be challenged by an induction effect from the contaminant. These results demonstrate that combined efforts of microbial metabolism and cometabolism have the potential to remediate hydrocarbon mixtures, but the rate and extent of remediation is dependent on the microorganisms and contaminants present.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 51 Poster Session I: Poster #I-25

Spatial and temporal distribution of surface water quality contaminants after the Intercontinental Terminal Company Fire

Suji Jang, Texas A&M University, College Station, TX Thomas J. McDonald, Texas A&M University, College Station, TX Sharmila Bhandari, Texas A&M University, College Station, TX Ivan Rusyn, Texas A&M University, College Station, TX Weihsueh A. Chiu, Texas A&M University, College Station, TX

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

After the fires at Intercontinental Terminal Company (ITC) in Houston, Texas in March 2019, Texas Commission on Environmental Quality (TCEQ) and Environmental Protection Agency (EPA) had collected water samples from 38 locations in Houston Ship Channel and Galveston Bay for up to two months, which were chemically analyzed for 433 hazardous substances. Although these data were displayed on the ITC- response website, no further analyses have been reported. In this study, we used these data to evaluate the time- and location-dependent patterns of chemical concentrations, as well as to compare with EPA water quality criteria. After web-scraping 77,841 rows of data, we selected for evaluation seven analytes of common concern for human health risk – benzene, ethylbenzene, toluene, xylene, oil and grease, total suspended solids, and total petroleum hydrocarbons – substances for which temporal and spatial analysis could be performed. Temporal analysis indicated that concentrations of hazardous substances dissipated rapidly, with most analytes declining to steady-state or below detectable levels after 1-2 weeks. Spatial analysis demonstrated that substances were most concentrated near the ITC site, with substantial dilution at distances of 1 km or more. At locations near the site, levels of benzene initially exceeded water quality criteria by up to 50,000-times; while most locations quickly returned to acceptable levels, a few locations remained elevated even after 4 weeks. Overall, contamination analyzed by TCEQ and EPA showed high concentrations near the ITC site during the first week, after which they largely declined to levels below EPA water quality concern. However, these conclusions are limited to the small number of substances analyzed for which there was sufficient data to examine their spatial distribution and time-dependence. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

52 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-26

The SLIM1 transcription factor regulates arsenic sensitivity in Arabidopsis thaliana

Timothy O. Jobe, University of Cologne, Cologne, Germany Qi Yu, University of California, San Diego, La Jolla, California Felix Hauser, University of California, San Diego, La Jolla, California Qingqing Xie, University of California, San Diego, La Jolla, California Yuan Meng, University of California, San Diego, La Jolla, California Tim Maassen, University of Cologne, Cologne, Germany Stanislav Kopriva, University of Cologne, Cologne, Germany Julian I. Schroeder, University of California, San Diego, La Jolla, California

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California-San Diego Superfund Research Center

The transcriptional regulators that mediate rapid heavy metal- and arsenic-induced gene expression in plants remain largely unknown. However, cadmium and arsenic exposure rapidly deplete cellular glutathione levels increasing demand for thiol compounds from the sulfur assimilation pathway. Thus, sulfur assimilation is tightly linked with cadmium and arsenic detoxification. To explore the hypothesis that the key transcriptional regulator of sulfur assimilation, SLIM1, is involved in cadmium and arsenic- induced gene expression, we evaluated the response of two allelic slim1 mutants to cadmium and arsenic treatments. We found that slim1 mutants were more sensitive to arsenic than to cadmium in root growth assays. Furthermore, arsenic treatment caused high levels of oxidative stress in the slim1 mutants and slim1 mutants were impaired in both thiol and sulfate accumulation. We also found enhanced arsenic accumulation in the slim1 mutants, which we propose is partly the result of up-regulation of phosphate transporters in the slim1 mutants. Furthermore, microarray analyses identified a gene known to be involved in the epigenetic regulation of sulfate assimilation as being significantly up-regulated in the slim1-1 mutant in response to arsenic exposure. Our results suggest that the severe arsenic sensitivity of the slim1 mutants is a result of both altered redox status as well as mis-regulation of key genes involved in epigenetic regulation of sulfate assimilation.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 53 Poster Session I: Poster #I-27

Distributed Harmonization and Analysis Across Three SRP Cohorts

David Kaeli, Northeastern University Daniel R Beene, University of New Mexico Nicolas Bohm Agostini, Northeastern University Zlatan Feric, Northeastern University Yuliya Halchenko, Dartmouth College Margaret R. Karagas, Dartmouth College Johnnye Lewis, University of New Mexico Debra MacKenzie, University of New Mexico Justin Manjourides, Northeastern University Martha Powers, Northeastern University Antonio J. Signes-Pastor, Dartmouth College Deborah Watkins, University of Michigan

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Northeastern University Superfund Research Center

In this presentation we discuss the results to date of our External Use Case project that studies environmental exposures across three SRP pregnancy and birth cohorts. The three cohorts include the University of New Mexico (METALS), Dartmouth College (New Hampshire Birth Cohort) and Northeastern University (PROTECT) centers. The key research questions we are addressing include whether biomonitoring data and other demographic and exposure data of three SRPs can be harmonized leveraging FAIR principles, enabling us to jointly study the effects of environmental exposures on multiple outcomes. We identified common data elements collected across the three cohorts, as well as challenges and opportunities for harmonization. In order to maintain tribal sovereignty and data sharing agreements with the Navajo Nation, we built a completely secure and distributed analysis framework, where Northeastern and Dartmouth teams can securely send their data to UNM. We now can build a set of well- defined queries that can be carried out across multiple cohorts, while not allowing access to or sharing of individual-level tribal data. Today, each team can securely analyze this multi-cohort data on our secure server. Our tools are open-source and will be shared with the SRP community. To date, we have focused on analysis of arsenic exposure and birth outcomes, and will be reporting our findings in this presentation.

Email: [email protected] Presenter Status: Researcher

54 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-28

Development of an online decision support tool for gardeners to assess soil contaminant risks and take action to reduce exposure

Catherine Kastleman, Duke University, Durham, North Carolina Elizabeth Shapiro-Garza, Duke University, Durhamm, North Carolina Bryan Luukinen, Duke University, Durham, North Carolina Samuel Cohen, Duke University, Durham, North Carolina Charlotte Clark, Duke University, Durham, North Carolina Lucy Bradley, North Carolina State University, Raleigh, North Carolina

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Duke University Superfund Research Center

Community gardens and urban farms offer many public health benefits. Chemical contaminants in garden soils, however, may present health risks for gardeners: lead, arsenic, and pesticides have been widely detected in both urban and rural soils in the United States. Based on previous research conducted by the Duke University Superfund Research Center (DUSRC) CEC, gardeners face barriers to assessment of, and response to, potential risks, including inadequate knowledge of soil contaminants and their health impacts, a lack of resources to evaluate or address concerns, and difficulty translating soil test results into actions to reduce exposure. To address this deficit, the CEC collaborated with NC State Extension and North Carolina gardeners to develop and pilot an online decision support tool and map to help gardeners identify potential sources of contaminants, assess health risks, and develop an action plan to test soils and reduce potential exposure. The online tool is integrated with an ArcGIS map built by the DUSRC that displays publicly-available data on potential sources of contamination in proximity to a user-specified address. Within the tool, geospatial data is combined with gardeners’ responses to a series of survey questions about the demographics of garden users, management practices, land use history, and additional sources of contamination to produce a customized report that communicates information about specific contaminants, their sources, and health risks, as well as suggested activities for remediation or prevention. Thus, the tool incorporates community knowledge and experiences into a basic risk assessment process. This project provides a model for the development of interactive, participatory mapping tools that can support soil contaminant risk assessment and offer action steps to reduce or prevent exposure in impacted communities.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 55 Poster Session I: Poster #I-29

Single cell transcriptomics demonstrates reduction in expression of genes associated with ribosomal proteins and erythropoiesis with TCDD treatment during early stages of human B lymphocyte development

DM Isha Olive Khan, Michigan State University, East Lansing Michigan Norbert Kaminski, Michigan State University, East Lansing Michigan Peer Karmaus, Michigan State University, East Lansing Michigan, Natl Inst of Enviro Health Sciences

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

Persistent aryl hydrocarbon receptor (AHR) activation impairs B cell development by mechanisms not fully understood. Single cell RNA-Sequencing (scRNA-Seq) was employed to study the effect of AHR activation by 1 nM 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on early stages of B lymphocyte development. An in vitro culture system that drives B cell development from human cord blood derived CD34+ hematopoietic stem and progenitor cells (HSPCs) was used. We identified distinct cellular clusters in the scRNA-Seq dataset characterized by unique gene expression patterns, defined in part by expression of the AHR transcriptional target CYP1B1. CYP1B1 induction was 2-fold higher in TCDD treated vs vehicle (VH; 0.02% DMSO) group, but not all TCDD treated cells expressed CYP1B1, indicating cellular transcriptional heterogeneity. Interestingly, CYP1B1 expression was observed in 30% of the cells in VH group, suggesting endogenous AHR activation. Cells with higher CYP1B1 expression in both treatment groups were associated with higher (>3.5 fold) expression of CD14, suggesting CD14 as a surface marker for high CYP1B1 expression. In total, 90 genes were differentially regulated (log fold change difference > 0.25, p value < 0.01) by TCDD treatment. TCDD downregulated ribosomal protein encoding genes (11) and genes associated with erythropoiesis. Irrespective of treatment, most cells expressing genes associated with erythropoiesis and ribosomal proteins expressed 4-fold lower CYP1B1 induction, suggesting that AHR activity is antagonistic to expression of these genes. This study allows characterization of differentiating HSPCs that are heterogeneous in AHR activity and shows that genes associated with erythropoiesis and ribosome formation are suppressed by AHR activation during early B cell development. Further time course studies using scRNA-Seq will elucidate TCDD’s effect on the cellular developmental trajectory from HSPCs to B cells.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

56 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-30

Metabolic Reprogramming by In utero Maternal Benzene Exposure

Lisa Koshko, Wayne State University

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Unaffiliated

Environmental chemicals play a significant role in the development of metabolic disorders, especially when exposure occurs early in life. We have recently demonstrated that benzene exposure, at concentrations relevant to a cigarette smoke, induces a severe metabolic imbalance in a sex-specific manner affecting male but not female mice. However, the roles of benzene in the development of aberrant metabolic outcomes following gestational exposure, remain largely unexplored. In this study, we exposed pregnant C57BL/6JB dams to benzene at 50 ppm or filtered air for 5 days/week (6h/day from gestational day 1 to birth) and studied male and female offspring metabolic phenotypes in their adult life. While no changes in body weight or body composition were observed between groups, 4-month-old male and female offspring exhibited reduced parameters of energy homeostasis (VO2, VCO2, and heat production). However, only male offspring from benzene-exposed dams were glucose intolerant and insulin resistant at this age. By six months of age, both male and female offspring displayed glucose and insulin intolerance, associated with elevated expression of hepatic gluconeogenesis and inflammatory genes. Additionally, this effect was accompanied by elevated insulin secretion and increased beta-cell mass only in male offspring. A decreased fetal weight was also observed at E17.5, but this reduction was compensated later in life. Thus, gestational benzene exposure can reprogram offspring for increased susceptibility to the metabolic imbalance in adulthood with differential sensitivity between sexes.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 57 Poster Session I: Poster #I-31

Lung macrophages modulate alveolar barrier dysfunction in cadmium-mediated acute lung injury

Jennifer Larson-Casey, University of Alabama at Birmingham Linlin Gu, University of Alabama at Birmingham Veena B, University of Alabama at Birmingham A. Brent Carter, University of Alabama at Birmingham and Birmingham Veterans Administration Med Ctr

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Alabama Superfund Research Center

Acute lung injury (ALI) is a disorder of acute inflammation that causes disruption of the lung epithelial and endothelial barriers. Many treatments targeting currently used ALI-models have failed to increase patient outcomes. Environmental exposure to heavy metals is often overlooked in ALI and has not been well studied as a potential risk factor for ALI. Cadmium is widely distributed in the environment and environmental levels of cadmium are steadily rising. With no curative therapy for heavy metal-induced lung injury, understanding the pathogenesis and progression of cadmium-mediated ALI is essential. Macrophages switch between phenotypically distinct subpopulations, the classically activated, pro- inflammatory and the alternatively activated, anti-inflammatory macrophages. Macrophage polarization is tightly regulated and mixed populations exist in complex pathological conditions, such as ALI. Our data demonstrate macrophages polarize to a pro-inflammatory, classical phenotype after cadmium-mediated ALI. RNA sequencing revealed significant upregulation of TNF-α, IL-1β, and IL-6 in lung macrophages from cadmium-exposed mice. Metabolic adaptation provides energy for the diverse macrophage functions. Our data indicate that metabolic reprogramming of macrophages to glycolysis increases extracellular acidification rate and lactate excretion occurs in cadmium-mediated ALI. These classically activated macrophages contribute to alveolar epithelial cell (AEC) barrier dysfunction by preventing wound repair after ALI. Modifying the macrophage phenotype to alternative activation with expression of peroxisome proliferator-activated receptor ɣ (or neutralizing macrophage derived TNF-α from cadmium-exposed macrophages restored AEC barrier function. Our data implicates monocyte-derived macrophages contribute to AEC barrier dysfunction and increased permeability after cadmium-mediated ALI due to the persistence of classically activated lung macrophages. Therapeutic strategies to promote the alternative activation of lung macrophages enhanced AEC barrier function and reduced ALI after cadmium exposure.

Email: [email protected] Presenter Status: Researcher

58 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-32

Characterization of covalent and noncovalent interactions of diverse PFAS with proteins using suspect-screening and molecular docking

Wenting Li, University of California, Davis Heather N. Bischel, University of California Davis

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California, Davis Superfund Research Center

Aqueous film forming foams (AFFF) used for hydrocarbon fire suppression consist of a mixture of diverse poly- and perfluoroalkyl substances (PFAS). The assessment of bioaccumulation of PFAS sourced from commercial products has been insufficient and challenging. We exposed human serum albumin protein (HSA) to a series of AFFF (3M, 1999) dilutions in equilibrium dialysis assessed by High Resolution Liquid Chromatography Quadropole Time-of-Flight Mass Spectrometry (HPLC-QTOF-MS). From equilibrium dialysis experiments, we assessed the noncovalent associations of 26-targeted PFAS, which exhibited strong binding affinities with HSA, including C4-PFAS (log Ka= 4.1-4.9 M-1). Low levels (5-20%) of the three PFAS(PFBS, PFOS, and PFOA) exposed to HSA are retained via covalent binding, which might contribute to toxic effects. We further expanded our analysis by screening our samples against a PFAS library containing over 3,000 compounds, as over 91% of organic fluorine in AFFF cannot be assessed with target-analysis against 26-PFAS standards. We calculated pseudo-bioconcentration factors (BCFpseudo) for covalently bound and noncovalently bound PFAS that were qualified from suspect-screening. The short-chain PFAS have been considered to be less bioaccumulative, yet BCFpseudo for the C4 sulfonate in the covalent- bond fraction (BCFPFBS,pseudo = 9.88 ± 1.94) was an order of magnitude greater than that for L-PFOS. By simulating interactions between PFAS and HSA crystal structures via molecular dockings and molecular dynamics, we were not only able to validate the computational method for targeted PFAS but also to predict protein binding affinities of novel PFAS qualified from suspect-screening. The combination of experimental and modeling techniques provided value in rapidly assessing the bioaccumulate potential of emerging PFAS in commercial products and addressed the significance of overlooked PFAS bioaccumulation pathways.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 59 Poster Session I: Poster #I-33

Modeling Fate and Transport of Volatile Organic Compounds inside Sewer Systems

Ying Li, University of Kentucky, Lexington, Kentucky Mohammadyousef Roghani, ARCADIS, Inc. Ariel Robinson, University of Kentucky, Lexington, Kentucky Elham Shirazi, Geosyntec Consultants Kelly G. Pennell, University of Kentucky, Lexington, Kentucky

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Kentucky Superfund Research Center

Several field studies have shown that volatile organic compounds (VOCs) can be transported in sewer pipes and ultimately contaminate indoor spaces via plumbing fixtures. To date, there are no models available to describe the fate and transport of VOCs through sewer systems, and this poses a challenge when assessing and mitigating risks associated with this exposure pathway. As part of this research, we developed a numerical model that predicts concentrations of VOCs within a sewer system. Since every sewer system has a unique set of situations, the model allows various input parameters, including temperature, sewer liquid depth, groundwater depth, and sewer construction specifics. The model was calibrated to simulate TCE (trichloroethylene) fate and transport within a section of sewer system located near a TCE contaminated groundwater plume near a Superfund site in California. Overall, the model and the TCE field data compare well and provide insights about how site assessment and mitigation approaches can be improved.

Email: [email protected] Presenter Status: Researcher

60 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-34

Community Engagement in the Time of COVID-19: Virtual Strategies for the Metals and Metal Mixtures: Cognitive Aging, Remediation, and Exposure Sources (MEMCARE) Study, San Luis Valley, Colorado

Francesca Macaluso, University of Colorado, Boulder, Colorado Diana Jaramillo, University of Colorado, Boulder, Colorado Katherine A. James, University of Colorado, Boulder, Colorado

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Harvard University Superfund Research Center

The goal of the Community Engagement Core (CEC) of the MEMCARE study is to better understand the concerns and needs of community members regarding early-life exposures to metals and later-life cognitive health and promote widespread participation and investment in these issues. While traditional community engagement efforts often center around events like town halls, workshops, and other local gatherings, COVID-19 has made in-person event planning challenging. The CEC branch of MEMCARE in the San Luis Valley, Colorado has designed a more flexible engagement strategy that can be modified to be more targeted or more widely available as public health guidance around COVID-19 changes. This strategy includes: (1) a YouTube channel, where researchers will post videos discussing topics related to the theme of environmental metals exposure and cognitive health, (2) a twitter page, where researchers can link upcoming events and articles from scientific literature and other media, (3) a podcast about study activities, and (4) a public Facebook page that will feature posts and graphics about the MEMCARE study, links to YouTube and podcast content, and serve as a forum for community discussion around study activities or other issues of interest. Town halls will still take place virtually over Zoom, but by having multiple engagement sources on social media, the research team hopes to keep conversations around the MEMCARE study and other environmental issues of concern going over a longer time frame. Additionally, online content will allow for more extensive translation and content adaptation efforts to make relevant study information and materials accessible to a broader audience.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 61 Poster Session I: Poster #I-35

Mechanisms of Benzene-Induced Endothelial Injury: Role of Heat Shock Proteins

Samantha McFall, University of Louisville, Louisville, Kentucky Nalinie S. Wickramasinghe, University of Louisville, Louisville, Kentucky Wesley Abplanalp, University of Louisville, Louisville, Kentucky Marina v. Malovichko, University of Louisville, Louisville, Kentucky Daniel J. Conklin, University of Louisville, Louisville, Kentucky Timothy E. O’Toole, University of Louisville, Louisville, Kentucky Sanjay Srivastava, University of Louisville, Louisville, Kentucky

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Benzene, a ubiquitous environmental pollutant, is one of the top 20 chemicals produced in the United States. It is also abundant in automobile exhaust and cigarette smoke. Our studies have shown that in humans, exposure to benzene is positively associated with circulating levels of endothelial microparticles, the sub-clinical markers of endothelial injury. These microparticles are often shedded from activated and injured endothelial cells. Increased levels of endothelial microparticles are well documented in pathological and toxicology conditions such as diabetes and smoking. To test the plausibility that exposure to benzene is sufficient to increase the endothelial microparticle levels in the blood, we exposed male C57BL/6 mice to benzene (50 ppm, 6h/day, 5days/week) via inhalation for two weeks and analyzed the blood microparticle levels by flow cytometry. Our data show that benzene exposure increased the blood endothelial microparticle and activated endothelial microparticle levels by 2-4 fold (P<0.05). In vitro studies showed that benzene metabolite t,t,-muconaldehyde (10 micro molar) causes the apoptosis of human aortic endothelial cells (HAEC) as assessed by caspase-3 and caspase-7 activation. RNA-seq analysis of HAEC incubated with t,t,-muconaldehyde (10 micro molar) showed a robust upregulation (log 2-6-fold) of heat shock proteins (HSPs), especially HSPA1A, HSPA1B, HSPA6, and HSPA7. siRNA-mediated knockdown of HSPA1B augmented the cleavage of caspase-3 and caspase-7 in t,t,-muconaldehyde- treated HAEC, suggesting that HSPA1B is causally involved in endothelial cell apoptosis. Together, our data suggest that benzene induces endothelial injury by upregulating HSPs, especially HSPA1B.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

62 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-36

Changes in county level toxic releases are associated with increased circulatory disease mortality between 2002-2012

Katlyn McGraw, University of Louisville, Louisville, Kentucky Shesh N. Rai, University of Louisville, Louisville, Kentucky Daniel W. Riggs, University of Louisville, Louisville, Kentucky Aruni Bhatnagar, University of Louisville, Louisville, Kentucky

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Cardiovascular disease (CVD) is the leading cause of death from environmental exposures. Exposure to air pollution, specifically PM2.5, is a well-established risk factor for CVD. However, the contribution of other hazardous pollutant exposure to CVD is less clear. To assess the link between environmental exposures and CVD, we used the publicly available Toxic Release Inventory (TRI) data and age adjusted circulatory disease mortality rates from the Centers for Disease Control and Prevention. We collected data for the years 2002 to 2012. We built linear mixed models to longitudinally assess the link between changes in annual county level toxic releases and circulatory disease mortality rates. We used two predictor variables, the sum of total releases per county (i) per year (j) in pounds, and a county level risk score. The county level risk score is based on reportable quantities (RQ) or toxicity equivalent scores (TES) of specific chemicals. For each county (i), each year (j), we summed the product of each chemical release in pounds (m) multiplied by the inverse of the RQ or TES. Models were adjusted for demographic estimates of county percent male, percent White, percent Hispanic, and percent less than high school diploma earned, median household income, alcohol, smoking, frequency of Superfund sites, and PM2.5. For every 10% increase in toxic releases at the county level, we found a 1.1% (0.47, 1.73; p-value=0.0006) increase in circulatory disease mortality rate. Additionally, we found that for every 10% increase in county level risk score, there was a 1.2% (95%CI 0.54, 1.83; p-value=0.0003) increase in circulatory disease mortality. These results suggest that both the quantity and toxicity of county level releases may contribute to CVD mortality.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 63 Poster Session I: Poster #I-37

Identification of a cell-based osteoarthritis model for evaluating biological actions of xenobiotic exposures

Cindy McReynolds, University of California, Davis Christophe Morisseau, University of California, Davis Sung Hee Hwang, University of California, Davis Bruce Hammock, University of California, Davis

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California, Davis Superfund Research Center

Background: Epoxy-fatty acids (EpFAs) are natural lipid metabolites that have a regulatory function in maintaining cellular homeostasis. These lipids are degraded from cells through a variety of mechanisms, including metabolism by the soluble epoxide hydrolase (sEH) enzyme. The sEH catalyzes the hydrolysis of the EpFAs into its corresponding inflammatory or less-active diol; and because xenobiotics alter the expression and activity of sEH, there is on-going research efforts into understanding how environmental contaminants influence human health through modulation of sEH expression and activity. Equally important is understanding the biological role of EpFAs in cellular systems to understand the importance of how xenobiotic exposures affect the biological outcomes of altering this enzyme. Increased exposures to environmental chemicals, such as PCBs, have been linked to both increased sEH activity and increased OA. The purpose of this research is to develop a cell-based system to evaluate xenobiotic exposures on an in-vitro model of osteoarthritis (OA). Methods: Human chondrocyte cells were treated with IL-1β, an inflammatory cytokine identified in arthritis joints. Cells were treated with EpFAs and the corresponding diols to determine if alternations in lipid profiles would protect or worsen chondrocyte cytotoxicity. Results: Exposure to IL-1β increased chondrocyte cytotoxicity and both epoxides of omega-3 and -6 fatty acids were protective of chondrocytes while the diols significantly increased cytotoxicity compared to the vehicle controls. Conclusion: This in vitro model can be used to not only investigate the potential mechanisms of direct action on chondrocytes or indirect action through sEH activity but also screen other environmental contaminants on potential risks in promoting osteoarthritis.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

64 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-38

Solubility and Thermodynamic Measurements of Na- and K- Uranyl Arsenate Solids

Maria Meza, University of New Mexico Isabel Meza, University of New Mexico Jorge Gonzalez, University of New Mexico Peter Burns, University of Notre Dame Ginger Sigmons, University of Notre Dame Jennifer Szymanowski, University of Notre Dame Tori Forbes, University of Iowa Lindsey Applegate, University of Iowa Abdul-Mehdi S. Ali, University of New Mexico Peter Lichtner, University of New Mexico José M. Cerrato, University of New Mexico

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

This study aims to determine the thermodynamic properties of Na- and K-bearing uranyl arsenate solids (NaUAs and KUAs) integrating solid analyses, aqueous chemistry, calorimetry, spectroscopy, and modeling. The solubility of uranyl arsenate solids needs to be further investigated to predict soluble uranium (U) and arsenic (As) concentrations in water streams near sites affected by natural and anthropogenic processes. The stoichiometries of Na(UO2)(AsO4)(H2O)3 and K(UO2)(AsO4)(H2O)3 were obtained from synthetic solids based on X-ray diffraction, TGA, and acid digestions. The measured solubility product (Log Ksp) for the NaUAs ranged from -22.74 to -24.00, and for KUAs ranged from -23.71 to -24.19 obtained from solubility experiments from undersaturated conditions at pH 2. The standard- state enthalpy of formation from oxides of NaUAs is -357.73 kJ mol-1 and for KUAs is -458.74 kJ mol-1 measured by drop solution calorimetry. Supersaturated conditions and solubility measurements with a different pH are still in process. This work provides novel thermodynamic information that will be useful for reactive transport models that aim to interpret and predict the solubility of U and As in environmental systems.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 65 Poster Session I: Poster #I-39

Modulation of polycyclic aromatic hydrocarbon (PAH)-mediated pulmonary carcinogenesis in mice lacking genes for Cyp1a1/1a2, Cyp1a1/1a2/1b1, or Cyp1b1: Role of CYP1B1 in carcinogenesis

Bhagavatula Moorthy, Baylor College of Medicine, Houston, Texas Grady Gastelum, Baylor College of Medicine, Houston, Texas Chun Chu, Baylor College of Medicine, Houston, Texas Gudong Zhou, Texas A&M University Weiwu Jiang, Baylor College of Medicine, Houston, Texas Xia Guobin, Baylor College of Medicine, Houston, Texas Lihua Wang, Baylor College of Medicine, Houston, Texas

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Baylor College of Medicine Superfund Research Center

Polycyclic aromatic hydrocarbons (PAHs) are environmental contaminants that are present in superfund sites in the U.S. Human exposure to PAHs has been linked to increased risk of lung cancer. In this study, we tested the hypothesis that mice lacking the genes for Cyp1a1/1a2, Cyp1a1/1a2/1b1, or Cyp1b1 would lead to differential modulation of PAH carcinogenesis. Wild type (WT) (A/J), Cyp1a1/1a2, Cyp1a1/1a2/1b1, or Cyp1b1-null mice were treated a vehicle corn oil (CO) or the PAHs, benzo[a]pyrene (100 mg/kg) or 3-methylcholanthrene (MC) (25 mg//kg), once daily for 4 days, and mice were sacrificed at 1, 8, r 15 days after the last dose. For tumor studies, the mice were treated with a single dose of the PAH, and animals were terminated after 40 weeks. We found that Cyp1a1/1a2/1b1-/- mice had a significant reduction in hepatic DNA adducts, which was seen to an even greater extent in the lungs. While NQO1 RNA and protein expression were elevated in WT mice, male and female Cyp1a1/1a2/1b1-/- mice did not show this effect. Additionally, Cyp1a1/1a2/1b1-/- mice displayed high levels of Ahrr RNA, a negative regulator of the AHR receptor. Our tumor formation study found that both male and female Cyp1a1/1a2/1b1 or Cyp1b1-/- mice treated with MC harbored significantly fewer tumors than WT counterparts. Interestingly, Cyp1a1/1a2-/- mice showed a significant increase in tumor multiplicity after MC treatment. Together these findings suggest that CYP1B1 has the ability to metabolize MC into metabolites that have significant potential to induce pulmonary carcinogenesis. Further investigation into the mechanistic roles the CYP1 enzymes play in PAH-induced carcinogenesis will help inform efforts to develop prophylactic measures against PAH-induced carcinogenesis, such as dietary fish oil.

Email: [email protected] Presenter Status: Researcher

66 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-40

Investigation of Heavy Metal Biomarkers for the Assessment of Remediated Surface Waters

Juan C. Morales, Florida International University, Miami, Florida Leonel Lagos, Florida International University, Miami, Florida Lisa Bramer, Pacific Northwest National Laboratory, Richland, Washington Katrina Waters, Pacific Northwest National Laboratory, Oregon State University, Corvallis, Oregon

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Oregon State University Superfund Research Center

Heavy metals are ubiquitous environmental compounds and their prolonged exposure is known to cause deleterious effects in the environment and human health. Moreover, arsenic (As), cadmium (Cd) and mercury (Hg) can enter the food chain and accumulate in the liver of fish, but their combined impact on toxicity mechanisms is poorly understood. We propose, the identification of biomarkers to help discriminate heavy metals and understand their individual pathway-based toxicity mechanisms in response to exposure. We utilize a Random Forest (RF) algorithm, implementing decision trees and ranking subsets of genes using a variable of importance measure. We analyzed the transcriptional responses in zebrafish to heavy metals along with control samples using publicly available datasets in the Gene Expression Omnibus. With observed accuracy around 90%, the RF model results proved to be successful against the test data. We determined the top 25 subsets of biomarkers according to node purity based variable importance measure. Ranked by RF, heavy metal biomarkers were selected for functional and pathway analysis including Gene Ontology (GO) and KEGG. We show statistically significant response to inorganic substance, response to metal ions, cellular processes, positive regulation of protein kinase cascade, positive regulation of transcription from RNA polymerase II promoter, and regulation of protein kinase cascades. We revealed dysregulation of multiple pathways responsible in p53 signaling, apoptosis, cell cycle, fatty acid regulation, cardiac muscle contraction and ubiquitin mediated proteolysis. Random Forest provides evidence that heavy metals biomarkers can be used to understand toxicity pathways derived from the gene expression data. In conclusion, RF provides a pathway framework to identify biomarkers that can be applied to monitor resident species for exposure to accumulated metals in surface waters.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 67 Poster Session I: Poster #I-41

The Air that We Breathe: Volatile PFAS in Indoor Environments

Maya Morales-McDevitt, University of Rhode Island Simon Vojta, University of Rhode Island Jitka Becanoca, University of Rhode Island Tom Bruton, Green Science Policy Institute Arlene Blum, Green Science Policy Institute Rainer Lohmann, University of Rhode Island

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Rhode Island Superfund Research Center

Neutral, volatile perfluorinated alkyl substances (PFAS) precursors, such as fluorotelomer alcohols (FTOHs), perfluorooctane sulfonamides (FOSAs), perfluorooctane sulfonamide ethanol (FOSEs) are industrial biproducts. In the presence of oxygen, they are known to degrade into stable perfluoroalkyl sulfonates (PFSAs) and perfluoroalkyl carboxylates PFCAs. As such, detection of volatile PFAS has been problematic. In order to accurately measure these compounds, we propose the use of apolar polyethylene (PE) sheets as a passive detection tool. The presence of volatile PFAS in indoor air from carpeted classrooms was evaluated using pre-cleaned polyethylene (PE) passive samplers. Two types of PE passive samplers differentiated by thickness (25um and 50um) were used to determine the interactions of the volatile PFAS with these detection tools. The compounds of interest included 6:2 FTOH, 8:2 FTOH, 10:2 FTOH. 8:2 FTAcr, 10:2 FTAcr, MeFOSA, MeFOSE, EtFOSA, and EtFOSE. All samples were analyzed through GC-MS. Results from a previous kinetic study and from both types of passive samplers show similar amounts per gram of PE sheet and appear to be in equilibrium. This suggests that interactions of the passive samplers with the volatile PFAS are occurring at the surface, rather than being absorbed. To determine the exposure risk in indoor environments, air concentrations were paired with dust and carpet measurements taken at the same locations, the vapor pressure of each compound, and the ventilation rates of the indoor environments.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

68 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-42

Role of astroglial cells in neurotoxicity Of 2,2’,5,5’-Tetrachlorobiphenyl (PCB 52) and its metabolites

Neha Paranjape, University of Iowa, Iowa City, Iowa Brianna Cagle, University of Iowa, Iowa City, Iowa Ronald Tjalkens, Colorado State University Hans-Joachim Lehmler, University of Iowa, Iowa City, Iowa Jonathan Doorn, University of Iowa, Iowa City, Iowa

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of Iowa Superfund Research Center

Exposure to polychlorinated biphenyls (PCBs) correlates with developmental neurotoxicity, and non- dioxin-like, ortho-substituted PCBs are specifically thought to be the primary drivers of these adverse outcomes. The overall goal of our work is to determine the mechanism(s) of neurotoxicity of PCB 52, a non-dioxin-like, ortho-substituted PCB congener. Since neuronal health is greatly dependent on astroglia, a major glial cell type in the brain, we hypothesize that astroglia play an important role in mediating neurotoxicity of PCB 52 and its metabolites. C6 cells (rat glioma cell line) and primary glial cells (isolated from C57BL/6 mice) were exposed to varying concentrations of PCB 52 or two of its metabolites found in humans, namely, 4-OH-PCB 52 or 4-PCB 52-sulfate, for 24 hours. The C6 cells were similarly treated with three other PCB sulfates, namely, PCB 3-sulfate, PCB 11-sulfate, and PCB 25-sulfate. The MTT assay was employed to determine cell viability. Results indicate that, for C6 cells and primary glia, 4-hydroxy-PCB 52 was the most cytotoxic compound, followed by the parent compound, PCB 52. The PCB 52-sulfate was cytotoxic to C6 cells but not primary glia. This observation indicates that PCB 52, along with its metabolites, is toxic to astroglial cells with varying toxicity for sulfated PCB 52. However, when exposed to C6 cells, the three other PCB sulfates did not show cytotoxicity at any of the tested concentrations, indicating the importance of structure-activity relationship in PCBs mediated toxicity. Further studies are needed to elucidate the mechanism of toxicity of PCB 52 and its metabolites in astroglia and the role of astroglial cells in PCB mediated neurotoxicity. This work was supported by NIH P42 ES 013661 and R01 ES 129035.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 69 Poster Session I: Poster #I-43

Mapping Superfund Translational Stories using the NIEHS Translational Research Framework

Kristi Pettibone, National Institute of Environmental Health Sciences

Superfund Mandates: None

Superfund Center: Other: National Institute of Environmental Health Sciences

The National Institute of Environmental Health Sciences (NIEHS) published the Translational Research Framework (TRF) in 2018. The model re-envisions the basic science phase of translation to better capture the nuance of fundamental research. We also illustrate the framework using concentric circles representing each translational phase with nodes on each ring to describe activities that might be conducted as part of each translational phase. Finally, we define any research that either bridges nodes within a ring, or that crosses rings, as translational. The TRF reflects grantee input and builds on previous NIH models that focused on improving health via clinical strategies. To ensure broad applicability, the TRF includes environmental health prevention strategies, mechanistic and epidemiological research, engineering strategies, clinical research, community engagement and policy. As a portfolio analysis and evaluation tool, the TRF enables us to track and describe research as it moves through the translational research spectrum and to give “credit” to research that bridges nodes in the fundamental questions (basic science) ring. This poster will illustrate the TRF and provide an example of Superfund translational research project. We will also highlight the role that community partners can play in the translational research process. Finally, the poster will describe resources available to facilitate grantees use of the translational research framework to tell their own translational stories. These resources include case studies, translational research story templates, editable graphics, a criteria checklist, and a protocol for writing retrospective translational research stories. We will use the iposter features to add video presentations and link to these valuable resources.

Email: [email protected] Presenter Status: Evaluator

70 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-44

Per- and Polyfluoroalkyl Substances Inhibit the Phagocytic Respiratory Burst

Drake Phelps, North Carolina State University Anika Palekar, North Carolina State University Jacob Driggers, North Carolina State University Jeffrey A. Yoder, North Carolina State University

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: North Carolina State University Superfund Research Center

There are nearly 5,000 per- and polyfluoroalkyl substances (PFASs) registered globally that are used to produce non-stick cookware, food contact materials, hydro- and oleophobic textiles, fire-fighting foams, and more. PFASs are ubiquitous, persistent, and mobile in the environment, leading to commonplace exposure. It is estimated that more than 95% of all Americans have detectable serum levels of PFASs. Exposure to PFASs has been linked to cancer, thyroid disease, and immune system dysfunction. Previous research has focused largely on the effects of PFASs on the adaptive immune system, leaving a knowledge gap on what is known about the effects of these compounds on the innate immune system. To bridge this gap, we utilized an in vivo larval zebrafish model and an in vitro human neutrophil-like cell culture model to investigate innate immune function after exposure to environmentally relevant PFASs. First, range- finding studies were performed to determine concentrations at which there was no developmental toxicity or cytotoxicity. Using the established non-teratogenic or non-cytotoxic concentrations, the respiratory burst was measured as a functional readout of innate immune function. In phagocytes, the respiratory burst is the rapid induction of microbicidal reactive oxygen species in response to infection. Preliminary data revealed that three PFAS inhibited the respiratory burst in vitro. While experiments are ongoing, one of these PFASs also inhibited the respiratory burst in vivo. Potency, as measured by logistic regression, was similar between the two model systems, which may indicate evolutionary conservation between zebrafish and humans in their response to PFASs. Current studies are exploring this hypothesis, whether PFAS-induced immunotoxicity is differentially influenced by chronic or acute exposure, and whether exposure to PFASs confers susceptibility to infectious disease.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 71 Poster Session I: Poster #I-45

Stream Chemical and Biological Recovery Following Mine Water Treatment at the Central City Superfund Site

James F. Ranville, Colorado School of Mines Christopher Kotalik, Colorado State University William Clements, Colorado State University Evan Lloyd, Colorado School of Mines Joseph Meyer, Colorado School of Mines, and ALPs LLC

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: R01

Mineral extraction has resulted in widespread stream impairment due to habitat degradation and water quality impacts from acid mine drainage (AMD). The North Fork of Clear Creek (NFCC), Colorado, USA was impaired by AMD for over a century, with some stream segments devoid of aquatic life prior to remediation. Starting March 2017, a water treatment plant (NCCWTP) began diversion and treatment of AMD effluents entering NFCC. We examined both chemical and biological responses to the NCCWTP operations from 2017-2020. Elemental concentrations and mass loading were used to investigate water chemistry changes following treatment. The NCCWTP was successful at decreasing concentrations and loading of metals in NFCC. Total iron concentrations decreased by 95%, total copper by 84%, and total zinc by 64%. However, concentrations of aluminum, cadmium, copper, and zinc are all elevated above the Colorado aquatic life standards. To characterize biological recovery, we conducted biomonitoring of benthic communities using a Before-After Control-Impact (BACI) study design from 2011-2019. The NFCC stream community responded rapidly to improved water quality. Benthic algal biomass increased at impacted sites and macroinvertebrate surveys showed significant increases in abundance, taxa richness, and emerging adult aquatic insects. However, the dominant taxa colonizing downstream segments of NFCC differed considerably from those predicted based on previous field and mesocosm results. Although we expect continued biological recovery in NFCC, additional habitat restoration and remediation of residual sources of metals will be required to recover aquatic life to reference conditions.

Email: [email protected] Presenter Status: Researcher

72 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-46

Lack of Nrf2 Does Not Exacerbate the Detrimental Metabolic Outcomes Caused by In Utero PCB126 Exposure

Brittany B. Rice, University of Kentucky, Lexington, Kentucky Sara Ngo Tenlep, University of Kentucky, Lexington, Kentucky Obadah Tolaymat, University of Kentucky, Lexington, Kentucky Attaas Alvi, University of Kentucky, Lexington, Kentucky Fallon Slone, University of Kentucky, Lexington, Kentucky Claire Crosby, University of Kentucky, Lexington, Kentucky Stevi Howard, University of Kentucky, Lexington, Kentucky Hollie Swanson, University of Kentucky, Lexington, Kentucky Kevin Pearson, University of Kentucky, Lexington, Kentucky

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Kentucky Superfund Research Center

Environmental exposures to toxicants, such as polychlorinated biphenyls (PCBs), increase oxidative stress and disease susceptibility. Such exposures during pregnancy and/or nursing have been demonstrated to adversely affect offspring health outcomes. Nuclear factor erythroid-2-related factor 2 (Nrf2) regulates antioxidant response and is involved in the detoxification of coplanar PCBs, like PCB126. The purpose of this study was to investigate glucose tolerance and body composition in PCB-exposed offspring expressing or lacking Nrf2. We hypothesized that offspring lacking Nrf2 expression would be more susceptible to the long-term health detriments associated with perinatal PCB exposure. During gestation, Nrf2+/- and Nrf2- /- mice were exposed to vehicle or PCB126. Shortly after birth, litters were cross-fostered to unexposed dams to prevent PCB exposure during nursing. At two months of age, PCB exposure resulted in a significant reduction in body weight (p < 0.05) that was mostly made up by the decrease in lean body mass in both male and female offspring (p < 0.01). However, the lack of Nrf2 did not enhance the differences. Thirty minutes after an oral glucose challenge, both male and female PCB-exposed offspring exhibited significant impairments in glucose disposal irrespective of Nrf2 genotype (p < 0.05). These findings demonstrate that the lack of Nrf2 expression does not worsen the metabolic derangements caused by in utero PCB exposure and gives rise to a host of future directions independent of Nrf2 signaling.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 73 Poster Session I: Poster #I-47

Identifying Studies on the Key Characteristics of Carcinogens related to Chronic Inflammation and Immunosuppression

Gabrielle Rigutto, University of California Berkeley, Berkeley, CA Luoping Zhang, University of California Berkeley, Berkeley, CA Allen Louie, University of California Berkeley, Berkeley, CA Helen Guo, University of California Berkeley, Berkeley, CA Yun Zhao, University of California Berkeley, Berkeley, CA Sarah Dahlberg, University of California Berkeley, Berkeley, CA Michael Sholinbeck, University of California Berkeley, Berkeley, CA Martyn T. Smith, University of California Berkeley, Berkeley, CA

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of California‐Berkeley Superfund Research Center

Chronic inflammation and immunosuppression are two key characteristics (KCs) of carcinogens (Smith et al, 2016) as well as being two important forms of immunotoxicity caused by chemical exposures. Recently, we have systematically reviewed these two KCs of benzene, a known human leukemogen, and reported that benzene activates innate immunity by inducing pro‐inflammation and suppresses adaptive immunity via immunosuppression (Guo et al, 2020). PFASs (per‐ and polyfluoroalkyl substances) are manmade emerging environmental pollutants. Previously, the National Toxicology Program (NTP) evaluated immunotoxicity of PFOA (perfluorooctanoic acid) and PFOS (perfluorooctane sulfonate) – two commonly utilized PFASs (NTP, 2016). Here, we focused on creating the best set of search terms to evaluate the chronic inflammatory and immunosuppressive effects of potential carcinogens. We then compared our newly created search terms with the ones previously applied by IARC and NTP. A total of 1135 PFASs studies were identified using our new set of search terms and 311 of them were qualified for further detailed review (202 for chronic inflammation and 282 for immunosuppression). In comparison with IARC and NTP search terms, our new search strategy prevents exclusion of 84% and 78% of qualified PFASs studies for chronic inflammation from IARC and NTP, respectively. Similarly, we also increased number of qualified PFAS studies for immunosuppression by 65% and 85% when compared to IARC and NTP, respectively. Additionally, we applied the new search terms to benzene to compare with our previous results as well as for another human carcinogen, chromium. In conclusion, our new search strategy will greatly improve search results and to better evaluate all the studies qualified for these two important KCs.

Email: [email protected] Presenter Status: Trainee (pre‐/post‐doc)

74 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-48

University of New Mexico METALS SRP Center Responds to COVID-19 in Tribal Communities

Carolyn W. Roman, University of New Mexico, Alburquerque, New Mexico Chris Shuey, University of New Mexico, Alburquerque, New Mexico, Southwest Research and Info Center David Begay, University of New Mexico, Alburquerque, New Mexico Joe Hoover, Montana State University-Billings Daniel Beene, University of New Mexico, Alburquerque, New Mexico Mallery Quetawki, University of New Mexico, Alburquerque, New Mexico Romaisha Rahman, University of New Mexico, Alburquerque, New Mexico Johnnye Lewis, University of New Mexico, Alburquerque, New Mexico Matt Campen, University of New Mexico, Alburquerque, New Mexico Debra MacKenzie, University of New Mexico, Alburquerque, New Mexico Melissa Gonzales, University of New Mexico, Alburquerque, New Mexico

Superfund Mandate: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

The Navajo Nation had the highest per capita COVID-19 infection rate of any community in the U.S. in May 2020. This crisis intensified the decades-long environmental and health disparities faced by Navajo and other tribal communities. Lack of piped-in water limits access to safe water for drinking, hand-washing and cleaning for thousands of homes. At that time, METALS researchers were invited to join a federal- tribal task force charged with constructing 59 new clean water stations for communities with large numbers of COVID cases and homes without running water, and promote the program through navajosafewater.org and radio PSAs. METALS researchers also briefed the Navajo Nation Council on trends in COVID-19 cases rates and mortality, and advised health care professionals and tribal leaders of regional Pueblos on rapid testing opportunities. METALS Native Artist-in-Residence created widely- distributed materials promoting social distancing and mental health. Recognizing a PPE shortage on Navajo Nation and Zuni Pueblo hospitals due to delayed federal funding, METALS promoted an independent GoFundMe account to purchase masks, gowns, shields, sanitizer for tribal hospitals and clinics. Word of mouth about the program resulted in direct donations of PPE and basic living necessities to healthcare workers and families in remote areas. The pandemic has not only highlighted inequities in health care in tribal communities, but also the need to better understand links between environmental exposures and infectious disease. METALS investigators were highlighted in news reports on the link between metals exposures from hundreds of abandoned uranium mines to immune dysregulation that may increase susceptibility to COVID-19.

Email: [email protected] Presenter Status: RTC/CEC Administrator

NIEHS SRP 2020 Virtual Annual Meeting Program • 75 Poster Session I: Poster #I-49

Comparison of environmental metals and metal mixtures on oxidative stress and DNA damage in T-cells

Jodi Schilz, University of New Mexico, Alburquerque, New Mexico Erica J. Dashner-Titus, University of New Mexico, Alburquerque, New Mexico Karen Simmons, University of New Mexico, Alburquerque, New Mexico Tammi R. Duncan, University of New Mexico, Alburquerque, New Mexico Laurie G. Hudson, University of New Mexico, Alburquerque, New Mexico

Superfund Mandate: None

Superfund Center: University of New Mexico Superfund Research Center

Chronic mixed metal exposure from unremediated abandoned mines and waste piles pose unknown health risks to populations living in their proximity. Metals elevated in environmental or biological samples from people living near mine and milling sites include arsenic (As), cadmium (Cd), cobalt (Co), iron (Fe), manganese (Mn), uranium (U) and vanadium (V). There is a need to investigate the biological impacts of environmentally relevant mixed metal combinations. Metals are associated with immune dysregulation. Generation of oxidative stress and DNA damage are common cellular responses to metal exposure and may reflect shared mechanisms of toxicity. T-cells play a key role in immune system function and the effects of metal induced oxidative stress and DNA damage may alter lymphocyte function causing reactive oxygen species (ROS) mediated immunotoxicity. We compared the ability of relevant metals and metal combinations to alter viability, induce oxidative stress and DNA damage in Jurkat (human T-lymphocytic) cells. Cells were most sensitive to As toxicity (IC50 20 µM) compared to U and Co (IC50 > 100 µM). Cd plus As showed enhanced toxicity whereas Mn plus As was less toxic than each metal alone. Both As and Cd induced oxidative stress signaling as measured by increases in HMOX1 and NQO1 with enhanced HMOX1 expression when combined. In contrast, V induced expression of CAT but not HMOX1 potentially highlighting different mechanisms of ROS generation. Both As and V enhanced DNA damage as measured by pH2AX staining whereas Cd, Fe and U did not. Understanding how specific metals affect immune cell function may be useful for prioritizing metals and metal mixtures of concern for population-based studies.

Email: [email protected] Presenter Status: Researcher

76 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-50

Development of temperature responsive polymeric gels with intramolecular pi- pi interactions acting as a physical crosslinking site for water treatment applications

Rishabh Shah, University of Kentucky, Lexington, Kentucky Thomas Dziubla, University of Kentucky, Lexington, Kentucky Zach Hilt, University of Kentucky, Lexington, Kentucky

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Kentucky Superfund Research Center

Poly (N-isopropylacrylamide) (PNIPAAm) is a well-known temperature responsive polymer and has received extensive attention due to its sharp transition from hydrophilic to hydrophobic at its lower critical solution temperature (LCST). Development of temperature responsive polymers with PNIPAAm has been of great interest in recent years. In this research, novel non-covalent crosslinked polymers have been synthesized with biphenyl functionalities and are tested for their unique properties. PNIPAAm was polymerized with co-monomers containing a biphenyl moiety to create a unique thermoresponsive physically crosslinked system due to the presence of pi-pi interactions between the biphenyl moieties. The biphenyl monomers used were 2-phenylphenol monoacrylate (2PPMA) and 4-phenylphenol monoacrylate (4PPMA). These monomers were utilized to synthesize a set of polymers with biphenyl monomer (2PPMA/4PPMA) content from 2.5 to 7.5 mole percent and with initiator concentrations from 0.1 and 1.0 weight percent. The resulting polymers were characterized by various techniques, such as gel permeation chromatography (GPC), swelling studies and mechanical testing. These physically crosslinked polymeric systems with their unique properties have potential applications spanning environmental remediation/sensing, biomedicine, etc.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 77 Poster Session I: Poster #I-51

Nrf2-dependent upregulation of liver carbohydrate metabolism is a driver of arsenic-promoted diabetes

Aryatara Shakya, University of Arizona Pengfei Liu, University of Arizona Matthew Dodson, University of Arizona Donna D. Zhang, University of Arizona

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Arizona Superfund Research Center

The prevalence of type II diabetes continues to increase at an alarming rate. Among the major factors associated with enhanced diabetes risk, exposure to environmental toxicants continues to represent one of the least characterized drivers of diabetic outcomes. This is partly due to the broad scope of exposure contexts, including timing, dose, and route of exposure, as well as which tissues are most affected by a particular toxicant. However, despite these variables, a large body of epidemiological evidence indicates a clear link between chronic toxicant exposure and increased risk of developing diabetic phenotypes. In the case of arsenic, a toxic metalloid found ubiquitously throughout the environment, chronic exposure to >10 μg/L in drinking water significantly increases diabetes incidence, affecting an estimated 200 million people worldwide. Despite the clear epidemiological link between arsenic exposure and diabetes, very little is known regarding the molecular mechanisms that result in a pro-diabetic metabolic shift. A number of studies have shown that arsenic-induced autophagy dysfunction activates the Nrf2 signaling pathway in a p62-dependent manner. Furthermore, chronic activation of Nrf2 has been shown to have significant effects on adipogenesis, lipogenesis, insulin resistance, and obesity. In this study, wild type, but not Nrf2-/-, p62-/-, or Nrf2-/-;p62-/- mice exposed to arsenic for 20 weeks became insulin resistant and glucose intolerant. Analysis of significant transcriptomic and metabolomic changes revealed that arsenic induces a p62-Nrf2-dependent increase in fructose metabolism and gluconeogenesis. These findings indicate a critical role for the p62-Nrf2 axis in driving arsenic-induced changes to metabolism and highlight altered liver fructose metabolism as a possible novel therapeutic target to treat arsenic-promoted diabetes.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

78 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-52

The Aryl Hydrocarbon Receptor Suppresses Tumor Immunity Through Immune Checkpoint Regulation

David Sherr, Boston University, Boston, Massachusetts Jessica Kenison (Department of Pathology and Laboratory Medicine, Boston University) Zhongyan Wang (Department of Environmental Health, Boston University), Kangkang Yang (Department of Environmental Health, Boston University) Megan Snyder (Program in Genetics and Genomics, Boston University)

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Boston University Superfund Research Center

The aryl hydrocarbon receptor (AhR) has recently been identified as a driver of cancer progression and a master regulator of immune activation. AhR is both over-expressed and chronically active in many tumor types, and may serve as a mediator of tumor-specific immunosuppression through regulation of T cell subsets, macrophages and myeloid-derived suppressor cells (MDSCs). Here, we report that AhR expression in murine oral cancer cells (MOC1) is critical for the suppression of anti-tumor immunity and that deletion of the AhR (AhRKO) completely suppresses in vivo tumor growth in an immune system- dependent manner. Injection of AhRKO MOC1 cells decreases the expression of various markers of suppression on tumor-associated macrophages (TAMs), MDSCs, and CD4+ and CD8+ T cells in tumor draining lymph nodes, suppresses T cell exhaustion signaling pathways, and renders mice completely immune to a re-challenge with wild-type MOC1 cells. Furthermore, AhR controls the expression of suppressive Ido and PD-L1 in MOC1 tumor cells and plays a role in IFNγ-driven Ido and PD-L1 expression, suggesting that IFNγ enhances an immunosuppressive AhR-IDO-AhR ligand amplification loop in malignant cells. Taken together, these data suggest that the presence of AhR in malignant cells drives immunosuppression through the regulation of multiple immune checkpoints.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 79 Poster Session I: Poster #I-53

Relational Database for 2D Nanomaterial Environmental and Biological Interactions

Vidushi Shukla, Brown University, Providence, Rhode Island Aidan Lee Stone, Brown University, Providence, Rhode Island Robert H. Hurt, Brown University, Providence, Rhode Island

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Brown University Superfund Research Center

Two dimensional (2D) sheet-like nanomaterials are now the subject of a large international R&D effort targeted at technology development, but their safety assessment is challenged to keep pace with the large number and variety of new materials. In previous work, our lab put together a proposed framework for 2D material hazard screening based on degradation rates in the human lung and the chemical toxicity of the degradation products. To further develop this screening method, we have assembled an SQL-platform relational database that captures literature data on biodegradation rates, pathways, and products. The database is now being used for systematic review, incorporating meta- analyses and predictive Machine Learning algorithms to identify trends, assign hazard categories to a pilot set of new 2D materials, and explore predictive capabilities.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

80 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-54

Prenatal exposure to arsenic and lung function in children from the New Hampshire Birth Cohort Study

Antonio Signes-Pastor, Dartmouth College, Lebanon, NH Pablo Martinez-Camblor, Dartmouth College, Lebanon, NH Emily Baker, Dartmouth-Hitchcock Medical Center, Lebanon, NH Juliette Madan, Dartmouth College, Lebanon, NH Margaret F. Guill, Dartmouth-Hitchcock Medical Center, Lebanon, NH Margaret Karagas, Dartmouth College, Lebanon, NH

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Dartmouth College Superfund Research Center

Environmental exposure to toxicants during critical windows of fetal development may contribute to childhood and later-life respiratory health. Prenatal arsenic exposure from highly contaminated drinking water is associated with an increased risk of lung cancer along with multiple non-carcinogenic outcomes including respiratory diseases. However, few prospective studies have been conducted on prenatal arsenic exposure and children’s lung function particularly in the USA. We investigated the association between in utero arsenic exposure and lung capacity of children participating in the New Hampshire Birth Cohort Study. Urinary arsenic speciation including inorganic arsenic (inAs), monomethylarsonic acid (MMA), and dimethylarinic acid (DMA) and arsenobetaine was determined in maternal samples collected during pregnancy using HPLC coupled with ICP-MS. Spirometry was performed on a total of 358 children at a median age of 7.4 years. The FVC, FEV1, and FEF25-75 standardized z-scores were assessed in linear models as dependent variables. The log2-transformed sum of urinary arsenic (ΣAs) species: inAs, MMA and DMA (excluding arsenobetaine) corrected for specific gravity was examined after adjusting for maternal smoking status, children’s age, sex and height. We observed that urinary arsenic was inversely related to children’s FVC and FEV1 z-scores, but not to FEF25-75 z-scores. A doubling of ΣAs was related to a -0.8 decrease in FVC z-scores (95% confidence interval (CI): -0.14 to -0.01). A reduced FEV1 z-score was associated with ΣAs (ß = -0.10; 95% CI = -0.18 to -0.02). The inverse associations appeared stronger among participants with lower secondary methylation capacity (i.e., DMA/MMA). Our results suggest that in utero arsenic exposure at levels relevant to the general US population during the vulnerable period of lung formation may adversely effect lung function in childhood.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 81 Poster Session I: Poster #I-55

Associations between private well water and community water supply arsenic concentrations in the U.S.

Maya S. Spaur, Columbia University, New York, New York Melissa A. Lombard, US Geological Survey, Pembroke, Hew Hampshire Joseph Ayotte, US Geological Survey, Pembroke, Hew Hampshire Ana Navas-Acien, Columbia University, New York, New York David E. Harvey, US Public Health Service, Commissioned Corps, Rockville Maryland Anne E. Nigra, Columbia University, New York, New York

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Columbia University Superfund Research Center

Background: Many US aquifers contain elevated concentrations of geogenic arsenic. Groundwater is a major water source for many public water systems (PWSs) in the US. The US Environmental Protection Agency (EPA) regulates arsenic in community water systems (CWSs) by a Maximum Contaminant Level (MCL) of 10 µg/L, which does not apply to private wells. The association between private well and CWS arsenic concentrations has not yet been studied nationwide. Methods: We evaluated county-level 90th percentile probability of private well water arsenic exceeding 10 µg/L, and CWS average arsenic concentrations using linear regression. We stratified by compliance monitoring period (2006-2008 vs. 2009-2011), region, sociodemographic county cluster, and majority PWS source type for 2,231 counties. Results: Nationwide, county-level mean CWS arsenic concentrations were 8.4 (95% CI 7.7, 9.2) µg/L higher in 2006-2008 per unit (0 to 1) change in 90th percentile probability of private well arsenic exceeding 10 µg/L. This association decreased significantly between 2006-2008 and 2009-2011 nationwide and in the Southeast and Southwest. This relationship increased in the Central Midwest and Pacific Northwest, and in Semi Urban, Hispanic counties, though not significantly. Discussion: The probability of private well water arsenic exceeding 10 µg/L was associated with CWS arsenic concentrations nationwide. Regional differences in this association suggest that water sources used for private wells and CWSs differ regionally. The nationwide attenuation in 2009-2011 likely reflects Final Arsenic Rule implementation. Regional differences over time suggest that MCL interventions by State and Federal Drinking Water Programs were more pronounced in some regions. Strong associations in Rural, American Indian and Semi Urban, Hispanic counties suggest future research and regulatory support are needed to reduce water arsenic exposures in vulnerable subpopulations.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

82 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-56

Engineered heart tissues as a humanized platform to assess cardiotoxicity of Superfund chemicals

Yao-Hui Sun, University of California-Davis Padmini Sirish, University of California-Davis James Overton, University of California-Davis Deborah K. Lieu, University of California-Davis Christophe Morisseau, University of California-Davis Aldrin V. Gomes, University of California-Davis Bruce D. Hammock, University of California-Davis Xiao-Dong Zhang, University of California-Davis Nipavan Chiamvimonvat, University of California-Davis

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

At least 23% of cardiovascular diseases, the leading cause of morbidity and mortality in the United States, are related to environmental exposure of potential cardiotoxicants. Our investigative team have identified several compounds including ibuprofen (one of the most frequently prescribed nonsteroidal anti- inflammatory drugs) and dichlorophen (an ingredient in antimicrobial soaps and shampoos), as potent cardiotoxicants, as they significantly induced production of reactive oxygen species in cardiomyocytes. Further assessment of these chemicals on human cardiotoxicity and cardiac functionality, however, is hampered due to the lack of a suitable testing platform (traditionally in large animal models). Our goal was to develop humanized engineered heart tissues (EHT) as a physiologically relevant platform to evaluate these environmental chemicals in vitro. We show that such EHTs can be generated by reconstitution of decellularized extracellular matrix (ECM) from human left ventricular tissue, with human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes. The contractile cardiomyocytes, differentiated by stage manipulation of WNT signaling pathway, display ventricular phenotypes as revealed by immunostaining, gene expression, and electrophysiological characterization. These cardiomyocytes, when repopulated in left ventricle ECM, form muscle bundles that aligned with the collagen fiber scaffolds, mimicking the native ventricular myocardium. Additionally, left ventricle ECM- based EHTs with cardiomyocytes derived from hiPSCs, expressing genetically encoded calcium indicator GCaMP6f, enables the monitoring of spontaneous and dynamic calcium transients in response to pharmacological interventions. Thus, our humanized EHT model can be used to evaluate the cardiotoxicity of Superfund chemicals and drugs, and decipher the underlying molecular mechanisms. The novel insights are critical to the development of remediation for cardiotoxicity from Superfund chemicals.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 83 Poster Session I: Poster #I-57

Contribution of adenosine deaminases acting on RNA-1 on benzene metabolite t,t-muconaldehyde-induced endothelial activation

Breandon S. Taylor, University of Louisville, Louisville, Kentucky Nalinie S. Wickramasinghe, University of Louisville, Louisville, Kentucky Marina V. Malovichko, University of Louisville, Louisville, Kentucky Srinivas D. Sithu, University of Louisville, Louisville, Kentucky Shizuka Uchida, University of Louisville, Louisville, Kentucky Michael Nantz, University of Louisville, Louisville, Kentucky Sanjay Srivastava, University of Louisville, Louisville, Kentucky

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Volatile organic compounds (VOCs) are gaseous chemicals used as industrial solvents. They are also abundant in automobile exhaust, tobacco smoke, paints, wood preservatives, cleansers and disinfectants. High levels of VOCs are also present at various Superfund and hazardous waste sites. Our studies have shown that VOCs such as acrolein and benzene are associated with many adverse health outcomes, including cardiovascular disease (CVD). Our pre-clinical studies show that 20 weeks of exposure to benzene (10 ppm, 6h/day, 5days/week) augments atherosclerotic lesion formation (>25 %. P<0.05) in the aortic valves of male LDL receptor-KO mice as compared with corresponding air-exposed mice, without affecting plasma lipoproteins. Because endothelial cells play a critical role in leukocyte recruitment and initiation of atherosclerosis, we next examined the effect of benzene metabolite t,t- muconaldehyde on markers of endothelial activation. We observed that t,t-muconaldehyde (10 micro molar) increased the expression of adhesion molecules ICAM-1 and IL-8 on human aortic endothelial cells (HAEC; by 2-8-fold), adhesion of leukocytes to HAEC (1.8-fold), and transmigration of leukocytes through HAEC (1.4-fold). This was accompanied by an increased expression of ADAR1, a member of adenosine deaminases acting on RNA (ADAR) proteins which cataylze adenosine to inosine editing, in t,t- muconaldehyde-treated HAEC. siRNA-mediated knockdown of ADAR-1 augmented the t,t- muconaldehyde-induced ICAM-1 and IL-8 expression and leukocyte adhesion to HAEC. Together, these data suggest that ADAR-1 may be a critical regulator of benzene-induced endothelial activation.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

84 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-58

Internet of Things-based Edge Computing (IoTEC) for Rapid Response Vapor Intrusion Identification and Potential Mitigation

Chandra Mouli Tummala, Wayne State University, Detroit, Michigan Shariat Mobasser, Wayne State University Javad Roostaei, Wayne State University Yongli Zhang, Wayne State University Weisong Shi, Wayne State University Timothy M. Dittrich, Wayne State University Carol J. Miller, Wayne State University

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Unaffiliated

Many post-industrial urban centers in the U.S. contain numerous sites contaminated with volatile organic compounds (VOCs). These compounds can contaminate soils and shallow groundwater aquifers and, through vapor intrusion, negatively affect human health in exposed individuals. Various soil vapor extraction strategies can be used to remove soil vapors from below foundations before they enter a structure; however, these are generally costly and require significant time to deploy and run to completion. An affordable system that combines real-time sensing with air purification would be a helpful tool, especially for sensitive populations. Recent advances in sensor and communication technologies such as the Internet of Things (IoT) have allowed for the coupling of onsite contaminant monitoring with off-site data processing capabilities, permitting real-time data analysis and sharing of information for prompt decision-making. We present a novel environmental monitoring platform, IoT-based Edge Computing (IoTEC) that can be used for cost- and energy-efficient real-time monitoring. We integrate IoT sensing with edge computing (a distributed computing paradigm where computation is performed on distributed device nodes instead of in a centralized data processing warehouse) for energy- and cost- saving. Our results indicate that IoTEC could reduce >10% of data latency and save 40-60% of energy use and >50% of cost for one sensor. For a rapid response mitigation technique, we are developing modified air purifiers equipped with engineered sorbents optimized to remove VOCs from structures that have an immediate need based on high VOC concentration measurements from a sensor network. We evaluated the sorption capacity of traditional air purifier materials (activated carbon and zeolite clay) and a novel organosilica; preliminary indications suggest the filter operation period to be >3 months.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 85 Poster Session I: Poster #I-59

MIT DNA and Protein Models Adapt to Covid-times: Teaching Hands-On Exposure Biology via Online Instruction

Kathleen Vandiver, Massachusetts Institute of Technology, Cambridge, Massachusetts Amanda G. Mayer, Massachusetts Institute of Technology, Cambridge, Massachusetts

Superfund Mandates: None

Superfund Center: Massachusetts Institute of Technology Superfund Research Center

Public health professionals working with immigrants and people of color in environmental justice communities often overlook genetic susceptibility as a risk factor in health outcomes caused by environmental exposures. World populations differ in the frequency of gene variants for cytochrome P 450 proteins (CYP), the proteins responsible for metabolizing toxic substances. To help teach about genetic variability in CYP proteins, a new lesson was created for the MIT Edgerton Center DNA and Protein Model Sets in the “Genes and Health” booklet. Additionally, as a result of the Covid-19 pandemic, the MIT hands-on teaching method with the MIT Edgerton Center molecular models was adapted for online instruction. During the summer of 2020, instructors utilized the Zoom platform with Boston Public School high schoolers, as well as public health majors at Georgia State University in Atlanta, Georgia. We also led a teacher professional development workshop, at the Environmental Health Sciences Summer Institute for Teachers, in partnership with Texas A & M University. Teacher workshops were facilitated by online resources: 1) For teachers looking to deliver classroom lessons, we have videos that include best practices, pedagogical explanations, and examples showing students “learning by doing”. 2) For teachers looking to deliver hands-on lessons from a distance, we have resources for virtual instruction. The MIT Edgerton Center Youtube playlist includes 2-3 minute “How to” clips. Students received packages on loan with kits, booklets, and a web cam on a stand to provide a reliable way for teachers to view student work. Booklets and videos can support independent study. Classroom Sets of DNA and Protein are available for purchase. https://edgerton.mit.edu/DNA-proteins-sets.

Email: [email protected] Presenter Status: Other Community Engagement Core Lead

86 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-60

Investigating impact of ecological growth strategies on plasmid transfer and function to model and promote a precision bioremediation framework

Paige Varner, Duke University, Durham, North Carolina Claudia K. Gunsch, Duke University, Durham, North Carolina

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Duke University Superfund Research Center

Bioremediation is a sustainable treatment approach for the removal of harmful contaminants from the environment. This technology tends to be more cost effective and more environmentally friendly than alternative remediation methods such as dredging and excavation because it is much less intrusive. Genetic bioaugmentation is a targeted bioremediation method that uses horizontal gene transfer to introduce relevant genes to indigenous microorganisms already adapted to a polluted environment to increase in situ contaminant biodegradative capacity. In this project, we aim to apply this approach to the treatment of polycyclic aromatic hydrocarbons (PAHs), a class of ubiquitous and recalcitrant hydrocarbon contaminants that are carcinogenic to humans and toxic to the environment. As some bacterial genes for PAH degradation lie on plasmids, it is feasible to treat this class of contaminants via genetic bioaugmentation. However, one significant challenge to this approach is the lack of understanding surrounding the ecological growth strategies of indigenous microorganisms and the resulting impact on in situ plasmid transfer and functionality. For instance, many known degraders of the more recalcitrant high-molecular-weight PAHs grow slowly. Thus, the objective of this project is to identify the impact of bacterial growth strategy on plasmid transfer and functionality for the purpose of targeted bioremediation. To investigate this, we use two bacterial hosts harboring mobile PAH-degrading plasmids (NAH7 and pNL1) and six recipient strains of varying growth strategies. We describe here the methods developed to track plasmid transfer via conjugation and preliminary results surrounding conjugation of NAH7 and pNL1.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 87 Poster Session I: Poster #I-61

Resolution of eicosanoid/cytokine storm prevents aflatoxin-induced hepatocellular cancer progression

Weicang Wang, University of California-Davis, Davis, California Anna Fishbeina, Harvard Medical School, Boston, Massachusetts Jianjun Deng, Harvard Medical School, Boston, Massachusetts Jun Yang, University of California-Davis, Davis, California Victoria M Hallisey, Harvard Medical School, Boston, Massachusetts Haixia Yang, Harvard Medical School, Boston, Massachusetts Sanne M.L. Verheul, Harvard Medical School, Boston, Massachusetts Sung Hee Hwang, University of California-Davis, Davis California Allison Gartung, Harvard Medical School, Boston, Massachusetts Yuxin Wang, University of California-Davis, Davis, California Diane R. Bielenberge, Harvard Medical School, Boston, Massachusetts Sui Huang, Institute for Systems Biology Mark W. Kierang, Harvard Medical School, Boston, Massachusetts Bruce D. Hammock, University of California-Davis, Davis, California Dipak Panigrahy, Harvard Medical School, Boston, Massachusetts

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

Aflatoxins, a group of foodborne carcinogens, represent one of the critical causes of hepatocellular carcinoma (HCC), the second leading cause of cancer mortality worldwide. Aflatoxins induce lineage switching of hepatocytes to initiate HCC, yet the non-genetic mechanisms remain poorly characterized. Here, we demonstrate that aflatoxin B1 induces hepatocyte apoptotic cell death (“debris”), which triggers eicosanoid and cytokine storm in macrophage and stimulates tumor growth in both subcutaneous and orthotopic HCC tumor models in mice. Aflatoxin B1-generated debris upregulates cyclooxygenase-2 (COX- 2), soluble epoxide hydrolase (sEH), and endoplasmic reticulum (ER) stress response genes including BiP, CHOP, and PDI in macrophages. Thus, selective cytokine or eicosanoid blockade is unlikely to prevent aflatoxin B1-induced cancer. Pharmacological abrogation of both the COX-2 and sEH pathways by dual inhibitor PTUPB prevents the debris-stimulated eicosanoid and cytokine storm, reduces ER stress gene expression, and stimulates macrophage phagocytosis of debris. In murine HCC tumor models, treatment of PTUPB suppresses debris-stimulated subcutaneous and orthotopic tumor growth and improves survival rate in mice. Together, our study demonstrates the inflammation resolution via dual COX-2/sEH inhibition as a novel approach to prevent aflatoxin B1-induced HCC.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

88 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-62

Systems level analysis of non-genomic effects of exposure to Polycyclic Aromatic Hydrocarbons

Forest White, Massachusetts Institute of Technology, Cambridge, Massachusetts Ishwar Kohale, Massachusetts Institute of Technology, Cambridge, Massachusetts

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Massachusetts Institute of Technology Superfund Research Center

Polycyclic aromatic hydrocarbons (PAHs) are a class of chemicals that have long been known to have a range of toxic effects. Selected PAHs, including Benzo[a]pyrene (BaP), have been identified as Group 1 carcinogens. BaP is processed to a reactive epoxide that can bind to DNA and cause damage and mutations; at high exposures to BaP, DNA damage is evident from a variety of assays. Although BaP and other PAHs have been investigated in depth over the past several decades, the non-genomic effects of exposure to these contaminants are still poorly understood. Specifically, the pathways underlying cellular response to these contaminants, including pathophysiological response as well as cellular adaptation, are still poorly understood. Perhaps more importantly, the non-genomic cellular response to chronic, low- dose, exposures to BaP and mixtures of environmentally relevant PAHs have yet to be fully characterized. Here we have exposed cells derived from liver and lung with 2-200 ppb of Benzo[a]pyrene (BaP), and quantified protein phosphorylation at 1, 3, and 6 days after treatment. We also exposed cells to different dilutions of mixtures of PAHs derived from several environmentally relevant sources. For each exposure condition, cell phenotypic effects, viability and cell migration, were measured, along with signaling networks associated with cellular survival, migration, proliferation, and DNA damage response. We observed changes in signaling networks regulating cell proliferation, migration and DNA damage response (DDR) in a time and dose dependent manner. Overall, we observed quantitative rewiring of different pathways suggesting that cells are adapting to exposure over time even at low doses.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 89 Poster Session I: Poster #I-63

Zebrafish bioactivity screening identifies variable toxicity of the most commonly occurring chemicals from global wristband personal passive sampling studies

Lindsay B. Wilson, Oregon State University, Corvallis, OR Briana N. Rivera, Oregon State University, Corvallis, OR Kim A. Anderson, Oregon State University, Corvallis, OR Susan C. Tilton, Oregon State University, Corvallis, OR Robyn L. Tanguay, Oregon State University, Corvallis, OR

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Oregon State University Superfund Research Center

The development of small, personal passive sampling devices like silicone wristband samplers has revolutionized our ability to assess interindividual variability in personal chemical exposures. This technology gives researchers the ability to compare chemical exposure profiles and identify common exposures within and across populations. A recently published study utilizing wristband samplers identified a group of 14 chemicals detected in extracts from over 50% of samplers deployed across three continents. Of these common chemicals, most are present in consumer products, representing a large percentage of personal exposure across demographics. The high frequency of exposure to these chemicals elicits questions of their potential adverse human health effects. Additionally, the possibility for exposure to mixtures of these compounds is likely, due to their co- occurrence, so the potential for mixtures to induce differential bioactivity warrants further investigation. The zebrafish is an established model for human health-related research and chemical screening. Using early life-stage zebrafish, we assessed the bioactivity of the 14 compounds individually and in combination, assessing 22 morphological and 2 sensitive behavioral endpoints. Of the individual compounds, 10 exhibited bioactivity, but with variable potency based on benchmark concentration which induced a 50% effect (BMC50) relative to control groups for each chemical. Additionally, an equimolar mixture of all 14 compounds induced bioactivity at lower concentrations than any individual compound alone, indicating potential chemical interactions within the mixture. Future effects-directed analysis with permutations of sub-mixtures of the 14 chemicals will help to understand the observed mixture effects. Finally, this study offers exciting possibilities to broadly evaluate and communicate the hazards posed by the most relevant combinations of human chemical exposures by coupling passive chemical sampling with high-throughput toxicity screening in zebrafish.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

90 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-64

The role of environmental metal exposure and extracellular vesicles in the pathophysiology of Alzheimer’s disease

Charlotte Wirth, Harvard TH Chan School of Public Health Quan Lu, Harvard TH Chan School of Public Health

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Harvard Superfund Research Center

Advances in health sciences has resulted in an increase in the global population suffering from age related illnesses such as Alzheimer’s disease (AD). While genetic mutations are partially responsible for AD, metals are AD mediators through gene-environment interactions. AD is characterized by the progressive loss of structure and function of the nervous system. AD pathogenesis involves the aggregation of specific misfolded proteins in the brain: amyloid-beta and tau. The manner by which neurodegenerative proteins spread to or begin to form aggregates in a previously healthy cell, is not understood. Microglia, the innate immune cell of the brain, have been shown to aggregate around misfolded proteins such as amyloid-beta and tau in the brain and may be involved in their spread. We hypothesize that microglia are involved in AD pathogenesis through their extracellular vesicle (EV). EVs are bilipid bound particles which shuttle biologically active protein, lipid, and nucleic acid cargo from donor cells to recipient cells. We also hypothesize that metal exposure alters microglial EV cargo. Previous proteomic analysis of healthy human immortalized microglial EVs revealed the presence of numerous unfolded protein response proteins involved in AD. We will derive microglia from AD and healthy donor human induced pluripotent stem cells (iPSCs) then expose these cultures to Pb, As, Mn, Cd, Cr, or Se. EV cargo will be characterize via proteomics and RNA-seq. Pathway analysis and validation of EV cargo of interest in microglia-neuron co-cultures will provide evidence of microglia EV cargo’s role in AD pathogenesis. The characterization of microglia EVs cargo after metal exposure will elucidate how each metal affects cargo shed by healthy and AD microglia, narrowing targets for future treatment or therapies.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 91 Poster Session I: Poster #I-65

Development of A Rapid and Quantitative Toxicity Assessment Tool for Drinking Water Quality Evaluation and Identification of Drinking Water Toxicity Drivers

Jingyi Wu, Cornell University Josh M. Allen, University of South Carolina Susan D. Richardson, University of South Carolina Michael J. Plewa, University of Illinois at Urbana-Champaign Stuart W. Krasner, Metropolitan Water District of Southern California Akram N. Alshawabkeh, Northeastern University April Z. Gu, Cornell University

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of North Carolina at Chapel Hill Superfund Research Center

In Puerto Rico Testsite for Exploring Contamination Threats (PROTECT) center, one of the objectives is to develop technologies to eliminate the whole toxicity of drinking water at the end of pipeline. Currently, there is a lack of feasible and effective methodology to rapidly and quantitatively evaluate drinking water toxicity. Developing a relevant toxicity assessment tool for drinking water quality monitoring and toxicity driver identification is essential for efficacy evaluation of drinking water treatment systems. In this study, a recently established quantitative in vitro effect-based and omics-oriented yeast toxicity assay and a human cell RT-qPCR assay were employed for comparative toxicity evaluation of 30 water samples collected from raw water and distribution systems at seven drinking water treatment plants in the United States. Twelve key proteins with high predictability for phenotypic endpoints of genotoxicity and oxidative stress, and five key genes previously shown to be indicative of DNA damage, oxidative stress, chemical stress and apoptosis were selected as molecular toxicity biomarkers in the yeast and human cell assays, respectively. Targeted screening of 68 disinfection byproducts (DBPs) was performed to unravel the potential relationship between the DBP composition and the toxicity of disinfected drinking water. Concentration-dependent toxicity fingerprints and distinct pathway-level perturbations were observed among samples collected in raw water and distribution systems from different geographical locations and treatment processes. Correlation analysis between molecular toxicity responses and DBP concentrations were conducted to reveal the high-priority contaminants in the drinking water. This study proposes an innovative biomarker-based approach for the cost-effective and feasible evaluation of mixture toxicities of drinking water, and provides a promising direction of drinking water toxicity monitoring and risk assessment.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

92 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session I: Poster #I-66

A Preliminary Analysis of the Association between Arsenic Consumption and Diabetes Phenotypes in a Small Cohort of Diversity Outbred Mice

James G. Xenakis, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Christelle Douillet, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Timothy A. Bell, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Pablo Hock, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Joseph Farrington, University of North Carolina at Chapel Hil, Chapel Hill, North Carolina l Tianyi Liu, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Caroline E.Y. Murphy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Avani Saraswatula, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Ginger D. Shaw, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Qing Shi, Abhishek Venkatratnam, Fei Zou, Rebecca C. Fry, Miroslav Stylblo, Pardo Manuel de Villena,University of North Carolina at Chapel Hill, Chapel Hill, North Carolina

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

Inorganic arsenic (iAs) is a ubiquitous environmental contaminant poisoning hundreds of millions worldwide. Chronic exposure to iAs is associated with the risk of type 2 diabetes (T2D), and the ability to metabolize iAs into monomethyl-As (MMAs) and dimethyl-As (DMAs) critically drives this risk. Understanding of the genetic underpinning of the susceptibility to iAs-associated T2D remains incomplete. This research addresses the Superfund mandate to advance techniques for the assessment and evaluation of the effect of iAs on human health by examining a range of metabolic phenotypes in Diversity Outbred mice that were exposed to 100ppb of iAs for 28 weeks. At evenly-spaced timepoints, body weight and composition, blood glucose and plasma insulin levels were measured and used to calculate FBG, FPI, 15’PGC and 15’PI. We used these to derive the homeostatic model assessment measures of insulin resistance and beta-cell function. We fit linear mixed models to each longitudinal endpoint, adjusting for time, weight and body composition, the effective dose of arsenic each mouse consumed, and relevant interactions. Weight and/or its interaction with time had the most consistently significant effect in these models; an increase in body weight had a deleterious association with most phenotypes except 15’PGC. Although the marginal effect of effective arsenic dose was not statistically significant in most models, it exhibited compelling interactions with other variables. For example, the effective dose of arsenic was marginally associated with increased FBG for mice with high body fat. We also observed an association between total arsenic consumption and the distribution of arsenic species found in the liver at 28 weeks; mice that consumed the most water had low proportions of iAs and high proportions of DMAs.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 93 Poster Session I: Poster #I-67

In-situ degradation of Ibuprofen and DEET in a flow-through reactor

Yuwei Zhao, Northeastern University Stephanie Sarrouf, Northeastern University Akram N. Alshawabkeh, Northeastern University

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Northeastern University Superfund Research Center

The electrochemical systems can be used to induce in situ Fenton reaction: hydrogen peroxide (H2O2) forms through the catalyzed reaction between electro-generated oxygen and hydrogen and further activates to hydroxyl radicals (OH∙) via reaction with ferrous iron (Fe(II)). The goal of our studies focused on the continuous electrogeneration of hydroxyl radicals in the flow-through reactor to remove contaminants. A PTFE coated graphite felt with a PDMS dampproof coating was used for the production of H2O2. The iron anode or FeSO4 was chosen as the ferrous iron source for the Fenton reaction. The performance of each on removing ibuprofen and DEET was evaluated in this research. The result showed that 40 ppm of Fe2+ in the electrolyte has the best Ibuprofen removal under flow-conditions. The iron anode has better performance when it is located at the top of the reactor, compared to at the bottom of the middle.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

94 • NIEHS SRP 2020 Virtual Annual Meeting Program

Poster Session II: December 14, 2020 | 4:00 – 5:00 p.m.

Last Name First Name Poster Session ID Ashrap Pahriya II-1 Bako Christian II-2 Blomberg Annelise II-3 Blount-Pacheco Jessica II-4 Bolt Alicia II-5 Bozack Anne II-6 Brearley Adrian II-7 Brown Phil II-8 Connolly Morgan II-9 Cordova Alexandra II-10 Croisant Sharon II-11 Dekker Harrison II-12 Dodds James II-13 NIEHS Downey Autum II-14 Elkin Elana II-15 SRP Fawkes Leanne II-16 Foley Caredwen II-17 2020 Furlong Clement II-18 Furlow David II-19 Annual Meeting Ghetu Christine II-20 Guo Chuqi II-21 Halder Sujoy II-22 Hammond Charles II-23 Harmon Ashlyn II-24 Harris Sean II-25 Holloway Zade II-26 Jurgelewicz Amanda II-27 Kelly Jamie II-28 Klingelhutz Aloysius II-29 Leuthner Tess II-30 Li Fu Jun II-31 Lingappan Krithika II-32

96 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: December 14, 2020 | 4:00 – 5:00 p.m.

Last Name First Name Poster Session ID Liu Jamie II-33 Maier Monica II-34 Malecha Matthew II-35 Marsillach Judit II-36 Medina Sebastian II-37 Mills Rollie II-38 Moran Ian II-39 Moreno Ramirez Denise II-40 Nault Rance II-41 Obeidy Chelsea II-42 O’Leary Brendan II-43 Orr Asuka II-44 NIEHS Owiti Norah II-45 Powell Christian II-46 SRP Reif David II-47 Saktrakulkla Panithi II-48 2020 Sallach J. Brett II-49 Saxena Roheeni II-50 Annual Meeting Shaikh Nabil II-51 Shih Yu-Hsuan II-52 Sirish Padmini II-53 Sobel Marisa II-54 Sule Rasheed II-55 Suter Melissa II-56 Venkatratnam Abhishek II-57 Vickers Riley II-58 Wade Terry II-59 Whitt Aaron II-60 Wood Lindsey II-61 Yang Xiaojing II-62 Yang Yifei II-63 Young Jamie II-64

NIEHS SRP 2020 Virtual Annual Meeting Program • 97 Poster Session II: Poster #II-1

Psychosocial Status Modifies the Effect of Maternal Blood Metal and Metalloid Concentrations on Birth Outcomes

Pahriya Ashrap, University of Michigan, Ann Arbor, MI Amira Aker, University of Toronto Scarborough, Ontario, Canada Deborah J. Watkins, University of Michigan Ann Arbor, MI Bhramar Mukherjee, University of Michigan Ann Arbor, MI Zaira Rosario-Pabón, University of Puerto Rico, San Juan, Puerto Rico Carmen M. Vélez-Vega, University of Puerto Rico, San Juan, Puerto Rico Akram Alshawabkeh, Northeastern University, Boston, MA José F. Cordero, University of Georgia, Athens, GA John D. Meeker, University of Michigan, Ann Arbor, MI

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substance

Superfund Center: Northeastern University Superfund Research Center

Background: Metal exposure and psychosocial stress in pregnancy have each been associated with adverse birth outcomes, including preterm birth and low birth weight, but no study has examined the potential interaction between them. We examined the modifying effect of psychosocial stress on the association between metals and birth outcomes among pregnant women in the Puerto Rico Testsite for Exploring Contamination Threats (PROTECT) birth cohort study. Methods: In our analysis of 682 women in the PROTECT study, we measured 16 essential and non-essential metals in blood samples at two time points. We administered questionnaires to collect information on depression, perceived stress, social support, and life experience during pregnancy. Using K-means clustering, we categorized women into one of two groups: “good” and “poor” psychosocial status. We then examined associations between blood metal concentrations (geometric average) and adverse birth outcomes using linear (gestational age, birth weight z-score) and logistic (preterm birth, small for gestation [SGA], large for gestation [LGA]) regressions among the two clusters of women, adjusting for maternal age, maternal education, pre-pregnancy body mass index (BMI), and second-hand smoke exposure. Results: Blood manganese (Mn) was associated with an increased odds ratio (OR) of overall preterm birth (OR/interquartile range [IQR] = 2.76) and spontaneous preterm birth (OR/IQR: 3.68) only among women with “poor” psychosocial status. The association between copper (Cu) and SGA were also statistically significant only among women having “poor” psychosocial status (OR/IQR: 2.81). We also observed associations between nickel (Ni) and preterm birth and SGA that were modified by psychosocial status during pregnancy. Conclusions: Our findings provide evidence that prenatal psychosocial stress may modify vulnerability to metal exposure.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

98 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-2

Biodegradation of PCB Congeners in Wastewater Lagoon Sediment

Christian M. Bako, University of Iowa, Iowa City, Iowa Timothy Mattes, University of Iowa, Iowa City, Iowa Rachel Marek, University of Iowa, Iowa City, Iowa Keri Hornbuckle, University of Iowa, Iowa City, Iowa Jerald Schnoor, University of Iowa, Iowa City, Iowa

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Iowa Superfund Research Center

Experiments were conducted to measure biodegradation of polychlorinated biphenyl (PCB) congeners contained in mixture Aroclor 1248 and congeners present in wastewater lagoon sediment contaminated decades earlier at Altavista, Virginia. A well-characterized strain of aerobic PCB-degrading bacteria, Paraburkholderia xenovorans LB400 was incubated in laboratory bioreactors with PCB-contaminated sediment collected at the site. The experiments evaluated strain LB400’s ability to degrade PCBs in absence of sediment and in PCB-contaminated sediment slurry. In absence of sediment, LB400 transformed 76% of Aroclor 1248 within seven days, spanning all homolog groups present in the mixture. In sediment slurry, only mono-, di-, and trichlorinated PCB congeners were transformed. Furthermore, when bioreactors were amended with a phytogenic biosurfactant, saponin, biodegradation of di- and trichlorobiphenyls was significantly enhanced, relative to the unamended live cell treatment. These results show that LB400 is capable of rapidly biodegrading most PCB congeners when they are freely dissolved in liquid but cannot degrade PCB congeners having four or more chlorine substituents in sediment slurry. Finally, using GC/MS-MS triple quadrupole spectrometry, this work distinguishes between physical (sorption to cells) and biological removal mechanisms, and illuminates the process by which microorganisms with LB400-type congener specificity can selectively transform lower-chlorinated congeners over time.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 99 Poster Session II: Poster #II-3

Early-life associations between per- and polyfluoroalkyl substances and serum lipids in a longitudinal birth cohort

Annelise Blomberg, Harvard T. H. Chan School of Public Health, Boston MA Yu-Hsuan Shih, Harvard T. H. Chan School of Public Health, Boston MA Carmen Messerlian, Harvard T. H. Chan School of Public Health, Boston MA Pal Weihe, University of Southern Denmark, Odense DK Flemming Nielsen, University of Southern Denmark, Odense DK Philippe Grandjean, Harvard T. H. Chan School of Public Health, and University of Southern Denmark

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Harvard School of Public Health Superfund Research Center

Background: A growing body of literature has found that exposures to poly- and perfluoroalkyl substances (PFASs) may be associated with metabolic outcomes, including detrimental changes in lipid concentrations. However, few studies have evaluated potential longitudinal associations between PFAS and lipids. Objectives: We examined associations between PFAS and lipid concentrations at birth and several points in childhood. Methods: We studied 490 maternal-child pairs from a Faroese prospective birth cohort. Concentrations of five PFAS were measured in cord serum at birth and in serum collected at ages 18 months, five years and nine years. Total cholesterol (TC), low-density lipoprotein cholesterol (LDL- C), high-density lipoprotein cholesterol (HDL-C) and triglyceride (TG) concentrations were similarly measured in serum at birth, 18 months and nine years. We used multiple regression models to estimate associations between serum PFAS concentrations and lipids, and evaluated differences by sex using interaction models. Results: Childhood PFAS concentrations were associated with lipid concentrations measured at nine years. The cross-sectional associations between PFASs and lipids at nine years were strongest, with increases in PFDA, PFNA and PFOS associated with increases in TC, HDL-C and LDL-C. We also found statistically significant differences in estimated PFAS effects by sex. For girls, an increase in five- year and nine-year PFAS concentrations was associated with increased nine-year TC and LDL-C concentrations, while an increase in nine-year PFAS concentrations was associated with an increase in nine-year HDL-C concentrations in boys. Conclusions: Results from our study suggest that mid-childhood may be a vulnerable time for PFAS exposures. Understanding the impacts of PFAS on childhood lipids is critical, as a detrimental lipid profile in childhood is an important risk factor for atherosclerosis and CVD in adulthood.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

100 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-4

Novel method for assessing biological effects of volatile organic compound exposure in zebrafish

Jessica Blount-Pacheco, Wayne State University, Detroit, Michigan Tracie R.Baker; Wayne State University, Detroit, Michigan Camille Akemann, Wayne State University, Detroit, Michigan Jeremy Shields, Wayne State University, Detroit, Michigan Zoha Siddiqua, Wayne State University, Detroit, Michigan Shawn McElmurry, Wayne State University, Detroit, Michigan David Pitts, Wayne State University, Detroit, Michigan

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Unaffiliated

Anthropogenic volatile organic compounds (VOCs) and semi-VOCs have emerged as high priority environmental contaminants due to ubiquitous urban exposure and therefore play a pivotal role in several environmental and health related issues. Urban areas report increased levels of contamination due to industrial exhaust, fuel refineries, vehicle exhaust, especially in groundwater. Residents in urban areas are disproportionately exposed to potential VOC contamination as a result of indoor vapor intrusion, tap water, ingestion of contaminated fish, and ambient outdoor exposure, and at-risk communities are often chronically exposed to complex mixtures of VOCs. A wide variety of adverse health risks have been associated with VOC exposure, which involve respiratory, cardiovascular, renal, hepatic, endocrine, immune, and nervous systems, and include cancer and abnormal birth outcomes. Despite large-scale human and animal exposures to VOCs, the associated health risks remain unclear, especially during pregnancy and early developmental stages. To address this, we investigated the health impacts of exposures to VOCs in larval zebrafish. From 6 to 120 hours post-fertilization (hpf), developing zebrafish were exposed to a range of benzene and BTEX, and we evaluated dose-dependent accumulation, behavioral response, mortality, hatching rate, phenotypic changes, and transcriptomic analysis. To date, the biggest challenge in characterizing VOC toxicity has been technological limitations that create unsafe and unreliable exposure conditions. In collaborative efforts, we have designed an innovative sealed exposure system for safely containing a VOC and analyzing behavioral changes in aquatic organisms. This novel exposure system utilizes the zebrafish animal model for exposing and evaluating VOC toxicity. Using this model, we are able to elucidate health concerns that could potentially translate to humans. Future studies will explore the synergistic effects of VOC mixtures.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 101 Poster Session II: Poster #II-5

Inhalation exposure to tungsten particulates induces early markers of pulmonary injury following acute exposure

Alicia M. Bolt, University of New Mexico, Alburquerque, New Mexico Kara Burton, University of New Mexico Charlotte McVeigh, University of New Mexico Ed Barr, University of New Mexico Guy Herbert, University of New Mexico Russell Hunter, University of New Mexico Sebastian Medina, New Mexico Highlands University Selita Lucas, University of New Mexico Abdul-Mehdi S. Ali, University of New Mexico Matthew J. Campen, University of New Mexico

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

In the Southwestern US, elevated toxic metals are detected in air and water samples, due in part to increased mining activity. Tungsten is found elevated in environmental samples and individuals living near mine-waste sites. Inhalation to tungsten-alloys has been associated with increased incidence of lung pathologies, including interstitial lung disease and lung cancer. However, very little is known about the molecular mechanisms underlying these pathologies and the individual contribution of tungsten. We examined if inhalation exposure to tungsten metal particles causes acute damage to the lung environment that could contribute to these long-term pathologies. 24 female Balb/c mice were exposed to Filtered Air, 0.50 mg/m3 (low concentration), or 1.5 mg/m3 (high concentration) tungsten particles (< 2 µm) in a whole-body inhalation chamber for 4 hrs. Tissue samples were collected Day-1 and 7 post-exposure. A significant accumulation of tungsten was measured in the lungs and bone post-exposure. Multiple markers of acute lung injury were detected in the lung niche, including a significant infiltration of macrophages and neutrophils and elevated levels of the proinflammatory cytokine IL-1β, however no significant changes in peripheral blood counts were observed. Importantly, these inflammatory changes were associated with an increased percentage of activated fibroblasts (α-SMA+) in the lung in the high exposure group. These changes did not appear to be driven by sustained reactive oxygen species generation. These findings demonstrate that acute exposure to tungsten particles, at relevant human exposure levels, result in changes to the lung microenvironment that may promote the onset or exacerbation of these lung pathologies. Current work is focused on exploring these observations using in vitro multi-cell lung models and chronic exposure animal models. P20GM130422-01A1, IRG-17-178-22, INBRE P20GM103451.

Email: [email protected] Presenter Status: Researcher

102 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-6

Arsenic exposure at different life-stages and DNA methylation: Meta-analysis of differential methylation and differential variability in PBMCs and buccal cells

Anne K. Bozack, University of California-Berkeley Phillippe Boileau, University of California, Berkley Linging Wei, University of California, Berkley Martyn T. Smith, University of California, Berkley Craig M. Steinmaus, University of California, Berkley Ana Navas-Acien, Columbia University Mary V. Gamble, Columbia University Andres Cardenas, University of California, Berkley

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California-Berkeley Superfund Research Center

Background/Aims: Changes in DNA methylation (DNAm), may be involved in arsenic toxicity. Epigenome- wide association studies (EWAS) of arsenic exposure have been limited by sample sizes and methodological differences. In the current study involving four DNAm datasets, we use a harmonized data processing pipeline and combine results in a meta-analysis across studies in two continents. Methods: DNAm was measured among adults in Chile with and without prenatal and early-life exposure in PBMCs and buccal cells (N=40, 850K array) and among men in Bangladesh with high and low current exposure in PBMCs (N=32, 850K array; N=48, 450K array). Linear models were used to identify differentially methylated positions (DMPs) and differentially variable positions (DVPs). Probes common across EWAS were meta-analyzed using METAL, and differentially methylated and variable regions (DMRs and DVRs, respectively) were identified using Comb-P. Analyses were adjusted for age, smoking, cell type, and sex in the Chile cohort. Results: In a meta-analysis restricted to PBMCs, we identified one DMP and 23 DVPs associated with arsenic exposure; including buccal cells, we identified 3 DMPs, 19 DVPs (FDR<0.05). Using combined p-values, we identified 12 DMRs and 15 DVRs in PBMC samples, and 21 DMRs and 24 DVRs in PBMC and buccal cell samples. Arsenic-associated CpGs were annotated to genes involved in RNA binding (RBPMS), histone modifications (HDAC4), and p53 regulation (UBE3A and FAM129A) and insulin regulation (IGFALS). Conclusions: Using a two-step process of harmonized data processing and meta- analysis, we leverage four DNAm datasets from two continents of individuals exposed prenatally and during adulthood to identify DMPs and DVPs associated with arsenic exposure. Further research is needed to understand the role of arsenic-associated DMPs and DVPs in disease development.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 103 Poster Session II: Poster #II-7

Uranium-bearing nanoparticles derived from abandoned uranium mines: A previously unrecognized potential exposure risk from inhalation of windblown dust

Adrian J. Brearley, University of New Mexico, Alburquerque, New Mexico Chris Shuey, University of New Mexico, Alburquerque, New Mexico Kyle Swimmer, Pueblo of Laguna Jose Cerrato, University of New Mexico, Alburquerque, New Mexico Matthew Campen, University of New Mexico, Alburquerque, New Mexico Johnnye Lewis, University of New Mexico, Alburquerque, New Mexico

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

The numerous Abandoned Uranium Mines (AUM) in the Southwest US represent a potential source of airborne U-bearing dust that may disproportionally affect Native American communities. However, the potential health risks of such exposures are incompletely understood, because of the lack of knowledge of the carriers of U and other toxic metals. To address this issue, we have been investigating both potential source materials of U-bearing particulates from AUM and a legacy sample of airborne dust obtained from a site adjacent to the reclaim Jackpile Mine, Laguna Pueblo, NM. In particular, we have focused on the respirable component (<PM2.5) of these materials to determine the physical and chemical characteristics of the mineralogical carriers of U, etc. using transmission electron microscopy. Our studies of ore and mine waste from the Jackpile mine and Claim 28, Blue Gap/Tachee (BGT), Navajo Nation reveal that U occurs consistently as nanoparticles of U-bearing minerals in PM2.5 and rarely occurs as larger micron-sized particles. In the Jackpile and BGT samples, nanoparticles of the uranyl vanadates, carnotite, and tyuyamunite occur as clusters of nanoparticles up to 1 micron in size that show evidence of disaggregation. Each cluster consists of individual nanoparticles 20-50 nm in size that can easily undergo aeolian transport. In a legacy sample of windblown dust from Laguna Pueblo, U is dominantly present in submicron grains. These observations are of special concern, because bulk geochemical analyses of dust samples with low concentrations of U may indicate minimal exposure risk. However, if the U is dominantly present in nanoparticulate form, the potential exposure risks may be enhanced due to the ability of nanoparticles to penetrate into cell structures and disrupt their functions.

Email: [email protected] Presenter Status: Researcher

104 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-8

Chemical Class Consciousness: Scientific and Social Action on the PFAS Chemical Class

Phil Brown, Northeastern University, Boston Massachusetts Laurel Schaider, University of Rhode Island and Silent Spring Institute, South Kingstown, Rhode Island Rainer Lohmann, University of Rhode Island, South Kingstown, Rhode Island Carolyn Mattingly, North Carolina State University, Raleigh, North Carolina Detlef Knappe, North Carolina State University, Raleigh, North Carolina

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Northeastern University Superfund Research Center

In the last few years there has been a dramatic increase in research, policy, and social action on per- and polyfluoroalkyl substances (PFAS), even though PFAS were barely known to the public a few years ago, despite studies by academics go back to around 2000. Although industry, regulatory, and academic scientists have studied PFAS exposure and toxicity for decades, and contaminated communities have documented health concerns as a result of high exposure, PFAS use remains ubiquitous. Regulatory action has been limited, though an increasing number of states have set MCLs far below EPA’s health advisory. Congress’s last two omnibus defense bill have provided research funding and promoted restrictions on AFFF use. Dozens of bills are now in Congress and state legislatures. Northeast states, including MA and RI, supported by SRP scientists, are advancing regulating several PFAS as a group in drinking water. High- profile publications with SRP authors are also making the class-wise case for PFAS. A growing activist movement has had successes in research funding, litigation, regulation, policy, and business removal and reduction of PFAS. Scientists and activists recognize the need for class-based approaches, rather than a chemical-at-a-time approach, and also posit the “essential uses” concept. What we learn about PFAS may apply to other persistent organics in terms of the class-wide approach, the multi-sector response, and the essential use approach. STEEP, PROTECT and NC State are all working with states in their setting of MCLs for PFAS and for regulation concerning food packaging, are central organizers of the annual PFAS conferences in 2017, 2019 and upcoming 2021, and use the class concept in their research, research translation, and community engagement.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 105 Poster Session II: Poster #II-9

Engineering the Triazine Degradation Pathway for Improved Herbicide Bioremediation

Morgan Connolly, University of California, Davis Justin Siegel, University of California, Davis

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California, Davis Superfund Research Center

Application of bioremediation at contaminated sites is often limited by slow degradation rates and accumulation of intermediate products. Here we use genetic and enzyme engineering to increase flux through the triazine degradation pathway, which can break down various triazine herbicides, which are common contaminants of ground and surface water. The pathway has been introduced into E. coli and characterized to determine which step of degradation is most limiting flux through the pathway during degradation of atrazine. Engineering efforts are underway to increase expression and activity of the bottleneck enzyme. We hope to produce strains with increased flux through the triazine degradation pathway to improve their potential for use in bioremediation.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

106 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-10

Developing passive dosing methods to enable in vitro testing of environmental samples and complex mixtures

Alexandra Cordova, Texas A&M University, College Station, Texas Alina Roman-Hubers, Texas A&M University, College Station, Texas Alan Valdiviezo, Texas A&M University, College Station, Texas Thomas McDonald, Texas A&M University, College Station, Texas Weihsueh A. Chiu, Texas A&M University, College Station, Texas Ivan Rusyn, Texas A&M University, College Station, Texas

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

Environmental contaminants often occur in the form of complex mixtures of variable composition, earning them a reputation as “difficult-to-test” chemicals, both in terms of in vitro toxicity testing and analytical experimentation. In vitro toxicity testing of complex environmental samples and mixtures is traditionally achieved by extraction with a biocompatible solvent such as dimethyl sulfoxide (DMSO). However, extraction restricts sample chemical complexity and may not fully represent the bioavailable fraction of concern. OECD Test Guidelines detail methods for “difficult-to-test” substances as applied to ecotoxicology; still, these methods are not directly applicable to high-throughput in vitro tests that are performed in small-volume, multi-well plates. This study’s objective was to develop passive dosing methods that are suitable for testing mixtures and environmental samples in small-volumes. We hypothesized that passive dosing in vitro can be achieved using silicone micro-O-rings and molecular sieves, as well as a cell culture-compatible media accommodated fraction. To investigate this hypothesis, we tested a mixture of 25 polycyclic aromatic hydrocarbons (PAHs) with micro-O-rings and molecular sieves. The test mixture was also dissolved in cell culture media with and without proteins for media- accommodated fractions. Conventional DMSO extraction was used as a reference. In vitro experiments in EAHY926 cell line evaluated the recovery and bioavailable fraction of mixture components by gas chromatography-tandem mass spectrometry. We found consistent loading and recovery of the PAHs with micro-O-rings and the media-accommodated fraction. Dilution-dependent in vitro effects of the mixture were replicated with passive dosing and found to be an improvement over the DMSO extraction method. Our study therefore indicates the feasibility of passive dosing as an alternative, broad-scope method to gain insight into the bioavailable fractions of complex environmental mixtures. This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 107 Poster Session II: Poster #II-11

A Persistent Problem: PAH Superfund Site Pollutants and Community Engagement Programs to Reduce Risk of Exposure

Sharon Croisant, Baylor College of Medicine, Houston, TX Chantele Singleton, The University of Texas Medical Branch John Prochaska, The University of Texas Medical Branch Lance Hallberg, The University of Texas Medical Branch Cornelis Elferink, The University of Texas Medical Branch Jackie Young-Medcalf, Texas Health and Environment Alliance Hilton Kelley, Community In-Power and Development Association, Inc. Bhagavatula Moorthy, Baylor College of Medicine

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Baylor College of Medicine Superfund Research Center

Houston is the fourth-largest city in the US and its port one of the world’s busiest. Its Ship Channel is lined by port facilities, industry, refineries, and processing facilities established over the past 100+ years. Legacy pollutants, lack of zoning, and more than a dozen federal Superfund sites have long impacted surrounding communities. The Ship Channel includes two major Superfund sites, with many more highly toxic sites that are the source of significant levels of polycyclic aromatic hydrocarbons (PAHs), mercury, dioxins, furans, PCBs, etc. Within this heavily populated environment, PAHs constitute a persistent human health concern. The focus of the Baylor-Rice SRP on PAHs addresses a largely unmet need for identifying at-risk populations and developing innovative technologies for identifying and ameliorating risk. The overall goal of the Community Engagement Core (CEC) is to facilitate meaningful bidirectional communication and interactions among SRP stakeholders. Our mission is to work with residents and local leaders to facilitate prevention and intervention strategies to reduce community-level exposures to PAHs. We work with communities surrounding the Ship Channel and Superfund sites, many of which are impacted by Environmental Justice issues, heavy industrialization, poverty, and health effects of Superfund-related exposures. The CEC utilizes a variety of engagement mechanisms including initiatives such as an EJ “Encuentro”, which affords communications opportunities to share ideas, concerns, and exposure and risk assessment research data, outputs, and implications. They also enable SRP investigators to directly engage impacted communities. The CEC supports community projects such as education for pregnant women, community gardening activities including instruction to prevent exposure to harmful chemicals, and fish consumption education programs, which offer safe and healthy alternatives to contaminated local fish.

Email: [email protected] Presenter Status: Director, Community Engagement

108 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-12

A Workflow for the FAIRification of Environmental PFAS Data

Harrison Dekker, University of Rhode Island, South Kingston, Rhode Island Jitka Becanova, University of Rhode Island, South Kingston, Rhode Island Yana Hrytsenko, University of Rhode Island, South Kingston, Rhode Island

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Rhode Island Superfund Research Center

The URI STEEP SRP center is actively working to both enhance data interoperability between projects within the center and to promote reuse externally across scientific domains. One particular data interoperability challenge we are addressing is the effective documentation, management, and sharing of the data STEEP collects on PFAS concentrations in water and passive samplers generated by LC/MS method. To address this challenge, STEEP is attempting to adhere closely to the FAIR guidelines. Given the heavy emphasis FAIR places on metadata and machine actionability, new workflows are under development to ensure that these added requirements can be met efficiently and in accordance with existing standards. This poster will outline these new workflows with particular emphasis on the unique challenges inherent in the “ FAIRification” of data derived from mass spectrometry analyses.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 109 Poster Session II: Poster #II-13

Utilizing a Credentialing Strategy with IMS-MS for Untargeted Characterization of Xenobiotic Transformation Products

James N. Dodds, North Carolina State University, Raleigh, North Carolina Lingjue Wang, Washington University in St Louis, St Louis, Missouri Gary Patti, Washington University in St Louis, St. Louis, Missouri Erin Baker, North Carolina State University, Raleigh, North Carolina

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

The detection and quantitation of xenobiotics is a primary directive for characterizing environmental contamination, assessing mitigation efforts and evaluating remediation strategies. While current analytical strategies predominantly focus on the assessment of primary xenobiotics, evaluating the occurrence and fate of transformation byproducts in the environment is also of utmost importance. Thus, novel bioanalytical strategies are required to identify transformation byproducts of xenobiotics and assess their relative threat. In this work, we showcase how coupling an innovative analytical strategy known as credentialing with liquid chromatography and ion mobility spectrometry-mass spectrometry (IMS-MS) allows the annotation of new small molecules. Credentialing principally utilizes both 12C and 13C precursors and the observed 1:1 ratios enable the confident identification of molecules present and changing in various sample types. Our results highlight distinct advantages gained experimentally from the IMS separations and illustrate unique data acquisition and interpretation methods particularly suited for the characterization and assessment of xenobiotic transformation byproducts in various chemical classes (e.g. PFAS, PAHs, PCBs, etc.). This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

110 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-14

Assessing the variation in organic carbon and nitrogen concentrations within clastic cave sediments

Autum Downey, West Virginia University, Morgantown, West Virginia Dorothy Vesper, West Virginia University, Morgantown, West Virginia

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Northeastern University Superfund Research Center

The fate and transport of contaminants is closely tied to the presence and type of natural organic matter present in the system. This is particularly true in karst systems where voids and caves in the rock can store and transport large quantities of organic-rich sediments. To evaluate the importance of natural organic matter in a cave system, sediments from a cave in the northern karst region of Puerto Rico were analyzed for total organic carbon (TOC) and total nitrogen (TN). Sediments were collected both above and below water level that is typically present in the cave. Sediments collected below the water table (water saturated) contained higher concentrations and a greater range of TOC and TN values then did the samples collected from above the water table. The difference between the saturated and unsaturated sediments suggests that microbial reactions are more likely to occur in the saturated zone and generate TOC in-situ; the chemolithotrophic generation of TOC in caves has been demonstrated for water but not previously for the sediment phase. Horizontal redox banding, commonly present in the saturated sediments, supports the interpretation that significant chemical and microbial-driven reactions occur within this zone. These chemical processes have important implications for the transport and storage of organic contaminants as natural organic matter is a strong sorbate. These saturated sediments in the aquifer contribute to the retention of contaminants via sorption. In regions with frequent tropical storms and/or hurricanes, previously deposited sediment and associated organic compounds could remobilize and re-enter the groundwater, potentially entering public or private water supplies.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 111 Poster Session II: Poster #II-15

Sex-specific transcriptional responses to maternal trichloroethylene and/or N- acetyl cysteine exposure in Wistar rat placental tissue

Elana R. Elkin, University of Michigan, Ann Arbor, Michigan Anthony L. Su, University of Michigan, Ann Arbor, Michigan John F. Dou, University of Michigan, Ann Arbor, Michigan Justin A. Colacino, University of Michigan, Ann Arbor, Michigan Dave Bridges, University of Michigan, Ann Arbor, Michigan Rita Loch-Caruso, University of Michigan, Ann Arbor, Michigan Kelly M. Bakulski, University of Michigan, Ann Arbor, Michigan

Superfund Mandates: None

Superfund Center: Northeastern University

Numerous Superfund sites are contaminated with the volatile organic chemical trichloroethylene (TCE). Exposure to TCE in pregnant women is associated with reduced infant birth weight. Our previous study reported that TCE treatment in pregnant rats decreased fetal weight and elevated placental oxidative stress biomarkers, suggesting placental injury is a mechanism of TCE-induced adverse birth outcomes. Here, we investigate attenuation of TCE effects by testing TCE-co-treatment with the antioxidant N- acetylcysteine (NAC). Pregnant Wistar rats were exposed to 480 mg TCE/kg/day, 200 mg NAC/kg/day, or co-exposure with both chemicals via ingestion on gestation days 6-16 (tissue collection day). We evaluated TCE- and/or NAC-induced changes to gene expression and gene set enrichment in male and female rat placentae. In female placentae, exposure to TCE differentially expressed 129 genes and TCE+NAC differentially expressed 125 genes, both compared with controls (FDR<0.05 + Fold-change>2). In contrast, in male placentae, exposure to TCE differentially expressed 9 genes and TCE+NAC differentially expressed 35 genes, compared with controls (FDR<0.05 + Fold-change>2). NAC alone did not significantly alter gene expression in either sex. TCE treatment downregulated oxidative phosphorylation pathways in females, immune response pathways in males, and upregulated endoplasmic reticulum stress pathways in both sexes (FDR<0.01). In both sexes, TCE-treatment increased expression of genes regulated by the ATF4 and ATF6 transcription factors, indicating activation of the unfolded protein response. This study identified novel genes and pathways involved in mechanisms of TCE-induced placental injury and showed that NAC co-treatment did not attenuate TCE exposure effects.

Email: [email protected] Presenter Status: Trainee (pre-/post)

112 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-16

Exposure to Lead-Contaminated Drinking Water in Bryan and College Station Public Parks

Leanne Fawkes, Texas A&M University, College Station, Texas Anjali Katare, Texas A&M University, College Station, Texas Garett Sansom, Texas A&M University, College Station, Texas

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Texas A&M University Superfund Research Center

Safe drinking water is celebrated as a public health achievement and is a top priority for the Environmental Protection Agency. Yet today, lead (Pb) contaminated drinking water has the potential to be a public health crisis in the United States. Despite efforts to provide safe drinking water, update water infrastructure, and ensure strict drinking water regulations, there are incidents of unsafe lead levels and reports of associated adverse health effects. While there has been increased attention paid to the quality of drinking water within individuals’ homes, little research has examined the presence and concentration of lead in water from drinking fountain sources located in public parks. This study sampled drinking water from every accessible public park in the Bryan–College Station (BCS) metropolitan area (N=25). With a lower detection level of 2.0 ug/l this study discovered a mean lead concentration of 1.28 ug/l across all sites and a maximum of 8 ug/l. Further, neighborhoods below the median income for BCS were twice as likely to have detectable lead levels in their water and had 1.5 times the mean concentration. This research supports previous studies identifying a disparate burden to lead exposure among low socioeconomic populations within the United States. By examining the water quality in publicly accessible parks and drinking fountains, our study provides public health professionals with important information about how to improve the built environment to safeguard safe potable water. This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 113 Poster Session II: Poster #II-17

Towards an ontology for contaminant measurement in fish tissue samples

Caredwen Foley, Boston University, Boston Massachusetts Wendy Heiger-Bernays, Boston University, Boston Massachusetts Celia Chen, Dartmouth College

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Boston University Superfund Research Center

A thorough understanding of the magnitude and extent of the co-occurrence of contaminants in environmental media (sediment, water, fish) at a given Superfund site depends on the ability to unify multiple sources of data. In addition, efficacy of remediation at contaminated sites requires comparison of media pre-and post-remediation. We are combining existing datasets that contain heavy metals and polychlorinated biphenyls (PCBs) in fish tissue to meet our objective of querying the resultant dataset to answer questions about health risks of fish consumption. Disparities across datasets include variation in descriptions for metal and organic contaminants (e.g., PCB homologues vs. individual congeners; total mercury vs. methylmercury) and characteristics of the sample (fish species; geography; season and temporal variation; dry vs. wet weight; whole fish vs. filet; tissue/organ type; lipid fraction; etc.). The disparate nature of the datasets also derives from the source of the data: academic or contract laboratory. The lack of a uniform ontology poses significant challenges to researchers in centralizing data sources to assess contamination and remediation particularly as it relates to the co-occurrence of multiple contaminants. The difficulties this creates in examining PCB contamination—with only 209 congeners and ten homologues—suggests that this will become an even more pressing concern with respect to characterizing fish contaminated with per- and polyfluoroalkyl substances (PFAS), a class containing many thousands of individual contaminants. We are developing an approach for characterizing multiple contaminant measurements in fish tissue samples in order to enhance the compatibility of monitoring data from a variety of sources. Our ontology will employ Findable, Accessible, Interoperable, and Reusable (FAIR) principles and will provide researchers with a vital framework for making comparisons across datasets.

Email: [email protected] Presenter Status: Researcher

114 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-18

Development of a Multiplexed Generic Portable Surface Plasmon Resonance Detection System for Environmental Applications

Clement Furlong, University of Washington Scott Soelberg, University of Washington James Birch, Monterey Bay Aquarium Research Institute William Ussler, Monterey Bay Aquarium Research Institute Christina Preston, Monterey Bay Aquarium Research Institute Louise Lingerfelt, NOAA/National Ocean Service/CSS Greg Doucett, NOAA/National Ocean Service Michael Unger, Virginia Institute of Marine Science, William & Mary, Gloucester Point, VA

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Washington Superfund Research Center

Surface plasmon resonance systems provide an excellent means for following intermolecular interactions in real time. It is straightforward to follow interactions of proteins with other proteins, proteins with nucleic acids, nucleic acids with nucleic acids or nucleic acid analogs like morpholinos, enzymes with effector molecules and to detect other specific molecules, microbes or viruses using target-specific antibodies or receptors to name some of the possibilities. SPR instrumentation initially was large and expensive. This presentation describes the development of a portable SPR system based on a “chip” invented by Jose Melendez when he was at Texas Instruments. This robust chip made it possible to build a portable SPR sensor system around the chip. Further refinement of the portable system made it possible to incorporate the sensors into the long range underwater autonomous vehicle (LRUAV) developed at MBARI. Two examples of monitoring analytes with the portable system are presented. A monoclonal antibody specific for 3-5 membered rings was used to measure phenanthrene as an indicator of oil spills or seepage. Both polyclonal anti-okadaic acid antibodies and a very robust, high affinity anti-okadaic acid antibody were used to measure the shellfish toxin okadaic acid, known as a diarrhetic shellfish toxin (also a carcinogen). The recent version of the portable system fitted into the MBARI LRUAV is currently being used to monitor microcystin in Lake Erie and has also been used to monitor domoic acid, the amnesiac shellfish toxin, in the Pacific Ocean. Supported in part by the following grants: NIEHS-SRP grant RO1ES02424; NIEHS P50 ES012762–NSF OCE-0434087; NSF OCE 0961702; NOAA NA20OAR4320271 and Washington State Sea Grant.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 115 Poster Session II: Poster #II-19

Cells to systems screening of thyroid hormone disrupting chemicals: unexpected interactions with environmental and synthetic RXR ligands

David Furlow, University of California-Davis, Davis, California Brenda J. Mengeling, University of California-Davis, Davis, California

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

Precisely timed and regulated thyroid hormone (TH) levels are essential for vertebrate development and metabolism. To detect environmental chemicals that disrupt TH action, we developed an integrated luciferase reporter cell line in rat pituitary GH3 cells that is TH responsive through activation of endogenous thyroid hormone receptors (TRs). This cell line was used for quantitative high throughput screening of libraries of environmentally relevant compounds by the Tox21 consortium. We validated the reporter gene activity using endogenous gene responses, and identified many of the non-TH related positive hits as RXR agonists, a “silent” heterodimer partner for TRs. These data showed RXR agonists can affect TR signaling in pituitary cells. We extended these results to our sensitive and specific in vivo model for TH action, Xenopus laevis metamorphosis. RXR agonists like organotins and the pharmaceutical bexarotene strongly potentiate (and RXR antagonists inhibit) TH induced metamorphic programs, at the morphological, cellular (e.g. apoptosis or proliferation), and transgenic reporter gene levels. RNA-Seq data in the tadpole brain, a common target of TH in humans and frogs, revealed specific gene sets particularly affected by RXR ligands. The remarkable overlap between tributyltin and bexarotene regulated transcriptomes provides strong evidence that they have a common molecular target. We created germline mutations in all RXR family members in Xenopus tropicalis, and the TR beta gene in Xenopus laevis, allowing us to further investigate TR-RXR heterodimer function across tissues and developmental timeframes, and in response to known and suspected TR and RXR ligands. Our studies revealed an unanticipated degree of TR and RXR ligand interactions in vitro and in vivo, highlighting a surprising role of RXRs as avenues for TH endocrine disruption.

Email: [email protected] Presenter Status: Researcher

116 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-20

Predicting contaminant sources in environmental samples using GC-MS/MS alkylated PAH analysis and a comprehensive list of forensic ratios

Christine Ghetu, Oregon State University, Corvallis, OR Richard P. Scott, Oregon State University, Corvallis, OR Glenn Wilson, Oregon State University, Corvallis, OR Rachel Liu-May, Oregon State University, Corvallis, OR Kim A. Anderson, Oregon State University, Corvallis, OR

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Oregon State University Superfund Research Center

Parent and alkylated polycyclic aromatic hydrocarbons (PAHs) are present in a number of different sources in varying proportions depending on the source material and weathering. This range of PAH sources can make it difficult to determine the origin of exposure(s). Parent PAHs have been studied, and some are identified as toxic to humans and other organisms, but less is known about alkylated PAH toxicity. Ratios of alkylated and parent PAHs have been applied as a forensic tool to distinguish between different sources. However, few studies have examined PAH ratios comprehensively as indicators for sourcing beyond a single study area or matrix type. In this study we introduce a modified, expanded analytical method based on ASTM D7363-13a which we adapted for a gas-chromatography triple-quadrupole mass spectrometry instrument. The modifications increase selectivity and sensitivity compared to the ASTM method. By modifying and expanding the ASTM D7363-13a method to a gas chromatography triple- quadrupole mass spectrometry instrument, we obtained a calibration range that was two orders of magnitude larger and contained an additional six alkylation series. This method was then applied to 22 independent forensic ratios. We evaluated the method and the forensic ratios with five certified standard reference materials and three sets of known environmental samples. This analytical method and thirteen PAH ratios were found to accurately predict sources of PAHs.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 117 Poster Session II: Poster #II-21

Applying Phytosampling as a Passive Particulate Matter Sampling Method for Environmentally Persistent Free Radicals (EPFRs) and Polycyclic Aromatic Hydrocarbons (PAHs) Analysis

Chuqi Guo, North Carolina State University, Louisiana State University Farhana Hasan, Louisiana State University Dean Lay, Louisiana State University Slawo M. Lomnicki, Louisiana State University

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: North Carolina State University Superfund Research Center

Ambient particulate matter (PM) exposure has been attributed to cardiovascular health effects and mortality. Insufficient evidence exists about the health effects of PM components, although studies suggest that the presence of compounds such as environmentally persistent free radicals (EPFRs) and polycyclic aromatic hydrocarbons (PAHs) increase the health risk of PM exposure. This study evaluates a passive sampling method relying on the particle entrapment by the leaves to analyze PM-associated EPFRs and PAHs. Phytosampling allows for easy particle recovery, the collection under natural conditions, and the potential for increased spatial coverage due to its low cost. Comparison of the speciation and concentration of EPFRs and PAHs on PM collected by phytosampling and by active air sampling shows that sometimes pollutant speciation is different on PM samples collected by two methods. Phytosamples were collected near an intersection on the LSU campus during the years 2015-2018. The active air sampler was installed on the roof of a seven-story campus building and continuously ran for two weeks during each collection event. Vehicle emissions provide the main PM source. We detected a larger contribution of oxygen-centered EPFRs, different decay behavior, and more consistent PAH distribution among different PM sizes from phytosampling compared to the active sampler. These results indicate that the isolation of samples from the ambient air during active sampling and exposure to high volume airflow may alter the chemical composition of samples, while phytosamples could provide more details on the original speciation and concentration. However, phytosampling cannot evaluate PM concentration given the integrative collection design, so it should serve as a supplementary tool to active PM collection methods in future field studies.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

118 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-22

Fabrication and characterization of a sensor array to detect volatile organic compounds in environmental air

Sujoy Halder, University of Louisville, Louisville, Kentucky Prasadanie K. Adhihetty, University of Louisville, Louisville, Kentucky Zhenzhen Xie, University of Louisville, Louisville, Kentucky Michael H. Nantz, University of Louisville, Louisville, Kentucky Xiao-An Fu, University of Louisville, Louisville, Kentucky

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Louisville Superfund Research Center

This work reports on the fabrication and characterization of a chemiresistor sensor array chip that comprises four different sensor materials and performs simultaneous measurements of target hazardous volatile organic compounds (VOCs) in environmental air. The sensor array is manufactured on a silicon wafer using microelectromechanical systems (MEMS) technology. The detection is based on measuring the changes in resistance across a film comprised of gold nanoparticles (AuNPs) protected by a thiolate ligand monolayer. When gaseous molecules are introduced into a testing chamber containing the sensor array, the thin films of AuNPs differentially absorb the vapors and undergo swelling, which change sensor resistances accordingly. AuNPs can support surface modifications using a variety of thiols with tailored functional motifs. Moreover, thiol functionalized AuNP sensors provide advantages such as operation at ambient temperature and short recovery time. The sensor array chip is 1 cm x 1 cm, where Pt/Cr metals are deposited by sputtering. The fabrication is completed by a lift-off process. Four different thiols including dodecanethiol, 4-methoxy-toluenethiol, 4-mercaptobenzoic acid, and 11-mercaptoundecanoic acid were used to functionalize AuNPs and then drop-casted on interdigitated electrodes to form the sensor array. In this study, we will report the sensor array responses to different VOCs, such as formaldehyde, acetone, ethanol, pentane, benzene, toluene, ethylbenzene, and xylene. A mixture of different VOCs is introduced to test AuNPs sensor responses and to characterize the selectivity of each sensor for comparison to the response obtained with a single analyte. The responses are rapid, reversible, and linear for all of the tested target analytes. The sensor array exhibits good selectivity, has a potential for real-time measurement of toxic VOCs at ambient conditions.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 119 Poster Session II: Poster #II-23

White Rot Fungal Bioreactors for the Treatment of Xenobiotics

Charles R. Hammond, University of California-Davis, Davis, CA Frank J. Loge, University of California-Davis, Davis, CA

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

There are many xenobiotic organic contaminants found in wastewater (both municipal and industrial) that are not effectively removed with conventional treatment technologies. For example, conventional activated sludge, a common treatment process, is not effective at removing most pharmaceuticals. Wastewater treatment plant effluent is responsible for discharging a variety of chemicals, like endocrine disruptors, pesticides, and surfactants, to the environment, where they can accumulate, causing harm to aquatic life and affecting drinking water supplies. Alternative or additional treatment processes are needed to prevent these contaminants from reaching the environment, and White rot fungi (WRF) have the potential to fill this need. They are effective degraders of many recalcitrant organic contaminants due to the low-specificity oxidative enzymes they produce to degrade lignin, a plant-produced polymer of phenylpropanoid alcohols. The key enzymes, lignin peroxidase, manganese peroxidase, and laccase, can degrade many different contaminants. Bioreactors and treatment processes designed to optimize the activity and stability of WRF and their ligninolytic enzymes could provide a low-cost, effective treatment technology for broad classes of contaminants. The main challenges to be addressed in this research are maintaining long-term fungal dominance under non-sterile conditions, and immobilizing ligninolytic enzymes to both prevent washout and increase degradation of target contaminants.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

120 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-24

Vascular Endothelial Injury after Inhalation of Combustion-Derived Environmentally Persistent Free Radicals is Mediated via AhR Activation

Ashlyn C. Harmon, Louisiana State University, Baton Rouge, Louisiana Ankit Aryal, Louisiana State University, Baton Rouge, Louisiana Lavrent Khachatryan, Louisiana State University, Baton Rouge, Louisiana Merilyn Jennings, Louisiana State University, Baton Rouge, Louisiana Zakia Perveen, Louisiana State University, Baton Rouge, Louisiana Alexandra Noël, Louisiana State University, Baton Rouge, Louisiana Arthur L. Penn, Louisiana State University, Baton Rouge, Louisiana Tammy R. Dugas, Louisiana State University, Baton Rouge, Louisiana

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Louisiana State University Superfund Research Center

Environmentally persistent free radicals (EPFRs) are formed due to the incomplete combustion of organic pollutants. Poor mixing of gases and formation of oxygen-poor pockets in cool zones of the flame leads to chemisorption of organics with transition metals and forms stabilized complexes containing redox-active particles. To investigate the cardiopulmonary effects of EPFR exposure, combustion-derived particles were generated in a controlled laboratory setting. Two aliquots of particles with different radical concentrations were produced (EPFRlo: 1.5e16; EPFRhi: 1.0e18). Initial studies used EPFRlo and EPFRhi at 250 µg/m3 to investigate the effects of EPFRs on vascular endothelium and pulmonary oxidative stress in C57BL/6 male mice. These studies demonstrated that exposure to combustion-derived EPFRs results in an increase in endothelin-1 (ET-1) and intracellular adhesion molecule-1 (ICAM-1) at high, but not low radical concentrations. In addition, genes associated with AhR activation (Cyp1a1/1b1) and oxidative stress (NQO1) were upregulated in both the lungs and the aorta. Thus, we hypothesized that inhalation of EPFRs leads to pulmonary oxidative stress and vascular endothelial dysfunction via activation of AhR in a manner dependent upon radical concentration. To address our hypothesis, AhR was knocked-down in alveolar type 2 pneumocytes in male and female mice using the Cre/lox recombinase system prior to exposure to filtered air (FA), EPFRlo or EPFRhi for 4 hours. Plasma ET-1 was unchanged between KO and WT mice exposed to FA and EPFRlo; however, ET-1 was significantly decreased in AhR KO mice exposed to EPFRhi versus WT. Supporting our hypothesis, AhR deficient mice were protected from EPFRhi exposure. Together, these data suggest that EPFR exposure promotes vascular dysfunction via pulmonary oxidative stress and AhR activation.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 121 Poster Session II: Poster #II-25

A Data Mining Approach Reveals Chemicals Detected at Higher Levels in Non- Hispanic Black Women Target Preterm Birth Genes and Pathways

Sean M. Harris, University of Michigan Justin Colacino, University of Michigan Miatta Buxton, University of Michigan Lauren Croxton, University of Michigan Vy Nguyen, University of Michigan Rita Loch-Caruso, University of Michigan Kelly M. Bakulski, University of Michigan

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Northeastern University Superfund Research Center

Racial disparities exist in exposures to environmental contaminants and in rates of adverse pregnancy outcomes. In particular, preterm birth occurs disproportionately among Black women, who also face increased exposure to certain environmental toxicants. Challenges posed by the diverse set of Superfund toxicants require new approaches to understand more completely the role that environmental toxicants play in racial disparities in preterm birth. The goal of this study was to use publicly available toxicogenomic, exposure and disease databases to identify chemicals that contribute to preterm birth disparities. We tested 31 chemicals with differential exposure levels between non-Hispanic Black and White women. We obtained chemical-gene interactions from the Comparative Toxicogenomics Database and a list of genes involved in preterm birth from the Preterm Birth Database. We tested each chemical for a statistically high number of interactions with genes involved in preterm birth. We observed that 30 out of 31 chemicals had a significantly high number of interactions with preterm birth genes (FDR<0.05). Of these chemicals, 18 were detected at higher levels in non-Hispanic Black women, and 12 were detected at lower levels. Our approach demonstrates a method for evaluating a large set of environmental toxicants, such as those found at Superfund sites, for potential impacts on complex biological endpoints, like preterm birth. Future studies with our collaborators in the PROTECT Superfund Center will apply this methodology to the unique exposure scenarios faced by pregnant women in Puerto Rico. Through collaboration with PROTECT’s Research Translation Core, we will to coordinate the dissemination of findings to entities including local stakeholders in Puerto Rico, as well as the entire Superfund Research Program community.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

122 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-26

The use of tocofersolan as a rescue agent in larval zebrafish exposed to benzo[a]pyrene in early development

Zade Holloway, Duke University, Durham, North Carolina Andrew Hawkey, Duke University, Durham, North Carolina Helina Asrat, Duke University, Durham, North Carolina Nidhi Boinapally, Duke University, Durham, North Carolina Edward Levin, Duke University, Durham, North Carolina

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Duke University Superfund Research Center

Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants created by incomplete combustion. Benzo[a]pyrene (BaP), among other PAHs, is known to exert toxicity through oxidative stress which is thought to occur through inhibition of antioxidant scavenging systems. The use of agents that reduce oxidative stress may be a valuable route for ameliorating the adverse effects of PAHs on neural development and behavior. This study was conducted to determine if tocofersolan supplementation can prevent or reduce neurobehavioral deficits in zebrafish embryos exposed to BaP during early development. Newly hatched zebrafish were assessed on locomotor activity, light responsivity, and habituation. Zebrafish embryos were exposed to vehicle (DMSO), tocofersolan (0.3uM - 3uM), and/or BaP (5uM) from 5-120 hours post-fertilization. This dose range was below the threshold for producing overt dysmorphogenesis or increased survival. BaP (5uM) was found to cause locomotor hypoactivity in larval fish. Co-exposure of tocofersolan (1uM) led to a restoration of locomotor function, similar to that of the control group. Based on the findings of this study, this model can be expanded to assess the outcome of vitamin E supplementation on other potential environmental neurotoxins, and additionally lead to determination if this rescue persists into adulthood.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 123 Poster Session II: Poster #II-27

Genetics-based Approach to Identify Novel Genes Regulated by the Aryl Hydrocarbon Receptor (AHR) in Mice

Amanda Jurgelewicz, Michigan State University, East Lansing, Michigan John J LaPres, et al. Michigan State University, East Lansing, Michigan

Superfund Mandate: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Michigan State University Superfund Research Center

The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor in the PAS superfamily of environmental sensors that is linked to several metabolic diseases, including non-alcoholic fatty liver disease. Much remains unknown regarding the impact of genetic variation in AHR-driven disease, as past studies have focused on a small number of inbred strains. Recently, presence of a wide-range of interindividual variability amongst humans was reported in response to 2,3,7,8-tetrachlorodibenzo-p- dioxin (TCDD), the prototypical ligand of the AHR. In this study, a panel of 14 diverse mouse strains were exposed to TCDD for 10 days to better characterize the impact of genetics on AHR-mediated responses. Results indicate that responses to TCDD are heavily-dependent on genetic background. Mice strains are known to carry a diverse set of Ahr alleles that impact the response to the receptor’s ligands. While the Ahr allele significantly influenced hepatic accumulation of TCDD and the number of differentially expressed genes, we also observed significant intra-allelic variability suggesting the presence of novel genetic modifiers of AHR signaling. We used linear regression to scan for novel genes that are regulated by the AHR and/or associated with phenotypes that may not have been uncovered in past experiments with small numbers of in-bred strains. This approach identified seven genes associated with TCDD burden and, thus, are likely regulated by the AHR and one gene that was significantly associated with change in TCDD-induced change in body fat. The results not only suggest three novel genes that are regulated by AHR-mediated response, but also indicate the power of using mouse genetic-based approaches to characterize physiological mechanisms.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

124 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-28

Global Cancer Risk from Unregulated Polycyclic Aromatic Hydrocarbons

Jamie M. Kelly, Massachusetts Institute of Technology, Cambridge, Massachusetts Peter D. Ivatt, University of York, (UK) Mathew J. Evans, University of York (UK) Jesse H. Kroll, Massachusetts Institute of Technology, Cambridge, Massachusetts Amy I. H. Hrdina, Massachusetts Institute of Technology, Cambridge, Massachusetts Ishwar N. Kohale, Massachusetts Institute of Technology, Cambridge, Massachusetts Forest M. White, Massachusetts Institute of Technology, Cambridge, Massachusetts Bevin P. Engelward, Massachusetts Institute of Technology, Cambridge, Massachusetts Noelle E. Selin, Massachusetts Institute of Technology, Cambridge, Massachusetts

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Massachusetts Institute of Technology Superfund Research Center

Scientists and regulators commonly use benzo[a]pyrene (BAP) concentrations to assess and manage cancer risk from the complex mixture of atmospheric polycyclic aromatic hydrocarbons (PAH). Here, we show that BAP is a poor indicator of PAH risk distribution and management: nearly 90% of cancer risk worldwide results from other PAHs, including unregulated degradation products of emitted PAHs. Previous studies identified BAP as the dominant contributor to the human cancer risk of PAH mixtures, but they did not consider a comprehensive range of emitted PAHs and their atmospheric degradation products, especially those containing a nitro group (N-PAHs). We develop and apply a global-scale atmospheric model and conduct health impact analyses to estimate human cancer risk from 16 PAHs and their N-PAH degradation products. Our health impact analyses use two different methods, one using toxicity information on PAHs from epidemiological studies, and another using toxicity information from animal studies. Using the second method, which captures the relative risk of different compounds, we find that BAP is a minor contributor to the total cancer risks of PAHs (11%); the remaining risk comes from the other directly-emitted PAHs (73%) and N-PAHs (15%). We show that research and assessments that rely on BAP exposure provide misleading estimates of risk distribution, the importance of chemical processes, and the prospects for risk mitigation. We conclude that more accurate assessments of human health risks of PAHs could be achieved if scientific research, environmental guidelines and environmental monitoring are extended beyond BAP, and explicitly account for PAH degradation products. This project draws upon expertise from across the MIT SRP, and demonstrates the importance of collaboration between engineers and biologists in advancing our understanding of pollutants.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 125 Poster Session II: Poster #II-29

PCB52 and its metabolite, OH-PCB52, cause transcriptional changes in preadipocyte stem cells that are associated with obesity, inflammation, and insulin resistance

Aloysius J. Klingelhutz, University of Iowa, Iowa City, Iowa Francoise A. Gourronc, University of Iowa, Iowa City, Iowa James A. Ankrum, University of Iowa, Iowa City, Iowa

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of Iowa Superfund Research Center

PCBs are associated with the development of metabolic syndrome, yet their specific role in this cluster of conditions is unknown. PCBs are found at high levels in food and in the air of schools and old buildings. While there are over 200 different PCB congeners, lower-chlorinated congeners are serious air contaminants and exposure results in high levels of accumulation in adipose tissue. Adipose tissue is considered to be a major endocrine organ. Dysfunctional adipose tissue is known to result in obesity and insulin resistance. PCB52 is a lower-chlorinated PCB that has been associated with neurological toxicity but also with the development of diabetes. PCB52 can be metabolized to its hydroxylated form OH-PCB52 in liver cells, potentially making it a more reactive and toxic compound. To begin to understand how PCB52 and OH-PCB52 affect adipose tissue function, we exposed human preadipocyte stem cells to these compounds over a time course (9, 24, and 72 hours) and assessed transcript changes using RNAseq. Top biological pathways that were changed by exposure included PPAR signaling, insulin resistance, TGF-beta signaling, and fatty acid metabolism. Activation of inflammatory pathways was present at later time points. All of these pathways are known to play roles in the development of dysfunctional adipose tissue. While there was some overlap, many of the pathways affected by PCB52 and OH-PCB52 were different, indicating that the biological effects of PCB52 are altered by hydroxylation. Further studies will assess how changes in gene expression and perturbations of the affected pathways affect adipose function, potentially leading to obesity, insulin resistance and steatosis. (Supported by P42 ES013661)

Email: [email protected] Presenter Status: Researcher

126 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-30

Investigating the effects of chemicals on mitochondrial DNA mutagenesis using cross-species and ultra-sensitive sequencing approaches

Tess Leuthner, Duke University Laura Benzing, Duke University R. Nathan Keith, Lawrence Berkeley National Lab Craig E. Jackson, Indiana University Kim Young, Duke University Danielle F. Mello Trevisan, Duke University Richard Di Giulio, Duke University Jennifer Wernegreen, Duke University Scott R. Kennedy, University of Washington Joseph R. Shaw, Indiana University Joel N. Meyer, Duke University

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Duke University Superfund Research Center

Effects of chemical exposure on mitochondrial DNA (mtDNA) mutagenesis are poorly understood but is important to investigate, as mutations are implicated in many diseases. Here, we test the effects of several chemicals on germline mitochondrial DNA mutagenesis, and the processes involved across three different organisms and two sequencing approaches. First, we analyzed mtDNA sequences from a mutation accumulation experiment in which independent lineages of cadmium-sensitive and cadmium-tolerant populations of Daphnia pulex were exposed to an ecologically relevant concentration of cadmium for ~12,000 generations. After whole genome NGS, we observe variation in frequency, rate, spectrum, and location of mtDNA mutations between control and cadmium treated lines. We then applied the novel and ultra-sensitive Duplex Sequencing technique to two organisms, Caenorhabditis elegans and Fundulus heteroclitus. C. elegans mtDNA was sequenced in the wildtype strain with and without ultraviolet C irradiation, and two mitophagy deficient strains, pink-1 and dct-1. We also completed a mutation accumulation experiment in these strains of C. elegans in the presence of cadmium or Aflatoxin B1, and conducted Duplex Seq after ~500 generations of exposure to investigate the role of mitochondrial dynamic processes in chemical-induced, heritable mtDNA mutations. Finally, to test this tool as a field biomarker, we conducted Duplex Seq in F. heteroclitus collected from the Duke University Superfund Research site in Chesapeake, VA. mtDNA was sequenced from whole killifish embryos from a population adapted to PAHs, and a sensitive, reference population, after maternal exposure to the complex PAH mixture, Elizabeth River Sediment Extract. Overall, we strive to characterize the potential effects of Superfund chemicals on mutagenesis, while integrating and applying novel toxicogenomics approaches.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 127 Poster Session II: Poster #II-31

ERP57 mediates Cd-induced endoplasmic reticulum stress, citrullinated-vimentin secretion and subsequent lung fibrosis

Fu Jun Li, University of Alabama at Birmingham Ranu Surolia, University of Alabama at Birmingham Zheng Wang, University of Alabama at Birmingham Pooja Singh, University of Alabama at Birmingham Mohammad Athar, University of Alabama at Birmingham Veena B. Antony, University of Alabama at Birmingham

Superfund Mandates: None

Superfund Center: University of Alabama Birmingham Superfund Research Center

Cadmium (Cd), a common component of cigarette smoke and environmental particulate matter, has been demonstrated an increase in lung tissue from subjects with idiopathic pulmonary fibrosis (IPF) in our previous study. Cd correlates with citrullinated vimentin (Cit-Vim) and Cit-Vim correlates with lung function. Importantly, we demonstrated that Cit-Vim secreted from Cd-activated lung macrophages, activate fibroblasts through TLR4 signaling and contributes to the pathogenesis of IPF. However, how does Cit-Vim translocate to membrane and secrete outside of the cells has not been understood. Endoplasmic reticulum (ER) stress characterized as an accumulation of misfolded proteins in the ER has been involved in Cd-induced oxidative stress and cell death. However, whether ER stress involves in Cd-induced pulmonary fibrosis is unknown. Here, we found that Cd exposure mice exhibited increases in ER chaperone glucose-regulated protein (GRP) 78 as well as increases in downstream transcriptional effector, CCAAT/enhancer-binding protein-homologous protein (CHOP). Another two ER chaperone proteins, calreticulin (CRT) and ERP57 were also increased in a time- dependent manner after Cd exposure. Phenylbutyric acid (PBA), an ER stress inhibitor, prevented Cd-induced Cit-Vim production, ER stress proteins accumulation and lung fibrosis. The Cd-induced lung architectural destruction and collagen deposition were markedly recovered by PBA. We further demonstrated that Cd enhanced the binding of Cit-Vim with CRT and ERP57 by using mass spectrometry, solid-phase receptor binding assay and immunoblot analysis. By knocking down ERP57 using siRNA, we found that ERP57 was required for Cd-induced Cit-Vim membrane localization and secretion. ERP57 deficiency attenuated Cd-induced ER stress-related misfolded proteins accumulation, a-SMA activation and collagen deposition. Together, this studies support the role of ERP57 in Cd-induced ER stress, Cit-Vim secretion, and subsequent lung fibrosis.

Email: [email protected] Presenter Status: Researcher

128 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-32

Effect of antenatal Exposure to Polycyclic Aromatic Hydrocarbons on Fetal Lung Development, Gut Microbiome, and Postnatal Hyperoxic Lung Injury – Role of the Gut-Lung Axis

Krithika Lingappan, Baylor College of Medicine, Houston, Texas Guobin Xia, Baylor College of Medicine, Houston, Texas Lauren Swanson, Baylor College of Medicine, Houston, Texas Kristi Hoffman, Baylor College of Medicine, Houston, Texas Weiwu Jiang, Baylor College of Medicine, Houston, Texas Lihua Wang, Baylor College of Medicine, Houston, Texas Bhagavatula Moorthy, Baylor College of Medicine, Houston, Texas

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Baylor College of Medicine Superfund Research Center

Pregnant women living near Superfund sites are often exposed to polycyclic aromatic hydrocarbons (PAHs). In this investigation, we hypothesized that prenatal PAH exposure adversely affects fetal lung development, causes gut microbiome dysbiosis, and exacerbates oxygen mediated lung injury in newborn mice. Pregnant wild type (WT) (C57BL/6J) mice were treated with orally with benzo[a]pyrene (BP) (20 mg/kg) or vehicle corn oil (CO) on gestational days 16-19, and newborn mice were maintained in room air or exposed to hyperoxia for 4 days (P1-4). Intestinal tissue was collected on P4 and 15 and then subjected to 16S rRNA gene sequencing. The lungs of newborn WT mice exposed antenatally to the vehicle control, followed by postnatal hyperoxia showed lung injury, inflammation, and alveolar simplification, compared to animals maintained in room air. WT mice that were prenatally treated with BP, followed by postnatal hyperoxia showed attenuation of lung injury and abnormal alveolarization compared to those that were prenatally exposed to CO. The overall composition of the gut microbiome was analyzed by principle coordinates analysis (PCoA) of β-diversity at PND 4 and 15. Prenatal BP exposure was associated with statistically significant differences in bacterial community composition (Bray-Curtis beta diversity) among mice given postnatal oxygen but not mice exposed to room air. Difference were evident at PND 4, but was strongest at PND 15. Our results suggest greater similarity between control animals—those exposed to corn oil and room air—and BP-exposed mice treated with oxygen. Hyperoxia increased pulmonary IL-6 and TNF-α expression in the vehicle treated animals, while this was significantly attenuated in the BP-treated group. Our results suggest contribution of the gut-lung axis in neonatal hyperoxic lung.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 129 Poster Session II: Poster #II-33

Periods of Susceptibility: Exposure to Per- and Polyfluoroalkyl Substances and Children’s Body Composition

Jamie Liu, Brown University, Providence, Rhode Island Nan Li, Brown University, Providence, Rhode Island George D. Papandonatos, Brown University, Providence Rhode Island Antonia M. Calafat, Centers for Disease Control and Prevention, Atlanta, Georgia Aimin Chen, University of Cincinnati College of Medicine, Cincinnati, Ohio Bruce P. Lanphear, Simon Fraser University, Burnaby British Columbia, Canada Kim M. Cecil, Cincinnati Children’s Hospital Med Ctr, Univ of Cincinnati Coll of Med, Cincinnati, Ohio Heidi J. Kalkwarf, Cincinnati Children’s Hospital Med Ctr, Univ of Cincinnati Coll of Med, Cincinnati, Ohio Kimberly Yolton, Cincinnati Children’s Hospital Med Ctr, Univ of Cincinnati Coll of Med, Cincinnati, Ohio Joseph M. Braun, Brown University, Providence, Rhode Island

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Brown University Superfund Research Center

Background/Aim: Epidemiological studies suggest that per- and polyfluoroalkyl substances (PFAS) exposure may increase adiposity and obesity risk in children. However, we are unaware of any studies extending these findings into adolescence or identifying potential periods of heightened susceptibility. Thus, we estimated associations of five repeated measures of pre- and postnatal PFAS exposure with body composition in adolescence. Methods: We studied 212 mother-offspring pairs from the HOME Study, a prospective pregnancy and birth cohort study that enrolled pregnant women in Cincinnati, OH from 2003- 2006. We quantified serum perfluorooctane sulfonate (PFOS), perfluorooctanoate (PFOA), perfluorononanoate (PFNA), and perfluorohexane sulfonate (PFHxS) concentrations in mothers at ~16 weeks gestation and their children at birth and ages 3, 8, and 12 years. At 12 years, we assessed children’s adiposity using anthropometry and dual X-ray absorptiometry (DXA). We used multiple informant models to estimate covariate-adjusted associations of ln-transformed PFAS with adiposity measures for each exposure period, and tested the difference in these associations. Results: Prenatal serum concentrations of all four PFAS were positively associated with body fat, but there was no clear pattern for postnatal PFAS exposures. Associations were generally stronger for central adiposity measures compared to peripheral ones. In multipollutant models, prenatal PFOA and PFHxS were more strongly associated with body fat than PFOS and PFNA. We observed evidence suggesting that prenatal PFOA was more strongly associated with greater adiposity in girls, but not boys. Conclusions: Prenatal serum concentrations of PFAS, particularly PFOA and PFHxS, were positively associated with measures of central body fat in adolescence.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

130 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-34

Determination of Benzo[a]pyrene (BaP) Metabolism and Toxicokinetics in Humans using UPLC-Accelerator Mass Spectrometry and Oral Micro-Dosing with [14C]-BaP

Monica L.V. Maier, Oregon State University Lisbeth K. Siddens, Oregon State University Sandra Uesugi, Oregon State University Kim A. Anderson, Oregon State University Ted Ognibene, Oregon State University Kenneth Turteltaub, Oregon State University Jordan Smith, Oregon State University David E. Williams, Oregon State University

Superfund Mandates: None

Superfund Center: Oregon State University Superfund Research Center

Designated by the International Agency for Research on Cancer as a known class 1 human carcinogen, benzo[a]pyrene (BaP) is formed by incomplete combustion of organic materials. Though BaP is the most widely studied polycyclic aromatic hydrocarbon (PAH) risk assessment relies on toxicokinetics and cancer studies in rodents at doses 5-6 orders of magnitude greater than human populations. In an effort to provide human toxicokinetic profiles the sensitivity allowed by accelerator mass spectrometry (AMS) and the separation of liquid chromatography (UPLC-AMS) were used to follow uptake, metabolism and elimination of [14C]-BaP over 48 hours in plasma with increasing oral doses. Seven human volunteers were administered 25, 50, 100 and 250 ng (2.7-27 nCi) of [14C]-BaP) and quantification of parent BaP and metabolites in plasma measured over 48 hours. Marked inter-individual variability in metabolite profiles and toxicokinetics was observed. Independent of dose [14C]-BaP Tmax varied from 30-60 min; with Cmax and area under the curve (AUC) approximating dose-dependency. Metabolite profiles showed that, even at the earliest timepoint (30 min), a substantial percentage of total [14C] in plasma was the polar BaP metabolites which increased as parent [14C]-BaP concentrations fell. Tetrols and dihydrodiols at the bay region predominated, suggesting extensive first pass metabolism and bioactivation. Despite evidence of extensive metabolism, parent [14C]-BaP was observed at the final timepoint (48 h) following both the 100 and 250 ng doses. This data suggests potential bioaccumulation of BaP in humans as the average dietary consumption of BaP is 250 ng daily. AMS allows for study of environmental carcinogens in humans with de minimus risk, lending to important testing and validation of risk assessment and physiologically based pharmacokinetic models derived from animal data and interpretation of data from high-risk occupationally exposed populations.

Email: [email protected] Presenter Status: Trainee (pre- / -post doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 131 Poster Session II: Poster #II-35

Comparing Regional Drivers of Toxics Transferal Risk: Applying the Toxics Mobility Vulnerability Index in San Diego County, CA; Harris County, TX; and the State of Rhode Island

Matthew Malecha, Texas A&M University, College Station, Texas Galen D. Newman, Texas A&M University, College Station, Texas Jennifer A. Horney, University of Delaware, Wilmington, Delaware Ibraheem M. Karaye, University of Delaware, Wilmington, Delaware Scott Frickel, Brown University, Providence, Rhode Island Thomas W. Marlow, Brown University, Providence, Rhode Island Iiya Zaslavsky, University of California, San Diego, San Diego, California Keith Pezzoli, University of California, San Diego, San Diego, California

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

The convergence of natural disasters and environmental contamination from anthropogenic sources heightens the potential for toxicant mobility and transfer and increases the potential for subsequent negative health impacts. We developed and applied a Toxics Mobility Vulnerability Index (TMVI) to understand the contribution of various components of vulnerability to toxic materials transfer in three areas of the U.S.: San Diego County, California; Harris County, Texas; and the State of Rhode Island. Vulnerably components considered include % industrial land, % vacant land, % of land designated as floodplain, % of land designated as impervious surface, social vulnerability, and the prevalence of 13 health conditions. Findings indicate “hot spots” of vulnerability to hazard-induced toxics transfer in each study area, but the main drivers of increased risk differ across the three study areas. However, the most vulnerable U.S. Census tracts in all three study areas have overlapping exposure to a combination of vacant land, industrial land use, impervious surfaces, floodplains, and social vulnerabilities. Recommendations related to ways to expand the scope of the TMVI to the entire U.S. and house, maintain, and expand the dataset to ensure access by researchers, decision-makers, and the public will be provided. Applying tools such as the TMVI to highly vulnerable urban and coastal locations will help to identify changes needed to preparedness and mitigation planning and highlight areas where limited resources for investment- and policy-related remediation should be focused, both before and after disasters. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

132 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-36

Effects of manganese exposure in paraoxonase-2 deficient mice

Judit Marsillach, University of Washington, Seattle, WA Rebecca J. Richter, University of Washington, Seattle, WA Jacqueline M. Garrick, University of Washington, Seattle, WA Toby B. Cole, University of Washington, Seattle, WA Lucio G. Costa, University of Parma, Italy, University of Washington, Seattle, WA Clement E. Furlong, University of Washington, Seattle, WA,

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substance

Superfund Center: University of Washington Superfund Research Center

Environmental, nutritional and occupational chronic manganese exposures result in increased levels of oxidative stress and neuroinflammation, ultimately leading to development of several pathologic conditions. The paraoxonases (PONs; PON1, 2 and 3) are lipolactonases involved in modulating oxidative stress and inflammation. PON1 is mostly found in circulation bound to high-density lipoproteins, while PON2 is intracellular and ubiquitous, with high levels of expression in brain. Our aim was to understand how PON1 prevents systemic accumulation of oxidative stress, and to study our hypothesized protective role of brain PON2 in manganese-related neurotoxicity. PON1 activity was dramatically inhibited in vitro with increased concentrations of products of oxidative stress, such as 4-hydroxy-2-nonenal (4-HNE), malondialdehyde (MDA) and the atherogenic oxidized phospholipid OxPAPC, as well as with increased concentrations of manganese and other heavy metals. This would indicate that compounds such as heavy metals, that result in increased levels of oxidative stress, lead to pathogenesis due to inhibition of the potent antioxidant PON1. In vivo manganese exposure via drinking water of wild-type and Pon2-deficient mice resulted in a modest increase of oxidative stress (MDA) in plasma, brain and liver. In addition, Pon2- deficient mice exposed to manganese showed evidence of impaired memory, as assessed by the Object Location Task behavioral test, compared with wild-type mice. Manganese exposures also had an effect on body weight in all mice, but differences were only significant for the male Pon2-deficient mice. All Pon2- deficient mice, independent of Mn treatment, displayed motor deficits when tested on the rotarod. Our in vivo results are indicative of a neuroprotective role of PON2 in manganese exposures, as well as a motor deficit effect related to decreased Pon2 expression. Supported by NIEHS (P42ES04696).

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 133 Poster Session II: Poster #II-37

Exposures to uranium and arsenic alter intraepithelial and innate immune cells in the small intestine of male and female mice

Sebastian Medina, The University of New Mexico, Albuquerque, NM Fredine T. Lauer, The University of New Mexico, Albuquerque, NM Eliseo F. Castillo, The University of New Mexico, Albuquerque, NM Alicia M. Bolt, The University of New Mexico, Albuquerque, NM Abdul-Mehdi S. Ali, The University of New Mexico, Albuquerque, NM Ke Jian Liu, The University of New Mexico, Albuquerque, NM Scott W. Burchiel, The University of New Mexico, Albuquerque, NM

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

Human exposures to environmental metals, including uranium (U) and arsenic (As) are a global public health concern. Chronic exposures to U and As are linked to many adverse health effects including, immune suppression and autoimmunity. The gastrointestinal (GI) tract is home to many immune cells vital in the maintenance of systemic immune health. However, very little is known about the immunotoxicity of U and As at this site. The present study examined the burden of U and As exposure in the GI tract as well as the resultant immunotoxicity to intraepithelial lymphocytes (IELs) and innate immune cells of the small intestine following chronic drinking water exposures of male and female mice to U (in the form of uranyl acetate, UA) and As (in the form of sodium arsenite, As3+). Exposure to U or As3+ resulted in high levels of U or As in the GI tract of male and female mice, respectively. A reduction of small intestinal CD4+ IELs (TCRαβ+, CD8αα+) was found following As3+ exposure, whereas U produced widespread suppression of CD4- IEL subsets (TCRαβ+ and TCRγδ+). Evaluation of innate immune cell subsets in the small intestinal lamina propria revealed a decrease in mature macrophages, along with a corresponding increase in immature/proinflammatory macrophages following As3+ exposures. These data show that exposures to two prevalent environmental contaminants, U and As produce significant immunotoxicity in the GI tract. Collectively, these findings provide a critical framework for understanding the underlying immune health issues reported in human populations chronically exposed to environmental metals.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

134 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-38

Membranes for PFOA and PCB Water Contaminant Remediation to Nanosized Aerosol Capture

Rollie Mills, University of Kentucky A. Saad, University of University of Kentucky M. Bernard, University of Kentucky R. Vogler, University of Kentucky Y. Wei, University of Kentucky T. Hastings, University of Kentucky T. Dziubla, University of Kentucky D. Bhattacharyya, University of Kentucky

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Kentucky Superfund Research Center

Membranes with appropriate functionalization allow applications for toxic pollutant remediation from water to aerosol type virus capture from air media. Perfluorochemicals, such as PFOA, and chlorinated organic compounds, such as 2-chlorobiphenyl (PBC-1), have become emergent water pollutants in recent years, due to their high toxicity and their inability to break down naturally in the environment. A temperature responsive polymer, Poly-N-isopropylacrylamide (PNIPAm), was used to functionalize the pores of a polyvinylidene difluoride (PVDF) membrane for a reversible water pollutant adsorption/desorption cyclic process. This process utilizes the reversible change in the polymer exhibiting its hydrophilic or hydrophobic domain, which is due to the lower critical solution temperature (LCST) of PNIPAm, which is approximately 32°C. Above the polymer’s LCST, this membrane also exhibited a higher water permeability, thus allowing higher water volume throughput. One can also bring a catalytic domain with selective pollutant sorption by functionalizing the membrane with poly(methyl methacrylate) (PMMA) and PNIPAm to integrate iron-palladium nanoparticles in the membrane pores. With these catalytic particles, the membrane effectively degraded PCBs via the reductive pathway in an enhanced capacity than other methods, due to the PNIPAm’s higher adsorptive capacity above its LCST. We further extended the water-related work to air media for virus to nanoparticle air filtration. The air permeation of poly(methacrylic acid) (PMAA)-functionalized PVDF membranes was investigated, as well as this membrane’s ability to capture and/or filter aerosolized polystyrene latex (PSL) nanoparticles (to simulate airborne virus particles) of varying diameters. It was found that this functionalized membrane was able to selectively filter aerosolized particles with a tunable and precise particle diameter filtration cut-off with a high air flow (over 1×10^6 liters/m2/hour), even at pressures below 1 bar.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 135 Poster Session II: Poster #II-39

Role of Alkylated Polycyclic Aromatic Hydrocarbons in Mixture Toxicity from a Legacy Creosote Site

Ian Moran, Oregon State University Robyn L. Tanguay, Oregon State University Katrina M. Waters, Pacific Northwest National Laboratory, Oregon State University Kim A. Anderson, Oregon State University

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Oregon State University Superfund Research Center

Creosote is a wood treatment product, often derived from distillation of tars. However, as a complex mixture containing mostly polycyclic aromatic hydrocarbons (PAHs) and their derivatives, the toxic effects of legacy creosote contamination are poorly understood. In particular, alkylated PAHs are products of PAH weathering and are known to be abundant constituents of creosote and many petroleum products. Less is known about the toxicity of alkylated PAHs than their parent compounds. Despite this, alkylated PAHs have been shown to contribute substantially to the toxicity of PAH mixtures. In one study, 80% of the total toxicity to aquatic organisms from contaminated sediments was due to alkylated PAHs. The goal of this study is to understand the contribution of alkylated PAHs to the toxicity of a complex mixture from a legacy creosote site. This study utilizes low density polyethylene passive samplers deployed at a former wood treatment facility to accumulate freely dissolved organics in the surface water. Passive samplers are extracted and analyzed by gas chromatography – tandem mass spectrometry (GC-MS/MS) for over 60 alkylated and parent PAHs. To assess toxicity, embryonic zebrafish are exposed to PAH mixtures in 96- well plates and are observed at 24 and 120 hours post fertilization for a suite of behavioral and morphological endpoints. Zebrafish embryos are exposed to both passive sampling extracts from the contaminated site and representative mixtures from reference materials in order to quantitatively describe the contribution of alkylated PAHs to the toxicity of a complex mixture of weathered PAHs. Understanding the role of alkylated PAHs and their interactions with other mixture constituents can inform remediation efforts and improve our ability to protect human health and water quality.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

136 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-40

Voices Unheard: Arizona’s Environmental History Project

Denise Moreno Ramirez, University of Arizona, Tucson, Arizona Jamie Lee, University of Arizona, Tucson, Arizona Mark Nichter, University of Arizona, Tucson, Arizona Janick Artiola, University of Arizona, Tucson, Arizona Raina Maier, University of Arizona, Tucson, Arizona

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Arizona Superfund Research Center

It is necessary to listen to the stories of individuals and document local knowledge when it comes to environmental issues. The Voices Unheard: Arizona’s Environmental History project recovers the historical accounts, descriptions, and insights from the people living near two contaminated sites in Arizona. One being the Tucson International Airport Area Superfund site that has been impacted primarily by a metal degreaser called trichloroethylene; and the second is the Iron King Mine and Humboldt Smelter Superfund site that contains mine waste with arsenic and lead. The Voices Unheard project draws on lived experience together with historical archival documents to tell more complex stories of health impacts and community advocacy. The oral histories have been video and audio recorded, then these histories have been transcribed, and the interviewees have provided photographs and artifacts that illustrate their history. The project combines environmental science, medical anthropology, oral history, and qualitative methods to implement a mixed approach and create a community-engaged platform. By encapsulating this environmental history, the project creates an invaluable and rich archive for today and for the future by establishing primary sources of public history that will be accessible online and at the local libraries. Most importantly, it is adding to the dominant history narratives of both areas and amplifying the voices of those who often go unheard.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 137 Poster Session II: Poster #II-41

Application of Single Nuclei Transcriptomics to Assess the Hepatic Effects of Dioxin

Rance Nault, Michigan State University, East Lansing, Michigan Kelly Fader, Michigan State University, East Lansing, Michigan Sudin Bhattacharya, Michigan State University, East Lansing, Michigan Tim Zacharewski, Michigan State University, East Lansing, Michigan

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

Characterization of cell specific transcriptional responses to liver toxicants can be lost in the averages of bulk RNA-sequencing (RNA-seq). Emerging single cell/nuclei RNA-seq technologies now allows researchers to evaluate the transcriptomes of individual cell (sub)types from in vivo and in vitro models. To demonstrate the application of this technology for toxicological evaluation of xenobiotics, we performed single nuclei RNA-sequencing (snSeq) on frozen liver samples from male C57BL/6 mice gavaged with sesame oil vehicle or 30 µg/kg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) every 4 days for 28 days. A total of 19,907 genes were detected across 16,015 sequenced nuclei from control and TCDD treated livers. Eleven cell (sub)types were identified including distinct pericentral, midzonal, and periportal hepatocyte sub-populations, and reflected the expected cell diversity of the liver. TCDD altered relative proportions of hepatic cell types and their gene expression. For example, macrophages increased from 0.5% to 24.7%, while neutrophils were only present in treated samples, consistent with histological evaluation. The number of differentially expressed genes in each cell type ranged from 122 (cholangiocytes) to 7,625 (midcentral hepatocytes), and partially correlated with the basal expression level of Ahr, the canonical molecular target of TCDD. In addition to the expected functional enrichments within each cell (sub)types, RAS signaling and related pathways were specifically enriched in nonparenchymal cells while enrichment of metabolic processes occurred primarily in hepatocytes. snSeq also identified the expansion of a Kupffer cell subtype highly expressing Gpnmb, consistent with a population reported in a dietary NASH model. Overall, we show how snSeq can be used to decipher the heterogeneity of transcriptional changes and cell population shifts in response to chemical exposure.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

138 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-42

The influence of Critical Zone age on contaminant reactivity along a fluvial chronosequence in the Oregon Coast Range

Chelsea S. Obeidy, University of Oregon Matthew Polizzotto, University of Oregon

Superfund Mandates: None

Superfund Center: University of North Carolina - Chapel Hill

Critical Zone (CZ) processes are recognized as drivers on the sustainability of soil and water resources. However, how the stage of CZ development impacts contaminant reactivity and mobility in soils has yet to be investigated. The objectives of this study were to quantify (1) how CZ development and age of pedogenesis influence contaminant reactivity, and (2) determine solid-phase hosts and soil characteristics that govern contaminant binding and retention within soils from different stages of development. In order to achieve these objectives, soils from a fluvial chronosequence in the Oregon Coast Range with ages 20, 69, 140, 200, and 908 ky from two depths (30 and 100 cm) were subjected to arsenic (As) adsorption isotherms, with As removal from solution serving as a proxy for soil-contaminant reactivity. Sequential extractions and spectroscopic analyses, targeting amorphous and crystalline Fe- and Al-oxides, were conducted on the original and dosed soils to identify the contaminant host phases across the suite of soils. Data were subjected to multiple linear regressions to identify correlations between soil characteristics and derived As sorption capacities. Older soils (908 ky) at depth (100 cm) have the highest contaminant sorption capacities (5,494 mg kg-1) due to the pedogenetic accumulation of secondary minerals, and thus contaminant reactivity increases with age at depth. The second highest sorption capacities were observed in the youngest (20 ky) and shallowest (30 cm) soils (5,031 mg kg-1) due to the preponderance of high-surface-area amorphous Fe-oxides found in younger soils. The lowest sorption affinities were observed in the 69 ky soils from a depth of 30 cm. Crystalline forms of iron and aluminosilicates are expected to increase with pedogenesis and show a positive correlation with sorption, particularly at depth. This knowledge will assist in improving models for predicting CZ processes and how they govern the sustainability of soil and water quality.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 139 Poster Session II: Poster #II-43

Exploring mechanisms and pathways of VOC migration beneath Detroit's urban landscape

Brendan O’Leary, Wayne State University, Detroit, Michigan Carol J. Miller, Wayne State University, Detroit, Michigan

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Unaffiliated

Urban water movement plays an important role in urban systems from determining building structure, moving chemicals, facilitating urban utilities, to developing urban sustainability plans. However, anthropogenic influences on urban water systems are poorly understood and these influences can impact chemical exposure routes. Specifically, urban subsurface water movement provides a conduit for soil vapors and groundwater to negatively impact the health of communities in urban settings. Volatile organic compounds (VOCs) are a group of compounds that have high vapor pressure and low water solubility. This project seeks to understand anthropogenic influences on the VOC movement in the vadose zone. In this project, we collaborate with the Michigan Department of Environment, Great Lakes & Energy (EGLE) to investigate two example brownfield sites in Detroit neighborhoods for extensive subsurface testing and numerical modeling. The first site contains an abandoned dry-cleaning site with dense non-aqueous phase liquid (DNAPL) VOC plume and the second site comprises of an abandoned leaking underground storage tank with light non-aqueous phase liquid (LNAPL) VOC plume. Fieldwork includes hydraulic conductivity, temperature, moisture content, VOC concentrations and gradients, and flux. These parameters inform the development of numerical models for each location. HYDRUS 1-D is used to simulate both soil water and soil vapor movement in a discontinuous subsurface typical in urban field locations. The field and modeling work contribute to an analysis of understanding vertical gradients and the influence of anthropogenic modification to urban exposure pathways. The work will inform vulnerability and exposure metrics for urban residents living near known sites of vapor intrusion.

Email: [email protected] Presenter Status: Researcher

140 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-44

Prediction of Montmorillonite Clay Efficacy for Mitigating Chemical Exposure through a Hybrid Data-Driven and Molecular-Modeling Approach

Asuka Orr, Texas A&M University, College Station, Texas Meichen Wang, Texas A&M University, College Station, Texas Burcu Beykal, Texas A&M University, College Station, Texas Hari Ganesh, Texas A&M University, College Station, Texas Timothy D. Phillips, Texas A&M University, College Station, Texas Efstratios N. Pistikopoulos, Texas A&M University, College Station, Texas Phanourios Tamamis, Texas A&M University, College Station, Texas

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

People and animals can be exposed to hazardous chemicals, such as polyaromatic hydrocarbons, pesticides, and organic solvents, following natural disasters through contaminated drinking water, food, and feed. Montmorillonite clays have been identified as attractive, safe and effective sorbents of hazardous chemicals to mitigate human and animal exposures Despite their promise, there is a lack of computational methods to screen toxic chemicals for possible adsorption, and rapidly predict and select optimal clay-chemical combinations for further testing. This has limited our predictive ability of which toxic chemicals can be mitigated by montmorillonite clay, as each compound must be evaluated through experimental adsorption isothermal analysis. Here, we aimed to use simulations and parametric models to predict binding affinity of toxic compounds for clays. Using the properties of toxic compounds and the solvent used in experiments, enriched by computationally derived energetic data, we have been developing parametric, data-driven, hybrid models for the prediction of a toxic compound’s affinity to the clay. Such models can be used in the future to predict the efficacy of montmorillonite clay for new chemicals of interest using computationally inexpensive, short-duration molecular dynamics simulations, greatly increasing the number of candidate chemicals that can be screened. This research is supported by NIEHS Superfund Research Program P42 ES027704.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 141 Poster Session II: Poster #II-45

Effect of N-nitrosodimethylamine on Gene Expression in Mice with Varied DNA Repair Capacity

Norah Owiti, Massachusetts Institute of Technology Hee Jae Hong, Massachusetts Institute of Technology Jenny Kay, Massachusetts Institute of Technology Joshua Corrigan, Massachusetts Institute of Technology Bevin P. Engelward, Massachusetts Institute of Technology

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Massachusetts Institute of Technology Superfund Research Center

DNA alkylating agents are genotoxic and carcinogenic chemicals that can damage DNA and induce cytotoxicity and mutagenesis. N-nitrosodimethylamine (NDMA) is an example of SN1 DNA alkylating agents that can react with DNA after metabolic activation, leading to the formation of O6-Methylguanine (O6MeG) and N-methylated purines (e.g., N3-methyladenine [3MeA] and N7-methylguanine [7MeG]). O6MeG is repaired by direct reversal via the O6-methylguanine methyltransferase (MGMT) and 3MeA and 7MeG are removed by the alkyladenine DNA glycosylase (AAG) glycosylase. The objective of this work is to investigate the impact of Aag and Mgmt on the DNA damaging potential of NDMA and on gene expression. Mice lacking Aag (Aag-/-), overexpressing Aag (AagTg), and lacking Mgmt (Mgmt-/-) were either sham treated or exposed to NDMA, and livers were collected 24 hrs, 48 hrs, 5 days and 10 months post NDMA exposure. In vivo analysis of DNA damage in liver samples using the CometChip showed a significant induction of DNA damage following NDMA exposure. RNA was also collected from the liver samples and analyzed via 3’DGE RNA sequencing. NDMA exposure led to differential expression of multiple genes in each genotype. Gene ontology and gene set enrichment analyses revealed genotype- and sex-specific alterations in genes associated with inflammation, DNA repair, apoptosis, protein phosphorylation, and cell cycle, among others. Notably, interferon response was significantly activated in Aag-/- mice compared to WT and AagTg animals had the lowest interferon response, suggesting that AagTg mice are protected from an NDMA-induced interferon response. Taken together, results show that DNA repair is a susceptibility factor for NDMA-induced genotoxicity and a modulator of damage-induced inflammatory responses.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

142 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-46

The mwtab Python library for RESTful Access and Enhanced Deposition, Quality Control, and Curation of the Metabolomics Workbench Data Repository

Christian Powell, University of Kentucky, Lexington KY Hunter N.B. Moseley, Institute for Biomedical Informatics, and University of Kentucky, Lexington KY

Superfund Mandate: None

Superfund Center: University of Kentucky Superfund Research Center

Introduction: The Metabolomics Workbench is a public scientific data repository consisting of experimental data and metadata from metabolomics studies collected from mass spectroscopy (MS) and nuclear magnetic resonance (NMR) analyses. Metabolomics Workbench has been constantly evolving; updating their ‘mwTab’ text file format, adding a JavaScript Object Notation (JSON) file format, implementing a REpresentational State Transfer (REST) interface, and have seen the number of analyses hosted on the repository triple within the last three years. Objectives: In order to keep up with the ever- evolving state of the Metabolomics Workbench repository, the ‘mwtab’ Python library and package has also been continuously updated and is now officially released as version 1.0.0. The package has been updated to mirror the changes in the ‘mwTab’ file format and now contains enhanced validation features, methods for utilizing the Metabolomics Workbench REST interface, and additional features for parsing metabolite data and metadata from repository entries. Moreover, all of these facilities are available through an updated Command Line Interface (CLI) and as a Python application programming interface (API). Methods Development: of the ‘mwtab’ Python package has continued to use Git version control, Python unit-testing and code coverage of the current release and multiple back versions, and documentation available through Github, the Python Package Index (Pypi), and ReadTheDocs. Results/Conclusions: The ‘mwtab’ package continues to strive for promote the most FAIR utilization of the Metabolomic Workbench repository, coevolving with and continuing to improve alongside the repository. Source code is freely available on GitHub and PyPI under the Clear Berkeley Software Distribution (BSD) 3 license.

Email: [email protected] Presenter Status: Other Staff

NIEHS SRP 2020 Virtual Annual Meeting Program • 143 Poster Session II: Poster #II-47

The COVID-19 Pandemic Vulnerability Index (PVI) Dashboard: Monitoring county- level vulnerability using visualization, statistical modeling, and machine learning

David M Reif, North Carolina State University Skylar W Marvel, North Carolina State University John S House, National Institute of Environmental Health Sciences Matthew Wheeler, National Institute of Environmental Health Sciences Kuncheng Song, North Carolina State University Yihui Zhou, North Carolina State University Fred A Wright, North Carolina State University Weihsueh A Chiu, Texas A&M University Ivan Rusyn, Texas A&M University Alison Motsinger-Reif, National Institute of Environmental Health Sciences

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: North Carolina State University Superfund Research Center

While the COVID-19 pandemic presents a global challenge, the U.S. response places substantial responsibility for both decision-making and communication on local health authorities, necessitating tools to support decision-making at the community level. We created a Pandemic Vulnerability Index (PVI) to support counties and municipalities by integrating baseline data on relevant community vulnerabilities with dynamic data on local infection rates and interventions. The PVI visually synthesizes county-level vulnerability indicators, enabling their comparison in regional, state, and nationwide contexts. We describe the data streams used and how these are combined to calculate the PVI, detail the supporting epidemiological modeling and machine-learning forecasts, and outline the deployment of an interactive web Dashboard. Finally, we describe the practical application of the PVI for real-world decision-making. Considering an outlook horizon from 1 to 28 days, the overall PVI scores are significantly associated with key vulnerability-related outcome metrics of cumulative deaths, population adjusted cumulative deaths, and the proportion of deaths from cases. The modeling results indicate the most significant predictors of case counts are population size, proportion of black residents, and mean PM2.5. The machine learning forecast results were strongly predictive of observed cases and deaths up to 14 days ahead. The modeling reinforces an integrated concept of vulnerability that accounts for both dynamic and static data streams and highlights the drivers of inequities in COVID-19 cases and deaths. These results also indicate that local areas with a highly ranked PVI should take near-term action to mitigate vulnerability. The COVID-19 PVI Dashboard monitors multiple data streams to communicate county-level trends and vulnerabilities and facilitates decision-making and communication among government officials, scientists, community leaders, and the public.

Email: [email protected] Presenter Status: Researcher

144 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-48

PCBs in Food

Panithi Saktrakulkla, The University of Iowa Tuo Lan, The University of Iowa Jason Hua, The University of Iowa Rachel F Marek, The University of Iowa Peter S Thorne, The University of Iowa Keri C Hornbuckle, The University of Iowa

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Iowa Superfund Research Center

We measured the concentrations of 205 polychlorinated biphenyl (PCB) congeners in 26 food items: beef steak, butter, canned tuna, catfish, cheese, eggs, french fries, fried chicken, ground beef, ground pork, hamburger, hot dog, ice cream, liver, luncheon meat, margarine, meat-free dinner, milk, pizza, poultry, salmon, sausage, shrimp, sliced ham, tilapia, and vegetable oil. Using Diet History Questionnaire II, we calculated the PCB dietary exposure in mothers and children participating in the AESOP Study in East Chicago, Indiana, and Columbus Junction, Iowa. Salmon had the highest concentration followed by canned tuna, but fish is a minor contributor to exposure. Other animal proteins are more important sources of PCB dietary exposure in this study population. Despite the inclusion of few congeners and food types in previous studies, we found evidence of a decline in PCB concentrations over the last 20 years. We also found strong associations of PCB congener distributions with Aroclors in most foods and found manufacturing byproduct PCBs, including PCB11, in tilapia and catfish. The reduction in PCB levels in food indicates that dietary exposure is comparable to PCB inhalation exposures reported for the same study population.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 145 Poster Session II: Poster #II-49

Insights into the bioavailability of dioxins in soils and sediments

J. Brett Sallach, Michigan State University Qi Yuan, Creighton University Robert Crawford, Michigan State University Hui Li, Michigan State University Cliff Johnston, Purdue University Brian Teppen, Michigan State University Norbert Kaminski, Michigan State University Stephen Boyd, Michigan State University

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

Polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs) are a group of highly toxic persistent organic pollutants ubiquitous in soils and sediments throughout the world. While they can form naturally, hotspots of contamination have resulted from legacy anthropogenic activities including chemical manufacturing and waste incineration. Due to their recalcitrance in soils and sediments and their high toxicities, PCDDs are a priority for remediation throughout the world. Recently, remediation criteria have been adjusted (relaxed) based on measurements of site-specific bulk soil/sediment mammalian bioavailability studies. However, until recently little was known about how the interactions between PCDDs and specific component geosorbents comprising soils and sediments impact bioavailability, making site specific predictions of PCDD bioavailability impossible. In this context, we have evaluated the bioavailability of PCDDs using both a mammalian (mouse) model and microbial model utilizing the dioxin degrading bacteria, Sphingomonas wittichii strain RW1 (RW1). Using the mammalian model, we have shown that sorption to silicas, natural organic matter and intercalation by clays result in no reduction of PCDD bioavailability. However, the sequestration of PCDDs by activated carbons with a wide range of pore structures eliminated the bioavailability of PCDDs to mice. Consistent with the mammalian model, smectite clay intercalated PCDD was shown to facilitate RW1 growth and hence was bioavailable, whereas PCDD sequestration by AC eliminated its microbial bioavailability. In combination, this work has provided key insights into how the sequestration of PCDDs by the primary sorptive constituents of soils and sediments affects their bioavailability, with implications for both exposure and risk assessment as well as remediation alternatives. Overall pyrogenic carbonaceous materials, both naturally occurring and anthropogenic, have emerged as key components responsible for reducing PCDD/F bioavailability.

Email: [email protected] Presenter Status: Researcher

146 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-50

Mixed Metals Exposure and Cognitive Function in Bangladeshi Adolescents

Roheeni Saxena, Columbia University, New York, NY Mary Gamble, Columbia University, New York, NY Gail A. Wasserman, Columbia University, New York, NY Xinhua Liu, Columbia University, New York, NY Faruque Parvez, Columbia University, New York, NY Ana Navas-Acien, Columbia University, New York, NY Tariqul Islam, Columbia University Arsenic Project Office, Mohakhali, Dhaka Bangladesh Pam Factor-Litvak, Columbia University, New York, NY Mohammed Nasir Uddin, University of Chicago Research Office, Mohakhali, Dhaka, Bangladesh Marathi-Anna Kioumourtzoglou, Hasan Shahriar, Mohakhali Elizabeth A. Gibson, Nancy LoIacono, Mahbubul Eunus, Mohakhali Anjan Ghosal, Mohakhali Olgica Balac, and Joseph H. Graziano, Columbia University, New York, NY

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substance

Superfund Center: Columbia University Superfund Research Center

Background: Over 57 million Bangladeshis have been chronically exposed to water arsenic (As) and also face environmental exposure to cadmium (Cd), manganese (Mn), and lead (Pb). These metals have been linked to adverse neurocognitive outcomes in adults and children, though effects on adolescents are not fully characterized. Additionally, previous studies have linked selenium (Se) to protective effects against toxicity of these metals. Objectives: To examine the associations between metals mixture exposure and cognitive function in Bangladeshi adolescents. Methods: The Metals, Arsenic, & Nutrition in Adolescents Study (MANAS) is a cross-sectional study of 572 Bangladeshi adolescents aged 14-26 years. Biosamples were collected for measurement of blood levels of As, Cd, Mn, Pb, and Se. Participants completed an abbreviated Cambridge Neuropsychological Test Automated Battery (CANTAB), a test designed to assess cognitive performance. Results: Linear regression revealed negative associations of As and Mn with Spatial Working Memory (β=-2.40 and -5.31, both p<0.05, respectively). Negative associations were seen between Cd and Spatial Recognition Memory (SRM) (B=-2.77, p<0.05), and between Pb and Delayed Match to Sample (DMS), a measure of visual recognition and memory (B=-3.67, p<0.05). A positive association was seen between Se and Spatial Span Length (B=0.92, p<0.05). BKMR showed no overall effect of the mixture, but indicated that Pb was negatively associated with DMS and that Cd was negatively associated with SRM. Se was positively associated with Planning, Reaction Time, and Spatial Span. Posterior inclusion probability consistently rated Se as the most important mixture member. Conclusions: Overall, Se had a protective effect for cognitive outcomes, whereas Mn and As were linked to poorer working memory and Cd and Pb were linked to poorer visual recognition and memory.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 147 Poster Session II: Poster #II-51

Functionalized Electrospun Nanofiber Mats (ENMs) for uranium (U) remediation in contaminated waters

Nabil Shaikh, University of New Mexico, Alburquerque, New Mexico Margaret Carolan, University of Iowa, Iowa City, Iowa Andres Martinez Araneda, University of Iowa, Iowa City, Iowa Keri Hornbuckle, University of Iowa, Iowa City, Iowa David Cwiertny, University of Iowa, Iowa City, Iowa Jose Manuel Cerrato Corrales, University of New Mexico, Alburquerque, New Mexico

Superfund Mandate: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

UNM METALS Superfund trainee Nabil Shaikh received the 2019 K.C. Donnelly Externship Award Supplement to work with the Iowa Superfund Research Program (ISRP) under Drs. Keri Hornbuckle and David Cwiertny, at the University of Iowa. During his externship, Nabil worked on novel technique called electrospinning, that was used to make electrospun nanofiber mats (ENM). The ENM’s specific chemistry and surface morphology was modified for binding with uranium (U) using functionalized surfactants. The various ENMs were tested for their ability to capture and remove U in different water matrices. Uranium uptake experiments were carried out using 5 mg of phosphonate functionalized ENMs with 10 μM U at pH 6.8 in four reactors using different combinations of solutions containing 5 mM calcium and bicarbonate ions. U uptake was similar in control solutions containing no calcium and bicarbonate (resulting in 19 ± 3% U uptake), and in those containing only 5 mM calcium (resulting in 20 ± 3% U uptake). A decrease in U uptake (10 ± 4% U uptake) was observed in experiments with bicarbonate, indicating that UO2-CO3 complexes may increase uranium solubility. Negligible U uptake was observed in experiments with U, Ca, and HCO3 likely due to the formation of neutral and negatively charged U-Ca-CO3 ternary complexes. Additional results indicate that increasing polarity of the ENMs and modifying surface functional groups can improve the U uptake even in “harsher” water matrices. The knowledge derived from this study will help develop point-of-use treatment and detection strategies, that will protect human health and reduce associated risks near uranium-contaminated sites particularly in rural Native American communities in New Mexico.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

148 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-52

Serum vaccine antibody concentrations in adults exposed to perfluorinated compounds

Yu-Hsuan Shih, Harvard T. H. Chan School of Public Health, Boston MA Annelise J. Blomberg, Harvard T. H. Chan School of Public Health, Boston MA Dorte K. Holm, Odense University Hospital, Odense, Denmark Carsten Heilmann, National University Hospital, Copenhagen, Denmark Pal Weihe, University of the Faroe Islands, Torshavn, Faroe Islands Philippe Grandjean, University of Southern Denmark, Odense, Denmark

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Harvard Superfund Research Center

Background: Perfluorinated alkylate substances (PFASs) are highly persistent in the environment and may cause depressed immune function. Previous studies have linked PFASs exposure to lower vaccine responses in children. However, research in adults is sparse. Objectives: We evaluated the associations of lifetime exposure to PFASs with serum concentrations of hepatitis A and B antibodies among 399 vaccinated participants aged 28 years in the Faroe Islands. Methods: PFAS concentrations were determined from cord blood collected at birth and serum samples collected at ages 7, 14, 22, and 28 years. Serum concentrations of hepatitis A and B antibodies were analyzed from blood samples collected about six months after the second vaccine inoculation at age 28 years. Linear regression models were used to estimate the percent change (hepatitis B) or unit change (hepatitis A) and their confidence intervals for each doubling of the PFAS concentrations in the overall study population as well as among females and males. Results: Serum concentrations of perfluorooctanoate (PFOA) at age 28 years were associated with lower hepatitis A and B antibody concentrations, as revealed by a decrease of 0.24 signal-to-cutoff ratio (S/CO) and 16.77 mIU/mL for each doubling of exposure, respectively. Sex-specific associations were observed for cord blood perfluorooctane sulfonate (PFOS) and PFOA; inverse associations with hepatitis A antibody concentrations were observed only among females. Conclusion: Our findings suggest a reduced immune response to hepatitis A and B vaccination among adults with exposure to PFASs, and sex may modify these associations especially for prenatal exposures.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 149 Poster Session II: Poster #II-53

Chronic diclofenac exposure results in severe cardiotoxicity in mice

Padmini Sirish, University of California-Davis Phung N. Thai, University of California-Davis Lu Ren, University of California-Davis Wilson Xu, University of California-Davis James Overton, University of California-Davis Valeriy Timofeyev, University of California-Davis Carol E. Nader, University of California-Davis Jun Yang, University of California-Davis Bruce Hammock, University of California, Davis Nipavan Chiamvimonvat, University of California-Davis and Dept of Veterans Affairs, Northern CA HCS

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

Introduction: Acute and robust inflammatory responses occur with tissue injury, which is clinically treated with nonsteroidal anti-inflammatory drugs (NSAID). Diclofenac (DIC) is a commonly prescribed NSAID that increases the risk of cardiovascular diseases. However, its mechanistic cardiotoxic effects remain largely unelucidated. In this study, we tested the hypothesis that chronic exposure to DIC will increase oxidative stress, which will ultimately impair cardiovascular function. Methods and Results: Mice were treated with DIC for 4 weeks and subsequently subjected to functional cardiovascular assessments before cardiomyocytes were isolated. We found that chronic DIC exposure impaired cardiac function, as evident by a depressed fractional shortening, reduced mitral valve E/A ratio, and prolonged isovolumetric relaxation time. DIC treatment, however, did not alter blood pressure or electrocardiography recordings. Treatment with DIC significantly increased cardiac fibrosis as assessed by increase in cardiac fibroblast percentage, proliferation and activation. Cardiomyocytes isolated from DIC-treated mice exhibited reduced cell shortening, calcium transients, and sarcoplasmic reticulum load. Mechanistically, we found increased reactive oxygen species production, as well as greater oxidative stress in cardiomyocytes from DIC-treated mice, which resulted in more depolarized mitochondrial membrane potential and reduced energy production. Conclusions: Together, chronic treatment with Diclofenac resulted in severe cardiotoxicity, which was mediated, in part, by an increase in mitochondrial oxidative stress.

Email: [email protected] Presenter Status: Researcher

150 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-54

Relationship between well water arsenic and uranium in Northern Plains Native lands

Marisa Sobel, Columbia University Tiffany Sanchez, Columbia University Benjamin Bostick, Columbia University Brian Mailloux, Barnard College Martha Powers, Johns Hopkins University Joseph Yracheta, Missouri Breaks Industries Research David Harvey, Indian Health Service Lyle G. Best, Missouri Breaks Industries Research Annabelle Black Bear, Missouri Breaks Industries Research Khaled Hasan, Johns Hopkins University Rui Chen, Johns Hopkins University Elizabeth Thomas, Johns Hopkins University Marcia O’Leary, Missouri Breaks Industries Research Camille Morgan, Johns Hopkins University Christine Marie George, Johns Hopkins University Pablo Olmedo, Johns Hopkins University Ana Navas-Acien, Columbia University

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Columbia University Superfund Research Center

Background: Arsenic and uranium in unregulated private wells affect many rural populations across the US, including American Indian communities. The current EPA maximum contaminant level (MCL) is 10 µg/L for arsenic and 30 µg/L for uranium. Objective: Evaluate the association between drinking water As and U levels in three tribal regions in North/South Dakota participating in the Strong Heart Water Study. Methods: Well water samples were collected in 2014-2020. Metal concentrations were dichotomized by their MCL for a graduated visualization across the regions. Spearman correlations and region-specific semivariograms were computed. Spatial interpolation using inverse distance weighting was conducted by region. Results: 441 wells were included, primarily from Region 3 (n=373). The prevalence of metals above the MCL was 29% for arsenic and 7% for uranium. In Region 3, 36% of wells had uranium and/or arsenic MCL. In Regions 1 and 2, no wells had both metal concentrations MCL. The statistical correlation of both metals was weak (rs=0.06), however, there appeared to be a spatial correlation of wells co-contaminated by arsenic and uranium in Region 3, arching from the southwest towards the northeast. Conclusion: Elevated arsenic and uranium in drinking water are common across Northern Plains Tribal Lands. More than a third of all wells sampled contained either arsenic or uranium at levels exceeding the MCL, but co-contamination was more common in Region 3. These findings indicate the importance of measuring multiple metals in well water, and to understand underlying hydrogeological conditions. Translation: The findings of this study support the need for additional protection of private well users from metal exposure in the Northern Plains and to develop guidelines and treatment options for multiple toxicants.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 151 Poster Session II: Poster #II-55

The possible importance of Reactive Oxygen Species in Herbicide Action

Rasheed Sule, University of California-Davis, Davis, CA Naoki Matsumoto, University of California, Davis, CA Christophe Morisseau, University of California, Davis, CA Bruce D. Hammock, University of California, Davis, CA Nipavan Chiamvimonvat, University of California, Davis, CA Aldrin V. Gomes, University of California, Davis, CA

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

Herbicide contamination of surface waters has been well documented worldwide and constitute a major issue that gives rise to concerns at local, regional, national and global level. The usage of atrazine, prometryn, paraquat and other herbicides in agriculture has increased the levels of xenobiotic compounds in soil, water, and even in air near production or application sites. Atrazine is widely used in agriculture and has been previously associated with poor birth outcomes in humans. A major concern about atrazine is that it is persistent in the environment and was present in as much as 80% of drinking water samples obtained from 153 public water systems. Investigation of atrazine ability to cause oxidative stress in H9c2 cardiac cells showed that at a concentration of 10µM atrazine did not induce reactive oxygen species (ROS) or oxidative stress. However, atrazine mercapturate at the same concentration (10µM) induced ROS and oxidative stress in cardiac cells. Atrazine mercapturate is a breakdown product of atrazine and accounts for a major proportion of atrazine metabolites in human urine. Investigation of paraquat, cacodylic acid and prometryn (all used at 10µM) both resulted in increased ROS and oxidative stress in cardiac cells. Further investigation of prometryn (10µM) showed that this herbicide increases superoxide ions produced by H9c2 mitochondria and reduced the mitochondrial membrane potential in these cardiac cells. These results suggest that herbicides can alter mitochondrial function and that a significant proportion of the ROS induced by herbicides are from mitochondria. Reducing mitochondrial ROS or stabilizing mitochondria may be beneficial to reduce the toxic effects of some herbicides in animals.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

152 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-56

Specific polycyclic aromatic hydrocarbons elicit differential expression of CYP1A1 in trophoblast cells

Melissa A. Suter, Baylor College of Medicine, Houston Texas Min Hu, Baylor College of Medicine, Houston Texas Sohini Banerjee, Baylor College of Medicine, Houston Texas Bhagavatula Moorthy, Baylor College of Medicine, Houston Texas Kjersti Aagaard, Baylor College of Medicine, Houston Texas

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Baylor College of Medicine Superfund Research Center

Prenatal exposures to polycyclic aromatic hydrocarbons (PAHs) are associated with adverse outcomes including small for gestational age infants. We have previously reported differential levels of PAHs in the placenta associated with preterm delivery. We found that levels of a specific PAH, dibenzoanthracene (DBA), was significantly higher than levels of either benzo[a]pyrene (BaP) or Benzo[b]fluoranthene (BbF). Furthermore, we reported that placental DBA levels are significantly and negatively correlated with gestational age at delivery. In this study we aimed to determine if there is a differential response in trophoblast cells with distinct PAH treatments. Immortalized choriocarcinoma cells (BeWo) were treated with DMSO or 10 M of either BaP, BbF or DBA for 24 hours. Following treatment, RNA was extracted and used to make cDNA. TaqMan primers and probes were used to measure expression levels of CYP1A1 using RT-qPCR. GAPDH served as a control. While treatment with each PAH increased CYP1A1 expression compared with DMSO, DBA elicited the largest response, with a 54-fold increase in CYP1A1 expression (p<0.001). CYP1A1 expression was increased 16.5-fold with BbF treatment (p=0.001), and 7.4-fold with BaP treatment (p=0.006). In this study we find that DBA elicits a higher upregulation of CYP1A1 compared with BaP and BbF. Further experimentation is necessary using RNA-Seq analyses to help yield insight into pathways differentially responsive to DBA, yielding insight into its association with preterm delivery.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 153 Poster Session II: Poster #II-57

Sex-dependent effects of preconception exposure to arsenite on gene transcription in parental germ cells and on transcriptomic profiles and diabetic phenotype of offspring

Abhishek Venkatratnam, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

Chronic exposure to inorganic arsenic (iAs) has been linked to diabetic phenotypes in both humans and mice. However, diabetogenic effects of iAs exposure during specific developmental windows have never been systematically studied. The goal of the present study was to determine if preconception exposure alone can contribute to this outcome. This research is relevant to the development of methods to assess the risks to human health presented by hazardous substances. We have examined metabolic phenotypes in male and female offspring from dams and sires that were exposed to iAs in drinking water (0 or 200 ug As/L) for ten weeks prior to mating. The effects of iAs exposure on gene expression profiles in parental germ cells, and pancreatic islets and livers from offspring were assessed using RNA sequencing. We found that iAs exposure significantly altered transcript levels of genes, including diabetes-related genes, in the sperm of sires. Notably, some of the same gene transcripts and the associated pathways were also altered in the liver of the offspring. The exposure had a more subtle effect on gene expression in maternal oocytes and in pancreatic islets of the offspring. In female offspring, the preconception exposure was associated with increased adiposity, but lower blood glucose after fasting and after glucose challenge. HOMA-IR, the indicator of insulin resistance, was also lower. In contrast, the preconception exposure had no effects on blood glucose measures in male offspring. However, males from parents exposed to iAs had higher plasma insulin after glucose challenge and higher insulinogenic index than control offspring, indicating a greater requirement for insulin to maintain glucose homeostasis. Our results suggest that preconception exposure may contribute to the development of diabetic phenotype in male offspring, possibly mediated through germ cell-associated inheritance.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

154 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-58

Molecular dynamics simulations of water transport through crosslinked aromatic polyamide reverse osmosis membranes

Riley Vickers, University of North Carolina-Chapel Hill Timothy Weigand, University of North Carolina-Chapel Hill Cass T. Miller, University of North Carolina-Chapel Hill Orlando Coronell, University of North Carolina-Chapel Hill

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

In the United States, 14% of Americans use private wells as their drinking water source. Geogenic and anthropogenic contaminants such as inorganic arsenic, manganese, chromium, and vanadium can infiltrate these waters and lead to negative health outcomes. Reverse osmosis thin-film composite (RO TFC) membrane filtration is a US EPA recommended and energy efficient method to remove such contaminants from water at point of use and in centralized treatment plants. The work of separating contaminants from the influent water to produce a purified effluent water is performed by the extremely thin (~20-200 nm) crosslinked aromatic polyamide (XLPA) active layer. RO TFC membrane performance (water flux and contaminant rejection) is controlled by physicochemical properties of water, contaminants, and the XLPA active layer, and molecular-scale morphology of the XLPA active layer. However, none of these properties are explicitly accounted for in the most widely used method for analyzing membrane performance, the solution-diffusion model. In our work, we have found molecular dynamics (MD) simulations of water transport through XLPA active layers to be promising methods to develop a fundamental understanding of how the physicochemical properties of water and XLPA, and XLPA molecular morphology interact to determine the diffusive and advective transport of water through the membranes. Using these simulations, optimal polymer chemistries and morphologies can be rapidly identified as compared to the traditional build-and-test paradigm. We have validated MD methods to generate all-atom XLPA domains and measure pressure gradient behavior of transported water molecules during simulated operation. This approach provides a basis to simulate simultaneous water and contaminant transport—including inorganic arsenic, manganese, chromium, and vanadium—to aid in optimizing RO TFC membrane design for improved performance.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 155 Poster Session II: Poster #II-59

Intercalibration Comparison Study: Targeted GC/MS Compared to Untargeted Ion Mobility/MS

Terry Wade, Texas A&M University, College Station, Texas Gopal Bera, Texas A&M University, College Station, Texas James Dodds, North Carolina State University, Raleigh, North Carolina Erin S. Baker, North Carolina State University, Raleigh, North Carolina

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Texas A&M University Superfund Research Center

A study was designed with the objective of comparing the detection (presence/absence) and absolute quantification of environmental contaminants (especially PCB and PAH) between traditional targeted gas chromatography / mass spectrometry (GC/MS) and untargeted ion mobility – mass spectrometry (IM- MS). The study procedures included of the use NIST Standard Reference Material (SRM) 1944 which is a mixture of marine sediment collected near urban areas in New York and New Jersey. SRM 1944 has been used in evaluating analytical methods for the determination of selected polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyl (PCB) congeners, chlorinated pesticides, and trace elements in marine sediment and similar matrices. Reference values are also provided for selected polybrominated diphenyl ether (PBDE) congeners, selected dibenzo-p-dioxin and dibenzofuran congeners, total organic carbon, total extractable material, and particle size characteristics. All the constituents for which certified, reference, and information values are provided for SRM 1944 were naturally present in the sediment before processing. SRM 1944 and procedural blanks (with and without surrogates) were extracted using traditional methods with an Automated Solvent Extractor using dichloromethane, treated with copper to remove sulfur and further cleaned up using silica/alumina chromatography. The extract from each step were quantitatively split. All the extracts were then analyzed by IM-MS and the extract process through the entire clean up by GC/MS. The GC/MS analyses confirmed the certified concentrations for targeted PAH and PCB analytes that were present and will be compared to the results of untargeted IM-MS result. This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Researcher

156 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-60

Exploring the role of paraoxonase 2 in non-small cell lung carcinoma

Aaron G. Whitt, University of Louisville, Louisville, Kentucky Jin Jiu-Zhen, University of Louisville, Louisville, Kentucky Sengodagounder Arumugam, University of Louisville, Louisville, Kentucky Joseph Burlison, University of Louisville, Louisville, Kentucky Chi Li, University of Louisville, Louisville, Kentucky

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Paraoxonase 2 (PON2) is a ubiquitously-expressed intracellular enzyme with lactonase activity. Early studies into PON2 biology established a protective role against the development of atherosclerosis. More recent research has demonstrated PON2 is upregulated in various human tumors to promote survival and drug resistance. We recently observed that PON2 is upregulated in human NSCLC tumors. Thus, we hypothesized that PON2 may contribute to NSCLC survival and proliferation. We determined PON2’s role in cellular proliferation and metabolism in PON2-knockout (PON2-KO) mice, tissue lysates, primary cells, and NSCLC cell lines using traditional molecular biology, liquid chromatography-mass spectrometry (LC- MS), and high-resolution nuclear magnetic resonance (NMR). PON2-KO mice were generated using CRISPR/Cas9. PON2 status was confirmed by sequencing, western blot analysis, and enzymatic assay. Primary mouse embryonic fibroblasts (MEF) from wild type (WT) or PON2-KO animals were analyzed for PON2 activity using LC-MS. CRISPR/Cas9 was used to generate PON2-KO and vector control NCI-H1299 cells; intra/extracellular metabolites were quantified using NMR following 72h culture in uniformly-13C- labeled glucose. Statistical analysis was performed using Student’s unpaired t-test.PON2 status does not affect mouse body or organ weight. Sequencing, western blot, and enzymatic assay indicate loss of PON2 expression in PON2-KO mice. NCI-H1299 cells lacking PON2 expression exhibit decreased proliferation and reduced oxidative metabolism. Together, these findings indicate PON2 is dispensable for mouse development and normal cell proliferation, but required for NSCLC proliferation and oxidative metabolism. Thus, PON2 may serve as a selective therapeutic target against NSCLC.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 157 Poster Session II: Poster #II-61

Pandemic Possibilities: The Heart of the Matter

Lindsey Wood, University of Louisville, Louisville, Kentucky Madeline Tomlinson, University of Louisville, Louisville, Kentucky Delana Gilkey, University of Louisville, Louisville, Kentucky Jack A. Pfeiffer, University of Louisville, Louisville, Kentucky Savanna Kerstiens, University of Louisville, Louisville, Kentucky Joy L. Hart, University of Louisville, Louisville, Kentucky Kandi L. Walker, University of Louisville, Louisville, Kentucky Aruni Bhatnagar, University of Louisville, Louisville, Kentucky

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Physical distancing, mask wearing, and the host of actions necessary to prevent transmission of SARS-CoV- 2 are vital for public health, but present challenges in community engagement and research translation. Adhering to recommendations, the university paused clinical trials and research in communities. In this poster, we address methods—pandemic possibilities—employed to maintain contact with community members, translate research findings, and preserve interest in ongoing community collaborations. Prior to the pandemic, our RTC/CEC initiatives included extensions of previous work—The Heart of the Matter research translation program, which centers on cultivating deeper understanding of scientific findings, especially VOCs and cardiovascular health. This work ranged from elementary school educational programming to presentations at community meetings and production of a newsletter. Once pandemic prevention protocols were in place, we pivoted to preserve community partnerships. For example, we produced infographics (e.g., how COVID-19 was influencing planned research, VOCs and indoor air) that were distributed electronically. We participated in virtual neighborhood association and community meetings. Additionally, the Community Advisory Board converted to virtual meetings. This shift has gone so well that the Board is hosting a community-wide virtual meeting. Fortunately, the annual Art and Literature Showcase employed an online submission process, but, working with teachers and administrators, we recognized student winners virtually and through their schools. In collaboration with the Compassionate Schools Project, team members joined a virtual choir performance of Lean on Me. The quarterly newsletter, The Canopy, has taken on even more importance in conveying information and sustaining partnerships. Despite some cancellations (e.g., Green Screen community movie), the pandemic has led to new possibilities for community partnership, ultimately strengthening the work.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

158 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-62

Define Role of Peroxisome Proliferator-Activated Receptor Alpha (PPARα) in Arsenic Induced Toxicity

Xiaojing Yang, University of California, San Diego Elvira Mennillo, University of California, San Diego Andre Weber, University of California, San Diego Shujuan Chen, University of California, San Diego Robert Tukey, University of California, San Diego

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California, San Diego Superfund Research Center

Arsenic is ubiquitous environmental toxicant. Its ability to induce oxidative stress and subsequently initiate cellular and tissue damage is regarded as a major cause in its toxicity. The UDP- glucuronosyltransferase 1A1 (UGT1A1) is the sole enzyme responsible for the metabolism of bilirubin and its delay leads to the development of severe neonatal hyperbilirubinemia. Its expression is also closely associated with proliferation and maturation of intestinal epithelial cells (IECs) during the neonatal development. Peroxisome proliferator-activated receptor alpha (PPARα) is a nuclear receptor that serves as a xenobiotic and lipid sensor to regulate energy combustion, lipid homeostasis, and inflammation. The UGT1A1 gene can be transcriptionally induced by PPARα. In the present study, humanized UGT1 (hUGT1) neonatal mice, which display hyperbilirubinemia, were treated with a single dose of sodium m-arsenite (As3+) at 10 mg/kg. After 48 h, the length and weight of the small intestine was significantly increased in As3+ treated mice compared to vehicle group. The IEC specific maturation markers such as Sis and Akp3 were upregulated dramatically. Total serum bilirubin (TSB) levels were reduced to normal levels while intestinal UGT1A1 and Pparα were induced. To further investigate the potential crosstalk between As3+ generated intestinal maturation, UGT1A1 induction and Pparα gene activation, hUGT1/Pparα−/− mice were generated and treated with As3+ as described above. After 48 h, induction of gene expression demonstrated that the intestinal maturation marker gene Sis was significantly less in hUGT1/Pparα−/− mice compared to control mice. The expression of the UGT1A1 was also lower in hUGT1/Pparα−/− mice. These observations led us to speculate that induction of intestinal maturation and UGT1A1 expression is partially driven by activation of Pparα upon As3+ exposure.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 159 Poster Session II: Poster #II-63

Chronic arsenic exposure perturbs the gut microbiome and bile acid homeostasis in mice

Yifei Yang, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Liang Chi, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Chih-wei Liu, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Yun-Chung Hsiao, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Kun Lu, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

Gut microbiota affects human health through modulating host metabolism. Our previous studies have demonstrated that arsenic exposure can cause gut microbiota dysbiosis. However, it is unclear whether arsenic-induced gut microbiota dysbiosis plays a role in arsenic-perturbed metabolic imbalance in the host. Bile acids are important signaling molecules involved in numerous human disease, which are synthesized in host but also regulated by gut microbiota. In this study, we exposed arsenic (0.25 ppm and 1 ppm) to C57BL/6 mice via drinking water for 3 months, and investigated whether arsenic exposure affects the bile acid homeostasis and whether it is associated with the changes in gut bacteria. We found that arsenic exposure caused significant decrease of major unconjugated primary bile acids in serum. The ratios of unconjugated bile acids and taurine-conjugated bile acids were also decreased. In contrast, the hepatic level of several primary bile acids, including TMCA, MCA, TCA, was significantly increased in arsenic-treated mice. Additionally, secondary bile acids were consistently decreased in serum and liver by arsenic. Moreover, the relative abundance of Bacteroidetes and Firmicutes was differentially changed by arsenic exposure, which was strongly associated with the bile acid levels in serum. In summary, this study shows that arsenic exposure perturbs the bile acid homeostasis and arsenic-induced gut microbiota dysbiosis may impact bile acid metabolism, suggesting the potential role of arsenic-gut microbiota-bile acid interaction in arsenic-induced disease, including diabetes.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

160 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session II: Poster #II-64

Sexual Dimorphism of Cadmium-Exacerbated, Diet-Induced Liver Disease

Jamie Lynn Young, University of Louisville, Louisville, Kentucky Lu Cai, University of Louisville, Louisville, Kentucky

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Non-alcoholic fatty liver disease (NAFLD) affects more than 25% of the world’s population. Although obesity is a major risk factor for NAFLD, it does not account for all cases, suggesting the contribution of other factors such as sex and environmental exposures. Exposure to the non-essential metal cadmium is implicated in the development of NAFLD; however, the ability of early-life, in utero cadmium exposure to influence the development of obesity-induced NAFLD has not been studied. Additionally, studies do not take into account such environmental exposures may be life-long and multigenerational. Therefore, we developed an in vivo two-hit model to study the effect of whole life, low dose cadmium exposure on high fat diet (HFD)-induced NALFD. Adult male and female C57BL/6J mice fed normal diets (ND) were exposed to 0 or 5 ppm cadmium-containing drinking water for 14 weeks before breeding. At weaning, offspring were fed ND or HFD and continuously exposed to the same drinking water regiment as their parents. Water consumption and body weights were recorded weekly. Both male and female mice were sacrificed 24 weeks post-weaning. Exposure to cadmium altered HFD-associated weight gain in males, but not females. In males, exposure to cadmium altered HFD-associated changes in glucose tolerance and exacerbated HFD-induced liver disease, but not in females. These results are consistent with epidemiology showing female are less likely to develop NALFD, although they accumulate more cadmium in their body. Overall, this study provides insight into the role cadmium exposures during windows of susceptibility in the development of HFD-induced NAFLD and highlights the importance of considering sex as a risk factor in disease development.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 161 Poster Session III: December 15, 2020 | 12:00 – 1:00 p.m.

Last Name First Name Poster Session ID Abuawad Ahlam III-1 Aly Noor III-2 Armstrong Mikayla III-3 Ball Raymond III-4 Bera Gopal III-5 Bonsey Kendra III-6 Campbell Kyle III-7 Cardona Nancy III-8 Chen Celia III-9 Chernick Melissa III-10 Coarfa Cristian III-11 Crittenden Joshua III-12 Cruz Nathan III-13 Damond Jada III-14 NIEHS Dang Audrey III-15 Dean Rachel III-16 SRP Deng Pan III-17 Dias da Silva Priscilla III-18 2020 Ding Jiahui III-19 Dodson Jack III-20 Annual Meeting Eaves Lauren III-21 Ewald Jessica III-22 Fling Russell III-23 Foster MaKayla III-24 Gaither Kari III-25 Garrick Jacqueline III-26 George Andrew III-27 Ghetu Christine III-28 Gripshover Tyler III-29 Kaushal Simran III-30 Koempel Annie III-31 Kohale Ishwar III-32 Koike Shinichiro III-33 Lafarga Previdi Irene III-34

162 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: December 15, 2020 | 12:00 – 1:00 p.m.

Last Name First Name Poster Session ID LaRosa-LoPresti Savannah III-35 Li Hui III-36 Luo Yu-Syuan III-37 Martin Shannon III-38 Matsushita Megumi III-39 Moorthy Bhagavatula III-40 Morris Andrew III-41 Neville Amber III-42 Nigra Anne III-43 Odera Matilda III-44 Orlowska Karina III-45 Pettibone Kristi III-46 Qi Meng III-47 NIEHS Rivera Brianna III-48 SRP Roman-Hubers Alina III-49 Rudel Holly III-50 2020 Schroeder Julian III-51 Sethupathy Praveen III-52 Annual Meeting Shuey Chris III-53 Singh Nalin III-54 Stapleton Heather III-55 Sun Mohan III-56 Surolia Ranu III-57 Tachachartvanich Phum III-58 Tang Peixin III-59 Tao Zhihan III-60 Toyoda Jennifer III-61 Wang Meichen III-62 Weber Andre III-63 Yang Jun III-64 Zhang Chunyun III-65 Zhang Duo III-66 Zychowski Katherine III-67

NIEHS SRP 2020 Virtual Annual Meeting Program • 163 Poster Session III: Poster #III-1

Association between body mass index and arsenic methylation in three studies of Bangladeshi adults and adolescents

Ahlam Abuawad, Columbia University, New York, NY Miranda Spratlen, Columbia University, New York, NY Faruque Parvez, Columbia University, New York, NY Vesna Slavkovich, Columbia University, New York, NY Vesna Ilievski, Columbia University, New York, NY Angela M. Lomax-Luu, Columbia University, New York, NY Roheeni Saxena, Columbia University, New York, NY Hasan Shahriar, Columbia University, New York, NY Mohammad Nasir Uddin, Columbia University, New York, NY Tariqul Islam, Columbia University, New York, NY Joseph H. Graziano, Columbia University, New York, NY Ana Navas-Acien, Columbia University, New York, NY Mary Gamble, Columbia University, NY, NY

Superfund Mandates: Advanced techniques for detection, assessment, and eval. of effect on human health of hazardous substances

Superfund Center: Columbia University Superfund Research Center

Arsenic (As) exposure is a global health problem. Ingested inorganic As is methylated to mono-methyl (MMA) and dimethyl (DMA) arsenicals via one-carbon metabolism (OCM). People that inefficiently methylate As (higher relative% of urinary MMA), have been shown to have a higher risk of cardiovascular disease and several cancers but appear to have a lower risk of diabetes and obesity. Our objective is to characterize the association between As methylation and BMI in Bangladeshi adults and adolescents participating in the Folic Acid and Creatine Trial (FACT); Folate and Oxidative Stress (FOX) study; and Metals, Arsenic, and Nutrition in Adolescents Study (MANAS). The association between As species and concurrent BMI were analyzed using linear regression models, adjusting for nutrients involved in OCM such as choline. Mean BMIs were 19.2/20.4, 19.8/21.0, and 17.7/18.7 kg/m2 in males/females in FACT, FOX, and MANAS, respectively. BMI was associated with As species in female but not in male participants. In females, after adjustment for total urine As, age, and plasma folate, the adjusted mean differences (95% confidence) in BMI (kg/m2) for a 5% difference in MMA% and DMA% in urine were -0.79 (-1.29,- 0.30) and 0.51 (0.23,0.78), respectively in FACT, -0.45 (-1.07,0.17) and 0.29 (-0.05,0.62) in FOX, and -0.41 (-0.83,0.01) and 0.13 (-0.01,0.28) in MANAS. BMI was negatively associated with MMA% and positively associated with DMA% in females; associations were attenuated after plasma choline adjustment. These findings may be related to the role of body fat on estrogen levels that can influence OCM. Research is needed to determine whether the associations between BMI and As species are causal and their influence on As-related health outcomes.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

164 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-2

Environmental impacts of Hurricane Florence flooding in Eastern North Carolina: Temporal analysis of contaminant distribution and potential human health risks

Noor Aly, Texas A&M University, College Station, TX Gaston Casillas, Texas A&M University, College Station, TX Yu-Syuan Luo, Texas A&M University, College Station, TX Thomas J. McDonald, Texas A&M University, College Station, TX Terry L. Wade, Texas A&M University, College Station, TX Weihsueh A. Chiu, Texas A&M University, College Station, TX Ivan Rusyn, Texas A&M University, College Station, TX

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Texas A&M University Superfund Research Center

Hurricane Florence made landfall in Wilmington, North Carolina, on September 13, 2018 and torrential rains continued in the area for 5 days causing extensive flooding. There are a number of potential sources of hazardous substances and Superfund sites in Eastern North Carolina, many of these sustained water damage and contaminants may have been released into the environment. The objective of this study was to conduct temporal analysis of contaminant distribution and potential human health risks from Hurricane Florence-associated flooding. Soil samples were collected from 12 sites across four counties in Eastern North Carolina in September 2018, immediately after Hurricane Florence, as well as at two 4-month intervals thereafter (January and May 2019). Chemical analyses were performed for polycyclic aromatic hydrocarbons (PAHs), pesticides, polychlorinated biphenyls and other industrial chemicals by gas chromatography-mass spectrometry. Metals were analyzed using inductively coupled plasma mass spectrometry. We calculated hazard index and cancer risk at each site using EPA Regional Screening Level Soil Screening Levels (SSL) for residential soil. Elevated levels of PAH and metals, indicative of presence of a pyrogenic source of contamination (e.g., coal ash), were detected in the locations downstream from the coal ash storage pond that is known to have leaked due to Hurricane Florence-associated flooding. Importantly, levels of PAHs at these sites are of human health concern because cancer risk values exceeded 1×10-6 threshold. Levels of other contaminants measured across other sites, or corresponding hazard index or cancer risk, did not exhibit spatial or temporal differences. This work shows the importance of rapid exposure assessment following natural disasters that may cause re-distribution of hazardous substances. It also establishes baseline levels of contaminants for future comparisons. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 165 Poster Session III: Poster #III-3

Modifying commercial reverse osmosis membranes with solvent pre-treatment and additional polymerization can enhance water productivity and salt rejection

Mikayla D. Armstrong, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Orlando Coronell, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

In the US, 14% of Americans rely on private wells as their drinking water source. In NC, contaminants (e.g., inorganic arsenic, iAs) infiltrate private wells, placing ~3 million residents at risk for adverse health outcomes. The US EPA and NC-DHHS recommend using reverse osmosis (RO) membrane filtration to treat contaminated well-water. RO membranes are polymeric materials made from an interfacial reaction between monomers, trimesoyl chloride (TMC) and m-phenylenediamine (MPD). An important challenge in membrane technology is to advance the trade-off between water productivity and contaminant rejection, particularly for small, neutral contaminants like iAs(III) and for house-hold systems that need high flow rates. Treating RO membranes with solvent has been shown to enhance their water productivity but at the expense of contaminant rejection. We hypothesize that solvent pre-treatment followed by additional polymerization may result in a membrane that exhibits enhanced water productivity and contaminant rejection (e.g., iAs). In our work, we subjected RO membranes to three sequential modification steps: 1) pre-treatment with an organic solvent, 2) exposure to TMC in an organic solvent, and 3) exposure to MPD in water. The first step swells the membrane, and the second step impregnates it with TMC, facilitating additional polymerization during the third step. We evaluated the solvent used in the solvent pre-treatment, polymerization time, and monomer concentrations. Initial results show that certain combinations of modification parameters resulted in membranes with higher water permeance and salt rejection than the unmodified RO membranes. Future experiments will focus on testing the promising modified membranes for the rejection of iAs. Our work builds on the concept of solvent pre- treatment to develop membranes with improved water productivity and iAs rejection.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

166 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-4

Combined In-Situ and Ex-Situ Remediation of PFAS at Hazardous Waste Sites

Raymond Ball, EnChem Engineering, Inc. Dr. Thomas Boving, University of Rhode Island

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: None: EnChem Engineering, Inc.

The objective of our technology demonstration is to show that in-situ flushing of PFAS from a contaminated aquifer source area with XCT® (extra contact technology) solution can reduce PFAS soil and groundwater concentrations. Extracted PFAS are destroyed ex-situ and on-site. The treatment process consists of an enhanced foam fractionation step that concentrates the PFAS by 1 to 2 orders of magnitude, producing clean water for discharge. The destruction of PFAS is facilitated by UV enhanced oxidation- reduction. Recovered and treated XCT flushing solution can be recycled by reinjection into the treatment zone. Ongoing laboratory tests of our XCT® process, which relies on a complex sugar and salts to desorb PFAS from soil for subsequent extraction and ex-situ treatment, show the effectiveness of this approach and its advantages. Specifically, the complex sugar is non-toxic and biodegradable under either aerobic or anaerobic conditions and there are no toxic by-products remaining in the subsurface after soil flushing. Further, any potential of PFAS migration away from the source area can be controlled using the push-pull method. Importantly, this in-situ flushing technology can be used to remove PFAS in saturated zone soil under buildings or high traffic areas (e.g., airports), allowing operations to continue, which is very attractive to the site owners. Together with speediness of the clean-up process, our approach saves energy and total project costs. Our treatment system is designed to be transportable and modular, such that it can be quickly mobilized and de-mobilized from one site to another. We anticipate performing our demonstration test at a military base with PFAS in the subsurface.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 167 Poster Session III: Poster #III-5

Post-Harvey: Status of sediment contaminants along the upper Houston Ship Channel, Texas

Gopal Bera, Texas A&M University, College Station, TX Tim Dellapenna, Texas A&M Galveston, TX Jose Sericano, Texas A&M University, College Station, TX Kusumica Mitra, Texas A&M University, College Station, TX Terry L. Wade, Texas A&M University, College Station, TX Anthony Knap, Texas A&M University, College Station, TX

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Texas A&M University Superfund Research Center

The Houston Petrochemical complex is located along the upper Houston Ship Channel (HSC), within the lower Buffalo Bayou and the San Jacinto Estuary. The petrochemical complex became established and experienced rapid expansion beginning in the early 1930’s. As a direct consequence, by the late 1960’s, these water bodies were among the top ten most polluted water bodies in U.S. Historically, HSC area contains elevated concentrations of polychlorinated biphenyls (PCBs) when compared with similar size industrial areas worldwide. The results from post Hurricane Ike study (2010-2012) found PCBs and PCDDs/PCDFs concentrations above Texas DSHS guidelines (for human consumption) in blue crabs and fish from the HSC while organochlorine pesticides have a declining trend. Hurricane Harvey (2017) produced an estimated input of 14 × 109 m3 of freshwater and deposited 9.9 × 107 metric tons of sediments to Galveston Bay. Flood from Hurricane Harvey inundated a large number of refineries, chemical plants and legacy waste sites and released 0.57 × 106 tons of raw sewage and about 22,000 barrels of oil, refined fuels and chemicals to the Galveston Bay estuary. Nonetheless, it is obvious that Hurricane Harvey has caused a significant loading of pollutants and redistribution in the region of upper the HSC area. This in turn has provided an opportunity to investigate the present status of sediment contaminants along the HSC area. Surface sediments were collected in 2019 from 20 outfall stations along the upper HSC. High concentrations of total PCBs (highest 2414 ng/g), total PBDEs (60 ng/g), total PAHs (74 µg/g) and mercury (3.6 µg/g) were found in some these outfall stations. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Researcher

168 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-6

Influence of the gut microbiome on in vivo bioavailability and transformation of particle-bound arsenic

Kendra Bonsey, University of Arizona, Tucson, Arizona Rob Root, University of Arizona, Tucson, Arizona Pawel Kiela, University of Arizona, Tucson, Arizona John Chorover, University of Arizona, Tucson, Arizona

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of Arizona Superfund Research Center

Arsenic (As) is a toxic and carcinogenic metalloid implicated in the development of a wide variety of diseases. Legacy mine tailings enriched in As pose a human health risk, particularly in (semi)-arid environments where tailings lack vegetative cover and are susceptible to wind dispersion, which results in inhalation and ingestion exposure to As-bearing geodusts in proximal communities. In order to accurately determine the human health risk posed by inhalation or ingestion of airborne particle-bound As, reliable site-specific bioavailability assessments for ingested As are needed. Arsenic bioavailability, which controls its relative toxicity, is dependent upon its molecular form (i.e., speciation) in the ingested material; therefore, equivalent dosages of As ingested from different environmental media may not pose equal health risk. While bioavailability measurements rely on the use of animal models, in vitro bioaccessibility assays (IVBAs) using a simple gastrointestinal (GI) fluid simulant have been shown to be a valuable predictive measure of bioavailability. However, emerging evidence indicates that the gut microbiome performs key transformations of As that impact its speciation in the GI system and, therefore, traditional abiotic IVBA may not accurately predict As bioavailability and toxicity. Our project seeks to improve GI As transformation models by performing in vivo bioavailability assays using both germ-free and intact microbiome mice exposed to particle-bound As in the form of mine tailings and As-bearing minerals. These assays will inform our understanding of the impact of the gut microbiome on As biotransformation and bioavailability. Ultimately, this information will aid in the optimization of predictive IVBA for site-specific As bioaccessibility determinations, which will improve the efficacy of human health risk assessment for mining impacted communities.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 169 Poster Session III: Poster #III-7

Placental villous tissue analysis for toxicology and epidemiology: complementary single-cell and sorted cell type-specific gene expression profiling for estimation of cell type abundance

Kyle Campbell, University of Michigan Justin A. Colacino, University of Michigan Muraly Puttabyatappa, University of Michigan Sue Hammoud, University of Michigan Steven E. Domino, University of Michigan Dana C. Dolinoy, University of Michigan Vasantha Padmanabhan, University of Michigan Jaclyn Goodrich, Rita Loch-Caruso, and Kellly M. Bakulski, University of Michigan

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Northeastern University Superfund Research Center

Placental development is sensitive to environmental exposures and critical for fetal-maternal health. Approaches that account for placental cell-type abundance will advance understanding of toxicological impacts on placental and pregnancy health. We dissociated placental villous tissue into cell suspensions for unbiased single-cell RNA-sequencing and targeted fluorescence-activated cell sorting of cell types (cytotrophoblast, endothelial, extravillous trophoblast, fibroblast, Hofbauer cells, leukocyte, and syncytiotrophoblast-enriched). We sequenced 8,325 single-cell libraries from two placentas in replicate. We observed six lymphocyte and five placental cell clusters. Cytotrophoblasts overexpressed 78 genes (padj1) relative to other placental cells and were enriched for epithelial cell differentiation pathway (p=1.3×10-6). Among 24 sorted-cell samples across four placentas, Hofbauer cells (placental resident macrophages) overexpressed 1,310 genes (padj1) and were enriched for defense responses (p=4.0×10-6). Expression profiles from single-cell profiling and cell sorting were highly correlated (e.g., Hofbauer cells, ρ=0.76). Next, we applied our results to model cell type abundance in previously published studies of placenta-related conditions. Using bulk placental tissue expression from 157 preeclamptic mothers and 173 controls, we estimated cell type proportions and tested for differences using multivariate beta regression. Fibroblasts (odds ratio = 0.77, p<.0001) and Hofbauer cells (odds ratio = 0.82, p<.0001) were less abundant in placental tissue from pre-eclamptic patients compared with controls, whereas extravillous trophoblasts were more abundant (odds ratio = 1.65, p<.0001). In term placental villous tissue, using complementary technologies, we provided robust cell type-specific gene expression profiling. We applied these profiles to deconvolute an epidemiologic bulk placental tissue preeclampsia dataset and identified biologically important cell type abundance differences, which may underscore exposure and disease mechanisms in the placenta.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

170 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-8

Multi-agency methodology approach to reduce phthalate exposure in the gestational period: An ECHO-PRO study

Nancy R. Cardona, University of Puerto Rico C.M. Velez-Vega, University of Puerto Rico J.F. Cordero, University of Georgia A. Alshawabkeh, Northeastern University J. Meeker, University of Michigan

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Northeastern University Superfund Research Center

In a developing world, reducing gestational exposure to consumer products ingredients associated with altered neurodevelopmental effects during infancy and childhood is a public health challenge locally and globally. Pound for pound, children breathe more air, drink more water, and eat more food than adults; therefore, children are often more exposed to chemicals of concern than adults. Likewise, children’s behavior patterns and activities could increase their exposure to harmful chemicals. The Environmental Protection Agency (EPA) provides steps to develop human health risk assessments to estimate the nature and probability of adverse health effects in humans who may be exposed to chemicals in contaminated areas. The Environmental Influences in Child Health Outcomes Program in Puerto Rico (ECHO-PRO) through this project aims to identify predictors of phthalate exposures associated with adverse effects in infant and children necessary for the development of strategies towards minimizing them. Currently we have data on phthalates levels at three times during pregnancy for 1,090 participants. As part of our ongoing study we are identifying sources and pathways of aggregate exposure by evaluating lifestyle factors including consumer products, food consumption and phthalate urine metabolite levels in participants. A risk assessment will be developed using phthalate-containing consumer products to provide a baseline to compare among participants using phthalates-containing consumer products and associated levels of exposure. Looking at the combined burden of these products will enhance environmental health risk assessments, a first step in gestational exposures to phthalates aiming at policies that will protect and promote children’s health.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 171 Poster Session III: Poster #III-9

Mercury concentrations in commercially important fisheries species in the Gulf of Maine, USA

Celia Chen, Dartmouth College Superfund Research Center K. L. Buckman, Dartmouth College Superfund Research Center V. F. Taylor, Dartmouth College Superfund Research Center

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Dartmouth College Superfund Research Center

The Gulf of Maine is a complex ecosystem host to numerous productive commercial fisheries. In the Gulf of Maine, data for mercury (Hg) in sediments and in mussels have been documented along the coastline but very little is known about the concentrations in the offshore food webs of either the inshore or offshore regions of the Gulf. To date there are no publicly available data on Hg levels in commercial species for offshore regions where most of the productive fisheries are located. The current study examined total Hg, selenium (Se), and total lipid concentrations in filets from six commercial species: Atlantic cod (Gadus morhua), spiny dogfish (Squalus acanthias), Atlantic herring (Clupea harengus), red hake (Urophycis chuss), silver hake (Merluccius bilinearis), and skates (Leucoraja spp.) captured in both nearshore and offshore locations. Individuals of Atlantic cod, spiny dogfish, and skate presented tissue concentrations above the US EPA 300ppb regulatory threshold. Most species indicated a strong positive relationship of Hg with length (width for skates), as anticipated, with species-specific differences in Hg concentration observed. Se concentration was less variable among species than Hg. Relationships with catch location and feeding mode for Hg, Se, and lipids will be discussed.

Email: [email protected] Presenter Status: Researcher

172 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-10

Heart Development in Two Populations of Atlantic Killifish (Fundulus heteroclitus) Following Exposure to Elizabeth River Sediment Extract

Melissa Chernick, Duke University Tara Burke, Duke University, and EPA Noah Lieberman, Duke University Daniel R. Brown, Western Carolina University, Cullowhee Richard T. Di Giulio, Duke University David E. Hinton, Duke University

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Duke University Superfund Research Center

Atlantic killifish (Fundulus heteroclitus) inhabiting the Atlantic Wood (AW) Industries Superfund site in the Elizabeth River, Virginia have adapted to historically high levels of PAHs. Embryos have proved highly resistant to cardiac abnormalities indicative of PAH toxicity. In this study, embryos spawned from adults collected at AW and King’s Creek (KC), a reference site, were exposed at 24 hours post fertilization (hpf) to Elizabeth River Sediment Extract (ERSE) that comprised a range of PAH concentrations (0, 5.04, 50.45, 100.90, 151.35, or 252.25 µg/L total PAHs). Embryos were processed for histology at 144 hpf for evaluations of hearts at tissue and cellular levels. Morphometry and severity scoring were used to evaluate the extent of alterations. Unexposed embryos were similar in both populations. ERSE exposure resulted in multiple changes to hearts of KC embryos but not AW. Alterations were particularly evident in KC embryos exposed to concentrations above 1% ERSE, which had thinner ventricular walls and larger pericardial edema. Individuals with moderate pericardial edema maintained arrangement and proximity of heart chambers, but ventricular myocytes were altered. Severe pericardial edema was prevalent in exposed KC embryos and typically resulted in tube heart formation. Ventricles of tube hearts had very thin walls with small, basophilic cells and lacked trabeculae. Edematous pericardial fluid contained small amounts of proteinaceous material, as did controls, and was free of cells. This fluid was primarily unstained, suggesting water influx due to increased permeability. The use of histological approaches provided more specific detail for tissue and cellular effects in hearts of embryos exposed to PAHs and enabled understanding of potential links to later life effects of early life exposure.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 173 Poster Session III: Poster #III-11

A web-based framework for analyses and interpretation of polycyclic aromatic hydrocarbon (PAH) transcriptomic profiles

Christian Coarfa, Baylor College of Medicine, Houston, Texas Matthew J. Robertson, Baylor College of Medicine, Houston, Texas Tanmay Gandhi, Baylor College of Medicine, Houston, Texas Sandra L. Grimm, Baylor College of Medicine, Houston, Texas Heidi Dowst, Baylor College of Medicine, Houston, Texas Krithika Lingappan, Texas Children’s Hospital, Houston, Texas Bhagavatula Moorthy, Texas Children’s Hospital, Houston Texas Susan G. Hilsenbeck, Baylor College of Medicine, Houston, Texas

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Baylor College of Medicine Superfund Research Center

Proximity to Superfund sites in the Greater Houston area contribute to increased risks of PAH exposures, which may lead to COPD, lung cancer, and liver cancer. The availability of low-cost RNA-Sequencing makes it more affordable than ever to generate transcriptomic profiles from models of exposure to PAHs, or from residents exposed to PAH. Whereas basic analysis of RNA-Seq can be done effectively using off-the- shelf tools, interpretation of the findings in the context of existing knowledge is challenging. To enable effective analysis and interpretation of PAH transcriptomic results, and by leveraging the wide-range of capabilities at the Data Analysis and Management Core at the BCM/Rice Superfund Center, we are adopting a multi-pronged approach by (1) compiling gene signatures from PAH exposures in lung and liver, both in vitro and in vivo; (2) compiling clinical cohorts of COPD, lung and liver cancer; (3) implementing a set of tools, via R/Shiny, and enabling users to interrogate publicly available data using their own PAH transcriptomic data. Using our platform, users can upload a gene signature generated from analysis of their own transcriptomic data or qPCR validation, selecting the tissue (lung or liver currently) and species. Our platform would then provide similarity scores and rankings with the compendium of PAH signatures, correlation of user signatures with existing PAH signatures over healthy or diseased tissue cohorts, and clinical association with the disease cohorts. Our platform to be scalable so it can be extended as needed to other tissues or exposure models and clinical endpoints. Our overall goal is to help integrate each of our projects and cores, so new collaborations can be fostered, ultimately leading to novel research strategies.

Email: [email protected] Presenter Status: Researcher

174 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-12

Identification and Isolation of Polycyclic aromatic hydrocarbon (PAH) degrading fungi and bacteria in estuarine-influenced creosote contaminated sediment, Elizabeth River, VA

Joshua A. Crittenden, Duke University, Durham, North Carolina Savannah J. Volkoff, Duke University, Durham, North Carolina Daniel Raudabaugh, Duke University, Durham, North Carolina David Singleton, Duke University, Durham, North Carolina Claudia K. Gunsch, Duke University, Durham, North Carolina

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Duke University Superfund Research Center

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants and are notorious for their toxicity and carcinogenicity. Bioremediation using microorganisms is an economically feasible and environmentally sustainable process for PAH remediation projects globally but is limited by an incomplete understanding of the biology behind the process. We examined PAH-contaminated sites along the Elizabeth River (Virginia, USA) and used a variety of approaches to identify and isolate PAH-degrading bacteria and fungi from creosote-contaminated sediments. Indigenous fungi were isolated and examined for extracellular enzymatic production using a variety of selective media. PAH-degrading bacteria from the Republic Creosoting site were directly identified through the use of DNA stable-isotope probing (DNA- SIP) with phenanthrene (PHE) and fluoranthene (FLA). DNA-SIP associated sequences were primarily associated with the genera Croceicoccus (PHE), Azoarcus (FLA), and Hydrogenophaga (FLA). Targeted isolation of PAH-degrading bacteria on PHE, FLA, and pyrene resulted in recovery of strains primarily associated with the genera Sphingobium, Novosphingobium, and Hydrogenophaga, the last of which was also detected in the DNA-SIP experiments. Fungal enzymatic assays resulted in additional screening of root and soil fungal isolates, which were later incubated in the presence of PAHs and tested for transformation capability by measuring parent compound disappearance by high-performance liquid chromatography (HPLC). Isolated organisms and information obtained from these analyses will be used to develop a bioaugmentation strategy for cleanup of a portion of the Republic Creosoting site.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 175 Poster Session III: Poster #III-13

Barrier and Wound Healing Impacts of Polystyrene Microplastic Particles on an Airway Epithelial Model (A549)

Nathan Cruz, University of New Mexico, Alburquerque, New Mexico Guy Herbert, University of New Mexico, Alburquerque, New Mexico Matthew Campen, University of New Mexico, Alburquerque, New Mexico

Superfund Mandate: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

Microplastic particles have been identified in the most remote regions of the planet, representing an emerging environmental health concern. While limited data suggest that microplastics pose a health hazard for humans, concern remains due to 1) a lack of focused research on the topic and 2) the exponential rate of global plastic waste accumulation. Importantly, microplastics are observed in water, food, soil, and even air, threatening multiple routes of human exposure. The present study examined the effects of controlled delivery of manufactured polystyrene microspheres, as an initial laboratory-based model, on the barrier integrity and wound healing capacity of adenocarcinomic human alveolar basal epithelial cells (A549). We hypothesized the presence of MP of all sizes would present adverse effects on reestablishing electrical resistance in a dose-dependent manner. A549 cells were grown to confluence on an electric cell-substrate impedance (ECIS; Applied Biophysics) plate and treated with 1µm, 5µm, or a mix of 1 and 5µm microspheres at concentrations ranging from 0.1 to 10 µg/ml. Alveolar epithelial monolayer resistance was recorded continuously for all wells. After 24h, cells were subjected to an electric current over a limited portion of the ECIS well to induce injury; the reestablishment of electrical resistance after this “wounding” was then assessed to determine the impacts of microplastic exposure on cellular regrowth. Results illustrated both barrier function and wound healing were significantly compromised, with the mixture of 1 and 5 µm microspheres having an additive negative effect. Interestingly, a dose- dependent response was not observed in cell membrane integrity. These data suggest that microplastics can have a direct impact on normal airway epithelial homeostasis.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

176 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-14

Novel Equilibrium Passive Sampling Device for the Measurement of Methylmercury in Water and Porewater of a Sediment Microcosm

Jada Damond, University of Maryland, Baltimore County, Baltimore Spencer Washburn, Smithsonian Environmental Research Center Cynthia Gilmour, Smithsonian Environmental Research Center Upal Ghosh, University of Maryland, Baltimore County, Baltimore

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: R01

Mercury is a global pollutant that can undergo methylation in natural systems to form the more toxic and bioaccumulative form, methylmercury (MeHg). MeHg can biomagnify within aquatic biota to levels unsafe for human consumption, making accurate determination of its concentration across aquatic environments crucial for adequate risk assessment. However, MeHg concentrations in waters and porewaters can be low (concentrations in unimpacted waters at the pM level) and highly variable through time, rendering accurate measurement with traditional field sampling methods challenging. Passive sampling approaches for predicting methylmercury concentrations, which can reflect a time-averaged concentration of MeHg in water and porewater, have thus been explored. The present work describes the development of a novel equilibrium-based passive sampling device comprised of activated carbon suspended in agarose gel and demonstrates the ability of this novel sampler to measure water and porewater MeHg concentrations in a sediment microcosm. Additionally, it shows the ability of a single passive sampler to measure MeHg concentrations across a sediment-water interface. Passive samplers inserted partially in the water column and partially in the sediment column successfully measured the gradient of dissolved MeHg concentration demonstrating the ability to measure concentration gradients critical for flux calculations. The results of this experiment demonstrate the capability of this material to predict water and porewater MeHg concentrations in an environmentally relevant setting.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 177 Poster Session III: Poster #III-15

Developing a multi-channel gas chromatograph – mass spectrometer for mobile measurements of volatile organic compounds (VOCs) in ambient air

Audrey Dang, Washington University in St Louis, St Louis Missouri Nathan Kreisberg, Aerosol Dynamics Jay Turner, Washington University in St Louis, St Louis Missouri Brent Williams, Washington University in St Louis, St Louis Missouri

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Louisville Superfund Research Center

Emitted by diverse anthropogenic and biogenic sources and subsequently subjected to dilution, transport, and chemical reaction, volatile organic compounds (VOCs) exhibit spatial and temporal variability. One approach to describing such spatial variability is land use regression (LUR), which relates how concentration depends on land use variables. While spatially distributed networks of passive samplers have been used to generate datasets for LUR modeling of VOCs, other air pollutants have also been characterized by time-resolved mobile measurements with in situ instruments. Such in situ instruments have also been used for contemporaneous measurements of outdoor and indoor air, which both influence exposure and are subject to dynamic changes. In this project, we are developing a gas chromatography – mass spectrometry (GCMS) instrument suitable for in situ, mobile measurements of VOCs with lesser operational complexity than required for costly, state-of-the-art instruments (e.g. proton transfer reaction – mass spectrometer). More modest cost and physical footprint are achieved with compact, lower cost residual gas analyzers and miniaturized gas chromatographs (GC). Higher time resolution (5-10 minutes_ for mobile measurements is enabled by incorporating multiple channels (4 collection channels, 4 GC separation channels, 2 MS detection channels) in a single instrument. The multiple channel design is also well-suited to simultaneous, time-resolved measurements of indoor and outdoor air. We report laboratory characterization of the measurement capabilities of the instrument. Development of this new analytical instrument (Tool Testing, Application and Synthesis) is intended to address resource limitations in current Research Practice for estimating exposure to VOCs (Observation, Group and Population, Human) and for evaluating interventions (Intervention Pilot Testing, Application and Synthesis).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

178 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-16

In search of a dioxin dehalogenase – voyage from the Passaic River

Rachel K. Dean, Michigan State University, East Lansing, Michigan Gerben Zylstra, Rutgers University, New Brunswick, New Jersey Donna Fennell, Rutgers University, New Brunswick, New Jersey

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

The lower Passaic River in New Jersey is highly contaminated with toxic chemicals called dibenzo-p- dioxins, which can persist in the environment and cause serious health problems to exposed individuals for many years. It is the location of the Diamond Alkali Superfund site, so named for one of the major polluters of the river. We recently showed that bacteria in sediments from the Passaic River were capable of dechlorinating the most toxic dibenzo-p-dioxin congener, 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8- TeCDD). This activity was only observed when sediment bacteria were first enriched with 1,2- dichlorobenzene and also co-amended with 1,2-dichlorobenzene during incubation with 2,3,7,8-TeCDD. 16S rRNA gene sequencing indicated that the most abundant phylotype in this culture belonged to the class Dehalococcoidia, to which the known dechlorinating bacterial genera Dehalococcoides and Dehalogenimonas belong. We shogtun-sequenced the metagenome of this 2,3,7,8-TeCDD-dechlorinating culture and identified putative reductive dehalogenase (RdhA) gene sequences. Reductive dehalogenases are the class of enzymes which carry out the removal of chlorines from chlorinated compounds under anoxic conditions. Thirteen high-coverage RdhA genes were identified from our metagenome. Our next steps are to attempt heterologous expression of top RdhA candidates and test the ability of these enzymes to dechlorinate 1,2-dichlorobenzene and/or 2,3,7,8-TeCDD. Biochemical knowledge about these enzymes will contribute to the growing database of functionally-characterized RdhA, which can help inform the field of organohalide bioremediation and may provide future solutions for the Passaic River. Given that this site is at high risk to be impacted by climate change-related severe storms and sea level rise, timely progress with this research is crucial.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 179 Poster Session III: Poster #III-17

Dietary fiber consumption protects against dioxin-like PCB126-induced ceramide production in hyperlipidemic mice liver

Pan Deng, University of Kentucky, Lexington, Kentucky Jianzhong Chen, University of Kentucky, Lexington, Kentucky Jessie B. Hoffman, University of Kentucky, Lexington, Kentucky Andrew J. Morris, University of Kentucky, Lexington, Kentucky Bernhard Hennig, University of Kentucky, Lexington, Kentucky

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Kentucky Superfund Research Center

Polychlorinated biphenyls (PCBs) are persistent environmental pollutants, and exposure to PCBs promotes the production of bioactive lipids, which increases the risk of developing cardiometabolic diseases. Ceramides are important pro-inflammatory lipids which are drivers of tissue dysfunction that underlies cardiometabolic diseases. Diets high in soluble fibers have proven effective at modulating lipid metabolism, and especially of ceramides. In this study, the effects of the most potent dioxin-like pollutant PCB126 and dietary inulin intervention on lipid homeostasis were examined in an atherosclerosis mouse model. Four groups of male Ldlr-/- mice were fed a high cholesterol diet containing 8% inulin or 8% cellulose (control) for 12 weeks. At weeks 2 and 4, mice were exposed to PCB 126 or vehicle (control). Liver samples were collected at the end of the study and analyzed using LC-MS lipidomic method. Compared with controls, PCB126 exposed mice showed significant elevated hepatic ceramide levels; on the contrary, there were no changes in levels of phospholipids and neutral lipids. The elevated production of ceramides induced by PCB126 was ameliorated in inulin-fed mice. Liver qPCR analysis of genes responsible for biosynthesis of ceramides revealed that Smpd3, a gene that encodes neutral sphingomyelinase (NSMase), was upregulated after PCB126 exposure, and this effect was counteracted by inulin-feeding. Taken together, these results showed that inulin can protect against PCB126 induced production of ceramides through the NSMase pathway, suggesting a new mechanism by which dietary fiber could reduce environmental pollutant induced cardiometabolic disease risk. (Supported by NIEHS/NIH grants P42ES007380).

Email: [email protected] Presenter Status: Researcher

180 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-18

Pyrolytic Treatment of Soils Contaminated with Polyaromatic Hydrocarbons: Kinetics and Reactor Design

Priscilla Dias da Silva, Rice University, Houston, Texas Ye Gao, Rice University, Houston, Texas Pedro J.J. Alvarez, Rice University, Houston, Texas Kyriacos Zygourakis, Rice University, Houston, Texas

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Other: Rice University

Soils contaminated with persistent PAH priority pollutants represent a long-term source of contaminant release into surface water, groundwater, edible biota and air exposure pathways. Current remediation methods are either relatively slow or have unintended consequences such as soil damage, high-energy usage or generation of more noxious PAH derivatives. Thus, there is a pressing need for more efficient and sustainable remediation of soils contaminated with PAHs and other recalcitrant lipophilic organic pollutants. Our group has developed a novel pyrolytic method for ex-situ remediation of soils contaminated with heavy petroleum hydrocarbons that contain PAHs. This approach can rapidly and reliably remove total petroleum hydrocarbons (TPH) and PAHs with lower energy requirements and better post-treatment soil fertility than other ex-situ thermal remediation approaches. However, our studies also revealed potential tradeoffs between treatment intensity, soil detoxification efficacy and soil fertility restoration. Using the Distributed Activation Energy kinetic model for the pyrolysis of petroleum- contaminated soils, we simulated the operation of a non-isothermal rotary kiln reactor that may be used for ex situ treatment of soils contaminated with PAHs. Model predictions were compared to experimental data obtained by treating two contaminated soils. The overall TPH and PAH reductions predicted by the model agreed well with the experimental data for all the conditions considered. More importantly, our model was able to explain the trade-offs observed in our earlier study between pyrolysis treatment intensity, soil detoxification and soil fertility restoration. We will finally present results that extend our theoretical models to full-scale kiln reactors. Our focus here will be on providing guidelines for estimating the optimal operating conditions (pyrolysis temperature and residence time) for contaminated soils with various soil properties and pollutant content.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 181 Poster Session III: Poster #III-19

Benzene Exposure During Pregnancy Leads to Fetal Resorption and Intrauterine Growth Retardation in Mice

Jiahui Ding, Wayne State University, Detroit, MI Anthony Maxwell, Wayne State University, Detroit, MI Marianna Sadagurski, Wayne State University, Detroit, MI Lucas Debarba, Wayne State University, Detroit, MI Douglas M Ruden, Wayne State University, Detroit, MI Michael C Petriello, Wayne State University, Detroit, MI Gil Mor, Wayne State University, Detroit, MI

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Unaffiliated

Problem: Benzene is a colorless, flammable liquid with a gasoline-like odor, which is a major source of environmental pollution. Benzene exposure during pregnancy in humans is associated with several adverse outcomes, such as preterm birth, low birth weight and childhood leukemia in the offspring. Several studies have shown the direct effect of benzene in various fetal organs such as the brain. However, the mechanism associated with these detrimental outcomes induced by benzene exposure during pregnancy is poorly understood. Here we tested the hypothesis that exposure to benzene induces maternal inflammation, which will lead to intrauterine growth restriction (IUGR) and abnormal placental function. Methods of Study: Female C57BL/6 pregnant mice were exposed to benzene (50 ppm) for 5h/day from E0.5 to E17.5 using inhalation chambers. On E17.5, mice were sacrificed and tissues for RNA sequencing were collected. Pregnancy outcome and fetal parameters were evaluated, and cytokine expression was determined by Luminex. Results: Exposure of benzene during pregnancy leads to maternal inflammation characterized by increased concentrations of circulating pro-inflammatory cytokines, including IL-1α, IL-1β, IL-10, IL-12 (p70), Eotaxin, TNFα, IL-17 and IFN-gamma. Fetal resorption, placental hemorrhaging, and IUGR were all observed in our model. These phenotypes correlated to our RNA sequencing data, which showed a severe inflammatory response in the placenta. Conclusions: We report the characterization of a mouse model of maternal inflammation associated with the exposure to benzene during pregnancy. Our findings correlate with human studies describing the association between maternal exposure to benzene and pregnancy complications, such as preterm birth and IUGR. Our data suggests that benzene-induced maternal and placental inflammation may be the mechanism responsible for these pregnancy complications.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

182 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-20

Development of an interactive, R-based application for analysis of toxicogenomic data

Jack P. Dodson, Michigan State University, East Lansing, Michigan Rance Nault, Michigan State University, East Lansing, Michigan Tim Zacharewski, Michigan State University, East Lansing, Michigan

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

Initiatives that promote Findable, Accessible, Interoperable, and Reusable (FAIR) data principles are becoming a key criteria of public research funding agencies. FAIR principles enable new knowledge to be derived from existing data with minimal additional investment in data generation. The Michigan State University (MSU) Superfund Research Center (SRC) has produced vast amounts of data characterizing the mechanisms and impact of exposure to aryl hydrocarbon receptor (AHR) ligands. To promote their FAIRness, we have developed FAIRTox, an open-source web-based data exploration, visualization, and analysis application for toxicogenomic datasets. FAIRTox is built using a R Shiny framework, chosen due to its wide use as a tool for omics data analysis, and the availability of other R packages for enrichment and multidimensional analyses such as principal component analysis (PCA). Unprocessed and analyzed datasets are stored in a SQL relational database enabling querying through a user-friendly interface. Metadata filters allow users to visualize and compare gene expression responses to various experimental factors such as zeitgeber time, dose, and duration of exposure to various environmental contaminants. Enrichment and advanced analysis features also facilitate the integration of datasets furthering the development of novel hypotheses. In addition, the implementation of FAIRTox makes publicly available MSU SRC toxicogenomic data more accessible to researchers without transcriptomic expertise. FAIRTox is implemented as a Docker container ensuring portability and reproducibility for groups looking to execute their own local version. Ultimately, the goal of FAIRTox is to improve data sharing, reuse, and reproducibility through an intuitive interface that serves bioinformaticians and novices alike. Funded by the Superfund Research Program P42ES04911 and NHGRI R21HG010789.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 183 Poster Session III: Poster #III-21

Multi-metal analysis of private well water in North Carolina: implications for public health

Lauren A. Eaves, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Alexander P Keil, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Rebecca C Fry, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina l

Superfund Mandate: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

Background. In North Carolina (NC), nearly a quarter of the population relies on wells as their primary drinking source. Well water quality is not regulated under the Safe Drinking Water Act, thus well water users are vulnerable to metal contamination. In this study, we set out to comprehensively examine statewide trends in both toxic and essential metals, arsenic, cadmium, lead, mercury, chromium, manganese, copper and zinc in private wells to identify areas of public health concern in NC. Methods. We compiled and geocoded using ArcGIS well water tests conducted by the Department of Health and Human Services from an over twenty-year period (October 19, 1998 and 20th May 2019), totaling n=122,130 test reports. We calculated county-level distributions and assessed multi-metal co-exposures through pairwise correlations and clustering assessments. Results. Contamination of private wells by toxic metals was common, with over 2,500 reports over the EPA Maximum Contaminant Level (MCL) for arsenic, cadmium and lead. Manganese, which can be toxic at high levels, was detected over the NC Groundwater Standard in 25% of wells. These distributions varied notably by county, where counties such as Anson have over 20% of well water tests reporting over the EPA MCL for arsenic yet many counties having no over MCL reports. Many metals were significantly correlated, including arsenic and manganese, cadmium and lead, cadmium and zinc, chromium and copper and lead and zinc. Clustering analysis is currently underway. Conclusions. Metal contamination of private wells in NC is common as is co- occurrence of these metals. Metal exposure via well water should remain a top state public health concern and assessments considering multi-metal exposure should be a priority.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

184 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-22

A standardized approach to 16S rRNA amplicon data analysis that emphasizes data interoperability and reusability

Jessica M. Ewald, University of Iowa, Iowa City, Iowa Daniel Rodriguez, Duke University Claudia Gunsch, Duke University Timothy Mattes, University of Iowa, Iowa City, Iowa

Superfund Mandate: None

Superfund Center: University of Iowa Superfund Research Center

High throughput sequencing of genomic markers represents a powerful tool to characterize and engineer microbiomes. For example, in contaminated environments where environmental engineers seek to remediate pollution with microorganisms, 16S rRNA amplicon sequencing allows us to characterize the microbial community and develop site-specific remediation strategies. Analysis of these data sets requires that we assign taxonomic classifications to genomic sequences, and researchers must choose an analysis approach from a long list of available software. The incongruency between available analysis methods negatively affects the reusability and interoperability of the data. To improve the interoperability and reusability of these data sets, we developed and tested a data analysis pipeline that provides a standardized approach to 16S amplicon data analysis and visualization. The pipeline includes DADA2, a software that infers taxonomy from amplicon sequence variants, packaged alongside powerful visualization tools aimed to elucidate relationships between the microbial community composition and physical and chemical site characteristics. To enhance interoperability, we packaged these robust analysis tools within an easily deployable and computationally light container. Analysts first download the container from a publicly available repository and then executes the analysis within that computing environment. Finally, we plan to release analysis scripts and protocols that detail container use on the users choice of hardware (i.e. lab server, computer clusters, laptop computer). Ultimately, we anticipate that uniform 16S rRNA amplicon analysis within and between superfund centers will provide a greater capacity for microbiome driven scientific discovery.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 185 Poster Session III: Poster #III-23

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) elicits changes in gut microbiome consistent with progression of steatosis to steatohepatitis with fibrosis in mice

Russell R. Fling, Michigan State University, East Lansing, Michigan Rance Nault, Michigan State University, East Lansing, Michigan Kelly Fader, Michigan State University, East Lansing, Michigan Tim Zacharewski, Michigan State University, East Lansing, Michigan

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

Dysbiosis of the gut microbiota is commonly associated with development and progression of non- alcoholic liver disease (NAFLD). 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), the most potent AhR agonist, elicits NAFLD pathologies as well as gut dysbiosis in mice. To investigate gut microbiota changes in response to TCDD, male mice were orally gavaged with sesame oil vehicle control or 30 µg/kg TCDD every 4 days for 28 days. Shotgun metagenomic sequencing of the cecal contents was conducted to investigate shifts in taxa and microbial metabolic pathways. Taxonomic analysis revealed a 2-fold increase in taxa associated with the Lactobacillales order. Among the 22 significant species identified, 18 were from the Lactobacillus genus with the largest increase observed by Lactobacillus reuteri (90-fold) mirroring aberrant levels of L. reuteri in NAFLD patients. To further investigate genes and metabolic pathways associated with NAFLD, metagenomic functional analysis was performed. Many Lactobacillus species possess bile salt hydrolases (Bsh) producing secondary deconjugated bile acids. Increased Bsh levels were attributed to L. reuteri and Lactobacillus johnsonii in TCDD treated mice supporting previously reported increases in serum secondary bile acids in TCDD treated mice. Although major shifts in microbial community were not observed at higher taxonomic levels, i.e. the phylum or family level, our results suggest TCDD elicited increases in Lactobacillus species resulting in microbial metabolic pathway changes associated with NAFLD progression. Russell Fling is supported by the National Institute of Environmental Health Sciences (NIEHS) Multidisciplinary Training in Environmental Toxicology (NIEHS EHS T32ES007255). This work was funded by the NIEHS Superfund Research Program (NIEHS SRP P42ES04911), and the NIEHS R01ES029541 to Tim Zacharewski.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

186 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-24

Detecting Novel Per- and Polyfluoroalkyl Substances in Water Supplies and Human Biofluids Using LC-IMS-MS

MaKayla Foster, North Carolina State University, Raleigh, North Carolina Markace Rainey, Georgia Institute of Technology, Atlanta, Georgia Chandler Watson, Georgia Institute of Technology, Atlanta, Georgia Facundo M. Hernandez, Georgia Institute of Technology, Atlanta, Georgia Erin Baker, North Carolina State University, Raleigh, North Carolina

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: North Carolina State University Superfund Research Center

PFAS are environmental contaminants that are chemically inert, thermally stable, and hydrophobic. These properties make PFAS suitable for many household and industrial applications, but also cause bioaccumulation and toxicity. Therefore, with each environmental spill or release, public health concerns rise, particularly with regards to drinking water safety. Since advised concentrations are extremely low (ppb to ppt), analytical instrumentation with low limits of detection and high selectivity and sensitivity are needed. Liquid chromatography coupled with mass spectrometry (LC-MS) has become an essential PFAS analysis technique. However, since PFAS contain thousands of compounds and new molecules are regularly detected, untargeted analysis methods are essential. To address this need, we utilized simultaneous LC, ion mobility spectrometry and MS (LC-IMS-MS) to evaluate PFAS standards. Initially, standards were assessed by their LC retention time, IMS collision cross section (CCS) and m/z information. Due to fluorination occurring in every PFAS, we also calculated CF2-based kendrick mass defects for each of the m/z values. CCS versus m/z plots illustrate distinct trendlines between different PFAS subclasses. The 3D mass defect, CCS and m/z plots easily differentiated fluorinated and nonfluorinated species. We are currently extending these plots to include ~4000 PFAS species of EPA interest by utilizing machine learning predictions for CCS values. This initial data will be applied to water and serum samples. Since LC- IMS-MS provides a rapid multi-dimensional assessment of each molecule present in complex samples, known PFAS subclasses will be characterized and currently unknown PFAS will be discovered, ultimately enabling a better understanding of contamination and exposure.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 187 Poster Session III: Poster #III-25

Activity-Based Protein Profiling Identifies Key P450 Enzymes in PAH Metabolism in Human Liver Microsomes

Kari A. Gaither, Pacific Northwest National Laboratory, Richland, Washington Whitney Garcia, Pacific Northwest National Laboratory, Richland, Washington Kimberly Tyrrell, Pacific Northwest National Laboratory, Richland, Washington Aaron T. Wright, Pacific Northwest National Laboratory, Richland, Washington, Jordan N. Smith Oregon State University, Corvallis, Oregon

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Oregon State University Superfund Research Center

Phase I cytochrome P450s commence the metabolism of polycyclic aromatic hydrocarbons (PAHs), catalyzing chemical reactions to enhance solubility, while Phase II enzymes conjugate polar metabolites to further enhance solubility and elimination. Bioactivation of PAHs can also occur, resulting in metabolites with increased toxicity over the parent compound. To better understand PAH metabolism, we have developed a method using activity-based proteomics to identify important enzymes involved. Our objective herein is to identify specific P450 enzymes active in the metabolism of benzo[a]pyrene (BaP) in human hepatic microsomes at multiple substrate concentrations. We co-incubated BaP (6.25 and 20 µM) with 2-ethynylnaphthalene (2-EN), a probe specific for P450s, in pooled human liver microsomes using a kinetic metabolism assay developed in our laboratory. We enriched probe-labeled, active P450 enzymes and measured proteins using liquid chromatography-mass spectrometry. When BaP was co- incubated with 2-EN, we observed a decrease in enriched enzymes compared to probe-only controls, allowing the identification of specific enzymes involved in BaP metabolism. Preliminary data suggest CYPs 1a2, 2c19, 2d7, 4f2, and 51a1 are the key enzymes active in BaP metabolism in human liver microsomes. A kinetics assay of BaP (5 µM) co-incubated with 2-EN (10 µM) revealed no inhibition of BaP metabolism by the probe in human liver microsomes at the concentrations tested. We are currently evaluating other substrate concentrations to identify important enzymes throughout the Michaelis-Menten curve. This powerful new approach will help us identify P450 enzymes involved in human metabolism of additional PAHs and probe Phase II enzyme families of interest. The ability to identify specific enzymes active in PAH metabolism will greatly aid our efforts to determine the health risks of PAHs found at Superfund Sites.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

188 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-26

Characterizing PON2 Deficiency in the Brain: Transcript and Behavioral Assessment

Jacqueline Garrick, University of Washington Toby B. Cole, University of Washington Theo K. Bammler, University of Washington James W. MacDonald, University of Washington Judit Marsillach, University of Washington Clement E. Furlong, University of Washington Lucio G. Costa, University of Washington and University of Parma, Italy

Superfund Mandates: None

Superfund Center: University of Washington Superfund Research Center

Paraoxonase 2 (PON2) is a ubiquitously expressed, intracellular antioxidant enzyme shown to mitigate oxidative stress in the brain. Upon oxidant exposure, mouse Pon2-deficient neurons display higher reactive oxygen species (ROS) levels and cellular death compared to neurons from wildtype (WT) mice in- vitro, demonstrating that PON2 is important for protecting against exogenous oxidant damage. Considering oxidative stress is a primary mechanism by which many environmental toxicants cause cellular injury, PON2 may play an important role in protecting the brain against these toxicants. To further understand the consequences of Pon2 deficiency, we evaluated behavior and regional transcript changes in the brain of mice. We found changes in home-cage behavior, where Pon2-deficient mice moved around the cage more and spent significantly more time in the center of the arena. On the rotarod, Pon2-deficient mice had a shorter latency to fall compared to WT, suggesting impaired motor coordination. Finally, when looking at transcriptional changes with RNA-Seq, we found highly regional and sex-specific changes. Many of the observed expression changes were in RNA polymerase and processing pathways, suggesting Pon2 deficiency may indirectly influence a multitude of pathways in the brain by altering transcriptional machinery, although the mechanism by which this occurs is unclear. When comparing the mouse Pon2 deficient transcriptional profile to known toxicant profiles, Pon2 deficiency causes transcriptional changes like those observed in mice exposed to aflatoxin B1 and dibutyl phthalate, potentially leaving these mice more susceptible to these neurotoxic compounds. These findings support PON2 playing an important neuroprotective role in the brain, with deficiency consequences at both the behavioral and transcriptional level. (Supported in part by grants from NIEHS (P42ES04696, P30ES07033, T32ES007032, R01ES028273) and NICHD (U54HD08091)).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 189 Poster Session III: Poster #III-27

Well Water Engagement Starts Flowing: UNC SRP Community Engagement & Research Translation Activities

Andrew George, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Megan Rodgers, University of North Carolina-Chapel Hil, Chapel Hill, North Carolina l Crystal Lee Pow Jackson, NC Department of Health and Human Services Sarah Yelton, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Kathleen Gray, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina

Superfund Mandate: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

Over 3 million North Carolina (NC) residents—nearly a third of the state’s population—get their drinking water from private wells; this is one reason the Community Engagement Core (CEC) and the Research Translation (RT) team in the UNC Superfund Research Program support NC communities concerned about toxic metals contamination of well water to develop locally relevant solutions. This year, the UNC SRP CEC and RT team have joined public health and community leaders in facilitating dialogue about well testing and well treatment options in communities with identified contamination. In September 2020, the CEC coordinated a statewide gathering of more than 40 well water scientists, practitioners, and community- based organizations to develop strategies for increasing well water testing, implementing necessary treatment options, and securing funding to support these efforts. One of our primary community partners, Clean Water for NC, co-hosted the meeting and will conduct field work with us, as we build our research and policy goals to achieve our SRP aims. In November 2020, we convened the first meeting of the UNC SRP Public Health Advisory Committee (PHAC). Members include representatives from county environmental and public health departments, water quality and environmental justice organizations, and the NC Division of Public Health (DPH). UNC SRP Director Dr. Rebecca Fry briefed PHAC members on center research to understand mechanisms behind arsenic-induced diabetes and engaged them in brainstorming community-based strategies to prevent exposure to contaminated well water. The UNC SRP RT team is partnering with toxicologists in DPH to create an online well water treatment guide. Upgrading an existing treatment guide from brochure and fact sheet to an interactive, online database will enable enhanced accessibility and expanded content.

Email: [email protected] Presenter Status: Researcher

190 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-28

Assessing PAH diffusive and advective flux contributions and temporality in the environment

Christine Ghetu, Oregon State University, Corvallis, OR Jamie D. Minick, Oregon State University, Corvallis, OR Lane G. Tidwell, Oregon State University, Corvallis, OR Peter D. Hoffman, Oregon State University, Corvallis, OR Kim A. Anderson, Oregon State University, Corvallis, OR

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Oregon State University Superfund Research Center

We present the development of a field sampling approach to measure diffusive and advective chemical movement across a contaminated site. Diffusive flux describes the movement of chemicals from a compartment with a higher concentration to a compartment with a lower concentration. Advective flux describes the bulk movement of water and its transport of contaminants from one environmental compartment to another. Temporal impacts, such as seasons, likely also play a role in the direction and magnitude of chemical movement. We co-deployed passive sampling devices (PSDs), measuring advective and diffusive flux, with seepage meters at eleven locations at a former creosote site. PSDs were located at the sediment–water and water–air interface. Sampling took place in August, September and October of 2019. Samples were extracted then analyzed by gas chromatography triple-quadrupole mass spectrometry for 65 individual parent and alkylated polycyclic aromatic hydrocarbons (PAHs) and alkylated PAHs used for forensic source determination. Diffusive flux magnitude was, on average, three times larger than advective flux for all locations. Chemical movement of PAHs was dominated by water- porewater diffusive flux for all sampling time points. PAH air and sediment porewater concentrations increased 35% from summer to fall while water concentrations decreased 25%. Data also suggests that chemical movement direction and/or magnitude is temporally dependent.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 191 Poster Session III: Poster #III-29

Volatile Organic Compound Exposures Are Positively Associated with Liver Apoptosis in a Residential Cohort

T. C. Gripshover, University of Louisville, Louisville, Kentucky B. Wahlang, University of Louisville, Louisville, Kentucky D.W. Riggs, University of Louisville, Louisville, Kentucky S.E. Smith, University of Louisville, Louisville, Kentucky T.V. Krivokhizhina, University of Louisville, Louisville, Kentucky R.J. Keith, University of Louisville, Louisville, Kentucky C.J. McClain, University of Louisville, Louisville, Kentucky S. Srivastava, University of Louisville, Louisville, Kentucky M.C. Cave, University of Louisville, Louisville, Kentucky

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Background. The Health, Environment and Action in Louisville (HEAL) cohort was assembled by the NIEHS- funded University of Louisville Superfund Research Center to allow determination of associations between residential VOC exposures and cardiometabolic syndrome (inclusive of liver disease) in the Louisville community. The objective of this interim cross-sectional analysis of 679 HEAL subjects collected in 2018 and 2019 is to determine associations between urinary VOC metabolites and the circulating hepatocyte apoptosis biomarker, caspase-cleaved keratin 18 (CK18 M30). Methods. Serum CK18 M30 (normal cutoff <200 U/L) was determined by ELISA. 15 creatinine-adjusted urinary metabolites of 10 parent VOCs were measured by UPLC-MS2. Generalized linear models were used to test for associations between disease and exposure biomarkers. Final models were adjusted for race, BMI, alcohol use, cotinine-confirmed smoking status, and education level. Results are presented as % change (95% CI) per interquartile range of VOC metabolite. Results. The following results pertain to 391 samples collected in the first round of HEAL recruitment (2018) while analysis on the remaining samples are pending. Mean age was 50.3±12.0 (SD). Mean BMI was 29.9±6.1 kg/m2. The cohort was 57.8% female, 78.5% white and 22.0% diabetic. Elevated CK18 M30 was present in 12.3% of participants. CK18 M30 was positively associated with 4 urinary VOC metabolites corresponding to acrolein (3HPMA); acrylamide (AAMA); and 1,3-butadiene (DHBMA and HPMMA) exposures. These associations were modified by sex and race for three of the four significant VOC metabolites and were more positive in males and whites. Conclusions. This interim analysis demonstrated positive associations between specific VOC exposures and hepatocyte apoptosis in the residential HEAL cohort. The role of VOC exposures in metabolic liver diseases warrants further investigation.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

192 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-30

Creation of Polycyclic Aromatic Hydrocarbon Breakdown Mixtures and Investigation of their DNA Damaging Potential

Simran Kaushal, Massachusetts Institute of Technology, Cambridge, Massachusetts Amy Hrdina, Massachusetts Institute of Technology, Cambridge, Massachusetts James Rowe, Massachusetts Institute of Technology, Cambridge, Massachusetts Jesse Kroll, Massachusetts Institute of Technology, Cambridge, Massachusetts Bevin Engelward, Massachusetts Institute of Technology, Cambridge, Massachusetts

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Massachusetts Institute of Technology Superfund Research Center

Benzo-a-pyrene is an environmental contaminant found at many Superfund sites and is formed as a byproduct of incomplete combustion, such as in burning fossil fuel and cooking. Our research team is interested in the DNA damaging potential of polycyclic aromatic hydrocarbons (PAHs), since DNA damage is known to promote carcinogenic mutations. Interestingly, conditions in the atmosphere can lead to the formation of multigenerational breakdown products, and so we are interested in their biological impact. We are collaborating with atmospheric chemists to study benzo-a-pyrene oxidation product mixtures that are expected to occur in the atmosphere. Bioengineers are then testing the DNA damaging capacity of these oxidation products using the CometChip, a high-throughput comet assay that can be used to measure DNA damage. Our data will elucidate the impact of cell type and metabolism on the DNA damaging capacity of PAH breakdown products. We will also investigate the DNA repair pathways required to resolve damage caused by these contaminants, which will contribute to our understanding of genetic factors that impact susceptibility to exposure This study will ultimately contribute to risk assessment for populations exposed to benzo-a-pyrene and its oxidation products.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 193 Poster Session III: Poster #III-31

Reinforcing and Building New Community Relations in Appalachian Kentucky Amid the COVID-19 Pandemic

Annie Koempel, University of Kentucky, Lexington, Kentucky Gia Mudd-Martin, University of Kentucky, Lexington, Kentucky Dawn Brewer, University of Kentucky, Lexington, Kentucky

Superfund Mandates: None

Superfund Center: University of Kentucky Superfund Research Center

Health disparities in Appalachian Kentucky have possible links to environmental pollutants including sources from deteriorating infrastructure and contamination from mining and agricultural practices. The COVID-19 pandemic posed additional mental and physical stressors, particularly as food accessibility became more challenging as businesses and restaurants closed in spring 2020. In response, the University of Kentucky Superfund Research Community Engagement Core (UK-CEC) partnered with community collaborators across Appalachia to increase access to healthy foods through three major fruit and vegetable programs: (1) the Tanglewood to Table (T2T) Farmers’ Market Voucher and Walking program; (2) BerryCare: A Blackberry Club to Facilitate Community Engagement and Phytonutrient Intake; and (3) the Fruit and Vegetable Box to Produce Good Health program. Through T2T, Letcher County community members utilized appropriate methods to safely redeem their weekly $10 farmers’ market vouchers. Simultaneously, we expanded the BerryCare program into five new Appalachian Kentucky counties, including Harlan County, site of the Dayhoit Superfund site. Through a new partnership with Grow Appalachia, BerryCare provided 93 participants with blackberry plants, recipes, and information on how to reduce pollutant exposure. Finally, a new relationship with Red Bird Mission in Clay County resulted in distributing a box of fresh produce, nutrition and environmental pollution education, and recipes to 53 low-income families (16% of Red Bird’s clientele). Participants completed a survey that included an adapted version of the Environmental Health Engagement Profile-Kentucky Nutrition Version that assessed environmental health literacy. We discuss how the UK-CEC navigated these programs and strengthened community relations online, and include lessons learned and best practices for building community relations in rural areas amid a global pandemic. Finally, we present key preliminary outcomes of the programs.

Email: [email protected] Presenter Status: Other: CEC Program Manager

194 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-32

N-nitrosodimethylamine (NDMA) leads to activation of phosphorylation mediated signaling networks in mice

Ishwar N. Kohale, Massachusetts Institute of Technology, Cambridge, Massachusetts Jennifer E Kay, Massachusetts Institute of Technology, Cambridge, Massachusetts Joshua J Corrigan, Massachusetts Institute of Technology, Cambridge, Massachusetts Bevin P Engelward, Massachusetts Institute of Technology, Cambridge, Massachusetts Forest M White, Massachusetts Institute of Technology, Cambridge, Massachusetts

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Massachusetts Institute of Technology Superfund Research Center

N-nitrosodimethylamine (NDMA) poses a significant risk to human health as a probable human carcinogen. Contamination of NDMA has been discovered in processed food, water treatment facilities and even in commonly used drugs. Millions of gallons of poorly contained chemical waste has led to NDMA contamination of drinking water supply near the Olin Chemical Superfund site. Although genotoxic effects of NDMA are being studied more widely, the overall cellular response to NDMA is still poorly characterized. More specifically, how exposure to NDMA at early age would affect pathophysiological response is still not known. Additionally, how exactly the cellular response evolves over time is poorly understood. Here, we exposed mice with different DNA repair capabilities in a single split dose regimen of NDMA at 8 and 15 days old, and quantified protein phosphorylation at 24 hours, 10 weeks and 10 months after treatment. We quantified ATM/ATR kinase mediated DNA damage response as well as phosphotyrosine based signaling cascades in wild type, Mgmt knockout, Aag overexpressing and Aag knockout mice. We observed changes in signaling networks in a time dependent manner with additional differences in mice with different genetic backgrounds. Surprisingly, the activation of ‘oncogenic’ pathways was observed 24 hours after the treatment suggesting early changes in cellular state that would lead to tumor formation after 10-months post exposure. Overall, the data suggest strong and persistent activation of signaling networks as a mechanism of toxicity to NDMA exposure.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 195 Poster Session III: Poster #III-33

Variation of vitamin D metabolites in acute pancreatitis patients, and the potential role as a biomarker

Shinichiro Koike, University of California-Davis, Davis, California Debin Wan, University of California-Davis, David California Enrique de-Madari, Alicante University General Hospital Bruce D. Hammock, University of California-Davis, David California Fawaz G. Haj, University of California-Davis, David California

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

Acute pancreatitis (AP) is characterized by unpredictable local inflammation in the pancreas and is the most common gastrointestinal disease causes for hospitalization in the United States. The overall mortality of AP is 5%, but in severe AP with multisystemic organ failure or necrotizing acute pancreatitis (NAP), it reaches 20%. The mechanism of AP is not fully understood given the difficulty of examination during pancreatitis and acute progress of the disease, and there is no specific therapy. Vitamin D is mainly synthesized in the skin from 7-dehydrocholesterol and plays an important role in bone metabolism and calcium homeostasis. Additionally, Vitamin D is implicated in systemic inflammation and might downregulate inflammation by inhibiting dendritic cell differentiation and pro-inflammatory cytokines. Notably, decreased Vitamin D levels were reported in the diseased pancreas, including AP patients. In this study, we assessed plasma Vitamin D metabolite levels in healthy control and AP patients with different stages (mild, moderate, and severe) using an improved method to comprehensively evaluate alterations in vitamin D in AP. We demonstrated a negative correlation between vitamin D metabolites and previously reported AP markers such as C reactive protein, creatinine, and urea in AP patients. In addition, our observations suggested further reduction of vitamin D metabolites in the presence of persistent organ failure and NAP. This study suggested that the potential value of vitamin D metabolites as a multi- functional marker for AP severity and participant organ failure and NAP.

Email: [email protected] Presenter Status: Researcher

196 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-34

The Impact of COVID-19 on Maternal Health in Puerto Rico: A NIMHD RCMI Administrative Supplement

Irene Lafarga Previdi, Northeastern University, Boston, Massachusetts, University of Puerto Rico, Carmen M. Vélez Vega- University of Puerto Rico, Mayra Rivera Robles- University of Georgia Natacha Ivette Guilloty Rodríguez- University of Georgia Zulmarie J. Diaz Reguero- University of Georgia Chrystal Galán Rivera- Northeastern University Ishwara Ayala Ortiz- University of Georgia Zaira Yadira Rosario Pabón- University of Georgia

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Northeastern University Superfund Research Center

According to literature pregnant women are at a greater risk for exposure and death during emergencies. Pregnant women are at greater risk for influenza associated illnesses and death. The purpose of this study will be to assess and describe how the isolation and physical distancing measures related to the current Coronavirus pandemic have affected pregnant women and mothers of children 12 months or younger in Puerto Rico. The specific aims of the project are: 1) Examine the impact of the COVID-19 outbreak in pregnancy related experiences and outcomes; 2) Examine the mental health impact of the COVID-19 outbreak in pregnant women and mothers of children 12 months or younger; and 3) Identify risk and protective factors of pregnant women and mothers of children 12 months or younger in the COVID-19 outbreak. Participants will be recruited from the Puerto Rico Test- site for Exploring Contamination Threats (PROTECT) consortium cohort which is composed of pregnant women and mothers from the northwestern region of Puerto Rico that studies the effects of contaminant exposure and preterm birth and other birth outcomes. The research will have a mixed methods approach with a quantitative survey and semi-structured qualitative interviews. Results from the study will identify risk and protective individual and contextual factors among pregnant women and mothers of children 12 months or younger. The study findings will provide greater insight into the experiences and needs of Latino Hispanic pregnant women and mothers of children 12 months or younger during pandemics. We expect that the findings can lead to the development of interventions for local community health centers and also to the creation of evidence-based protocols aimed at the Puerto Rican population.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 197 Poster Session III: Poster #III-35

Windblown Transport of Toxic Metals in Airborne Particulate Matter Near the Jackpile-Paguate Uranium Mine on Laguna Pueblo, New Mexico

Savannah LaRopsa-LoPresti, University of New Mexico, Alburquerque, New Mexico Joseph Galewsky, University of New Mexico, Alburquerque, New Mexico Adrian Brearley, University of New Mexico, Alburquerque, New Mexico Melissa Gonzales, University of New Mexico, Alburquerque, New Mexico Luna Natoli, University of New Mexico, Alburquerque, New Mexico Kyle Swimmer, University of New Mexico, Alburquerque, New Mexico, Pueblo of Laguna Abdulmehdi Ali, University of New Mexico, Alburquerque, New Mexico Chris Shuey, University of New Mexico, Alburquerque, New Mexico, Southwest Research and Info Ctr

Superfund Mandate: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of New Mexico Superfund Research Center

The Jackpile-Paguate Uranium Mine on the Pueblo of Laguna, New Mexico, has had a significant environmental impact on several local communities. Potential exposures to uranium and other toxic metals derived from the mine as a result of inhalation of airborne particulate matter (PM) are of major concern for the local communities. In this study, we are using a combination of meteorological, mineralogical, and geochemical analyses to assess the local transport of PM in the area, determine the concentration of U and other toxic metals, and analyze the physical characteristics of U-bearing particles in airborne PM. Meteorological stations at 4 sites on Laguna Pueblo suggest relative humidity and wind speed are the dominant atmospheric parameters controlling transport of inhalable PM in a semi-arid region. Additionally, we have compared the local meteorological data to regional data from NOAA to examine the advantages of collecting local weather data, due to highly variable topography in the region that likely affects local wind patterns. Tisch high volume air samplers have been deployed at 2 locations on the Pueblo of Laguna to collect airborne PM over periods of several months. Preliminary mineralogical analyses of these samples by scanning electron microscopy have revealed the presence of U-bearing particles <PM10 in size, including uranyl-vanadate minerals that are associated with mining in the area. Geochemical analyses of the PM samples are currently in progress to determine concentrations of toxic metals including U, V, Pb, As and other elements previously identified in the area at levels exceeding safety regulations. This combination of different analytical methods will provide essential data that are necessary for communities at Laguna Pueblo to assess current inhalation exposure risks.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

198 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-36

Differential effects of arsenic exposure on hepatic and intestinal heme oxygenase 1 expression

Hui Li, University of Arizona, Tucson, Arizona Xiaoyu Fan, University of Arizona, Tucson, Arizona Xiangmeng Wu, University of Arizona, Tucson, Arizona Weiguo Han, University of Arizona, Tucson, Arizona Pengfei Liu, University of Arizona, Tucson, Arizona Donna Zhang, University of Arizona, Tucson, Arizona Xinxin Ding, University of Arizona, Tucson, Arizona Qing-Yu Zhang, University of Arizona, Tucson, Arizona

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Arizona Superfund Research Center

Populations residing near mining sites may be exposed to mine tailings through the inhalation of dust particles or ingestion of contaminated water or food. For arsenic exposure through drinking water, the intestine is the first site of contact, and absorbed arsenic is further transported to the liver via the portal vein. Thus, it is important to identify and compare tissue responses to arsenic exposure in liver and intestine. Previous studies reported that the expression of several stress-responsive genes, particularly heme oxygenase-1 (HO-1), can be dramatically induced in mouse liver by a bolus administration of arsenic at a high dose. However, it is unknown whether HO-1 induction occurs in either liver or intestine upon exposure to arsenic via drinking water, at relatively low doses that approach human exposure conditions in highly contaminated regions. Thus, the aim of this study was to examine effects of oral arsenic exposure via drinking water on the expression of HO-1 in the intestine and liver. Initial studies showed that a one- week exposure to sodium arsenite (25 ppm) in drinking water failed to induce hepatic HO-1 expression; but intestinal HO-1 expression was induced substantially. The apparent tissue-specificity of HO-1 induction in the intestine by drinking-water arsenic was observed in additional experiments with various arsenic doses (2.5-25 ppm) or treatment periods (1 day to 4 weeks). The intestinal HO-1 induction was detected at both mRNA and protein levels and was accompanied by increased enzymatic activity for metabolizing heme. These findings support the need to identify mechanisms as well as functional consequences of the tissue specific HO-1 induction in the intestine by arsenic in drinking water. (Supported by the Superfund Research Program, ES004940-31)

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 199 Poster Session III: Poster #III-37

Quantitative in vitro-to-in vivo extrapolation for mixtures: A case study of Superfund Priority List pesticides

Yu-Syuan Luo, Texas A&M University, College Station, Texas Alan C. Valdiviezo, Texas A&M University, College Station, Texas Zunwei Chen, Texas A&M University, College Station, Texas Weihsueh A. Chiu, Texas A&M University, College Station, Texas Ivan Rusyn, Texas AM University, College Station, Texas

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

In vitro cell-based toxicity testing methods generate large amounts of data informative for risk-based evaluations. To allow extrapolation of the quantitative outputs from cell-based tests to the equivalent exposure levels in humans, in vitro-to-in vivo extrapolation (IVIVE) method is used for predicting the in vivo toxicokinetics (TK) from in vitro data. IVIVE data for the individual environmental chemicals can be derived using established experimental or modeling approaches. However, the potential implications of chemical-to-chemical interactions in mixtures on IVIVE remain unexplored. We hypothesized that chemical co-exposures could modulate both protein binding efficiency and hepatic clearance of the individual chemicals in a mixture, which would in turn affect the quantitative IVIVE parameters. To test this hypothesis, we used 20 pesticides from the Superfund Priority List, both individually and as an equimolar mixture, and investigated the concentration-dependent effects and chemical interactions on in vitro TK parameters. Protein binding efficiency was determined by using ultracentrifugation, and hepatocyte clearance was estimated in suspensions of cryopreserved primary human hepatocytes. We found that for single chemicals, the protein binding efficiencies were similar at different testing concentrations. In a mixture, however, both protein binding efficiency and hepatocyte clearance were affected. Applying IVIVE using mixture-derived TK data resulted in more conservative estimates of Activity to Exposure Ratios (AERs) compared to using data from single chemical experiments. As human exposures always occur as mixtures of multiple chemicals, these results show the importance of incorporating mixture interactions in IVIVE in vitro bioactivity data in order to accurately prioritize risks and facilitate scientific-based decision-making. This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

200 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-38

Perfluorooctane Sulfonate (PFOS) Induced Microglial Hyperresponsiveness is Modulated by Neuronal Activity

Shannon E. Martin, Brown University, Providence, Rhode Island Nathan R. Martin, Brown University, Providence, Rhode Island Jessica Plavicki, Brown University, Providence, Rhode Island

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effects on human health

Superfund Center: Brown University Superfund Research Center

Per- and Polyfluoroalkyl Substances (PFAS) are a class of manufactured organic compounds present in repellant and stain-resistant products. These ubiquitous toxicants do not readily degrade and, consequently, are environmentally persistent. Perfluorooctane sulfonate (PFOS), a prevalent PFAS congener, is immunotoxic and known to disrupt the function of the adaptive immune system. However, it is not known if PFOS affects the function of microglia, the innate immune cells of the brain, which are essential for responding to physical and pathogenic insults as well as proper neuronal development. Here, we examined whether prior PFOS exposure disrupts microglial responses to brain injury. PFOS exposure caused a significant increase in microglial recruitment and prolonged microglial presence at and beyond the injury site. PFOS exposure did not cause an upregulation of inflammatory cytokine mRNA in the brain, nor was there increased cell death at the injury site. We therefore asked whether an altered neuronal environment could be modulating the observed microglial responses in PFOS-exposed embryos. Using the transgenic zebrafish line CaMPARI, which distinguishes active neurons from inactive neurons following a brief photoconversion, we found that PFOS exposure caused regional-specific increases in brain activity in 3-day-old embryos exposed to 32µM PFOS and in 5-day-old embryos exposed to 8µM PFOS. PFOS exposure also increased release of synaptic vesicles, providing further evidence of increased neuronal activity. We were able to recapitulate the hyperresponsive microglial phenotype by pharmacologically increasing neuronal excitation using the GABA antagonist pentylenetetrazol. Correspondingly, the hyperresponsive microglial phenotype observed in PFOS-exposed zebrafish was rescued by silencing neurons using optogenetics. Together, these data reveal that neuronal hyperactivity following PFOS exposure exacerbates microglial responses to injury in embryonic zebrafish.

Email: [email protected] Presenter Status: Researcher / Environmental Health

NIEHS SRP 2020 Virtual Annual Meeting Program • 201 Poster Session III: Poster #III-39

Inducible and conditional activation of adult neurogenesis rescues cadmium- induced hippocampus dependent memory deficits in a mouse model of Alzheimer’s disease

Megumi Matsushita, University of Washington, Seattle Zhengui Xia, University of Washington, Seattle

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of Washington Superfund Research Center

Elucidating mechanisms of effects of genetic factors and environmental exposures in model organisms is critical to understand the impact of gene-environment interactions (GxE) on human health. The apolipoprotein E (ApoE) gene is a genetic risk factor for late-onset Alzheimer’s disease, in which ε4 allele carriers have increased risk compared to ε3 carriers. Cadmium (Cd) is a toxic heavy metal that is a potent neurotoxicant. Our previous study found a GxE effect between ApoE4 and Cd that accelerates or increases the severity of cognitive decline in ApoE4-knockin (ApoE4-KI) mice exposed to 0.6 mg/L CdCl2 through drinking water compared to control ApoE3-KI mice. However, the mechanisms underlying this GxE effect is not yet known. Here we investigate the causal relationship between Cd-induced memory deficits and impairment of adult hippocampal neurogenesis in the observed GxE effect. We crossed the ApoE-KI strains to the caMEK5 mouse strain, which enables the stimulation of adult neurogenesis through genetic and conditional activation of the endogenous ERK5 MAP kinase signaling pathway in adult neural stem/progenitor cells with tamoxifen administration. We investigated whether stimulation of adult neurogenesis can rescue Cd-induced cognitive impairment in the respective multi-transgenic lines (simplified ApoE3-KI, ApoE4-KI). ApoE3-KI and ApoE4-KI mice were exposed to 0.6 mg/L CdCl2 throughout the experiment and tamoxifen was administered to stimulate adult neurogenesis once consistent impairment in working spatial memory was observed after Cd exposure. Cd exposure impaired working spatial memory earlier in ApoE4-KI than ApoE3-KI (onset week 16 vs week 28). In both strains, these deficits were rescued by tamoxifen-induced stimulation of adult neurogenesis. Our study suggests a causal link between the GxE effect of ApoE4 and cadmium on cognition and impairment in adult neurogenesis.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

202 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-40

Prenatal Exposure to Polycyclic Aromatic Hydrocarbons (PAHs) Augments Neonatal Hyperoxic Lung Injury and Alters the Gut Microbiome in Mice: Mechanistic Role of Cytochrome P450 (CYP)1A1, 1A2, and 1B1

Bhagavatula Moorthy, Baylor College of Medicine, Houston, Texas Lauren Swanson, Baylor College of Medicine, Houston, Texas Guobin Xia, Baylor College of Medicine, Houston, Texas Weiwu Jiang, Baylor College of Medicine, Houston, Texas Lihua Wang, Baylor College of Medicine, Houston, Texas Xanthi Couroucli, Baylor College of Medicine, Houston, Texas Kristi Hoffman, Baylor College of Medicine, Houston, Texas Krithika Lingappan, Baylor College of Medicine, Houston, Texas

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Baylor College of Medicine Superfund Research Center

Pregnant women exposed to high levels of polycyclic aromatic hydrocarbons (PAHs) are at increased risk for premature delivery. Preterm infants often require supplemental oxygen, a known risk factor for the development of bronchopulmonary dysplasia (BPD). Both prenatal BP and postnatal hyperoxia exposure alter the neonatal gut microbiome, which may modulate neonatal hyperoxic lung injury via the gut-lung axis. In this investigation, we tested the hypothesis that prenatal administration of PAHs [i.e. benzo[a]pyrene (BP) or a mixture of BP and benzo(b)fluoranthene (BbF)] differentially exacerbates lung injury and alveolar simplification in neonatal mice following postnatal hyperoxia., and this effect is altered in mice lacking the gene for cytochrome P450 (Cyp)1a1,1a2, or 1b1. Timed pregnant WT (C57BL/6J), Cyp1a1-null, Cyp1a2-null, and Cyp1b1-null mice (n=4) treated orally with vehicle corn oil (CO) or a mixture of PAHs BP and BbF (7.5 mg/kg each) on gestational days 16-19. Newborn mice (term, day 21) were exposed to hyperoxia or room air for 14 days. Hyperoxic lung injury was augmented in a dose-dependent manner following prenatal PAH exposure. PAH exposure differentially altered lung injury in WT, Cyp1a1- null and Cyp1a2-null, but not Cyp1b1-null mice. PAH administration resulted in induction of CYP1A1 expression. Hyperoxia and PAH alone caused modest increase in CYP1B1 expression but together caused significant induction, suggesting that CYP1B1 may contribute to the potentiation of lung injury by PAH. Prenatal PAH exposure causes significant alterations in neonatal intestinal microbiome compared with corn oil (CO). The microbiome difference observed between PAH and control groups in WT mice was no longer significant in Cyp1a1-null mice, suggesting that CYP1A1 metabolism of PAH plays a role in altering the microbiome. (Supported by 1P42

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 203 Poster Session III: Poster #III-41

Direct injection analysis of per- and polyfluoroalkylated substances (PFAS) from surface water by sample filtration and UPLC-tandem mass spectrometry

Andrew J. Morris, University of Kentucky, Lexington, Kentucky M. Abdul Mottaleb, University of Kentucky, Lexington, Kentucky Qunxing X. Ding, Kent State University, Kent, Ohio Kelly G. Pennell, University of Kentucky, Lexington, Kentucky Erin M. Haynes, University of Kentucky, Lexington, Kentucky Andrew J. Morris, University of Kentucky, Lexington, Kentucky

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Kentucky Superfund Research Center

Per- and polyfluoroalkylated substances (PFASs) are widely used in consumer products and PFASs are highly stable in various environments. Human exposures are widespread and have been associated with increased risk of cardiovascular and metabolic diseases, cancer and impaired immune function. Drinking water is an important source of PFAS exposure. PFAS contamination in groundwater and other drinking water sources is a particular concern. Methods for measurement of PFAS in environmental water employ pre concentration by solid phase extraction prior to analysis by HPLC coupled electrospray ionization tandem mass spectrometry. We developed an validated a method for PFAS measurement without preconcentration which offers a simplified and more efficient approach. Samples are centrifuged, supernatants passed through an Acrodisc Filter (GXF/GHP 0.2 um, 25 mm diameter), reconstituted in 20 mM ammonium acetate and analyzed by UPLC-MS/MS using an AB Sciex 6500 plus Q-Trap mass spectrometer operated in negative multiple reaction-monitoring (MRM) mode. A delay column between the pumps and autosampler mitigates interference from background PFAS. The method monitors eight short-/long-chain PFAS which are quantified using isotope mass-labeled internal standards. Average recoveries (n = 8) of target analytes ranged from 84 to 110% with 4 – 9% RSD. Mean spiked recoveries (n = 8) of four surrogates were 94 – 106% with 3 – 8% RSD. For laboratory control samples (LCS), average spiked recoveries (n=8) for analytes ranged from 88 to 114% with 4 – 11% RSD and for surrogates were 104 – 112% with 3 – 11% RSD. The limit of detection for the target analytes was between 0.02 and 0.05 ng/mL with S/N=3.

Email: [email protected] Presenter Status: Researcher

204 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-42

STEEP (Sources, Transport, Exposure and Effects of PFAS) SRP: Communicating PFAS medical concerns and related health implications in the time of COVID

Amber Neville, University of Rhode Island, Coastal Institute at University of Rhode Island Laurel Schaider, Silent Spring Institute, Judith Swift, Coastal Institute at the University of Rhode Island Alyson McCann, University of Rhode Island Nathan Vinhateiro, Coastal Institute at University of Rhode Island Emily Diamond, University of Rhode Island

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Rhode Island Superfund Research Center

STEEP focuses on Cape Cod, MA as a primary study site with PFAS-contaminated groundwater. Two key activities are 1) lead a private well water-testing program and 2) educate residents about the harmful health effects of PFAS. Following the completion of a first phase study, STEEP had one-on-one discussions with participants to hear concerns and provide information. Through a series of focus groups, STEEP will learn what actions participants have taken since receiving their results. STEEP is also developing community templates that highlight the water quality status of each town. To complement a range of PFAS reduction tip cards, STEEP is creating information sheets for PCPs alerting them to PFAS’s long-term adverse health impacts as well as two-pagers for placement in healthcare facilities. STEEP is producing a range of podcasts, a second video series, and mini popup videos to guide website users. COVID-required social distancing protocols and methods are under development to allow this work. STEEP’s second video series will focus on potential human health effects related to PFAS exposure. Using multifaceted distribution modes these videos will reinforce information learned through a COVID-inspired virtual series presented as an extended Science Day experience. The series of monthly online presentations and discussions, called “PFAS In Your Community,” will bring together STEEP researchers and community members including the Mashpee Wampanoag Tribe. Roundtable and Q&A sessions will continue bidirectional communication through virtual public exchanges covering a broad range, including PFASs in drinking water, emerging adverse health impacts, essential uses, and consumer goods. In addition, RTC- CEC will collaborate with the training core and local high school teachers to integrate PFAS into high school biology, chemistry, and environmental science curricula.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 205 Poster Session III: Poster #III-43

Inequalities in public water arsenic concentrations in counties and community water systems across the United States, 2006-2011

Anne Nigra, Columbia University, New York, NY Qixuan Chen, Columbia University, New York, NY Steven N. Chillrud, Columbia University, Palisades, NY Lili Wang, Office of Water, United States Environmental Protection Agency, Washington D.C. David Harvey, United States Public Health Service, Rockville, MD Brian Mailloux, Columbia University, Palisades, NY, and Barnard College, Columbia University Pam Factor-Litvak, Columbia University, New York, NY Ana Navas-Acien, Columbia University, New York, NY

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Columbia University Superfund Research Center

Background: In the US, nationwide estimates of public drinking water arsenic exposure are not readily available. We used the US Environmental Protection Agency’s (EPA) Six Year Review contaminant occurrence dataset to estimate public water arsenic exposure. We compared community water system (CWS) arsenic concentrations during (2006-2008) versus after (2009-2011) the initial monitoring period for compliance with EPA’s 10 µg/L arsenic maximum contaminant level (MCL). Our objective was to characterize potential inequalities in CWS arsenic exposure over time and across sociodemographic subgroups. Methods: We estimated three-year average arsenic concentrations for 36,406 CWSs (98%) and 2,740 counties (87%) and compared differences in means and quantiles of water arsenic (via quantile regression) between both three-year periods for US regions and sociodemographic subgroups. We assigned CWSs and counties MCL compliance categories (High if above the MCL; Low if below) for each three-year period. Results: From 2006-2008 to 2009-2011, mean and 95th percentile CWS arsenic (µg/L) declined by 10.3% (95% CI 6.5%, 14.1%) and 11.5% (8.3%, 14.8%) nationwide, by 11.4% (4.7%, 18.1%) and 16.3% (8.1%, 24.5%) for the Southwest, and by 36.8% (7.4%, 66.1%) and 26.5% (12.1%, 40.8%) for New England, respectively. CWSs in the High/High compliance category (not MCL compliant) were more likely in the Southwest (61.1%), served by groundwater (94.7%), serving smaller populations (mean 1,102 persons), and serving Hispanic communities (38.3%). Discussion: Larger absolute declines in CWS arsenic concentrations at higher water arsenic quantiles indicate declines are related to MCL implementation. CWSs reliant on groundwater, serving smaller populations, located in the Southwest, and serving Hispanic communities were more likely to continue exceeding the arsenic MCL, raising environmental justice concerns.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

206 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-44

A Mixed Methods Qualitative and Geospatial Approach to Community-engaged Research

Matilda Odera, Louisiana State University, Baton Rouge, Louisiana Jennifer Richmond-Bryant, Louisiana State University, Baton Rouge, Louisiana All others employed by North Carolina State University and affiliated with LSU SRP

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Louisiana State University Superfund Research Center

A community-engaged information-gathering effort is being conducted to inform air quality sampling strategy in Colfax, Louisiana, which hosts a hazardous materials thermal treatment (TT) facility that destroys waste from Superfund sites, explosives, military ordnances, and propellants through open burning or detonation. Two-thirds of residents are Black, and the median income is approximately $26,000 per year. Our mixed-methods approach synthesizes information on exposures within Colfax and the nearby community of The Rock, which are proximal to the TT facility. We combined a review of public comments submitted to the Louisiana Department of Environmental Quality over permit deliberations with 17 oral history interviews of residents. We focused on themes of Colfax’s changing community fabric, resident health, and air pollution. We recruited participants of the oral history interviews through community contacts. Interviews were conducted through Zoom’s telephone feature, and transcripts are being analyzed using the NVivo software to code and mine the interview text for emergent themes. The review of public comments procured self-reported concerns about health, smoke, noise, vibration, and damage to homes, which were then mapped. Several interviewed residents reported seeing smoke from the facility, hearing loud noises from open burning, smelling electrical odors, and experiencing health issues, including thyroid disease, skin rashes, and respiratory diseases. Interview participants also shared that small churches dominate the social fabric of The Rock and Colfax, and these community centers have been instrumental in mobilizing residents against the TT facility’s practice of burning hazardous waste in the open. Insights gathered during the public comment and interview stage of this research is informing our air sampling strategy with respect to sampling locations and chemical composition of sampled particulate matter.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 207 Poster Session III: Poster #III-45

Metabolic reprogramming following transsulfuration pathway inhibition by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to support glutathione biosynthesis in mice

Karina Orlowska, Michigan State University, East Lansing, Michigan Russell R. Fling, Michigan State University, East Lansing, Michigan Rance Nault, Michigan State University, East Lansing, Michigan Tim Zacharewski, Michigan State University, East Lansing, Michigan

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

Activation of the aryl hydrocarbon receptor (AhR) by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces steatosis and its progression to steatohepatitis with fibrosis. Oxidative stress is a key contributor in the etiology of these pathologies that is counteracted by antioxidant defenses such as glutathione. To examine the effect of TCDD on transsulfuration pathway and glutathione metabolism, C57BL/6 mice were orally gavaged with TCDD (0.01-30 µg/kg) every 4 days for 28 days. We show TCDD dose-dependently repressed the transsulfuration pathway, specifically cystathionine beta-synthase (CBS) and cystathionine gamma-lyase (CTH), by 2.6- and 2.1-fold at the mRNA level, and 4.1- and 8.7-fold at the protein level, respectively. Accordingly, cystathionine levels were decreased at 1, 3 and 10 μg/kg TCDD but recovered at 30 µg/kg TCDD, while cysteine levels were dose-dependently increased consistent with the 924.5-fold induction of Slc7a11 which encodes for the cystine/glutamate transporter. In addition, the glutamate- cysteine ligase catalytic subunit (Gclc) and glutathione synthetase (Gss), were dose-dependently induced 4.9- and 3.8-fold, respectively. The GSH/GSSG ratio decreased from 80 to 4 at 30 µg/kg TCDD consistent with increased levels of oxidative stress and protein glutathionylation. These results suggest cysteine transport was induced in response to the inhibition of the transsulfuration pathway by TCDD to support glutathione biosynthesis in an oxidative environment. Funded by Superfund Research Program P42ES04911, NIEHS grant (R01ES029541) and NIEHS Multidisciplinary Training in Environmental Toxicology (NIEHS P2T32ES007255).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

208 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-46

Tell us about your Data Science Achievements!

Kristi Pettibone, National Institute of Environmental Health Sciences Chris Duncan, National Institute of Environmental Health Sciences Amanda Garton, National Institute of Environmental Health Sciences Nidhi Gera, National Institute of Environmental Health Sciences

Superfund Mandates: None

Superfund Center: Other: National Institute of Environmental Health Sciences

NIEHS and the Superfund program have been making significant investments in data science infrastructure and activities. We’ve also been developing a framework for tracking NIEHS data science achievements and would love to hear about your accomplishments. Perhaps you are creating data sets that can be used by others, developing new ways to manage or share data or creating data quality control plans? Or maybe you are developing new software or applying new methodologies for interoperability? How are you applying the FAIR principles? What data have you disseminated, what ontologies have you developed or contributed to? Have you developed any data science training courses or materials? We want to hear about it all. Join us for an interactive poster presentation about where and how to report your data science successes as well as challenges. We’ll also give you a sneak peak of the resources that will be available on the soon to be published NIEHS Data Science webpage and get your input on other resources we should include.

Email: [email protected] Presenter Status: Evaluator

NIEHS SRP 2020 Virtual Annual Meeting Program • 209 Poster Session III: Poster #III-47

On-spot quantitative analysis of Dicamba in field waters using a lateral flow immunochromatographic strip with smartphone imaging

Meng Qi, Hebei Agricultural University, University of California-Davis Cong He, Hebei Agricultural University Hui Zhou, Hebei Agricultural University Dongyang Li, University of California-Davis Yuxin Wang, University of California-Davis Jingqian Huo, Hebei Agricultural University Jinlin Zhang, Hebei Agricultural University Bruce D. Hammock, University of California-Davis

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of California-Davis Superfund Research Center

Dicamba, one of the old and most widely used selective herbicides is increasingly used in the world, in particular with the widespread cultivation of genetically modified dicamba-resisted crops. However, the drift problem in the field has caused phytotoxicity against naive, sensitive crops, raising legal concerns. Thus, it is particularly timely to develop a method that can be used for the on-spot rapid detection of dicamba in the field. In this paper, a lateral flow immunochromatographic strip (LFIC) was developed. The quantitative detection can be conducted by an App in the smartphone, named “Color Snap”. The tool reported here provides results in 10 mins and can detect dicamba in water with a LOD (detection limit) value of 0.1 mg/L. The developed LFIC shows excellent stability and sensitivity appropriate for field analysis. Our sensor is portable and excellent tool for on-site detection with smartphone imaging for better accuracy and precision of the results. This tool could provide guidance on the rational use of dicamba in the field. It also provides a tool to deal with legal concerns about dicamba drift.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

210 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-48

A Novel Framework to Form Sufficiently Similar Mixtures from Environmental Exposure Data

Brianna Rivera, Oregon State University, Corvallis, OR Christine Ghetu, Oregon State University, Corvallis, OR Kim A. Anderson, Oregon State University, Corvallis, OR Susan C. Tilton Oregon State University, Corvallis, OR

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Oregon State University Superfund Research Center

Sufficiently similar mixtures are used as a whole mixture approach to conducting safety assessments of complex chemical mixtures. Currently, there is a lack of an established framework for forming sufficiently similar mixtures, from environmental exposure data, to characterize hazard of exposure at a contaminated site. In this study, several approaches are explored to form sufficiently similar mixtures that can be used for a toxicological evaluation. Existing toxicity values and potential for exposure were incorporated to establish an overall rank to identify drivers of toxicity from an environmental mixture. Coal Tar Extract (CTE), a well-studied complex mixture will serve as the “test mixture,” and data from air sampling at a legacy creosote site will be the “real-world mixture.” Representative mixtures of the test and real-world mixture were formed using several approaches based on relative abundance (ExpoMix), average toxicity values (AVG-ToxMix), and a weighted approach incorporating toxicity and abundance (WAVG-ToxMix). Representative mixtures were made using mixing proportions based on the concentrations found in the test or real-world mixture. Mixtures were compared using 3D normal human bronchial epithelium. Differences in bioactivity between ExpoMix and AVG-ToxMix were evaluated using biomarkers of xenobiotic metabolism, oxidative stress, tight-junction integrity, and DNA damage. Results show that AVG-ToxMix was more bioactive (q<0.05) than ExpoMix confirming that less abundant, more potent chemicals are driving toxicity. Future directions involve comparing benchmark dose responses of ExpoMix, AVG-ToxMix, and WAVG-ToxMix to their respective whole mixtures to test for sufficient similarity. Results from this study will help inform regulatory agencies of the most effective ways in which sufficiently similar mixtures can be formed and evaluated in order to conduct a safety assessment.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 211 Poster Session III: Poster #III-49

Identification of polycyclic aromatic hydrocarbons (PAHs) in complex mixtures using non-targeted high-throughput analysis using ion mobility spectrometry- mass spectrometry (IMS-MS)

Alina T. Roman-Hubers, Texas A&M University, College Station, Texas Alexandra Cordova, Texas A&M University, College Station, Texas Arlean Rohde, Texas A&M University, College Station, Texas Thomas McDonald, Texas A&M University, College Station, Texas Erin Baker, North Carolina State University, Raleigh, North Carolina Dillon Lloyd, North Carolina State University, Raleigh, North Carolina Fred Wright, North Carolina State University, Raleigh, North Carolina Weihsueh A. Chiu, Texas A&M University, College Station, Texas Ivan Rusyn, Texas A&M University, College Station, Texas

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Texas A&M University Superfund Research Center

PAHs are a class of environmental contaminants that occur in multi-component mixtures. Current gas chromatography-mass spectrometry-based analytical methods for PAHs detection are time-consuming and have limited resolving power. Recent developments in IMS-MS, a high-resolution post-ionization separation technique, show its advantages for PAHs exposure assessment based on high mass accuracy and resolution, and an ability to distinguish molecules based on their collisional cross-section (CCS). However, non-targeted methods such as IMS-MS require verification of the molecular identity of the chromatographic features, a process that may be time-consuming. We hypothesized that high-resolution IMS-MS data can be deconvoluted through Kendrick Mass Defect (KMD)-based analysis of m/z and CCS to generate confident compositional information on PAHs in mixtures. We analyzed individual representative PAHs to demonstrate that CCS data improves the evaluation of homologous series based on both molecular composition (KMD vs. m/z) and structural (KMD vs. CCS) information. KMD analyses were conducted using a number of hydrocarbon-specific functional groups (CH2, C4H2, C6H6, H), as well as C and H trend-lines. Two-dimensional KMD analysis allowed for distinguishing isomeric structures (e.g., 1-methylnaphthanlene and 2-methylnaphthanlene). We then assembled KMD analysis steps into a workflow that confirmed each feature’s identity using multiple lines of evidence. This computational analysis workflow was applied to confidently identify PAHs (>90% of the features) in a number of hydrocarbon-containing mixtures of varying complexity. Overall, our results show that IMS-MS coupled with atmospheric photooxidation ionization (APPI) is a promising non-targeted method for analysis of hazardous PAHs. Together with a data analysis workflow for confident identification of PAHs demonstrated herein, IMS-MS is suitable for rapid exposure assessment of hazardous substances in the environment following natural disasters. This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

212 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-50

Resolving the roles of exposed crystal facet and surface organic groups in the sorption behavior of water contaminants on nano metal oxides

Holly E. Rudel, Yale University Julie B. Zimmerman, Yale University

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Unaffiliated

Nano metal oxides (NMOs) have recently garnered significant research attention in a variety of water treatment applications including catalysis and direct sorption. With this increased interest in the applications of NMOs comes an increased need to understand the inherent structural functionality that gives rise to enhanced sorption performance. Strategically controlling the exposed crystal facet, and consequently exposed surface atoms, of NMOs is a promising way to increase the efficiency and selectivity of these sorbents in treating contaminated water. This control is typically achieved by employing hydro- and solvo-thermal synthetic methods that utilize surface directing agents, or capping agents (CAs), which mediate the growth direction of the nucleated nanoparticles. However, CAs can remain persistent on the surface of the NMO, even after post-synthesis treatment, and can hinder adsorbate access to active sorption sites. Single crystalline iron oxide nanoparticles with different exposed crystal facets were synthesized in the presence of organic CAs and characterized using SEM, (HR)-TEM, XRD, XPS, ATR-FTIR and TGA. Batch adsorption experiments with clean-surface- and capped-NMOs were performed to elucidate the convoluted roles of crystal facet and CAs in sorption performance.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 213 Poster Session III: Poster #III-51

Functional Genomics Discovery of “Hidden” Genes in Heavy Metal Tolerance and Accumulation in Plants

Julian Schroeder, University of California, San Diego Jazmin Rio, University of California, San Diego Qingqing Xie, University of California, San Diego, and Salk Institute for Biological Studies La Jolla, CA Joy Guo, University of California, San Diego Qi Yu, University of California, San Diego Timothy O. Jobe, University of California, San Diego, and University of Cologne, Germany Yasman Zarabi, University of California, San Diego Paulo H. O. Ceciliato, University of California, San Diego Felix Hauser, University of California, San Diego

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California, San Diego Superfund Research Center

Heavy metal and arsenic contamination in soils at Superfund sites is an urgent problem. Plants have the potential of providing a low cost bioremediation platform for removal of toxic heavy metals and arsenic from soils. While previous studies mainly concentrated on the identification of the function of single genes for enhancing remediation, identification of functionally overlapping genes that mediate rapid detoxification in plants has not been previously feasible. Plant genomes have large gene families, causing redundancies. Artificial microRNAs (amiRNAs) are small non-coding RNA molecules that down-regulate gene expression in plants by targeting mRNAs for repression. AmiRNAs provide a powerful strategy to alleviate the limitations of functional redundancy in traditional screening methods in plants. Therefore, our laboratory has computationally designed genome-wide subclade-specific amiRNA libraries with a high-performance computing cluster, for genome-wide knock-down of homologous gene family members (F. Hauser, et al. Plant Cell). We have now generated over 14,000 individual plant T2 lines expressing these amiRNAs (F. Hauser, P. H. Ceciliato, et al. J. Exp. Bot. 2019). AmiRNA lines have been screened in response to cadmium or arsenic treatment to identify new genes that affect the responses to these toxicants. We report here new genes that are key to cadmium and arsenic resistance and accumulation. We have identified amiRNA lines targeting closely homologous ERF transcription factors, protein kinase superfamily members, stress-dependent transcription factors, phosphate transporters and many other interesting homologous gene sets that affect arsenic and cadmium sensitivity and toxicant accumulation. In summary, our results provide novel insights and uncover novel genes and mechanisms that mediate heavy metal tolerance and accumulation in plants. These newly discovered genes can be targeted for their potential for improved bioremediation.

Email: [email protected] Presenter Status: Researcher

214 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-52

Mapping the chromatin and transcriptional regulatory landscape of arsenic- exposed beta cells

Praveen Sethupathy, Cornell University Rowan Beck, University of North Carolina Chapel Hill, Chapel Hill, North Carolina Kieran Koch-Laskowski, Cornell University Mohit Chandi, University of North Carolina Chapel Hill, Chapel Hill, North Carolina Matt Kanke, Cornell University Ramja Sritharan, Cornell University Miroslav Stýblo, University of North Carolina Chapel Hill, Chapel Hill, North Carolina Praveen Sethupathy, Cornell University

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of North Carolina at Chapel Hill Superfund Research Center

Type 2 diabetes is a complex metabolic disorder characterized by insulin resistance and beta cell dysfunction. Our studies have shown arsenite (iAsIII) and its methylated trivalent metabolite, MAsIII, can impair insulin secretion by beta cells, but the underlying mechanisms remain uncharacterized. Transcriptional regulatory elements (TREs) that play a major role in transcriptional regulation of gene expression, such as enhancers, have emerged as potential targets for iAs exposure. Enhancers are defined as stretches of DNA that promote gene transcription by serving as binding sites for transcription factors (TFs), transcriptional coactivators, and RNA polymerase. In this study, we exposed rat insulinoma pancreatic beta-like cells (INS-1 832/13) to non-cytotoxic concentrations of iAsIII (1 µM) or MAsIII (0.5 µM) that inhibited glucose-stimulated insulin secretion. RNA sequencing was then used to investigate changes in gene expression. Matched chromatin run-on sequencing (ChRO-seq) was performed to identify differentially expressed TREs in the exposed cells. The high-throughput sequencing revealed distinct gene expression signatures specific to each exposure and also distinct TRE profiles. Further analysis of the TREs lost upon exposure indicated an enrichment for binding sites of PDX1 and RFX6, which are transcription factors critical for proper pancreatic beta cell maintenance and function. These altered TRE profiles may represent an important mechanistic link between iAs exposure and beta cell dysfunction that merits further investigation.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 215 Poster Session III: Poster #III-53

UNM METALS Community Engagement: Citizen and Indigenous Science in Assessments of Agricultural Lands Near Abandoned Uranium Mines

Chris Shuey, University of New Mexico, Alburquerque, New Mexico Paul Robinson, University of New Mexico, Southwest Research and Information Center Mae Franklin, Shawn Newell, Stanley Robbins and Tommy Rock, Jacques Seronde, Rona Williams, Cameron Agricultural Ad Hoc Committee Jeff Gaco, and Leon Kie, Pueblo of Laguna Vince Rodriquez, Pueblo of Laguna Kyle Swimmer, University of New Mexico, Southwest Research and Information Center, Pueblo of Laguna Joseph Hoover, Montana State University-Billings Dana Clashin and Jani Ingram, Northern Arizona University Robert Metzger, Radiation Safety Engineering, Inc. Evan Kipnis, University of Utah and Luna Natoli, University of New Mexico Savannah LaRosa-LoPresti, Eliane El Hayek, Adrian Brearley, José Cerrato, Matthew Campen, and Johnnye Lewis University of New Mexico

Superfund Mandate: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of New Mexico Superfund Research Center

Tribal communities in the Southwest are aiming to restart or expand traditional crop-growing to address food security and food sovereignty objectives. The presence of abandoned uranium mines (AUMs) near existing and proposed farmlands raises questions about the safety of the lands and the crops grown on them. Two indigenous communities working with the University of New Mexico METALS Center are taking part in community-engaged soil and plant sampling programs to assess possible impacts on farmland integrity and food safety of releases of metals mixtures and radioactive elements from adjacent AUMs. On the Pueblo of Laguna (NM), residents expressed concern about possible impacts of deposition of dusts from the massive Jackpile Uranium Mine on local farmland. More than 20 farms and gardens were sampled by teams of community members and METALS investigators in summer 2020. Results are pending. In the Cameron Chapter on Navajo Nation (AZ), community members sought collaboration with UNM and other universities to assess contaminant concentrations on 400 acres of fallow lands slated for development of commercial-scale farming. Teams of community members and researchers conducted surface radiation surveys and collected soil and plant samples according to established protocols. Preliminary soil and plant-sampling data showed that concentrations of radioactive substances and metals in soils exhibited little if any demonstrable effects of releases from 27 AUMs that border the lands. Members of both communities embraced the challenges of conducting rigorous environmental health science while applying traditional ecological knowledge related to soil properties and crop production. Results of these collaborations will help inform policies on AUM remediation and expanding healthy food production in remote and underserved areas.

Email: [email protected] Presenter Status: Researcher

216 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-54

Application of epoxyeicosatrienoic acids and soluble epoxide hydrolase inhibitors towards reducing cadmium and mercury induced toxicity in proximal tubular cells

Nalin Singh, University of California – Davis, Davis California Bruce D. Hammock, University of California – Davis, Davis California

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California-Davis, Davis California

Cadmium and Mercury are widespread environmental toxins and exposure to these heavy metals poses significant adverse health risks. Their inorganic salts (i.e. HgCl2 and CdCl2) are highly water soluble and are therefore especially toxic to the kidney, in particular the proximal tubules. Selective uptake into proximal tubular cells triggers cellular dysfunction and subsequent death and implicated modes of toxicity include mitochondrial disruptions, generation of reactive oxygen species (ROS), induction of Endoplasmic Reticulum (ER) stress and a robust inflammatory response. Currently, no safe or efficacious agents exist to preserve renal health against heavy metal toxicity. Polyunsaturated fatty acids are oxidized by Cytochrome P450 to epoxy fatty acids (EpFAs), mono-epoxide signaling molecules with various, promising beneficial bioactivities. Arachidonic acid derived Epoxyeicosatrienoic acids (EETs) are the most well studied group of EpFAs. Furthermore, soluble epoxide hydrolase (sEH) inhibitors are pharmacological tools that block the key degradation pathway of endogenous EpFAs, thereby facilitating their bioavailability and function. Through anti-inflammatory action and stabilization of the mitochondrial dysfunction-ROS-ER stress axis, EETs and sEH inhibitors have shown to ameliorate a wide range of pathologies and toxicities. Hence, in this study, the therapeutic activity of EET regioisomers and an sEH inhibitor towards inorganic cadmium and mercury toxicity was assessed in LLC-PK1 cells (an in vitro proximal tubules model) to investigate a novel pathway that could be utilized to shield human health from heavy metal toxicity.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 217 Poster Session III: Poster #III-55

The Human Health Analysis Resource (HHEAR): An Analytical Chemistry Resource for your Superfund Project

Heather Stapleton, Duke University, Durham, North Carolina P. Lee Ferguson, Duke University, Durham, North Carolina Heileen Hsu-Kim, Duke University, Durham, North Carolina Manish Arora, Icahn School of Medicine at Mount Sinai, New York City, New York

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Duke University Superfund Research Center

The Human Health Environmental Analysis Resource (HHEAR) program seeks to advance understanding of the role of the environment on human health over a lifetime. HHEAR was designed to provide researchers access to high-quality, exposure-assessment services at no cost to the investigator. Starting in 2019, the HHEAR program expanded to include the Duke Environmental Analysis Laboratory (DEAL). The Mount Sinai Untargeted Hub was part of the CHEAR program, the predecessor to HHEAR. Both hubs provide NIEHS and Superfund researchers with support for analytical services to help detect and measure organic contaminants (e.g., flame retardants, polyfluoroalkyl substances, phthalates, pesticides) and inorganic chemicals (e.g. major and trace metals/metalloids, as well as speciation and measures of bioaccessibility) from diverse sampling media. DEAL analyzes external environmental samples including air, water, soil, dust, and personal samplers (e.g. silicone wristbands). The Mount Sinai hub provides exposomic analysis of biological matrices, including blood, urine, dry blood spots, and teeth. The Hubs provide advice to applicants and support for sample preparation, extraction and quantitative analysis of organic and inorganic contaminants. Both hubs utilize the newest cutting-edge approaches for suspect- screening and non-targeted analyses of chemicals using high resolution mass spectrometry. We provide high-resolution, accurate-mass, mass spectrometry services for use in developing exposomics approaches and methodologies. HHEAR’s high-resolution mass spectrometry capability and technical expertise will be used to help measure the totality of human exposure to chemicals, (i.e., the human exposome, consisting of thousands of endogenous and exogenous chemicals). This presentation will provide an overview of the analytical services that are available to Superfund researchers through the HHEAR program.

Email: [email protected] Presenter Status: Researcher

218 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-56

Temporal effects of salt perturbation on the structural dynamics of a defined Dehalococcoides mccartyi-containing dechlorination consortium in completely mixed flow reactors

Mohan Sun, University of California-Berkeley Jae Ho Bae, University of California-Berkeley Eric Troyer, University of California-Berkeley Tong Liu, University of California-Berkeley Sara Gushgari, University of California-Berkeley Shan Yi, University of California-Berkeley, University of Auckland Lisa Alvarez-Cohen, University of California-Berkeley, Lawrence Berkeley National Laboratory

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California-Berkeley Superfund Research Center

Groundwater salinization due to climate change and human activities presents emerging challenges to in situ bioremediation due to elevated osmopressure imposed on microbial communities. Our previous batch culture study unveiled a cascade of responses and osmoadaptation mechanisms of a defined consortium consisting of Dehalococcoides mccartyi str. 195 (Dhc195), Desulfovibrio vulgaris str. Hildenborough (DvH), and Pelosinus fermentans str. R7 (PfR7) during trichloroethene (TCE) dechlorination under increased salinity. In this study, we further investigated the performance and interactions of the consortium members under high salinity using completely mixed flow reactors (CMFRs) that better simulate in situ bioremediation sites. Our results indicate that the consortium maintained robust dechlorination activity under a salinity up to 315 mM Na+ without exogenous osmoprotectant amendment. Intracellular accumulation of osmoprotectants, such as beta-mannosylglycerate and glutamate, might be responsible for the tolerance of high salinity. Although each member within the consortium demonstrated distinguished growth patterns, network analyses using random matrix theory show higher interactions between (1) transcriptomes of Dhc195 and DvH due to their syntrophic relationship and (2) intracellular metabolites and gene expression of Dhc195 and DvH. Our study provides a fundamental explanation of the high salt tolerance of the defined consortium in a CMFR setting, which can be extrapolated to understand in situ bioremediating communities that possess similar metabolic structures.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 219 Poster Session III: Poster #III-57

PAD4 induced NETosis and decreased effrocytosis attributes to cadmium induced emphysema

Ranu Surolia, University of Alabama at Birmingham, Birmingham, Alabama Fu Jun Li, University of Alabama at Birmingham, Birmingham, Alabama Tejaswini Kulkarni, University of Alabama at Birmingham, Birmingham, Alabama Pooja Singh, University of Alabama at Birmingham, Birmingham, Alabama Zheng Wang, University of Alabama at Birmingham, Birmingham, Alabama Veena B. Antony#, University of Alabama at Birmingham, Birmingham, Alabama

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Alabama at Birmingham Superfund Research Center

Cadmium (Cd) is a major component of cigarette smoke and an environmental pollutant that can cause emphysema. Neutrophils exposed to Cd contribute to neutrophil extracellular trap formation (NETosis). Macrophages clear NETs and dead neutrophils by the process of ‘efferocytosis’. Macrophages from COPD and emphysema patients have decreased capacity for efferocytosis. In this study, we examined the role of Cd and Cd-induced NETs on macrophage efferocytosis capacity. We found that the direct exposure (of what) to Cd results in inhibition of NF-KB pathways and efferocytosis. Furthermore, NETs isolated from Cd treated neutrophils also inhibit the NF-KB pathway and diminish the efferocytosis of NETs. Besides, the oxidized mitochondria bound to Cd-induced NETs have inhibitory effects on macrophages efferocytosis ability. The prolonged presence of Cd-induced NETs due to lack of efferocytosis leads to the worsening of emphysema. Our in vivo data demonstrated that treatment with PAD4 inhibitors protected from Cadmium-induced emphysema. Overall, the inhibition of PAD4 protects from Cd-induced emphysema. # Corresponding author

Email: [email protected]; [email protected] Presenter Status: Researcher

220 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-58

Identification of per- and poly-fluoroalkyl substances (PFASs) as novel androgen receptor antagonists and potential endocrine disruptors

Phum Tachachartvanich, University of California-Davis, Davis, CA Azhagiya Singam Ettayapuram Ramaprasad, University of California, Berkeley, Berkeley, CA Kathleen A. Durkin, University of California, Berkeley, Berkeley, CA J. David Furlow, University of California-Davis, Davis, CA Martyn T. Smith, University of California, Berkeley, Berkeley, CA Michele A. La Merrill, University of California-Davis, Davis, CA

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

Perfluoroalkyl and polyfluoroalkyl substances (PFASs) pose a substantial threat as endocrine disruptors, and thus early identification of those that may interact with steroid hormone receptors, such as the androgen receptor (AR), is critical. We screened 5,206 PFASs from the CompTox database against the different binding sites on AR using both molecular docking and machine learning techniques. The combination of docking-based screening and machine learning models identified 23 PFASs with strong potential for activity against AR. We attempted to procure these chemicals for biological validation of the computational screen and were able to obtain 5 commercially with a purity of 90%. We assessed androgenic and antiandrogenic activities of these chemicals in vitro. Of the five candidate AR ligands, 3 of them significantly modified AR transactivation. These 3 PFASs elicited an antagonistic effect at AR with 20% relative inhibitory concentrations of 10.5, 3.1 and 2.8 μM, respectively. Furthermore, we investigated the mechanism of AR inhibition of these 3 chemicals and found that all acted via a competitive antagonism at the ligand binding site. One chemical was found to be inactive because of potent cytotoxicity and we suspect the other PFAS may not reside in the ligand binding site for long periods. We are unaware of any information regarding the use, toxicity, or fate of these PFASs.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 221 Poster Session III: Poster #III-59

Super-adsorptive flexible porous organic cages for vaporous pesticides adsorption and detoxification

Peixin Tang, University of California, Davis Gang Sun, University of California, Davis

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California, Davis Superfund Research Center

Highly porous nucleophilic organic cages (Nu-POC) were in-situ synthesized on cotton fibers by a condensation reaction between cyanuric chloride and melamine, and the products can be employed as robust and flexible decontamination and detoxifying material (denoted as POCotton) against vaporous pesticides. The covalent growth of Nu-POC particles on surfaces of cotton fibers retained the physical characteristics of Nu-POC to the greatest extend, including specific surface area and porosity, while the cotton fabrics still remained flexible. Here, fumigants were used as the toxicant representatives to evaluate the protection efficiency of the POCotton. The resultant POCotton can repeatedly adsorb fumigant vapors instantly (i.e., equilibrium reached in one minute) and massively (i.e., the adsorption capacity of methyl iodide at 596.88 mg g-1). The nitrogen atoms in triazine rings of Nu-POC on POCotton are nucleophilic to detoxify sequestered fumigants or other electrophilic toxicants during long-term uses and storage. A colorimetric signal, which was formed by the formation of imine groups in the Nu-POC structure via fumigant detoxifications, could reveal the failure of the POCotton functions. The successful formation of Nu-POC particles on cotton fibers with retention of Nu-POC performance in terms of rapid fumigant adsorption and detoxification, provides great potentials as chemical protective materials. Acknowledgment: The research was partially supported by UCD Superfund (NIEHS Grant No. 5P42ES004699).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

222 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-60

Integrating Prediction Models and Urban Analytics into Scenario-based Resilient Design

Zhihan Tao, Texas A&M University, College Station, Texas Galen Newman, Texas A&M University, College Station, Texas Karishma Joshi, Texas A&M University, College Station, Texas Jiali Liu, Texas A&M University, College Station, Texas Kevin Fries, One Concern

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

Urban expansion can worsen climate change conditions and enlarge hazard zones. The use of land change prediction modelling to inform scenario-based planning can help increase capabilities when dealing with uncertainties in urbanization such as urban growth and flood risk. This research uses the Land Transformation Model (LTM) to predict three different urban growth scenarios for a highly contaminated site in Tampa, FL. The LTM is used to predict potential future urban growth according to the following scenarios: 1) Business as usual –growth based on current growth patterns; 2) Growth as planned –growth based on the current land use plan; and 3) Resilient growth –growth based on all future development occurring outside of the floodplain. The site comprises a coastal neighborhood which is both socially and physically vulnerable. It is heavily affected by flooding and characterized by industrial land uses, brown fields, and sites listed by the US Environmental Protection Agency’s Toxic Release Inventory. The research asks, how effective is the current comprehensive plan in adapting urban growth to decreasing flood risk and pollutant load? To achieve this, we develop master plans according to each predicted urban growth scenario then assesses their probable impacts of each using the Long-Term Hydrologic Impact Analysis Low Impact Development Spreadsheet as a performance model. Findings show that the current future land use plan for Tampa, while it appears to be better than current patterns of development, has higher flood exposure, stormwater runoff, and pollutant discharge than current conditions, but much higher conditions than resilient growth. All 14 pollutants examined decrease significantly in the resilient growth scenario, compared to other scenarios, and nearly 30 % lower than the current conditions. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee

NIEHS SRP 2020 Virtual Annual Meeting Program • 223 Poster Session III: Poster #III-61

Hexavalent Chromium Decreases Securin Expression and Increases Separase Substrate Cleavage in Human Lung Cells

Jennifer H. Toyoda, University of Louisville, Louisville, Kentucky Julieta Martino, University of Louisville, Louisville, Kentucky Rachel Speer, University of Louisville, Louisville, Kentucky John Pierce Wise, Sr., University of Louisville, Louisville, Kentucky Lu Cai, University of Louisville, Louisville, Kentucky

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Hexavalent chromium [Cr(VI)] is a well-known lung carcinogen with environmental and occupational exposure risks. Despite decades of industrial use and widespread environmental contamination, its carcinogenic mechanism is not well understood. Cr(VI) induces chromosome instability (CIN), including changes in chromosome number which may be explained by its ability to cause centrosome amplification. Centrosome amplification is observed in tumors and is an early event after Cr(VI)-exposure. This study investigates the mechanism of Cr(VI)-induced centrosome disruption in human lung cells. Our hypothesis is Cr(VI) alters activity of separase, the enzyme that cleaves centriole linkers, by decreasing its inhibitor, securin, thus leading to premature centriole disengagement and centrosome amplification. In Cr(VI)- exposed human lung cells, centriole engagement was analyzed by fluorescent immunostaining of centriole markers, CNAP1 and centrin. In normal interphase cells, centriole engagement blocks premature centriole duplication. Our findings show increasing concentrations and prolonged exposure to Cr(VI) increased centriole disengagement in human lung cells. Separase is the key enzyme responsible for cleaving centriole linkers and causing centriole disengagement. Separase activity is tightly controlled by its inhibitor, securin. We show Cr(VI) exposure caused significant loss of securin protein compared to untreated cells, and furthermore mRNA analysis shows Cr(VI) reduced securin expression. Kendrin is a key protein in stabilizing centriole engagement, and a substrate of separase cleavage. Protein analysis shows full length kendrin is reduced after Cr(VI) exposure. These data support the conclusion that Cr(VI) decreases securin protein levels by reducing securin expression, which disrupts separase inhibition. Premature centriole disengagement is a potential key to Cr(VI)-induced numerical CIN and may be due to increased kendrin cleavage by aberrant separase. This work was supported by NIEHS grant R01ES016893 (J.P.W.) and T32-ES011564 (J.H.T.).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

224 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-62

Testing the Efficacy of Broad-Acting Sorbents for Environmental Mixtures using Isothermal Analysis, Mammalian Cells, and H. vulgaris

Meichen Wang, Texas A&M University, College Station, Texas Zunwei Chen, Texas A&M University, College Station, Texas Ivan Rusyn, Texas A&M University, College Station, Texas Timothy Phillips, Texas A&M University, College Station, Texas

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

The hazards associated with frequent exposure to polycyclic aromatic hydrocarbons (PAHs), pesticides, Aroclors, plasticizers, and mycotoxins are well established. Adsorption strategies have been proposed for the remediation of soil and water, although few have focused on the mitigation of mixtures. This study tested a hypothesis that broad-acting sorbents can be developed for diverse chemical mixtures. Adsorption of common and hazardous chemicals was characterized using isothermal analysis from Langmuir and Freundlich equations. The most effective sorbents included medical-grade activated carbon (AC), parent montmorillonite clay, acid-processed montmorillonite (APM), and nutrient-amended montmorillonite clays. Next, we tested the ability of broad-acting sorbents to prevent cytotoxicity of class- specific mixtures using 3 mammalian in vitro models (HLF, ESD3, and 3T3 cell lines) and the hydra assay. AC showed the highest efficacy for mitigating pesticides, plasticizers, PAHs, and mycotoxins. Clays, such as APM, were effective against pesticides, Aroclors, and mycotoxins, while amended clays were most effective against plasticizers. Finally, a sorbent mixture was shown to be broadly active. These results are supported by the high correlation coefficients for the Langmuir model with high capacity, affinity, free energy, and significant protection in cell and hydra models (p < 0.05). The protection percentages in 3T3 cells and hydra models showed the highest correlation as suggested by both Pearson and Spearman with r = 0.84 and rho = 0.73, respectively, (p < 0.0001). Collectively, these studies showed that broad- acting sorbents may be effective in preventing toxic effects of chemical mixtures and provided information on the most effective sorbents based on adsorption isotherms, and in vitro and aquatic organism test methods. This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 225 Poster Session III: Poster #III-63

Triclosan exposure through lactation in neonatal mice leads to an ATF4 dependent lipid accumulation in liver

Andre Weber, University of California, San Diego Elvira Mennillo, University of California, San Diego Jeffrey Ding, University of California, San Diego Mohit Jain, University of California, San Diego Shujuan Chen, University of California, San Diego Robert Tukey, University of California, San Diego

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of California, San Diego Superfund Research Center

Triclosan (TCS) is an antimicrobial agent used in personal care products and is represented as an emerging environmental contaminant worldwide. Recent findings have demonstrated that TCS can accumulate in human breast milk. Neonatal C57BL/6 mice were breastfed from nurturing adults fed with a diet containing 120 ppm TCS. LC-MS/MS analysis in the breastmilk revealed a concentration of 291 ± 172 nM of TCS at day 14 and the serum concentration at 21 days of 527 ± 420 nM. By Western blot analysis we observed an increase in hepatic ATF4 in TCS breastfed neonatal mice, indicating that TCS exposure is activating the PERK-elf2α-ATF4 pathway. Histological sections and cryosection staining with Red Oil O showed accumulation of lipid droplets in the cytoplasm of hepatocytes in TCS breastfed mice at 21 days of age. Related to this finding, genes linked to de novo lipogenesis (Scd1, Scd2 and Srebp1-c) and fatty acid synthase (FASN) protein levels are increased in liver of these mice. RT-PCR analysis revealed lower expression of several genes associated with lipid metabolism like fatty acid synthase (Fasn) and stearoyl- CoA desaturase 1 and 2 (Scd1 and Scd2) in targeted liver ATF4-/- neonatal mice when compared with ATF4F/F. RNA sequencing analysis revealed that pathways associated with PPARα signaling, fatty acid metabolism and ketogenesis are upregulated in TCS treated mice. In contrast, pathways involved in the regulation of hormone levels and additional responses associated with glucocorticoids are downregulated in neonatal mice exposed to lactational transfer of TCS. In summary, these results confirm that TCS in breastmilk is causing fatty liver in neonatal mice, an event that can be linked to ATF4 driven lipid metabolism and ER stress.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

226 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-64

A Comprehensive Profiling Method for Regulatory Lipid Mediators Using UHPLC timsTOF

Jun Yang, University of California-Davis, Davis, California Xuejun Peng, Bruker Scientific LLC Debin Wan, University of California Bruce D Hammock, University of California-Davis, Davis California

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

Regulatory lipid mediators are a class of fatty acids derived from polyunsaturated fatty acids through the enzymatic processes. They played very important roles in various physio-pathological processes such as inflammation, blood pressure regulation, angiogenesis, pain perception. These lipid mediators often coordinate each other to participate in the biological processes mentioned above. Therefore, profiling of these lipid mediators provides the insight of their roles in vivo. Currently, the profiling method for these lipid mediators is dominated by the triple-quadrupole instruments due to the high sensitivity for these low abundance metabolites in the body. In this study, we utilize the high resolution of the TOF MS with ion mobility to achieve higher selectivity and sensitivity for profiling these lipid mediators in vivo.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 227 Poster Session III: Poster #III-65

Metabolite Profiles of a set of hydroxylated metabolites facilitate the identification of the metabolic pathways of 4-chlorobiphenyl (PCB 3) in HepG2 cells

Chunyun Zhang, University of Iowa, Iowa City, Iowa Susanne Flor, University of Iowa, Iowa City, Iowa Gabrielle Ludewig, University of Iowa, Iowa City, Iowa Hans-Joachim Lehmler, University of Iowa, Iowa City, Iowa

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Iowa Superfund Research Center

The elucidation of the metabolic pathways of lower chlorinated PCB congeners, such as PCB 3, is challenging because of the complex metabolite mixtures that are formed both in vitro and in vivo. We investigated the hypothesis that the metabolic pathways of PCBs can be elucidated by preforming parallel metabolism studies with PCBs and selected hydroxylated metabolites. To test this hypothesis, we employed nontarget high-resolution mass spectrometry (Nt-HRMS) to characterize the metabolism of PCB 3 in HepG2 cells, a human-relevant cell line. Briefly, HepG2 cells in the minimum essential medium were exposed for 24 h to PCB 3 and a series of hydroxylated metabolites, including 4’-chlorobiphenyl-2-ol (2’- OH-3), 4’-chlorobiphenyl-3-ol (3’-OH-3), 4’-chlorobiphenyl-4-ol (4’-OH-3), 4-chlorobiphenyl-2-ol (2-OH-3), 4-chlorobiphenyl-3-ol (3-OH-3), 3-chlorobiphenyl-4-ol (4-OH-2), 4’-chlorobiphenyl-2-ol (2’-OH-3) and 4’- chloro-3,4-dihydroxybiphenyl (3’,4’-di-OH-3), in DMSO or DMSO alone. Cell culture media were analyzed with liquid chromatography coupled with Nt-HRMS. Six classes of metabolites were identified in the cell culture medium exposed to PCB 3, including 3’-OH-3 or 4’-OH-3, sulfates of 3’-OH-3, 4’-OH-3, 3-OH-3, and 4-OH-2 (1,2-shift product), one glucuronide of 4’-OH-3, two hydroxylated PCB 3 sulfate, two isomers of methoxylated PCB 3 sulfates (4’-chloro-3-methoxy-4-sulfooxy-biphenyl and 4’-chloro-4-methoxy-3- sulfooxy-biphenyl), and one methoxylated PCB 3 glucuronide. The catechol metabolite, 3’,4’-di-OH-3, was extensively methylated, and the resulting hydroxylated-methoxylated metabolites underwent further sulfation and, to a lesser extent, glucuronidation. Overall, PCB 3 is preferentially oxidized to mono- and di-hydroxylated metabolites in the meta- and para-positions and the highly toxic PCB catechol metabolite, 3’,4’-di-OH-3 is readily transformed to methylated metaboliztes. These results demonstrate that parallel metabolism studies with the parent PCB and selected metabolites facilitate the elucidations of the complex PCB metabolic pathways likely present in the human liver [ES027169, ES013661, and ES005605].

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

228 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session III: Poster #III-66

Identification of Polychlorinated Biphenyl Sulfates in Human Serum

Duo Zhang, University of Iowa, Iowa City, Iowa Panithi Saktrakulkla, University of Iowa, Iowa City, Iowa Kristopher Tuttle, University of Iowa, Iowa City, Iowa Rachel F. Marek, University of Iowa, Iowa City, Iowa Hans-Joachim Lehmler, University of Iowa, Iowa City, Iowa Kai Wang, Dept of Biostatistics, University of Iowa, Iowa City, Iowa Keri C. Hornbuckle, University of Iowa, Iowa City, Iowa Michael W. Duffel, University of Iowa, Iowa City, Iowa

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Iowa Superfund Research Center

Polychlorinated biphenyls (PCBs) are a class of persistent legacy pollutants and are also current inadvertently produced toxic chemicals found in building materials and consumer products. PCB sulfates are metabolic products derived from hydroxylated metabolites of PCBs (OH-PCBs). Both OH-PCBs and PCB sulfates exert multiple toxicological effects on human health such as disrupting thyroid hormone transport and inhibiting steroid sulfotransferases (SULT1E1 and SULT2A1). Although PCB 11 sulfate has been previously detected in human serum samples, the lack of a generally applicable method for a broad range of PCB sulfates in human serum has limited our understanding of their prevalence and importance. We have now developed a method for extraction of PCB sulfates from serum followed by differential analysis with, and without, sulfatase-catalyzed hydrolysis to OH-PCBs. A sulfatase from Helix pomatia was purified by affinity chromatography, and it displayed broad specificity for PCB sulfates without contaminant glucuronidase activity. Following the sulfatase-catalyzed hydrolysis of the PCB sulfates that were extracted from serum, the corresponding OH-PCBs were derivatized to methoxy-PCBs and quantitated by GC-MS/MS. In a pooled sample of human serum, we identified 13 PCB sulfates, and those present in the highest concentrations were: 4-PCB 2 sulfate (4100 pg/g), 4-PCB 11 sulfate (1600 pg/g), and 2’-PCB 3 sulfate (1300 pg/g). All of the PCB sulfate congeners that were identified in a pooled sample of human serum contained lower numbers of chlorine atoms, and this is consistent with the known properties of PCB metabolism. Although this method for PCB sulfates in serum was developed with 74 OH-PCB standards, its usefulness can be readily expanded as more OH-PCB standards become available. [Supported by NIH: P42 ES013661]

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 229 Poster Session III: Poster #III-67

Particulate Matter Exposure from Abandoned Uranium Mines Exacerbates Inflammation in an Autoimmune Mouse Model

Katherine E. Zychowski, University of New Mexico, Alburquerque, New Mexico Alexis Wilson, University of Texas at El Paso, El Paso Texas Raul Salazar, University of New Mexico, Alburquerque, New Mexico Guy Herbert, University of New Mexico, Alburquerque, New Mexico Selita Lucas, University of New Mexico, Alburquerque, New Mexico Matthew Campen, University of New Mexico, Alburquerque, New Mexico

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

From the 1940s–1980s, uranium mining occurred throughout tribal lands in Southwestern United States leaving behind residual metal-based mine waste. There is an association between heavy metal exposure and autoimmune development. Navajo communities residing near abandoned uranium mines may be affected by inhaled metal-rich, windblown particulate matter (PM). Therefore, we examined effects of mine site-derived dust exposure in an autoimmune disease model. Lupus-prone mice (NZBWF1/J) were exposed to PM (100µg/50µL), from different mine sites, Claim 28 and St. Anthony mine. Mice were dosed with either dispersion media (control), Claim 28 mine PM or St. Anthony PM via oropharyngeal aspiration. After 24 weeks, lungs and brains were probed for inflammatory biomarkers and serum biomarkers of autoimmunity were assessed. Immunohistochemistry of brain tissue sections revealed significant staining including Iba-1 and GFAP in Claim 28 and St. Anthony groups. Learning/memory deficits were evident in Claim 28-exposed mice, as determined by decreased Y-arm frequency. There was a significant increase in serum neutrophil elastase in St. Anthony’s group. Pulmonary inflammation was observed by bronchoalveolar lavage neutrophil infiltration in both PM-exposed groups and lung CCL2 and IL-1 gene expression was significantly upregulated in St. Anthony’s exposure group. Interestingly, several serum autoantigens and biomarkers remained unchanged. In a second set of experiments, we determined that lung inflammation following aspirated PM, may be mediated by peptidyl arginine deiminase type IV (PAD4). Further research is warranted to fully decipher the mechanism driving this effect.

Email: [email protected] Presenter Status: Researcher

230 • NIEHS SRP 2020 Virtual Annual Meeting Program

Poster Session IV: December 15, 2020 | 1:00 – 2:00 p.m.

Last Name First Name Poster Session ID Abbott Derek IV-1 Alghzawi Ma’in IV-2 Banerjee Sohini IV-3 Beard Jessica IV-4 Bhandari Sharmila IV-5 Blevins Lance IV-6 Brisolara Kari IV-7 Buchholz Bruce IV-8 Bullert Amanda IV-9 Campagna Shawn IV-10 Cholico Giovan IV-11 Compton Patrick IV-12 Connolly Craig IV-13 Corless Elliot IV-14 NIEHS De Pree Thomas IV-15 SRP Debarba Lucas IV-16 Deng Pan IV-17 2020 Dunn Matt IV-18 Annual Meeting Eaton Jarrod IV-19 El Hayek Eliane IV-20 El-Moghazy Ahmed IV-21 Farrell Matthew IV-22 Frazar E. Molly IV-23 Gosline Sara IV-24 Guan Xia IV-25 Hahn Mark IV-26 Haine Dana IV-27 Harper Stacey IV-28 Hawkey Andrew IV-29 Hemond Harry IV-30 Hudson Laurie IV-31 Jahnke Jacob IV-32 Jasperse Lindsay IV-33

232 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: December 15, 2020 | 1:00 – 2:00 p.m.

Last Name First Name Poster Session ID Kaihatu James IV-34 Kamal Abu Hena Mostafa IV-35 Leniz Francisco IV-36 Li Nan IV-38 Li Dongyang IV-37 Li Xueshu IV-39 Matsumoto Naoki IV-40 McHale Cliona IV-41 McKenna Anissa IV-42 Mennillo Elvira IV-43 Middleton Beth Rose IV-44 Moorthy Bhagavatula IV-45 NIEHS Nghiem Athena IV-46 SRP Orlowska Karina IV-47 Orr Asuka IV-48 2020 Papuga Shirley IV-49 Annual Meeting Pardo-Manuel de Villena Fernando IV-50 Park Hae-Ryung IV-51 Peel Hannah IV-52 Prossner Kristen IV-53 Santos Rivera Amailie IV-54 Shankar Prathana IV-55 Simonsen Derek IV-56 Taylor Vivien IV-57 Valdiviezo Alan IV-58 Volk Lindsay IV-59 Wang Yuxin IV-61 Wang Hao IV-60 Wise James IV-62 Yamamoto Gen IV-63 Zhu Rui IV-64

NIEHS SRP 2020 Virtual Annual Meeting Program • 233 Poster Session IV: Poster #IV-1

A single cell, multi-parametric high content assay to quantify and classify endocrine disrupting chemicals in environmental contaminants that affect ERβ functions

Derek Abbott, Baylor College of Medicine, Houston Texas Adam Szafran, Baylor College of Medicine, Houston Texas Fabio Stossi, Baylor College of Medicine, Houston Texas Michael Mancini, Baylor College of Medicine, Houston Texas

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Texas A&M University Superfund Research Center

Endocrine disrupting compounds (EDCs) mimic or disrupt the actions of hormone signaling through nuclear receptors. The highly industrialized Galveston Bay/Houston Ship Channel (GB/HSC) and other Superfund sites produce complex, environmentally-relevant chemical mixtures that require rapid, robust, and cost-effective assays to determine EDC potential. To understand the risk to human health due to exposure to chemical toxicants and EDCs, the US EPA has been pursuing new high throughput in vitro assays and computational models to characterize effects on estrogenic signaling. While great effort and numerous assays have been produced to evaluate EDC impact on ERα, the number of assays and data available on ERβ is severely limited. ERβ signaling has been shown to play a prominent role in numerous diseases including infertility, endometriosis, prostate and ovarian cancers. Here, we present an engineered, dox-inducible GFP-ERβ:PRL-HeLa assay to asses direct visualization of the receptor binding to DNA responsive elements, effect upon chromatin binding and transcription function. This assay has been tested against an EPA-designed 45 compound reference set comprised of known ER-modulators (agonists, antagonists, EDCs). Treatment for 2 hours with the EPA45 shows the ability of the assay to distinguish ERβ agonists and antagonists, specificity for ERβ ligands, and robust detection of numerous EDCs. In conjunction with the bioinformatics and data science cores this data set may be used to model EDC activity against ERβ. This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

234 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-2

Investigating the Transformations of Arsenic and Toxic Metals Speciation in Mine Tailings Under Different Climatic Conditions

Ma’in Alghzawi, University of Arizona, Tucson, Arizona Brian Moravec, University of Arizona, Tucson, Arizona Rob Root, University of Arizona, Tucson, Arizona Mark Brusseau, University of Arizona, Tucson, Arizona Jon Chorover, University of Arizona, Tucson, Arizona

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Arizona Superfund Research Center

High concentrations of toxic metal/oids (e.g., As, Pb, Zn) in mine tailings present significant health risk to neighboring communities and ecosystems. These toxic elements are transported in dissolved or colloid form by natural waters or as geodust by wind. Understanding the behavior of these toxic metals and the factors controlling their molecular speciation in tailings sites is crucial to identifying effective tailings management and remediation plans. The climate and tailings primary mineral composition are two important factors that control the rate of the tailings weathering reactions and the types of the reaction products formed in the tailings sites. Furthermore, the transformations of arsenic and toxic metals caused by weathering processes can change their bioavailability and yield particles of higher or lower toxicity than particles originally present in mine tailings. We are aiming to evaluate the weathering processes in different mining sites spanning different states in the southwestern United States. Our hypothesis is that climate controls the distribution and molecular speciation of toxic elements by controlling the depth of the reaction front. We also hypothesize that bioavailability and bioaccessibility of toxic elements are predictable by quantifying element speciation. This project will enable the collection of a reactive transport dataset to identify the biogeochemical reactions controlling element transformations under different environmental conditions. The data collected will be used in implementing a reactive transport and fate model of tailings diagenesis that will serve as a predictive tool for accurate health risk assessment.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 235 Poster Session IV: Poster #IV-3

Environmental exposures of pregnant women to Superfund sites is associated with altered placental metabolomic signatures

Sohini Banerjee, Baylor College of Medicine, Houston, TX Melissa A. Suter, Baylor College of Medicine and Texas Children’s Hospital, Houston, TX Cristian Coarfa, Baylor College of Medicine, Houston, TX Matthew Robertson, Baylor College of Medicine, Houston, TX Nagireddy Putluri, Baylor College of Medicine, Houston, TX Bhagavatula Moorthy, Baylor College of Medicine and Texas Children’s Hospital, Houston, TX Kjersti M. Aagaard, Baylor College of Medicine and Texas Children’s Hospital, Houston, TX

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Baylor College of Medicine Superfund Research Center

Harris County, Houston, has 15 active and 11 remediated Superfund sites which includes a range of contaminants including polycyclic aromatic hydrocarbons, metals, sludge and petroleum waste and has a preterm birth rate that exceeds the U.S. rate (11.4% vs. 9.6%). In addition to factors relating to health disparities and genetic polymorphisms, the role of environmental exposures has been hypothesized as being a potential underlying contributor to preterm birth. Because biochemical and molecular activities of the placenta respond to and are modified by environmental insults, the placenta may be considered to contain a metabolic footprint of the in utero exposures the fetus experiences. The present work is a pilot study to identify metabolic pathways that may be differentially regulated by virtue of proximity of residence near a Superfund site. Untargeted metabolomics was performed using LC-MS placental samples from n=22 women, of which 11 subjects resided in zip codes corresponding to Superfund sites matched to 11 residing in zip codes not near a Superfund sites. The placental samples used are from subjects enrolled in the Bacteria and Birth Study, Aagaard, PI, Clinicaltrials.gov identifier: NCT02392650. Preliminary metabolomic analyses using partial least squares discriminant analysis of the identified features revealed a clear separation between the samples from Superfund and non-superfund sites. Pathway analyses show 49 major pathways of which tryptophan metabolism and cytochrome P450 metabolism pathways were amongst the top 5 identified and were found to be differentially regulated by virtue of residence near a Superfund Site. These findings inspire further large scale investigations to uncover the relation between Superfund sites, metabolites found in the placenta, and their correlation to adverse perinatal outcomes.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

236 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-4

Leveraging the Photochemistry of N-Nitrosamines for their Aqueous Detection

Jessica C. Beard, Massachusetts Institute of Technology, Cambridge, Massachusetts Timothy M. Swager, Massachusetts Institute of Technology, Cambridge, Massachusetts

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Massachusetts Institute of Technology Superfund Research Center

N-Nitrosamines are a class of compounds known to be highly toxic and carcinogenic; the best-known of these is N-nitrosodimethylamine (NDMA). These hazardous chemicals can be found as pollutants in air, water, and soil, but efforts to understand their formation and spread in the environment are hindered by a lack of inexpensive, portable methods for their detection. Our work here focuses on the creation of new in-field detection methods for N-nitrosamines in water by leveraging their well-known photolytic behavior to produce an optical response. Under UV light, N-nitrosamines undergo nitrogen-nitrogen bond fragmentation to produce electrophilic species. Subsequent reaction of those fragments with a small- molecule indicator can produce a colored or fluorescent product, thereby allowing optical detection of the parent N-nitrosamines.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 237 Poster Session IV: Poster #IV-5

Evaluating vulnerability to flood-induced chemical contaminant releases through integration of geospatial data and hydrodynamic simulations

Sharmila Bhandari, Texas A&M University, College Station, TX Kevin Fries, OneConcern, Inc., Menlo Park, CA Feyera Hirpa, OneConcern, Inc., Menlo Park, CA Weihsueh A. Chiu, Texas A&M University, College Station, TX

Superfund Mandates: None

Superfund Center: Texas A&M University Superfund Research Center

Houston has experienced three ‘500-year’ floods 3 years in a row (Memorial Day Flood in 2015, Tax Day Flood in 2016, and Hurricane Harvey in 2017), and chemical contamination after flooding events remains a major concern. This collaborative work from Texas A&M University and One Concern combines flood modeling simulations with GIS data to evaluate the potential risk of chemical releases and contamination due to flooding, and to integrate data into holistic metrics for assessing population vulnerability/resilience. Harvey, Memorial Day Flood and Tax Day Flood scenarios were run to represent three representative “samples” of storm-related vulnerability, the potential risk of flooding was evaluated at 722 chemical facilities in the Houston-Galveston-Brazoria area. Additional baseline metrics of chemical release vulnerabilities were added based on data from U.S. EPA’s Toxic Releases Inventory as well as incidents reported in facility Risk Management plans. Baseline vulnerability and flooding vulnerability data were integrated using ToxPi visualization. Simulation results indicated that while most chemical facilities were not affected in the simulation, the affected facilities had predicted water depths and velocities up to 6.90 m and 2.42 m/s in Harvey, 10 m and 2.1m/s in Memorial Day, and 4.09 m and 2.57 m/s in Tax Day Flood event, respectively. Flood prone facilities tended to concentrate near to Houston Ship Channel area. Low correlation (0.43) was found among baseline and flooding risks, suggesting they are measuring independent vulnerabilities when aggregated and prioritized using ToxPi. These results demonstrate the feasibility of identifying facilities with simultaneously high potential risks from flooding and high risks of chemical releases, and can be used to identify facilities what would benefit from improve resiliency planning. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

238 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-6

Regulation of STAT3 but not STAT1 signaling in human naïve B cells by TCDD- mediated alteration of the SHP-1/PP2a signaling axis

Lance K. Blevins, Michigan State University, East Lansing, Michigan Robert B. Crawford, Michigan State University, East Lansing, Michigan Norbert E. Kaminski, Michigan State University, East Lansing, Michigan

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a persistent environmental contaminant formed as a by- product of organic synthesis and incineration of organic materials. TCDD has potent immunotoxic effects in B lymphocytes resulting in decreased cellular activation and suppressed IgM secretion following activation with CD40 ligand. Previous work from our lab demonstrated that TCDD treatment of naïve human B cells resulted in significant increases in the levels of the tyrosine phosphatase SHP-1 which, corresponded with suppression of IgM secretion. STAT proteins are critical mediators of immune cell activation and effector function via their phospho-regulation. STAT3 is critical for B cell activation and secretion of immunoglobulins (Ig). STAT3 can form homodimers or heterodimers with STAT1, another critical mediator of interferon-mediated immune cell signaling and translocate to the nucleus following phosphorylation as a result of cytokine receptor signaling. We hypothesized that TCDD-mediated increases in SHP-1 could result in decreased STAT3 tyrosine phosphorylation. While we found no alteration of STAT3 or STAT1 tyrosine phosphorylation, there were significant reductions in levels of STAT3 but not STAT1 serine phosphorylation as early as 12 hours following activation. These results corresponded to decreased STAT3-mediated gene transcription and increased PP2a activity, a serine specific phosphatase, which is known to be regulated by SHP-1. Lastly, pharmacological inhibition of SHP- 1 phosphatase activity restored IgM secretion and STAT3 serine phosphorylation in TCDD-treated B cells while decreasing PP2a activation. Together, these data highlight a potential mechanism for TCDD suppression of Ig secretion and demonstrate the potential of inhibition of SHP-1 phosphatase activity as a means to reverse this effect in primary human B lymphocytes. (Support in part by P42 ES004911 and R01ES002520)

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 239 Poster Session IV: Poster #IV-7

ÉCOLE: Interprofessional Approach to Developing a Diverse Workforce Ready to Address Emerging Technologies in Occupational Health and Safety

Kari F Brisolara, Louisiana State University Health Sciences Center, New Orleans, Louisiana Stephania Cormier, Louisiana State University, Baton Rouge, Louisiana Margaret Reams, Louisiana State University, Baton Rouge, Louisiana

Superfund Mandates: None

Superfund Center: R25 award

ÉCOLE, an academic, government and industry occupational health collaborative developed to address emerging technologies in occupational health and safety was created in 2017 with the primary goal to enhance training efforts in the area of emerging technologies in occupational health and safety through interprofessional instruction in the form of workshops. The importance of the occupational health workforce has come to the forefront during the current COVID-19 outbreak as the need to protect workers and the public from infections affects a broad spectrum of industries. This issue has been recognized by the Occupational Safety and Health Administration (OSHA) as having direct impacts to the workplace and occupational health (OSHA 2020). Workshops have been conducted in-person, online synchronous, and online asynchronous. These workshops provide continuing education as required by Certified Safety Professionals (CSP) and Certified Industrial Hygienists (CIH). With a focus on interprofessional education (IPE) methods, teams of students learn about, from and with each other to broaden students’ perspectives on occupational health and develop novel solutions to emerging issues. The core competencies of the Interprofessional Education Collaborative (IPEC) encompass four domains: Values/Ethics for Interprofessional Practice; Roles/Responsibilities; Interprofessional Communication; and Teams and Teamwork (IPEC 2011). These four domains represent the cornerstone of the learning outcomes. Despite the growing body of evidence that interprofessional teams can improve health outcomes, our team is conducting a unique opportunity to broaden the IPE experience outside the healthcare realm.

Email: [email protected] Presenter Status: Administrator

240 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-8

Metabolite Quantitation and Biomarker Development of Toxicants Using Accelerator Mass Spectrometry

Bruce Buchholz, Lawrence Livermore National Laboratory Ted Ognibene, Lawrence Livermore National Laboratory Graham Bench, Lawrence Livermore National Laboratory Ken Turteltaub, Lawrence Livermore National Laboratory

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Other: Lawrence Livermore National Laboratory

Accelerator mass spectrometry (AMS) was originally applied in the life sciences to overcome limitations in detection sensitivity for studying the molecular damage caused by exposure to low levels of environmental carcinogens and pollutants. For example, AMS can be used to conduct metabolite analysis at the picomole to the attomole level and is also being used to identify macromolecular targets for drugs and toxic compounds. Studies using 14C-labeled agents show that activities as low as 5 nCi/person can be used to assess metabolism, and activities as low as 100 nCi/person can be used to address macromolecular binding in the study of environmental toxicants. In most cases the radiation dose is less than that received during a cross-country commercial airline flight. The high sensitivity of AMS allows use of small samples of exfoliated tissues, isolated cell subpopulations, and precious tissues of human or animal origin. Sensitivity also enables quantitative study of ligand—macromolecule interactions at physiologically relevant concentrations and allows studies of toxic substances in humans at exposures at or below environmentally acceptable levels. The increased sensitivity also facilitates the use of compounds that are difficult to synthesize at high specific activity or cannot be used in large amounts. Dermal exposure to permethrin, oral ingestion of aflatoxin and polyaromatic hydrocarbons, and respiratory exposure to naphthalene will be described. This work performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA2734. LLNL-ABS-815330.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 241 Poster Session IV: Poster #IV-9

The effects of polychlorinated biphenyls on the adolescent nervous system, a systematic review

Amanda J. Bullert, University of Iowa, Iowa City, Iowa Hans-Joachim Lehmler, University of Iowa, Iowa City, Iowa

Superfund Mandate: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Iowa Superfund Research Center

Polychlorinated biphenyls (PCBs) can be found in any population as exposure comes from our environments, e.g., soil, air, and building materials. PCBs are present at high concentrations in the indoor air of older school buildings in the United States because of their use in caulking, light ballasts, and floor resins. Exposure to PCB is implicated in many adverse outcomes in humans, including effects on the nervous system; however, the impact of PCBs on the adolescent nervous system has received little attention to date. We conducted a systematic review to identify papers that study neurotoxic outcomes following PCB exposure during the adolescent period in rats (postnatal days 21 to 60) and, subsequently, evaluated the quality of relevant papers using the toxRtool. The exclusion/inclusion criteria for the systematic review were defined ahead of time to avoid bias. After the consultation of a librarian, a literature search was conducted using three different databases, including Pubmed, Scopus, and Embase. Keywords and phrases were entered into each database to ensure no peer-reviewed papers were missing. This search identified 1,598 citations that were imported into EndNote. A total of 600 duplicate results were removed, and the remaining 998 articles were evaluated based on their titles, abstracts, and methods to identify manuscripts that had neurological findings and PCB exposure during the adolescent period. A total of five papers were found to meet all inclusion criteria and, based on the toxRtool score, report rigorous experimental results. This systematic review demonstrates that more research is needed to characterize how PCBs present in the air of schools adversely affect the adolescent nervous system [ES027169, ES013661, and ES005605].

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

242 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-10

Monitoring of bacterial reductive dechlorination processes with metabolomics

Shawn Campagna, University of Tennessee, Knoxville Frank E. Löffler, University of Tennessee, Knoxville Amanda L. May, University of Tennessee, Knoxville Samuel Rosolina, Microbial Insights, Inc., Knoxville, TN Dora Taggart, Microbial Insights, Inc., Knoxville, TN

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: R01/SBIR

Chlorinated ethenes are common contaminants impacting Superfund sites, and detoxification can be achieved by organohalide-respiring bacteria (e.g., Dehalococcoides mccartyi, Dhc) via stepwise reductive dechlorination to harmless ethene. Enhancing the activity of native or bioaugmented Dhc has emerged as a viable strategy for detoxification and reducing exposure risks. Recent advances in liquid chromatography–mass spectrometry (LC–MS) have allowed metabolomics to be a viable monitoring approach for such processes. Not only can LC-MS based metabolomics discover biomarkers for specific metabolic processes, the technique also detects thousands of molecules in a single analysis that can inform about the global metabolic state of the system. Despite the power of this approach, technical challenges have hindered the use of metabolomics to interrogate environmental microbial communities sampled in the field. Herein, efforts to circumvent these challenges and apply this emerging systems biology technique to natural microbiomes relevant for contaminant biodegradation will be discussed. An UPLC-Orbitrap MS was employed to measure the metabolome of the SDC-9™ bioaugmentation consortium during cis-1,2-dichloroethene conversion to vinyl chloride and ethene. An average of ~220 known metabolites and ~16,600 spectral features of unknown structure over the course of dechlorination. Patterns of spectral features (peak patterns) that correlated with dechlorination (in)activity were identified. In order to identify specific dechlorination biomarkers or biomarker patterns from these studies that would be applicable to the field, microbial biomass was collected from groundwater wells across multiple contaminated sites, and these samples yielded a range of~9,000-25,000 spectral features. This application of metabolomics has identified biomarkers/patterns that can potentially inform about the state of reductive dechlorinating activity, highlighting metabolomics’ value as an innovative and affordable site assessment and bioremediation monitoring tool.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 243 Poster Session IV: Poster #IV-11

Genome-wide interactions of AhR, COUP-TF, and HNF4 in TCDD-treated mouse liver

Giovan N. Cholico, Michigan State University, East Lansing, Michigan Rance Nault, Michigan State University, East Lansing, Michigan Tim Zacharewski, Michigan State University, East Lansing, Michigan

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor (TF) most prominently known for mediating the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. The canonical mechanism following ligand binding involves AhR heterodimerization with the aryl hydrocarbon receptor nuclear translocator, translocation into the nucleus, and binding to dioxin response elements leading to differential gene expression. Genome wide AhR chromatin immunoprecipitation assays identified the enrichment of non-canonical enhancer sequences including consensus motifs for COUP transcription factor (COUP-TF) and hepatocyte nuclear factor 4 (HNF4). Moreover, we have reported repression of liver specific gene expression in livers of male mice gavaged with TCDD. The present study examines putative interactions between AhR, COUP-TF, and HNF4 in differential gene expression in male C57BL/6J mice 2 hours after a single bolus dose of 30 μg/kg TCDD. ChIP-seq data were integrated with dose-response bulk RNA-seq data to assess differential gene expression. Results indicate AhR, COUP-TFII, and HNF4α were bound in the genomic region of 6,376 genes, with at least one TF showing differential enrichment. 2,680 of these genes showed differential expression (median fold- change = 0.58) following treatment with 30 µg/kg TCDD for 28 days. Functional analysis of these differentially expressed genes identified enrichment of liver-specific genes suggesting TCDD elicits the loss of liver-specific functions. TCDD increased AhR and, in general, also increased HNF4α binding in the genomic regions of liver-specific genes while decreasing COUP-TFII. This suggests repression of liver- specific gene repression by TCDD is dependent on the loss of COUP-TFII binding and an increase in AhR binding, which interferes with HNF4α regulation of liver-specific genes. GNC is supported by the NIEHS Training Program (T32ES007255). This project is funded by the SRP (P42ES004911).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

244 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-12

Optimizing Fenton-like oxidation of legacy munitions/insensitive high explosives utilizing a cost-effective MnO2-doped activated carbon catalyst in the presence of H2O2

Patrick Compton, Northeastern University, Boston, Massachusetts Akram Alshawabkeh, Northeastern University, Boston, Massachusetts Phil Larese-Casanova, Northeastern University, Boston, Massachusetts

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Northeastern University Superfund Research Center

The effective and efficient degradation of persistent, recalcitrant pollutants by emerging oxidation processes is vital to both reduce hazardous waste and remediate polluted waters. One such advanced oxidation process is the implementation of Fenton chemistry, which can be optimized by the use of heterogeneous catalysts. However, in order to make this AOP viable over conventional treatment methods, the technology needs to be approached from both a technical and economic standpoint. From a heterogeneous catalyst optimization perspective, varying the surface chemistry of activated carbon for use as a catalyst removes precipitation complications associated with utilizing traditional iron species in Fenton chemistry while introducing possible adsorption/regeneration capabilities if combined in a novel electrochemical system. Utilizing an additive-doped activated carbon catalyst in the presence of H2O2, the formation of hydroxyl radicals and degradation of a model compound RDX is hypothesized. Comparing various additives, raw activated carbon doped with 5% MnO2 in the presence of H2O2 realized a high concentration of hydroxyl radical formation while maintaining low cost and relative ease of synthesis. This AC-Mn5 catalyst performed effectively over a wide pH range, with the highest radical formation at acidic pH values. Further optimization can be achieved by determining the most effective and economical concentration of influent H2O2 and its delivery method (external addition vs. in-situ generation). With these optimized processes, the goal of designing a full set of oxidation functions towards RDX removal while maintaining cost-effectiveness and scalability is proposed. It is anticipated this platform holds great promises to eliminate analogous contaminants.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 245 Poster Session IV: Poster #IV-13

Flooding Regulates Groundwater Arsenic Contamination in Southeast Asia

Craig T. Connolly, Columbia University, New York Mason O. Stahl, Union College Beck DeYoung, Union College, Benjamin C. Bostick, Columbia University

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Columbia University Superfund Research Center

Chronic exposure to arsenic (As) from the consumption of groundwater poses a significant health risk for over 200 million people worldwide, especially for those living on floodplains in South and Southeast Asia (S-SE). Levels of groundwater As often range from safe to very dangerous over small spatial scales in S-SE Asia, making it remarkably challenging to identify the principal environmental conditions and their interactions that control groundwater As contamination and heterogeneity. Despite decades of research on the origin of As contamination, we are still unable to predict groundwater As concentrations in most environments and with enough confidence to make effective management decisions that reduce the risk of chronic As exposure. Here we demonstrate that surface flooding is a master variable that integrates the key environmental conditions that regulate groundwater As contamination in S-SE Asia. We develop a machine learning model that relies solely on remotely-sensed measures of flooding frequency and duration along with a parsimonious set of geomorphic variables to predict groundwater As concentrations at the household scale (< 100m)—allowing for the differentiation between areas of low (< 10 ugL-1) and high As at spatial scales consistent with those at which groundwater As heterogeneity is observed. We demonstrate that our approach performs well in Cambodia, Vietnam, and Bangladesh and suggest that it can be reliably applied to similar aquifer systems worldwide to estimate levels of groundwater As exposure and threats to public health with improved accuracy as well as identify safe drinking water sources.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

246 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-14

Leveraging machine learning to develop bias-free CometChip image analysis software

Elliott I Corless, Massachusetts Institute of Technology, Cambridge, Massachusetts Simran Kaushal, Massachusetts Institute of Technology, Cambridge, Massachusetts Norah A. Owiti, Massachusetts Institute of Technology, Cambridge, Massachusetts Le P. Ngo, Massachusetts Institute of Technology, Cambridge, Massachusetts Kemo BS Jammeh, Massachusetts Institute of Technology, Cambridge, Massachusetts Cheng Zheng, Massachusetts Institute of Technology, Cambridge, Massachusetts Peter So, Massachusetts Institute of Technology, Cambridge, Massachusetts Bevin P. Engelward, Massachusetts Institute of Technology, Cambridge, Massachusetts

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Massachusetts Institute of Technology Superfund Research Center

Accumulation of DNA damage can result in mutations leading to development of cancer. A widely used assay to quantify DNA damage in cells is the comet assay, whereby DNA damage is detected by electrophoresing lysed cells. Large intact fragments or chromosome(s) are essentially static under comet assay conditions. DNA damage-induced loops and fragments migrate more readily, producing an image that resembles a comet. The CometChip platform was developed to increase throughput and reduce variation by micropatterning cells in a grid to allow equidistant spacing of cells in a single focal plane. This greatly increases throughput and facilitates automated image analysis. One challenge is to discern the difference between DNA damage-induced comets and comets that arise as a consequence of apoptosis. Apoptotic comets have broad tails, and so users often manually remove them from the analysis. However, the removal process is subjective, and so when performed by different scientists, there can be different results. This manual curation process not only leads to inconsistencies between laboratories, but it is also time consuming. We will gather input from various laboratories, and then create a ground-truth set of training images. We will then leverage machine learning to create open-source, free software so that apoptotic comets can be removed in an automated fashion. This approach will normalize the selection process across all laboratories that use the software, while simultaneously increasing the throughput of analysis by automating the rejection process.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 247 Poster Session IV: Poster #IV-15

The Politics of Baselining in the Grants Uranium Mining District of Northwestern New Mexico

Thomas A. De Pree, University of New Mexico, Alburquerque, New Mexico

Superfund Mandate: None:

Superfund Center: University of New Mexico Superfund Research Center

From 1948 to 1970, the “Grants uranium district” of northwestern New Mexico served as the primary production site for one of the world’s largest nuclear arsenals, in addition to becoming a source for commercial nuclear energy from the late 1960s to the 1990s. By the twenty-first century, after a prolonged period of economic decline that began in the late 1970s, all uranium mining and milling in New Mexico had ceased, leaving a legacy of environmental health impacts. What was once referred to as “The Uranium Capital of the World” now encompasses over a thousand abandoned uranium mines and seven massive uranium mill tailings piles, which are associated with airborne and soil contamination, as well as groundwater plumes of uranium and other contaminants of concern. Drawing from over two years of ethnographic research, I will describe the diverse forms of expertise involved in monitoring and managing the mine waste and mill tailings, and the relationship between different stakeholders from local communities, academic institutions, government agencies, and transnational mining corporations, as they deliberate about the possibility of cleaning up the former mining district. My thesis is that the possibility of cleaning up the Grants district hinges on the “politics of baselining”—a term I introduce to describe the relationship between stakeholders and their competing environmental models and hydrogeological theories; each accounts for a different geological past prior to mining that can be deemed “natural,” as the background against which to measure the anthropogenic impacts from mining.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

248 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-16

Benzene exposure induces metabolic imbalance via central mechanisms in a sex- specific manner in mice

Lucas K. Debarba, Wayne State University, Detroit, Michigan H.S.M Jayarathne, Wayne State University, Detroit, Michigan L. Koshko, Wayne State University, Detroit, Michigan J.B.M. Lima, Wayne State University, Detroit, Michigan M. Sadajurski, Wayne State University, Detroit, Michigan

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Unaffiliated

Benzene that is one of the major components of air pollution is a well-known human carcinogen. Several epidemiological studies suggested an association between adverse health effects of benzene at low doses and the development of insulin resistance Sources of benzene in ambient air include cigarette smoke, e- cigarettes vaping, and evaporation of benzene containing petrol processes. While the carcinogenic effects of benzene exposure have been well studied, less is known about the metabolic effects of benzene exposure. We show that chronic exposure to benzene at 50 ppm induces a severe metabolic imbalance in a sex-specific manner and is associated with hypothalamic inflammation and endoplasmic reticulum (ER) stress. Benzene exposure rapidly activates hypothalamic ER stress and neuroinflammatory responses in male mice, while pharmacological inhibition of ER stress response by inhibiting the IRE1α-XBP1 pathway significantly alleviates benzene-induced glial inflammatory responses. Acute, short-term benzene exposure rapidly increases neuronal activity in the hypothalamus, while the responses to glucose stimulation are blunted within glucose-sensing neurons in male mice. These data imply that hypothalamic dysfunction precedes and mechanistically contributes to whole-body metabolic imbalance and Type 2 Diabetes upon benzene exposure.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 249 Poster Session IV: Poster #IV-17

Untargeted stable-isotope probing of the gut microbiota metabolome using 13C-labeled dietary fibers

Pan Deng, University of Kentucky Taylor Valentino, University of Kentucky Michael D. Flythe, University of Kentucky Hunter N.B. Moseley, University of Kentucky Jacqueline R. Leachman, University of Kentucky Andrew J. Morris, University of Kentucky Bernhard Hennig, University of Kentucky

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Kentucky Superfund Research Center

The gut microbiome generates numerous metabolites that exert local effects and enter the circulation to affect the functions of many organs. Despite extensive sequencing-based characterization of the gut microbiome, there remains a lack of understanding of microbial metabolism. Here, we developed an untargeted stable isotope-resolved metabolomics (SIRM) approach for the holistic study of gut microbial metabolites. Viable microbial cells were extracted from fresh mice feces and incubated anaerobically with 13C labeled dietary fibers including inulin or cellulose. High resolution mass spectrometry was used to monitor 13C enrichment in metabolites associated with glycolysis, the Krebs cycle, the pentose phosphate pathway, nucleotide synthesis, and pyruvate catabolism in both microbial cells and the culture medium. We observed differential use of inulin and cellulose as substrates for biosynthesis of essential and non- essential amino acids, neurotransmitters, vitamin B5, and other coenzymes. Specifically, use of inulin for these biosynthetic pathways was markedly more efficient than use of cellulose, reflecting distinct metabolic pathways of dietary fibers in the gut microbiome, which could be related with host effects. This is the first study utilizing SIRM technologies to investigate gut microbiota metabolism. This technology facilitates deeper and holistic insights into the metabolic function of the gut microbiome.

Email: [email protected] Presenter Status: Researcher

250 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-18

A Laboratory Validation of a Novel Passive Sampler for PFAS

Matt Dunn, University of Rhode Island Dr. Jitka Becanova, University of Rhode Island Christine Gardiner, University of Rhode Island Dr. Rainer Lohmann, University of Rhode Island

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Rhode Island Superfund Research Center

As worldwide concern grows over the potential health effects of perfluorinated alkyl substances (PFAS), so too does the need for a simple, reliable detection method for these compounds in the environment. As part of the Sources, Transport, Exposure, and Effects of PFAS (STEEP) Superfund Center, a novel tube passive sampler was proposed to detect PFAS in surface waters. Being able to predict a passive sampler’s sampling rate (Rs) allows for back calculation of the environmental water concentration without the need for grab water sampling and laborious water extraction. Sampling rates, liters sampled per day by the passive sampler, were calculated as the mass of contaminant in the sampler (ng) divided by the measured tank water concentration (ng/L) and deployment time (days) across four flow rates ranging from 0-60 cm/s at an average water temperature of 24 degrees Celsius. For many perfluoroalkylcarboxylates, Rs values generally increased with flow, while the other major group, perfluoroalkylsulfonates, displayed a Langmuir-type trend of Rs versus flow. One of the most common PFAS, PFOA, showed a linear increase ranging from 0.011 L/d at no flow up to 2.0 L/d at 60 cm/s flow. PFOS, another common legacy sulfonate, showed an increase from 0 cm/s to 10 cm/s, before leveling off to around 0.30 L/d at the highest flows. Further trials are needed to confirm these trends in sampling rates across other temperature regimes more common in colder New England surface waters. Once validated, this novel passive sampler could provide strong predictive power as an affordable and simple detection tool needed to address PFAS contamination research and remediation.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 251 Poster Session IV: Poster #IV-19

The association between urinary glyphosate metabolites and biomarkers of oxidative stress among pregnant women in PROTECT birth cohort study

Jarrod Eaton, University of Michigan, Ann Arbor, MI Amber Cathey, University of Michigan, Ann Arbor, MI Deborah Watkins, University of Michigan, Ann Arbor, MI Carmen Velez-Vega, University of Puerto Rico, San Juan Puerto Rico Zaira Rosario, University of Georgia, Athens, GA Jennifer Fernandez, University of Michigan, Ann Arbor, MI Jose Cordero, University of Georgia, Athens, GA Akram Alshawabkeh Northeastern University, Boston, MA, John Meeker, University of Michigan, Ann Arbor, MI

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Northeastern University Superfund Research Center

Background: Glyphosate is a widely used herbicide in global agriculture. Glyphosate and its metabolite, aminomethyl phosphonic acid (AMPA), have been shown to disrupt endocrine function and induce oxidative stress within in vitro and animal studies. To our knowledge, these relationships have not been characterized in epidemiological settings. Elevated exposure to glyphosate and AMPA may impact the health of vulnerable populations such as pregnant women via mechanisms like oxidative stress. Methods: Glyphosate and AMPA were measured in 347 urine samples collected at multiple visits from pregnant women in the Puerto Rico Testsite for Exploring Contamination Threats (PROTECT) birth cohort. Urinary biomarkers of oxidative stress, 8-isoprostane-prostaglandin-F2a (8-iso-PGF2a), its metabolite 2,3-dinor- 5,6-dihydro-15-F2t-isoprostane (8-iso-PGF2a metabolite) and prostaglandin-F2a (PGF2a), were also measured. One-way ANOVA tests evaluated differences in exposure and oxidative stress concentrations between study visits. Linear mixed effect models assessed associations between exposures and oxidative stress adjusting for maternal age, smoking status, alcohol consumption, household income and specific gravity. Results: No significant differences in exposure or oxidative stress biomarker concentrations were observed between study visits. An interquartile range (IQR) increase in AMPA was associated with 9.5% (95% CI: 0.5%–19.3%) higher 8-iso-PGF2a metabolite. Compared to low exposure, the second and third tertiles of AMPA were significantly associated with 12.8% (0.6%-26.5%) and 15.2% (1.8%-30.3%) higher 8- iso-PGF2a metabolite, respectively. An IQR increase in glyphosate was suggestively associated with 4.7% (-0.9%–10.7%) higher 8-iso-PGF2a. Conclusions: Urinary concentrations of AMPA were associated with higher levels of oxidative stress. Associations with glyphosate reflected similar trends, although findings were not as strong. Additional research is required to better characterize the association between glyphosate exposure and biomarkers of oxidative stress, as well as potential downstream health consequences.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

252 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-20

Uranium and arsenic interactions with the rhizosphere: impact of water chemistry and fungi-plant symbiosis

Eliane El Hayek, University of New Mexico, Albuquerque, NM Cherie De Vore, University of New Mexico, Albuquerque, NM Taylor Busch, University of New Mexico, Albuquerque, NM Benson Long, University of New Mexico, Albuquerque, NM Mehdi Ali, University of New Mexico, Albuquerque, NM Jennifer Rudgers, University of New Mexico, Albuquerque, NM Patrick Hudson, University of New Mexico, Albuquerque, NM Tamara Howard, University of New Mexico, Albuquerque, NM Adrian Brearley, University of New Mexico, Albuquerque, NM Michael Spilde, University of New Mexico, Albuquerque, NM Jose Manuel Cerrato, University of New Mexico, Albuquerque, NM

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

Understanding the biogeochemical processes affecting the bioaccumulation of metals in the rhizosphere is important to determine environmental exposure pathways and to manipulate the biogeochemistry to drive uptake under appropriate phytoremediation strategies. The environmental project 1 of the METALS Superfund Research center has separately investigated uranium (U) and arsenic (As) bioaccumulation mechanisms under the effect of water chemistry (i.e. concentrations of U, calcium (Ca), carbonate and phosphate) and fungi-plant symbiosis at environmentally relevant conditions. We have used hydroponic experiments, microscopy, spectroscopy, culturing and molecular biology, and water chemistry analytical methods to evaluate the uptake and distribution of U and As in plants. Our results suggest that: 1) high Ca concentration (6 mM) in carbonate water can modulate the uptake pathway of U by interrupting U apoplastic precipitation with endogenous phosphorus and enhancing U intracellular translocation towards the shoots; 2) precipitation of Ca phosphate minerals on roots surface can facilitate As adsorption in the cell walls through surface complexation and co-precipitation, enhancing As accumulation in the roots; and 3) roots symbiosis with endophytic fungi supported better external and vascular cellular structures that facilitate As intracellular uptake. Additionally, the surface precipitation of hydroxyapatite in the roots resulted in As adsorption and co-precipitation. Our results provide relevant insights about the optimal conditions for uptake by plants to immobilize metal mixtures in contaminated sites. Ongoing and future work is focused on investigating the impact of the co-occurrence of U and As contamination on the bioavailability of these metals, the role of diverse root colonizing fungi on metal uptake, and the involved toxicity mechanisms in metals accumulation.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 253 Poster Session IV: Poster #IV-21

Ultrasensitive electrochemical immunosensor for point-of-care detection of amanitin in human fluids

Ahmed El-Moghazy, University of California-Davis, Davis, CA Gang Sun, University of California, Davis, CA

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of California-Davis Superfund Research Center

α-Amanitin is one of the deadliest toxins from mushrooms, present in the deadly mushroom species Amanita phalloides. It is a bicyclic octapeptide and represents up to 40% of the amatoxins in mushrooms, damaging the livers and kidneys. Current methods of detecting amatoxins are time-consuming and require the use of expensive equipment. A novel electrochemical label-free immunosensor was successfully developed for rapid detection of α-amanitin, which was fabricated by immobilization of anti-α-amanitin antibodies onto functionalized cellulose nanofibrous membrane-modified carbon screen-printed electrode. An oxidation peak of the captured amanitin on the tethered antibodies was observed at 0.45 V. The performance of the nanofibrous membrane on the electrode and necessary fabrication steps were investigated by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Due to their unique structural features and properties such as high specific surface areas and microporous structure, the nanofibrous membrane as the immunosensor matrix for the antibody tethering exhibited improved electrochemical performance of the electrode by more than 5 times comparing with the casted membranes. Under the optimal conditions, the assembled immunosensor exhibited high sensitivity toward α-amanitin detection in the range of 0.009-2 ng mL-1 with a limit of detection of 8.3 pg mL-1. The results clearly indicate that the fabricated nanofibers-based-immunosensor is suitable for point-of-care detection of lethal α-amanitin in human fluids without any pretreatment within 30 min.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

254 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-22

Effects of PFAS on development and gene expression of zebrafish larvae

Mathew R. Farrell, North Carolina State University, Raleigh, North Carolina Antonio J. Planchart, North Carolina State University, Raleigh, North Carolina

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: North Carolina State University Superfund Research Center

Poly and perfluoroalkylated substances (PFAS) are a class of synthetic chemicals with a wide range of industrial applications. They are characterized by a terminal group connected to a highly stable fluorinated carbon chain, causing them to be extremely persistent in the environment. This study examines the acute toxicity of multiple perfluorocarboxylic acids (PFCAs), perfluorosulfonic acids (PFSAs) and perfluoroether carboxylic acids (PFECAs) on zebrafish larvae. Larvae will be subjected to 120 hour static aqueous exposures and then examined for developmental defects. Changes in gene expression will also be measured, with particular focus on immune response and apoptotic pathways. Preliminary results suggest an uninflated swim bladder and spinal malformations as common effects at higher exposure concentrations, as well as higher acute toxicity of PFSAs compared to other groups. These tests will help establish possible modes of action and outcomes for environmental exposures of PFAS in wildlife and humans.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 255 Poster Session IV: Poster #IV-23

Recyclable Cationic Polymer Composites as PFAS Removal from Contaminated Water Systems

E. Molly Frazar, University of Kentucky, Lexington, KY Thomas D. Dziubla, University of Kentucky, Lexington, KY J. Zach Hilt, University of Kentucky, Lexington, KY

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Kentucky Superfund Research Center

The widespread over-use of per- and polyfluoroalkyl substances (PFAS) in the past several decades has led to an overwhelming amount of soil and water contamination by these compounds. The chemical and thermal stability of PFAS have proved them to be an especially daunting challenge from an environmental remediation standpoint. Presently, the only full-scale water treatment separates via sorption and uses non-selective materials such as activated carbon (AC) or mineral media. However, research focused on selective and easily regenerable materials is becoming a more practical route for capture and removal from contaminated water systems. This research focuses on the development of magnetic, cationic co- polymers that can be regenerated through application of an alternating magnetic field (AMF). PFAS capture has been shown to most effectively occur through ionic exchanges between positively charged sorbent materials and the negatively charged “heads” of various PFAS. The materials synthesized within this work are based on a quaternary ammonium polymer, which can be used at varying pH values without significant change in binding efficiency. Inclusion of iron oxide nanoparticles provide and interesting route of separation and regeneration through application of a stationary magnet and an AMF source, respectively. Recent interest and growth in the area of cationic polymers as PFAS removal agents has led to an array of systems with numerous applications. In this work, however, key issues such as pH dependence and labor/energy intensive or expensive regeneration techniques, are addressed and considered. Polymers were synthesized via free radical polymerization reactions and various characterization techniques were utilized. PFAS detection was conducted via LC-MSMS.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

256 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-24

The OSU/PNNL Superfund Research Program Analytics Portal: linking environmental chemicals to biological phenotype through data science and visualization

Sara Gosline, Oregon State University, Corvallis, OR Paritosh Pande, PhD, Pacific Northwest National Laboratory, Richland, WA Dennis Thomas, PhD, Pacific Northwest National Laboratory, Richland, WA Lisa Truong, PhD, Oregon State University, Corvallis, OR Michael Barton, PhD, Oregon State University, Corvallis, OR Kim Anderson, PhD, Oregon State University, Corvallis, OR Robyn L. Tanguay, PhD, Oregon State University, Corvallis, OR Katrina Waters, PhD, Pacific Northwest National Laboratory, Richland, WA, and Oregon State University

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substance

Superfund Center: Oregon State University Superfund Research Center

The OSU/PNNL superfund Research Program represents a longstanding collaboration to quantify Polycyclic Aromatic Hydrocarbons at various superfund sites in the Pacific Northwest and assess their potential impact on human health. To link the chemical measurements to biological activity, we describe the use of the zebrafish as a high-throughput developmental model of human exposure that provides quantitative measurements of the biological ramifications of exposure to toxicants. Toward this end, we have identified more than 1000 chemicals found at Superfund sites and measured the effect of these same chemicals in zebrafish, creating a rich dataset that links environmental exposure to phenotype. We have implemented a dose-response modeling pipeline to calculate benchmark dose parameters that enable the comparison of potency across chemicals and phenotypes. Our portal provides public access to this dataset and an interactive web site designed to enable exploration and re-use of this data by the scientific community at http://srp.pnnl.gov.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 257 Poster Session IV: Poster #IV-25

Vertebrate Developmental Hazard of Environmentally Persistent Free Radicals and Protective Effect of Tempol in Zebrafish Model

Xia Guan, Louisiana State University, Baton Rouge, LA Lisa Truong, Oregon State University, Corvallis, OR Slawomir M. Lomnicki, Louisiana State University, Baton Rouge, LA Robyn L. Tanguay, Oregon State University, Corvallis, OR Stephania A Cormier, Louisiana State University, Baton Rouge, LA

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Louisiana State University Superfund Research Center

The vertebrate developmental hazard of Environmentally Persistent Free Radicals (EPFRs) in zebrafish embryo was investigated and protective effect of tempol was discovered. EPFRs-containing particles were acquired by dosing dichlorobenzene (DCB) vapor on the Cab-o-sil/5%CuO particles at 230 °C in vacuo (referred as DCB-230). Three controls were used in the experiments: Cab-s-sil particles, cab-o-sil-5%CuO and water. The particles were suspended in ultrapure water to make 1 mg/mL of stock solutions from which series dilution was made to obtain 10, 20, 30, 40, 50, 60, 80 and 100 µg/mL of solutions for direct addition to the test plate where 4 hpf zebrafish embryo were placed. Plates were run in duplicate to obtain 24 animals per concentration; 1 embryo was exposed per well, 12 embryos exposed per concentration per plate. Statistical analysis of the morphology endpoints was performed. We investigated overt toxicity responses to DCB-230 in a 22 end-point battery that encompasses zebrafish development from 24 hpf- 120 hpf. Exposure to 60 µg/mL of DCB -230 or above was associated with high mortality of zebrafish. No significant results were observed for three controls. The exposures to EPFRs were developmentally hazardous in the zebrafish model. Then the potential protective effects of tempol against EPFRs’ cytotoxicity in zebrafish were investigated. Exposure to EPFRs plus tempol shifted EC50 (µg/mL) from 23.6 to 30.8 which verifies tempol’s protective effect against EPFRs in the early development phase of zebrafish. Utilizing zebrafish as an animal model to study EPFRs’ cytotoxicity and antioxidants’ reaction mechanism helps us understand mechanism of disease in the early development phase of the zebrafish with the goal to translate the discoveries to the clinic and protect our communities and the environment.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

258 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-26

SuperFunBase: Integrating population genomic data to understand mechanisms of chemical susceptibility and resistance

Mark E. Hahn, Boston University, Boston, Massachusetts Filisia Agus, Boston University School of Medicine, Boston, Massachusetts Noah M. Reid, Univ of Connecticut, Thomas F. Schultz, Duke University, Durham, North Carolina Sibel I. Karchner, Woods Hole Oceanographic Institution, Boston University, Boston Massachusetts Joshua S. Osterberg, Duke University, Durham, North Carolina Mark DeLong, Duke University, Durham, North Carolina David Corcoran, Duke University, Durham, North Carolina Neel Aluru, Woods Hole Oceanographic Institution, Boston University, Boston Massachusetts Wesley Warren, University of Missouri Andrew Whitehead, University of California, Davis Adam T. Labadorf, Boston University School of Medicine, Boston University, Boston Massachusetts Richard T. Di Giulio, Duke University, Durham, North Carolina Mark E. Hahn, Woods Hole Oceanographic Institution, Boston University, Boston Massachusetts

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Boston University Superfund Research Center

Evolutionary adaptation of animal populations after long-term exposure to chemicals can reveal the pathways, genes, and genetic variants that influence chemical sensitivity. Several populations of Atlantic killifish (Fundulus heteroclitus) have evolved heritable resistance to polychlorinated biphenyls (PCBs) or polynuclear aromatic hydrocarbons (PAHs) at Superfund sites. In this SRP data supplement, we seek to integrate complementary genomic data from two SRP centers investigating the molecular mechanisms of this resistance, making them Findable, Accessible, Interoperable, and Reusable (FAIR). These data include Restriction site Associated DNA sequencing (RAD-seq), whole-genome sequencing (WGS), and RNA- sequencing (RNA-seq) data obtained from multiple populations of PAH- or PCB- sensitive and resistant fish. We established harmonized, best-practice pipelines using standard genomics data formats and re- mapped all of the data to a new killifish reference genome assembly. We then loaded the data and associated metadata to an instance of the open source genome browser software JBrowse, which allows the data to be visualized and queried. To facilitate comparison of killifish and human genetic variation, we mapped orthologous human genes onto the killifish genome with the ultimate goal of overlaying human variants onto killifish genes. To provide broad access to our results, we are constructing an Open Science Framework (OSF.io) Project Page, which, along with the JBrowse platform SuperFunBase, will become freely available. These resources will make the data more accessible, allow reuse by others, and facilitate comparison of environmental science and biomedical data, enhancing the use of wildlife as sentinels and models for human health. [P42ES007381-23S1, P42ES010356]

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 259 Poster Session IV: Poster #IV-27

Building the capacity of life science teachers to incorporate COVID-19 related SRP research into instruction

Dana B. Haine, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Kathleen Gray, PhD, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Fernando Pardo-Manuel de Villena, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Ilona Jaspers, PhD, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Beverly Koller, PhD, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Mirek Styblo, PhD, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina Rebecca Fry, PhD, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

In recent years, the UNC SRP CEC expanded its teacher professional development offerings to build the capacity of biology teachers to incorporate research findings into life science instruction by providing up- to-date resources for teaching required curriculum content. This poster will highlight strategies for effectively engaging researchers in developing scientifically rigorous, STEM-focused educational activities that prepare secondary science teachers to incorporate timely biomedical research topics and relevant environmental health issues into their teaching while aligning to state and national science standards. Using cutting-edge, interdisciplinary research uniquely suited for studies of the interaction between environmental inorganic arsenic (iAs) exposure and SARS-CoV-2 infection, research made possible with novel laboratory models only available at UNC, the CEC is collaborating with SRP researchers to develop timely educational materials for the life science classroom. Numerous curriculum connections can be addressed when describing how UNC SRP researchers are testing the hypothesis that chronic exposure to iAs enhances susceptibility to SARS-CoV2 infection and severity of COVID-19. From the use of genetic engineering to develop a humanized mouse model that metabolizes arsenic like humans, to evaluating changes in gene expression in response to iAs exposure and SARS-CoV2 infection, there are ample opportunities to reinforce required curriculum content by providing current examples from the biomedical research sector. Partnering with SRP scientists to develop standards-aligned educational materials that showcase the role of emerging science in solving problems and protecting public health promotes student engagement with current biomedical research approaches and data, while fostering an increased awareness of relevant biomedical careers and a greater understanding of how environmental contaminants influence human health.

Email: [email protected] Presenter Status: Other: CEC

260 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-28

Oregon State University Superfund Research Program on PAHs

Stacey Harper, Oregon State University, Corvallis, OR Diana Rohlman, PhD, Oregon State University, Corvallis, OR Molly Kile, ScD, Oregon State University, Corvallis, OR Katrina Waters, PhD, Pacific Northwest National Laboratory, Richland, WA Robyn Tanguay, PhD, Oregon State University. Corvallis, OR

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substance

Superfund Center: Oregon State University Superfund Research Center

The mission of the Oregon State University Superfund Research Program (OSU SRP) is to discover new pollutants, assess their toxicity, translate and apply our findings to improve public health. The OSU SRP studies the compositions of complex PAH mixtures, the ways in which those compositions change over time and during remediation, and the implications of PAH mixtures for human health. The OSU SRP aims to supply EPA and other regulatory agencies with actionable information and with tools that enable them to detect PAHs in the environment, to measure PAH concentrations, and to evaluate remediation. Our research translation and community engagement leaders will be sharing the resources and tools developed in the OSU SRP. Information will be available on passive sampling, human health models of toxicity, and Tribal engagement. We will share data visualization tools, OSU’s Disaster IRB materials and report-back tools, as well as research translation materials (i.e., fact sheets, videos, infographics, community resources, and Tribal resources and training materials) for your use.

Email: [email protected] Presenter Status: RT/CEC poster

NIEHS SRP 2020 Virtual Annual Meeting Program • 261 Poster Session IV: Poster #IV-29

Neurobehavioral effects of embryonic neurotoxicant exposures across the lifespan in zebrafish

Andrew B. Hawkey, Duke University, Durham, North Carolina Zade Holloway, Duke University, Durham, North Carolina Jonna Boyda, Duke University, Durham, North Carolina Reese Koburov, Duke University, Durham, North Carolina Edward D. Levin, Duke University, Durham, North Carolina

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Duke University Superfund Research Center

Many environmental contaminants have neurotoxic potential, and species like zebrafish are frequently used to assess neurotoxicity and its underlying mechanisms. However, the nervous system develops progressively across the lifespan and questions remain as to whether assessments conducted at a specific age adequately predict risk overall. Relatedly, we have often observed that larval zebrafish data is a poor predictor for neurobehavioral deficits later in adulthood, suggesting that development may be an influential mediator of functional impairment or recovery. Further study of functional trajectories across the lifespan is needed to allow data from different stages of development to be integrated more informatively. To this end, we have completed broad neurobehavioral studies in zebrafish using three representative compounds: benzo-a-pyrene (0.3-1.0uM), diazinon (70-700nM), and triphenyl phosphate (0.1-0.3uM). Exposures took place from 5-120h post-fertilization (hpf) and neurobehavioral tests were conducted at larval (6dpf), adolescent (10 weeks), adult (8 months) and late adult (14 months) ages. The resulting effects were dose-dependent, selective, and variable from one stage of development to the next. In several cases, toxicant exposures produced effects which attenuated over time, including DZN- or BAP- induced hyperactivity in adolescence, DZN- or TPP-induced predator avoidance alterations in young adulthood, and BAP-induced shoaling deficits in young adulthood. Likewise, there were effects which emerged at later time points, including BAP-induced enhancements of startle responses and TPP-induced hypoactivity and enhanced diving responses in late adulthood. This work shows that early exposure to neurotoxicants may lead to functional behavioral impairments that are specific to certain stages of development, such as early maturation or aging, and that developmental age is an important parameter to consider for in vivo neurotoxicity testing.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

262 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-30

Field testing of sensors for a novel trimodal eddy correlation flux meter

Harry Hemond, Massachusetts Institute of Technology Irene Hu, Massachusetts Institute of Technology

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Massachusetts Institute of Technology Superfund Research Center

Contaminated sediments can be a source of chemicals to the water of lakes, rivers, and estuaries. The ability to measure such fluxes is currently limited, but improvements in this ability can lead to better and more cost-effective remediation, especially in complex heterogeneous aquatic environments. The present study is directed toward development of a fluxmeter to measure chemical fluxes from sediments using the principle of eddy correlation (EC) and employing a novel high-speed tri-modal sensor head containing a spectrofluorometer, fast thermistor, and integrated conductivity sensor. The prototype fluxmeter system consists of the aforementioned three water quality sensors for fluorescence, conductivity, and temperature, integrated into a single sensor head designed to make the measurements of each sensor correspond, as nearly as practicable, to the same volume of water. An acoustic doppler velocimeter (ADV) measures velocity of the same parcel of water. The sensors are held in an adjustable distance above the lake sediment on a benthic lander, which also holds electronics to control the several sensors. Tests in a laboratory flume previously demonstrated the ability of each type of water quality sensor to measure fluctuations at the required frequency of several Hz, and to work in concert with the acoustic doppler velocimeter to measure fluxes of tracer substances released at controlled rates in a laboratory tank. Recent field deployments at shallow depth in Upper Mystic Lake in eastern MA were carried out to test the overall operation of the EC fluxmeter under field conditions. Results demonstrate the ability of the spectrofluorometer to measure dissolved fluorescent substances against a worst-case condition of interference from both ambient light and wave action; the modulated spectrofluorometer, using photon counting at frequencies (photon incidence).

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 263 Poster Session IV: Poster #IV-31

Preliminary Metal Biomonitoring Findings from the UNM METALS Thinking Zinc Clinical Trial

Laurie Hudson, University of New Mexico Debra MacKenzie, University of New Mexico Esther Erdei, University of New Mexico Erica Dashner-Titus, University of New Mexico Christopher Shuey, Southwest Research and Information Center, UNM Metals Center Sarah Henio-Adeky, Southwest Research and Information Center Thinking Zinc Research Team

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

The presence of numerous abandoned uranium mines (AUMs) have long raised health concerns for UNM METALS partnering communities. There are limited options to mitigate risks associated with exposures to toxic metals linked to AUMs. Experimental evidence demonstrates that certain metals, including arsenic and uranium, bind to zinc finger domain proteins and disrupt protein function. Supplemental zinc protects against the adverse metal effects. Based on these findings and evidence for higher incidence of zinc insufficiency in the population compared to NHANES, we launched “Thinking Zinc”, a one-armed cohort intervention trial in May 2019. The study will test the hypothesis that zinc supplementation, at recommended adult dietary levels, can alleviate harmful effects of uranium and mixed metal exposures. Primary endpoints include urinary and serum metal levels and biomarkers of cellular changes reflecting immune dysregulation and autoimmunity. The communities participating in “Thinking Zinc” were selected based on concerns of exposure and proximity to legacy uranium mine sites. Biomonitoring reveals that greater than 10% of samples exceeded the NHANES 95th percentile urinary metal levels for antimony, beryllium, manganese, strontium, tin and uranium. Of particular concern are the values for uranium where 40.8% of samples exceeded the NHANES 95th percentile and 24.5% of samples exceeded the 95th percentile for participants in the Navajo Birth Cohort Study. Longitudinal biomonitoring indicates substantial variability in uranium levels for some Thinking Zinc participants based on values obtained from visits spaced 3 months apart. This observation suggests patterns of episodic higher dose exposures for certain individuals. These initial findings demonstrate differences in metal exposures between communities and in individuals over time that should be taken into consideration of studies involving chronic AUM exposures.

Email: [email protected] Presenter Status: Researcher

264 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-32

Identifying Sources of Polychlorinated Biphenyls Measured in School Air Using Positive Matrix Factorization

Jacob Jahnke, University of Iowa, Iowa City, Iowa Moala K. Bannavti, University of Iowa, Iowa City, Iowa Rachel F. Marek, University of Iowa, Iowa City, Iowa Craig L. Just, University of Iowa, Iowa City, Iowa Keri C. Hornbuckle, University of Iowa, Iowa City, Iowa

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Iowa Superfund Research Center

People spend a majority of their time indoors. In order to provide safe indoor environments, we want to remove pollutants from the air. Polychlorinated Biphenyls (PCBs) are essentially always present in indoor air and due to their negative health effects, we want to remove sources of them. PCBs measured in ambient air samples are generally thought to come from legacy sources such as Aroclors. However, some of the PCBs may be coming from modern sources like paint and sealants, and it is important to distinguish these sources when considering remediation methods. We hypothesize that there are PCB signatures that relate to legacy and modern sources, and that they can be recognized and distinguished in environmental samples. To accomplish this, various statistical methods including Positive Matrix Factorization (PMF) and Pearson’s Correlation Coefficient were used to identify signals in air. Data was selected from multiple studies conducted in our lab including data of school air and outdoor air and data of PCB emissions from paint colorants. Individual PCB profile factors were resolved from the data using PMF. The solution was stable and produced a reasonable and interpretable number of factors. Some were closely related to Aroclors and others were not, suggesting the influence of modern sources on the environmental samples. Furthermore, we were able to identify sources of PCBs to individual rooms in a school instead of assuming a single source for the entire school. This work will help identify sources of PCBs in environmental samples and can aid in remediation decisions to reduce exposure and risk to PCBs.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 265 Poster Session IV: Poster #IV-33

Atlantic killifish retain PAH-adapted phenotype following Elizabeth River remediation

Lindsay Jasperse, Duke University, Durham, North Carolina Rafael Trevisan, Duke University, Durham, North Carolina Rose Trimpey-Warhaftig, Duke University, Durham, North Carolina Melissa Chernick, Duke University, Durham, North Carolina Richard T. Di Giulio, Duke University, Durham, North Carolina

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Duke University Superfund Research Center

The Elizabeth River (ER), a tidal estuary in southeast Virginia, experienced decades of creosote pollution during the 1900s, resulting in sediments with significant polycyclic aromatic hydrocarbon (PAH) contamination. Three major ER wood treatment facilities utilized creosote for most of the 20th century: Atlantic Wood Industries (AW), Republic Creosoting (REP), and Eppinger and Russell (Money Point; MP). Due to chronic exposure to contaminated sediments, Atlantic killifish living in the ER demonstrate an adapted phenotype that protects from PAH toxicity but is associated with fitness costs. Killifish offspring with the adapted phenotype are resistant to a complex mixture of PAHs extracted from AW sediment (Elizabeth River Sediment Extract; ERSE), while non-adapted fish from a reference population in King’s Creek develop severe cardiac abnormalities. MP and the AW Superfund site were remediated approximately 5-10 years ago, while the REP site remains heavily contaminated with PAHs. We hypothesized that given the relatively short generation time of killifish (~1 year), the adapted phenotype of AW and MP killifish would dissipate as PAH levels at these sites declined following remediation. Exposures to ERSE demonstrate that offspring of AW and MP killifish collected post-remediation continue to exhibit the adapted phenotype, with PAH-resistance comparable to pre-remediation. Extract from sediment collected at AW in July 2020 exhibited minimal toxicity in non-adapted fish, indicating low PAH concentrations post-remediation. This suggests that while remediation may have effectively reduced PAH exposure in the ER, killifish at AW and MP remain adapted to pollution through multiple generations, despite costs to fitness. This work demonstrates the need for continued research investigating the effects of remediation on killifish populations in the ER in order to accurately assess ecosystem recovery.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

266 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-34

Transport and Deposition of Contaminated Sediment in the Upper Houston Ship Channel by Coastal Hazards

James Kaihatu, Texas A&M University, College Station, Texas Mikyoung Jun, University of Houston, Houston Texas Gopal Bera, Texas A&M University, College Station, Texas Terry Wade, Texas A&M University, College Station, Texas Anthony Knap, Texas A&M University, College Station, Texas

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

The Port of Houston, the sixteenth busiest port in the world, is served by the Houston Ship Channel, a federally-maintained waterway which is regularly dredged and widened to accommodate increasing traffic. The Houston Ship Channel traverses Galveston Bay, the largest estuary in Texas. The Bay is also the site of significant pollution (dioxins, PAHs, pesticides, PCBs, and metals), much of which is bound up in the fine bottom sediment. While dredging and other anthropogenic means of mobilization are generally quite controlled and governed by environmental regulations, there is a significant potential for mobilization of these sediments by natural hazards (primarily surge due to hurricanes). This can lead to deposition of contaminated sediment by surge in the industrial and residential neighborhoods surrounding the Channel, as well as re-deposition of this sediment within the Channel and away from monitoring locations. The redeposition potential of these sediments within the Channel is studied using a hydrodynamic model for tides, surge, waves, and sediment transport in Galveston Bay and the Houston Ship Channel. Historical hurricanes (notably Hurricane Ike) are used to force the model and determine the surge and resulting sediment redeposition. In addition, synthetic scenarios redolent of possible global warming trends and natural climatological variability are also used to simulate surge and sediment transport. Despite high surge potential in this region, it is anticipated that significant on-land deposition of sediment along the western branch of the Channel may not be a significant threat. However, mobilization of contaminated sediment away from present monitored locations to other (unmonitored) locations in the Channel and Bay is much more likely. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 267 Poster Session IV: Poster #IV-35

Analysis of polycyclic aromatic hydrocarbons and carcinogens using liquid chromatography-mass spectrometry: identification of carcinogens in bladder cancer urine

Abu Hena Mostafa Kamal, Baylor College of Medicine, Houston, TX Chandra Shekar R. Ambati, Baylor College of Medicine, Houston, TX Vasanta Putluri, Baylor College of Medicine, Houston, TX Nagireddy Putluri, Baylor College of Medicine, Houston, TX

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Baylor College of Medicine Superfund Research Center

Polycyclic aromatic hydrocarbons (PAHs) and carcinogens are referred to as a ubiquitous group of chemically-related environmentally persistent organic compounds of various structures and varied toxicity. Metabolites of PAHs and carcinogens are mainly excreted in urine, while a small percentage is excreted as unmetabolized compounds. Previous studies have shown that cigarette smoke with its main components’ nicotine, NNK, BBN, and PAHs are a major risk factor for bladder cancer. In the current study, we developed a novel LC/MS/MS method to accurately measure NNK, BBN, and PAHs in urine collected from bladder cancer patients. A novel LC-MS/MS (1290 Infinity HPLC, 6490 QQQ) method was developed using HILIC based chromatography to quantitate NNK, BBN, and > eight PAH compounds and their metabolites. Multiple reaction monitoring was performed in both positive and negative ion modes. MS parameters (including MRM transitions, fragmentor voltage, and collision energy) and LC conditions were first optimized. Calibration curves were analyzed for both accuracy and reproducibility over multiple runs and days. A novel LC-MS/MS method was developed to accurately measure BBN, NNK, and > eight PAHs. X Bridge Amide (3.5 µm column, 4.6*100 mm, Waters Corporation) was selected for adequate chromatographic separation. Water/ACN with 0.1% formic acid was used as mobile phase for the positive ion mode, while water with 5mM ammonium acetate/ 100% ACN was used as mobile phase for the negative ion mode. Limits of detection, limits of quantitation, and linear dynamic range were calculated for each compound and ranged from two to five log units with excellent inter- and intra-day reproducibility of retention time. This study developed a novel LC-MS/MS method to precisely determine BBN, NNK, and PAHs in bladder cancer patient urines.

Email: [email protected] Presenter Status: Researcher

268 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-36

Low-Pressure Membranes for Capture and Separation of Toxic Metals Ions

Francisco Leniz, University of Kentucky, Lexington, Kentucky Mohammad Saiful Islam Phillip Sandman Ronald J. Vogler Isabel C. Escobar Dibakar Bhattacharyya

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Kentucky Superfund Research Center

Pollution of water comes from both metals and organics, and thus, water remediation (and possibly recovery) requires a variety of approaches. The availability of low-pressure membranes can not only concentrate feed streams but also provides filtered water for reuse. Nanofiltration (NF) membranes, which are capable of selectively separate ions and small molecules, provide an energy-efficient option of separation. Another example is functionalized microfiltration membranes, with absorptive materials for high-capacity metal capture or with integrated catalytic materials for degradation. The goal of this research is to develop new functionalized polymeric membranes for selectively separate compounds via different mechanisms (rejection, sorption, and/or reaction), and thus removing heavy metals or recovering metals of high value. Current research results have proven the potential of NF membranes for the recovery of valuable lanthanides from aqueous solutions, including a synthesized NF membrane with weak basic functionalities from the addition of polyallylamine hydrochloride. Separations of neodymium and lanthanum chloride were tested under different operating conditions, with rejection around 99.3% in mixtures with high ionic strength (100 mM, equivalent to ~6,000 ppm), 94% rejection and high Na+/La3+ selectivity at 80% water recovery (with 0% Na+ rejection starting at 60% recovery), and stable lanthanum rejection and permeate flux at even pH 2.7. Moreover, wastewater containing HgS nanoparticles and dissolved Hg2+ was treated with a commercial UF membrane in combination with thiol-functionalized membranes created in house. The thiol membrane adsorbed as high as 97% of Hg2+ from a dilute, synthetic feed. The presence of Ca2+ in real wastewater reduced the adsorption efficiency of Hg2+, but this effect was mitigated by functionalizing with cysteamine instead of cysteine to incorporate thiol groups.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 269 Poster Session IV: Poster #IV-37

Development of Nanobody Based Sandwich ELISA for Mouse Soluble Epoxide Hydrolase Detection

Dongyang Li, University of California-Davis Yongliang Cui, University of California-Davis Christophe Morisseau, University of California-Davis Young Sik Cho, Keimyung University, Daegu, South Korea Bruce D. Hammock, University of California-Davis

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

Enzyme linked immunosorbent assays (ELISA) for the detection of soluble epoxide hydrolase (sEH), a key enzyme in the metabolism of fatty acids and a biomarker, may increasingly represent an important diagnostic tool and reflection of sEH expression affected by exposure to numerous xenobiotics. However, there is a lack of ELISA for mouse sEH quantification, thus resulting in a bottleneck in understanding the pathogenesis of many diseases related to sEH based on mouse models. In this work, nanobodies recognizing mouse sEH were obtained through re-biopanning against mouse sEH in the previous phage display library of human sEH. Later, we developed four ELISAs involving a combination of anti-mouse sEH polyclonal antibody (pAb) and nanobody. It was found that the double antibodies worked as dual filters and had a huge impact on both the sensitivity and selectivity of sandwich immunoassays. The switch of anti-human sEH pAb to anti-mouse sEH pAb led to over 100-fold increase in sensitivity and dramatic decrease of LOD to pg/mL range in format B (pAb/biotin-VHH/SA-PolyHRP). Moreover, we found the four sandwich ELISAs might demonstrate excellent selectivity to mouse sEH even though antibodies alone showing significant cross-reactivity to the matrix were used, indicating the enhanced selectivity of double antibodies as dual filters. Eventually, the ELISA (format B) was successfully used to measure the mouse sEH level in cancer cells with ultralow abundance for the first time. The ELISAs proposed here represents a sensitive tool for tracking the sEH in various biological processes and also provides deep insights for developing sandwich immunoassay against various targets in terms of both sensitivity and selectivity. The work demonstrates the promise of nanobody based ELISA as a diagnostic tool.

Email: [email protected] Presenter Status: Researcher

270 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-38

Gestational and Childhood Exposure to Per- and Polyfluoroalkyl Substances and Cardiometabolic Risk at Age 12 Years

Nan Li, Brown University, Providence, Rhode Island Yun Li, Brown University, Providence, Rhode Island George D. Papandonatos, Brown University, Providence, Rhode Island Antonia M. Calafat, Centers for Disease Control and Prevention, Atlanta, Georgia Charles B. Eaton, Brown University, Providence, Rhode Island Karl T. Kelsey, Brown University, Providence, Rhode Island Kim M. Cecil, Cincinnati Children’s Hospital Med Ctr, Univ of Cincinnati Coll of Med, Cincinnati, Ohio Heidi J. Kalkwarf, Cincinnati Children’s Hospital Med Ctr, Univ of Cincinnati Coll of Med, Cincinnati, Ohio Kimberly Yolton, Cincinnati Children’s Hospital Med Ctr, Univ of Cincinnati Coll of Med, Cincinnati, Ohio Bruce P. Lanphear, Simon Fraser University, Burnaby British Columbia, Canada Aimin Chen, University of Pennsylvania Joseph M Braun, Brown University, Providence, Rhode Island

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Brown University Superfund Research Center

Background: Per- and polyfluoroalkyl substances (PFAS) may adversely influence cardiometabolic risk by activating peroxisome proliferator-activated receptors. However, few studies have examined the timing of early life PFAS exposure in relation to overall cardiometabolic risk. We examined the influence of gestational and childhood PFAS exposure on adolescents’ overall cardiometabolic risk. Methods: We quantified concentrations of four PFAS (perfluorooctanoate [PFOA], perfluorooctane sulfonate, perfluorononanoate, and perfluorohexane sulfonate [PFHxS]) in sera collected during pregnancy, at birth, and at ages 3, 8, and 12 years from 221 mother-child pairs in the HOME Study (Cincinnati, Ohio). We measured cardiometabolic risk factors using physical examinations, fasting serum biomarkers, and dual- energy X-ray absorptiometry scans at age 12 years. Cardiometabolic risk summary scores were calculated by summing age- and sex-standardized z-scores for individual cardiometabolic risk factors. We used multiple informant models to estimate covariate-adjusted associations of serum PFAS concentrations (log2-transformed) at each visit with cardiometabolic risk scores and their individual components, and tested for differences in associations across visits. Results: The associations of serum PFOA concentrations with cardiometabolic risk scores differed across visits (P for heterogeneity=0.03). Gestational and cord serum PFOA concentrations were positively associated with cardiometabolic risk scores (βs and 95% confidence intervals [CIs]: gestational 0.8 [0.0, 1.6]; cord 0.9 [-0.1, 1.9] per interquartile range increase). These positive associations were primarily driven by homeostatic model assessment for insulin resistance index (β[95%CI]=0.3 [0.1, 0.5]) and adiponectin to leptin ratio (β[95%CI]=-0.5 [-1.0, 0.0]). Gestational and cord PFHxS were also associated with higher cardiometabolic risk scores (βs[95%CIs]: gestational 0.9 [0.2, 1.6]; cord 0.9 [0.1, 1.7]). Conclusion: In this cohort, fetal exposure to PFOA and PFHxS was associated with unfavorable cardiometabolic risk in adolescence.

Email: [email protected] Presenter Status: Researcher / Environmental Health

NIEHS SRP 2020 Virtual Annual Meeting Program • 271 Poster Session IV: Poster #IV-39

The gut microbiome is related with the disposition of polychlorinated biphenyls in mice

Xueshu Li, University of Iowa, Iowa City, Iowa Kai Wang, University of Iowa, Iowa City, Iowa Julia Yue Cui, University of Washington, Seattle, Washington Hans-Joachim Lehmler, University of Iowa, Iowa City, Iowa

Superfund Mandate: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of Iowa Superfund Research Center

Background. Polychlorinated biphenyls (PCBs) are a group of ubiquitous humanmade persistent organic pollutants (POPs) and are implicated in adverse environmental and human health effects. PCBs are metabolized in the liver by cytochrome P450 enzymes to hydroxylated PCBs. The gut microbiome contributions to human health and disease, possibly by altering the disposition of xenobiotics, such as PCBs. This study aimed to investigate how the presence or absence of the gut microbiome affects the distribution of PCBs in target tissues in mice. Method. Three-month-old female conventional (CV) and germ-free (GF) adult mice were exposed orally for three consecutive days to a low (6 mg/kg bw/day) or a high dose (30 mg/kg bw/day) of the Fox River Mixture, an environmental PCB mixture, in corn oil. Control animals were analogously exposed to vehicle (corn oil) alone. The animals were euthanized 24 h after the last PCB administration, tissues (brain, liver, adipose, and serum) were collected, and levels of PCBs were determined by GC-MS. Results. In both CV and GF mice, the total PCB levels followed the rank order adipose > liver > brain > serum and increased 3- to 10-fold from the low to the high dose group. Interestingly, the levels of total PCBs in CV mice were about 2-4 fold higher than in GF mice, independent of the tissue. Higher liver PCB levels result in the expression of CYPs, which in turn metabolizes some PCBs. Conclusions. These observations indicate that the gut microbiome plays a currently overlooked role in the disposition of PCBs [ES05605, ES013661, ES027169, and ES031098].

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

272 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-40

A search of cardioselective toxins in mitochondrial functional assays on H9c2 cardiomyocyte-like cells

Naoki Matsumoto, University of California, Davis Rasheed Olamilekan Sule, University of California, Davis Romulo Leao Silva Neris, University of California, Davis Christophe Morisseau, University of California, Davis Nipavan Chiamvimonvat, University of California, Davis Bruce D Hammock, University of California, Davis Aldrin V Gomes, University of California, Davis

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of California, Davis Superfund Research Center

Different organs have different sensitivities to xenobiotics. The ATP supply in the heart is highly dependent on the mitochondrial oxidative phosphorylation. Thus disruption of cardiac mitochondrial function potentially causes a fatal event. Here we screened our pesticide library for the toxicity on the mitochondrial function of H9c2 cardiomyocyte-like cell line and compared the tissue selectivity of the compounds using non-cardiac cell types (HUVEC and HEK293). We assessed the pesticide effects on two cellular functions: mitochondrial membrane potential and CHOP promoter activity. The mitochondrial membrane potential is the source of ATP production in mitochondria, and CHOP is a pro-apoptotic transcription factor whose expression is driven by mitochondrial disruption. We found some chlorophenol compounds were toxic in a relatively tissue-nonselective manner, whereas rotenone and some dithiocarbamate compounds were highly H9c2-selective. It is important to examine whether acute or chronic exposure to these pesticides damages the heart in vivo and elucidate the toxic mechanisms and ways to minimize the toxicity.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 273 Poster Session IV: Poster #IV-41

Key Characteristics Approach to Hazard Identification: Update on a Collaborative Effort

Cliona M. McHale, University of California Berkeley, Berkeley, CA Luoping Zhang, University of California Berkeley, Berkeley, CA Sarah E. Elmore, Office of Environmental Health Hazard Assessment, California EPA Lauren Zeise, Office of Environmental Health Hazard Assessment, California EPA Martyn T. Smith, University of California Berkeley, Berkeley, CA

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of California-Berkeley Superfund Research Center

With an IARC expert committee, Martyn Smith helped to develop the key characteristics (KCs) of human carcinogens as a basis for searching, organizing, and evaluating mechanistic evidence to support cancer hazard identification and he co-authored a 2016 paper. In 2018, Martyn Smith and Lauren Zeise (CalEPA) led an international effort to advance the KCs of carcinogens approach and expand the concept to other toxicity areas to support hazard identification by CalEPA, EPA IRIS and NTP. The Berkeley SRP Research Translation Core helped to convene expert committees comprising a range of stakeholders including government agencies, intergovernmental organizations, academic/research institutions and pharmaceutical companies, to gain varied perspectives and insights. Through in-person meetings and teleconference calls, we developed KCs for endocrine disrupting chemicals, male reproductive chemicals, female reproductive chemicals, and cardiovascular toxicants and are developing or helping to develop KCs of neurotoxicants, immunotoxicants and hepatotoxicants. Regarding the KCs of carcinogens, we showed that interrelationships among the KCs can strengthen their application, recommended current and emerging assays and biomarkers and improvements in coverage and robustness, and developed approaches that could be used to assess how chemical mixtures may produce cancer. In addition, we conducted a systematic review of benzene and two immune-related KCs, that illuminated mechanistic understanding. Multiple papers have been published or are forthcoming and the KCs have been applied in assessments by CalEPA, IARC, and NTP. We engaged key stakeholders throughout the process and shared the KCs with them and we recently launched a website with KC-related resources for assessors and scientists. Our collaborative efforts are generating important tools and information for regulators and scientists, and are supporting hazard identification and improving mechanistic insight into SRP chemicals.

Email: [email protected] Presenter Status: Researcher

274 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-42

In Vitro Bioaccessibility of Arsenic in Mine Tailings Related Minerals

Anissa McKenna, University of Arizona, Tucson, Arizona Rob Root, University of Arizona, Tucson, Arizona John Chorover, University of Arizona, Tucson, Arizona

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Arizona Superfund Research Center

Contamination from legacy sulfide mine tailings is a persistent environmental problem due to the presence of high concentrations of toxic metal(loids) like arsenic (As). This problem can be enhanced in semi-arid regions, where dry climate and lack of vegetative cover create favorable conditions for off-site transport of tailings as fugitive dusts. Mine tailings dust poses a significant risk to proximal communities who may experience exposure through incidental inhalation and ingestion of particulate matter. Arsenopyrite (FeAsS) is a common parent rock mineral for arsenic at many sulfide ore derived mine tailings sites. Initial in vitro bioaccessibility (IVBA) experiments – conducted under anoxic conditions, with rigorous cleaning of oxidized surficial precipitates, and utilizing LC-ICP-MS for aqueous speciation analyses – suggested that arsenopyrite dissolution in synthetic gastric fluid resulted in the release of arsenate (AsV) to solution. The apparent oxidation of As during FeAsS dissolution was unexpected, and could reflect the co-elution of thioarsenicals with AsV during LC-ICP-MS analysis. Therefore, new experiments are being conducted using synthetic thioarsenical standards, and testing their stability in the development of a new LC-ICP-MS analytical method. Relevance of this work is supported by the fact that there is limited knowledge surrounding the aqueous stability of thioarsenical compounds during FeAsS dissolution, despite their high toxicity.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 275 Poster Session IV: Poster #IV-43

Role of Intestinal UGT1A1 in Irinotecan-induced Toxicity: A Novel Tissue-specific Humanized Model

Elvira Mennillo, University of California, San Diego Andre Weber, University of California, San Diego Xiaojing Yang, University of California, San Diego Yoshihiro Maruo, Shiga University of Medical Science Olivier Barbier, Centre de Recherche du CHU de Québec Shujuan Chen, University of California, San Diego Robert Tukey, University of California, San Diego

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California, San Diego Superfund Research Center

The human UDP-glucuronosyltransferases (UGTs) represent an important family of drug-metabolizing enzymes. Among the UGT1A proteins, UGT1A1 is the isoform which has been studied the most, as it is involved in the conjugation and detoxification of several exogenous substances, including pharmaceutical drugs and environmental toxicants. Humanized UGT1A1 mice expressing the human UGT1A1 gene in either liver (Alb1A1) or small intestine (Vil1A1) were generated in a Ugt1-null background, thus enabling the examination of tissue-specific glucuronidation. Hepatic and intestinal tissue specific expression and function of UGT1A1 were demonstrated. Although the liver is considered a major organ for detoxification, intestinal UGT1A1 is also an important contributor for drug clearance. Mice were challenged with irinotecan (CPT-11), a prodrug hydrolyzed by carboxylesterases to form the active metabolite SN-38, that is detoxified mainly by UGT1A1. Alb1A1 mice, that have no intestinal UGT1A1, displayed higher lethality rate than Vil1A1 mice. When exposed to a low dose of CPT-11 (10 mg/kg), Alb1A1 mice showed more susceptibility to CPT-11 toxicity, as highlighted by higher inflammatory (il-1β and il-6) and p53-triggered apoptotic responses. In vitro studies with intestinal crypt organoids isolated from the same models and exposed to CPT-11 confirmed the results observed in vivo and indicated that CPT-11 significantly impacts stemness and self-renewal capacity of organoids. This study reveals the importance of intestinal UGT1A1 in preventing inflammation, apoptosis, and loss of intestinal stemness capacity upon systemic challenge with a common anti-cancer pharmaceutical. Lastly, we propose the use of these models as important tools for studying intestinal and hepatic toxicity of drugs and environmental toxicants that are metabolized by UGT1A1.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

276 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-44

Identifying and Assessing Harmful Contaminants on the Yurok Indian Reservation

Beth R. Middleton, University of California-Davis, Davis, California Suzanne Fluharty, Community and Ecosystems, Yurok Tribe Environmental Program Thomas Young, University of California-Davis, Davis, California Dalia Rakha, University of California-Davis, Davis, California Brittany Saleeby, University of California-Davis, Davis, California Tamara Solorzano, University of California-Davis, Davis, California

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of California-Davis Superfund Research Center

Through unique exposure pathways, Native American tribes face disproportionate environmental hazards that contribute to significant health disparities. In order to address observed increases in adverse human health conditions and declines in their fisheries, the Yurok Tribe has partnered with the UC Davis Superfund Research Program (SRP) to identify contaminants in their watershed that may be impairing human and ecosystem health. The SRP Community Engagement Core is working with the Yurok Tribe Environmental Program (YTEP), and SRP Project 1 (Optimizing Bioremediation) and Project 3 (Immunochemical Biomarkers) to identify contaminants in the Klamath River and tributaries and in retired lumber processing facilities in the watershed. In 2019, estrogen receptor screening of soil and sediment samples from mill ponds resulted in evidence of elevated estrogen receptor activity in mill pond samples. We are currently analyzing additional samples to identify whether this activity is naturally occurring, or is human caused. The CEC and Project 1 have also developed at time series of samples along the Klamath over the last three years of sampling. Our collaborative goal is to develop analysis and mitigation strategies that correspond to the extent, duration, and cumulative aspects of tribal members’ subsistence and cultural exposure. If we are able to identify particular chemicals of risk, UC Davis and YTEP scientists will work with Project to develop rapid detection methodologies that community members can use to assess risk.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 277 Poster Session IV: Poster #IV-45

Mice exposed to pyrolytically remediated soil extracts display attenuation of pulmonary DNA adducts compared to contaminated soils: Implications for remediation of Superfund site soils

Bhagavatula Moorthy, Baylor College of Medicine, Houston, Texas Goudong Zhou, Texas A&M University Weiwu Jiang, Baylor College of Medicine, Houston, Texas Chun Chu, Baylor College of Medicine, Houston, Texas Kyriacos Zygourakis, Rice University, Houston, Texas Pedro J. J. Alvarez, Rice University, Houston, Texas

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Baylor College of Medicine Superfund Research Center

Pyrolytic treatment offers the potential for rapid remediation of contaminated soils. We recently reported (Song et al., Environ. Sci. Tech., 53, 2045-2053, 2019) the first pilot-scale study of pyrolytic remediation of crude-oil contaminated soil using a continuously-fed rotary kiln reactor. Pyrolysis experiments were conducted at different temperatures and times to determine the optimal conditions to achieve efficient remediation of contaminated soils. In this study, we tested the hypothesis that treatment of mice with soils subjected to pyrolytic remediation at different temperatures and times would display differential attenuation of DNA adducts in vivo. Contaminated soils [14,000-18,000 mg/kg total petroleum hydrocarbon (TPH)] were subjected to pyrolysis at 370 0C for 30 min, 420 0C for 30 min, 420 0C for 15 min, or 470 0C for 30 min, and the soils were extracted with 1:1 acetone/hexane. Mice were treated i.p. with contaminated or remediated soils (1 g/kg), once every day for 4 days, and mice were sacrificed 24 h after the last treatment. Genomic DNA from lungs was extracted, and subjected to 32P-postlabeling.for analyzing DNA adducts. Mice treated with contaminated soils gave rise to PAH-DNA adducts (RAL, 80 x 109), while clean soil-treated mice were devoid of DNA adducts. Mice treated with remediated soils showed significant attenuation of DNA adduct levels (35-45%), with maximum effect (~45%) being observed when soils were remediated 470 0C for 30 minutes. These results are promising, and further studies could lead to more efficient remediation at lower temperatures and residence times. Scaling up of these approaches could lead to novel strategies for remediation of soils in Superfund sites. (Supported by 1P42

Email: [email protected] Presenter Status: Researcher

278 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-46

Quantitatively linking arsenic-contaminated aquifers via spectroscopy: from deltaic aquifers of Southeast Asia to glacial aquifers of the upper Midwestern United States

Athena Nghiem, Columbia University, New York Lamont-Doherty, Earth Observatory Melinda L. Erickson, United States Geological Survey Benjamin C. Bostick, Columbia University, Lamont-Doherty Earth Observatory

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Columbia University Superfund Research Center

Contamination of toxic arsenic in groundwater over the Environmental Protection Agency (EPA) Maximum Contaminant Level and World Health Organization limits of 10 μg/L often occurs naturally in the subsurface, across the United States and worldwide. Arsenic is highly ranked as a contaminant of concern on the Agency for Toxic Substances & Disease Registry (ATSDR) Substance Priority List, topping the list in 2019. Arsenic release into groundwater is commonly attributed to microbial reductive dissolution of arsenic-bearing iron(III) minerals in the subsurface. Here we address two gaps in understanding arsenic solid-solution partitioning: (i) there is a paucity of solid-phase aquifer redox compositions to relate to measured aqueous concentrations of groundwater and (ii) there are limited and namely qualitative links between intensely studied regions of arsenic release such as South and Southeast Asia and contaminated aquifers in the United States. We examine the relationship between paired aqueous concentrations and iron mineralogy by X-ray absorption spectroscopy in a redox gradient of increasing arsenic contamination in the Red River Delta. With cluster analysis, groups of sediments based on weathering and redox transformations can be distinguished, including a group with evidence of iron reduction before arsenic concentrations are measured. These paired observations are combined in a multinomial logistic regression model of deltaic aquifers to upscale the model and establish whether aqueous compositions are indicative of aquifer redox processes. Then, we incorporate spectroscopy from glacial aquifers of the upper Midwest United States to determine the similarities or differences between solid phase redox states in deltaic versus glacial aquifers, with implications for populations that rely on groundwater in these aquifers.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 279 Poster Session IV: Poster #IV-47

Induction of acrylyl-CoA levels is due to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) decreases in vitamin B12 and propionyl-CoA shunting to a β-oxidation-like pathway

Karina Orlowska, Michigan State University, East Lansing, Michigan Russell R. Fling, Michigan State University, East Lansing, Michigan Rance Nault, Michigan State University, East Lansing, Michigan Tim Zacharewski, Michigan State University, East Lansing, Michigan

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Michigan State University Superfund Research Center

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a persistent environmental contaminant, has been shown to reprogram central carbon metabolism in mice. Propionyl-CoA, an end product of odd carbon number fatty acids, branched chain amino acids, bile acids and microbiome digestion, can be further metabolized by a carboxylation pathway to supply succinyl-CoA to the TCA cycle involving a vitamin B12-dependent methylmalonyl-CoA mutase (Mut) reaction. Alternatively, propionyl-CoA is metabolized by a B12- independent β-oxidation-like pathway producing acrylyl-CoA, a toxic intermediate. Untargeted liquid chromatography tandem mass spectrometry of liver extracts from male C57BL/6 mice orally gavaged with 0.01-30 μg/kg TCDD every 4 days for 28 days detected the dose-dependent induction of acrylyl-CoA and evidence of 3-hydroxypropionate suggesting propionyl-CoA metabolism via a β-oxidation-like pathway. Conversely, TCDD repressed genes associated with the alternative B12-independent propionyl-CoA carboxylation pathway. Accordingly, TCDD was also found to dose-dependently decreased serum vitamin B12 levels causing propionyl-CoA to be redirected to the β-oxidation-like pathway. However, repression of enoyl-CoA hydratase 1 (Echs1) and competition with other medium and short chain enoyl-CoA species resulted in the accumulation of the acrylyl-CoA. These results suggest the decrease in vitamin B12 by TCDD redirects propionyl-CoA metabolism towards the B12-independent β-oxidation-like pathway resulting in accumulation of the toxic acrylyl-CoA intermediate due to the repression and substrate overload of Echs1. Funded by Superfund Research Program P42ES04911, NIEHS grant (R01ES029541) and NIEHS Multidisciplinary Training in Environmental Toxicology (NIEHS P2T32ES007255).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

280 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-48

Molecular Modeling and Simulations Uncovering the Sorption of Bisphenols and Phthalates on Unamended and Carnitine-Amended Montmorillonite Clays

Asuka Orr, Texas A&M University, College Station, Texas Shujun He, Texas A&M University, College Station, Texas Meichen Wang, Texas A&M University, College Station, Texas Sara E. Hearon, Texas A&M University, College Station, Texas Timothy D. Phillips, Texas A&M University, College Station, Texas Phanourios Tamamis, Texas A&M University, College Station, Texas

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: Texas A&M University Superfund Research Center

Soil and sediment contaminated with toxic compounds can be transported to areas with high levels of human contact during natural disasters such as flooding or hurricanes. Montmorillonite clays could be promising sorbents to mitigate toxic compound exposures. Bisphenols A (BPA) and S (BPS) as well as phthalates, dibutyl phthalate (DBP) and di-2-ethylhexyl phthalate (DEHP), are used in the production of plastics and are ubiquitous environmental contaminants linked to adverse health effects. Here, we modeled both unamended and carnitine-amended montmorillonite clay in the presence of BPA, BPS, DBP, and DEHP, independently, using multiple multi-ns molecular dynamics (MD) simulations in CHARMM. Following MD simulations, we performed energy calculations and structural analyses to elucidate and predict the binding of the toxic compounds to the unamended and carnitine-amended clays. We computationally calculated the binding propensities and binding persistence of the toxic compounds for the clays, the key binding modes of the different toxic compounds upon binding to the clays, and the energetic favorability of each of the toxic compounds binding to the clays. Our computational analysis predicted that the binding propensity of BPA and BPS was higher than for DBP and DEHP for unamended clay. The binding or BPA and BPS was also predicted to be improved for carnitine-amended clay. Importantly, the results derived from computational MD simulations were in line with in vitro experimental results from adsorption isotherms. This research was funded by a grant from NIEHS (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 281 Poster Session IV: Poster #IV-49

Toward improved use of aboveground plant material to monitor belowground contaminants and their movement in urban environments

Shirley A. Papuga, Wayne State University, Detroit, Michigan Glen R. Hood, Wayne State University, Detroit, Michigan Connor Socrates, Wayne State University, Detroit, Michigan Kennadi Rankin, Wayne State University, Detroit, Michigan Kyotaek Hwang, Wayne State University, Detroit, Michigan Mary Whitfield, Wayne State University, Detroit, Michigan

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Unaffiliated

In urban environments, addressing public health issues associated with organic and inorganic carcinogens is limited by our ability to identify belowground chemical contamination in a spatially comprehensive and cost-efficient way. Current monitoring is generally limited to sampling from wells which are sparse and not well-distributed, especially in urban areas. Because many cities have complex highly-altered hydrologic conditions, relying on these data to understand contaminant movement is insufficient. Further, volatile contaminants such as TCE and 1,4-dioxane are particularly problematic because concentrations of concern can be low enough to exceed limitations of current monitoring techniques. While using plant tissue to screen for belowground contaminants has been evolving as a robust alternative to relying solely on groundwater sampling, this technique is not trivial because (1) most laboratories are not prepared to work with plant tissue; and those that are have found that contaminants extracted from plant tissue is often below detection limits and (2) the extent to which urban plants use water from various sources is generally not well understood. Interestingly, previous studies have shown higher concentrations of heavy metals in resource-accumulating insect-induced plant galls than in other plant tissues. We build on this research by working with analytical chemistry laboratories to zero-in on a robust methodology for detecting organic and inorganic contaminants in galls and other plant tissues. Here we present that preliminary pilot research. We also present preliminary water stable isotope-derived source waters for urban plants using a simple mixing model. We highlight how understanding source waters for urban plants positions us to link carcinogenic contaminants detected in aboveground plant tissue to belowground sources of contamination.

Email: [email protected] Presenter Status: Researcher

282 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-50

A new state of the art resource to overcome limitations of using mouse models for translational research in the study of arsenic-obesity-diabetes interactions

Fernando Pardo-Manuel de Villena, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Ginger D. Shaw, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Pablo Hock, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Timothy A. Bell, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina James G. Xenakis, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Matt Blanchard, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Caroline E. Y. Murphy, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Rebecca C. Fry, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Beverly H. Koller, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Miroslav Styblo, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina

Superfund Mandate: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

Inorganic arsenic (iAs) is a ubiquitous environmental contaminant currently poisoning of millions of individuals around the world. Chronic exposure to iAs is associated with the risk of type 2 diabetes (T2D), and the ability to metabolize iAs into monomethyl-As and dimethyl-As is a critical driver of this risk. Until now the laboratory mouse had two major limitations as a model for humans: a radically different profile in arsenic metabolism and a lack of genetic diversity that would allow the investigation of genome-wide genetic underpinnings of the susceptibility to iAs-associated T2D. The first limitation has been addressed by the generation of an inbred mouse model in which gene encoding for the main enzyme for arsenic metabolism, AS3MT, has been replaced by the human gene, including the regulatory regions that control temporal and tissue specific patterns of gene expression. To address the second limitation, we used a new genotyping array combined with a custom breeding design to generate speed congenic lines in which the humanized AS3MT gene will be placed in the context of several vastly different genetic backgrounds. These backgrounds represent different Collaborative Cross (CC) inbred strains. The CC is a multiparental population with over 40 million segregating variants and with a wide range of phenotypic variation. These phenotypes include several metabolic traits such as baseline fasting blood glucose, body weight and composition and propensity to T2D. This new resource will improve the rigor of research based on mouse models for this critical and widespread contaminant and allow for faster and more robust translational research.

Email: [email protected] Presenter Status: Researcher

NIEHS SRP 2020 Virtual Annual Meeting Program • 283 Poster Session IV: Poster #IV-51

Roles of Placental Extracellular Vesicles in the Regulation of Fetal Brain Development during Pregnancy

Hae-Ryung Park, Harvard School of Public Health, Boston Massachusetts Charlotte Wirth, Harvard School of Public Health, Boston Massachusetts Quan Lu, Harvard School of Public Health, Boston Massachusetts

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Harvard School of Public Health Superfund Research Center

Neural stem cells (NSCs) play an essential role in shaping the developing brain. Recently, it’s been proposed that placenta plays a critical role in fetal neuroprogramming through the inter- relationships between neurotoxicants and the placenta. Placental cues can reach the fetal brain through the immature blood-brain barrier (BBB) inducing physiological or pathological changes with consequent neurodevelopmental outcomes. Placental trophoblasts actively release nano- sized membrane-bound extracellular vesicles (EVs). The EVs can cross the fetal BBB and shuttle cargoes of bioactive molecules, such as proteins, lipid, and nucleic acids. We hypothesize that placental EVs modify NSC functions by transferring cargoes of bioactive molecules to NSCs. We further hypothesize that environmental metal exposure such as arsenic (As) in utero alters fetal brain development by disrupting normal cargoes of placental EVs. To test these hypotheses, we first harvested EVs from the culture medium of a human first trimester placenta cell line HTR-8/SVneo using ultracentrifugation. Then, ReNcell Cx human neuroprogenitor cells were incubated with the harvested EVs in the absence or presence of As. We found treatment of placental EVs stimulated proliferation of ReNcell Cx cells. We further showed that pretreatment of ReNcell Cx cells with placental EVs protects against As-induced inhibition of cell proliferation. Proteomics analysis revealed that brain-enriched proteins have been detected in placental EVs. Our study proposes placental EVs as a novel signaling factor mediating the NSC responses to As exposure thereby linking environmental exposures to health trajectories regulated by the placenta. Further study will be warranted to elucidate the mechanisms by which placental EVs affect NSC proliferation against As exposure.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

284 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-52

Hydrogeochemical investigations on the distribution and cycling of redox- sensitive geogenic contaminants in NC Piedmont groundwater

Hannah R. Peel, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Owen Duckworth, North Carolina State University, Raleigh, North Carolina David Vinson, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Charlotte Matthew Polizzoto, University of Oregon, Eugene, Oregon Robert Austin, North Carolina State University, Raleigh, North Carolina Casey Miller, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina Elizabeth Batianis, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: University of North Carolina-Chapel Hill Superfund Research Center

In North Carolina, roughly three million people rely on well water as their, where the natural hydrogeology of the Piedmont results in widespread natural (geogenic) arsenic contamination of groundwater. As a result, an estimated 76,000–112,000 people consume well water with unsafe As concentrations. Elevated concentrations of other geogenic contaminants, such as Cr, V and Mn, are also found in clusters within the NC Piedmont, with distributions largely governed by geology and/or groundwater chemical conditions. The mobility and health impacts of these contaminants are largely dependent on their redox speciation, and therefore, a fundamental understanding these elements’ redox state is critical for predicting well-water concentrations and human exposure risks. In this study, groundwater’s dissolved speciation (e.g. As(V)/As(III), Cr(VI)/Cr(III), V(V)/V(III))will be paired to solid phase chemistry and speciation (e.g. X-ray spectroscopy, sequential extraction) measurements. The information gleaned as a part of this study will lay the foundation to not only to determine the concentration of arsenic in groundwater and the associated risk to human health, but also to begin to understand the chemical mechanisms controlling the speciation of As, Cr and V in an aquifer system, and potentially to create subsurface geospatial models to help predict where contamination may occur.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 285 Poster Session IV: Poster #IV-53

Antibody-based biosensor technology as a rapid PAH screening tool for seafood safety assessments of oysters

Kristen Prossner, Virginia Institute of Marine Science Michael Unger, Virginia Institute of Marine Science George Vadas, Virginia Institute of Marine Science Ellen Harvey, Virginia Institute of Marine Science

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: R01

Polycyclic aromatic hydrocarbons (PAH) are highly toxic contaminants found in creosote and oil that can bioaccumulate in bivalves, posing a hazard to human health. Risk assessments to guide reopenings of fishing sites after spills are often time-consuming and expensive. The Gulf of Mexico seafood industry lost billions of dollars due to closures after the Deepwater Horizon spill. The lengthy turnaround time required for complex analyses such as gas chromatography-mass spectrometry (GC-MS) delays information dissemination to the public. An initial screening method commonly employed after spills to rapidly detect petroleum-tainted seafood is sensory analysis (i.e. sniff-testing). Limitations in both methods contribute to ineffective risk communication and public distrust. Here, we present an alternative screening tool to rapidly quantify total PAH concentrations in adult oysters. Using a monoclonal antibody coupled with fluorescence detection, the biosensor method allows near real-time (7 minutes) measurement of the total 3- to 5- ring PAH concentration in the oyster mantle fluid at a low (sub-ppb) detection limit in small volume (1 to 5 mL) samples. Results from the biosensor method correlate to those of GC-MS analysis allowing prediction of whole animal tissue concentrations. This study also investigates the potential to calibrate the biosensor method to align with specific subsets of PAH compounds targeted for risk assessments, demonstrating its wide applicability. Rapid analysis of oysters throughout a watershed demonstrated that concentration gradients and areas of concern can easily be identified. Having access to a rapid and inexpensive quantitative screening tool will provide for more efficient post-spill seafood safety analysis and time-sensitive communication to the public.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

286 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-54

Community Engagement and Educational Efforts during SARS-COV2: The PROTECT Story

Amalie Santos Rivera, Northeastern University, Boston, Massachusetts N.R. Cardona Cordero, Northeastern University, Boston, Massachusetts N. Hernadez Otero, Northeastern University, Boston, Massachusetts C.M. Galan Rivera, Northeastern University, Boston, Massachusetts H.R. Torres Zayas, Northeastern University, Boston, Massachusetts Z.J. Dias Reguero, Northeastern University, Boston, Massachusetts C.M. Velez Vega, Northeastern University, Boston, Massachusetts J.F. Cordero, Northeastern University, Boston, Massachusetts A. Alshawabkeh, Northeastern University, Boston, Massachusetts

Superfund Mandates: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: Northeastern University, Boston, Massachusetts

In March 2020, the first positive case of SARS-COV2 was detected in Puerto Rico. The participants’ safety and adapting our research in accordance with the governmental executive orders was the main priority for the Puerto Rico Testsite for Exploring Contamination Threats (PROTECT). As uncertainty and local challenges approached, our team was interested in how to continue helping pregnant women and their families during this pandemic period. From March to May 2020, our first tactic was to contact over 2,000 participants through our call center and ask them how they wanted to receive information about prevention strategies to up front SARS-COV2 and environmental exposures. As a result, our cohort expressed interest in receiving materials through digital platforms with visual representations. Our second approach, from May to September 2020, the Community Engagement Core (CEC) team designed strategies in order to adapt the information to our new reality. We generated and disseminated numerous diverse virtual educational efforts like; webinars, Facebook lives, digital infographics, social media posts and audiovisuals. The infographics and videos focused on how to take care of their pregnancy stage during the COVID-19 pandemic and minimize exposures to Phthalates, Bisphenols, Triclosan, Triclocarban, Parabens and other chemicals of concern. As a third strategy, the CEC disseminated all educational materials through text-messages and social media, which gave the opportunity to amplify our capacity building. As an outcome, thousands of individuals were involved and connected in the health promotion and educational efforts. Our research team combines diverse public health specialties which were a center piece in developing and making accessible these public environmental health approaches providing prevention strategies to our cohort and their families.

Email: [email protected] Presenter Status: Other: Education Specialist

NIEHS SRP 2020 Virtual Annual Meeting Program • 287 Poster Session IV: Poster #IV-55

Identification and functional characterization of a novel AHR2-dependent gene in zebrafish

Prathana Shankar, Oregon State University, Corvallis, OR Jane K. LaDu, Oregon State University Gloria R. Garcia, Oregon State University Christopher M. Sullivan, Oregon State University Katrina M. Waters, Pacific Northwest National Laboratory Robyn L. Tanguay, Oregon State University (Corresponding author)

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: Oregon State University Superfund Research Center

The aryl hydrocarbon receptor (AHR) is required for vertebrate development and is activated by several chemicals including polycyclic aromatic hydrocarbons (PAHs) and TCDD. AHR activation is well understood, but the roles of downstream genes are largely unknown. Previously, we conducted transcriptomics in 48-hours post fertilization (hpf) zebrafish exposed to 16 PAHs. One of the most highly elevated transcripts by several PAHs was wfikkn1 (WAP, kazal, immunoglobulin, kunitz and NTR domain- containing protein), a potential novel biomarker for AHR activation. We used qRT-PCR to demonstrate wfikkn1 is expressed from 2.5 to 120 hpf. When we exposed AHR2 (ortholog of human AHR)-null and Wildtype zebrafish to DMSO or TCDD, and quantified the expression of wfikkn1 at 48 and 120 hpf, the significant induction of wfikkn1 was AHR2-dependent. To functionally characterize wfikkn1, we generated a wfikkn1 CRISPR-Cas9 mutant zebrafish line that produces a truncated protein. The mutant zebrafish appear morphologically normal, but have altered behavioral responses as larvae and juveniles. To identify genes/pathways associated with wfikkn1, Wildtype and mutant zebrafish were exposed to DMSO or TCDD, and mRNA from 48-hpf zebrafish was sequenced. We identified over 200 differentially expressed genes between each pair of treatment combinations (p2). The significant GO term processes associated with wfikkn1 mutants include ATP-binding, immunoglobulin, and iron binding processes. ATP-binding, transmembrane, and protein kinase processes were uniquely altered in TCDD-exposed wfikkn1 mutants compared to TCDD-exposed Wildtype zebrafish. We are currently conducting in-depth analyses of the RNA-sequencing data, and characterizing in situ RNA and protein expression. Understanding downstream transcriptional events that occur upon AHR activation and play a role in toxicity pathways is necessary to accurately guide remediation strategies.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

288 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-56

Monkey Liver Microsomes and Cytosol Enantioselectively Metabolize Benoxacor

Derek W. Simonsen, University of Iowa, Iowa City, Iowa David Cwiertny, University of Iowa, Iowa City, Iowa Jacob Heffelfinger, University of Iowa, Iowa City, Iowa Hans-Joachim Lehmler, University of Iowa, Iowa City, Iowa

Superfund Mandate: Methods to assess the risks to human health presented by hazardous substances

Superfund Center: University of Iowa Superfund Research Center

Safeners like benoxacor are inert ingredients of herbicide formulations; however, only little information is available about their metabolism and toxicity. The goal of this study was to elucidate the enantioselective metabolism of benoxacor in vitro. We studied the depletion of benoxacor in microsomes and prepared from female and male monkey livers. Benoxacor levels were determined by gas chromatography-mass spectrometry. Enantiomeric fractions (EF) were determined by enantioselective gas chromatography with electron capture detection. Benoxacor was readily metabolized in incubations with active microsomes and cytosol. After 30 minutes, 18±1% and 26±3% of benoxacor remained in incubations with microsomes, and 29±2% and 32±1% remained in incubations with cytosol obtained from female and male monkeys, respectively. Without the addition of NADPH in the microsomes, 68±1% and 63±1% of benoxacor remained in microsomes obtained from female and male monkeys, respectively, indicating metabolism by carboxylesterases. Without GSH, 78±1% and 78±1% of benoxacor remained in incubations with cytosol from female and male monkeys, respectively, indicating metabolism by cytosolic carboxylesterases. Microsomal incubations showed time-dependent enantiomeric enrichment of benoxacor. A more pronounced enrichment was noted at 30-minutes in incubations with microsomes from female than male monkeys (EF = 0.66±0.01 vs. EF = 0.54±0.01, respectively). Incubations with cytosol from both female and male monkeys resulted in the complete depletion E1-benoxacor, while levels of E2 benoxacor remained relatively unchanged. These results indicate that microsomal and cytosolic enzymes from monkeys enantioselectively metabolize benoxacor. Further research is needed to determine the effects of enantioselective metabolism on the toxicity of benoxacor (Supported by NIH P42 ES013661).

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 289 Poster Session IV: Poster #IV-57

Mercury cycling and bioavailability at two contaminated estuarine sites in the Northeast U.S.

Vivien F. Taylor, Dartmouth College, Hanover, NH Emily Seelen, University of Southern California Kate Buckman, Dartmouth College, Hanover, NH Rob Mason, University of Connecticut Celia Chen, Dartmouth College, Hanover, NH

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Dartmouth College Superfund Research Center

Similar concentrations of methylmercury (MeHg) were found in fish (Fundulus sp) downstream of two contaminated estuarine sites in the Northeast US: the Penobscot River, a remote site contaminated by historic inputs from a chemical plant in Orrington, Maine, and Berry’s Creek, which runs from a Superfund site of the same name to the Hackensack River, NJ. Despite the similar levels of MeHg in fish near these sources of contamination, the sites vary substantially in Hg loading (THg was 14 mg/kg and 0.8 mg/kg at the Berry’s Creek and Penobscot sites, respectively). The sites also differ in their ecological setting and degree of anthropogenic impact, indicating biogeochemical factors other than sediment and water MeHg levels alone control bioavailability of MeHg to the food web. Concentrations of THg and MeHg in the sediment and water column of Berry’s Creek, which is heavily impacted by urbanization and industrial effluent and is overrun with the invasive marsh grass, Phragmites, are much higher than in the Penobscot where Hg is the sole contaminant and the surrounding ecosystem is relatively pristine and dominated by the native marsh grass, Spartina. Concentrations of DOC and salinity gradients were comparable between sites, whereas DOC quality in Berry’s creek was significantly more degraded, based on δ13C and fluorescence measurements. These findings demonstrate much higher bioaccumulation in fish at the less impacted site, where carbon quality was less degraded. The findings of this work imply that parameters such as carbon quality and land use affect bioavailability in near-shore environments, suggesting that more biogeochemical factors need to be included in contaminated site assessments.

Email: [email protected] Presenter Status: Researcher

290 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-58

Analysis of Per- and Polyfluoroalkyl Substances in Houston Ship Channel Following the Intercontinental Terminals Company Fire Incident Using Ion- Mobility Mass Spectrometry

Alan Valdiviezo, Texas A&M University, College Station, Texas Noor Aly, Texas A&M University, College Station, Texas Yu-Syuan Luo, Texas A&M University, College Station, Texas Gaston Casillas, Center for Disease Control and Prevention, Atlanta, Georgia Erin Baker, North Carolina State University, Raleigh, North Carolina Ivan Rusyn, Texas A&M University, College Station Texas

Superfund Mandates: Methods and technologies to detect hazardous substances in the environment

Superfund Center: Texas A&M University Superfund Research Center

Per- and polyfluoroalkyl substances (PFAS) are persistent organic pollutants of emerging concern. Following the fires at the Intercontinental Terminals Company (ITC) in Deer Park, TX, very large quantities of PFAS-containing firefighting foams were deployed. The release of these potentially hazardous substances into waterways in the Houston Ship Channel (HSC) prompted concerns over the extent and level of environmental contamination. A liquid chromatography tandem mass spectrometry (LC-MS/MS)- based study of temporal and spatial patterns of PFAS associated with the ITC fire incident indicated extensive contamination immediately after the incident. PFAS levels gradually decreased over a 6-month period. While highly informative, the LC-MS/MS “targeted” approaches are limited to a narrow range of analytes which do not account for the diversity of PFAS in firefighting foams. Therefore, we tested a hypothesis that an untargeted liquid chromatography, ion mobility and mass spectrometry (LC-IM-MS)- based analysis of environmental samples with known presence of PFAS will provide a comprehensive profile of the contaminants and improve exposure assessment. We analyzed 31 samples from 9 sites in the HSC that were collected from April through August 2019. We used unique feature identification capabilities of LC-IM-MS to identify dozens of previously undetected PFAS in HSC. Our data showed that PFAS presence in HSC decreased gradually in concordance with LC-MS/MS analysis. The abundance of PFAS features detectable by LC-MS/MS were significantly correlated with those identified by LC-IM-MS; however, LC-IM-MS provided information on PFAS species not evaluated with traditional targeted analysis. In conclusion, these findings illustrated that LC-IM-MS enables a comprehensive and informative technique for rapid screening of environmental samples to facilitate exposure assessment during emergencies and provides guidance for subsequent focused targeted chemical analyses. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 291 Poster Session IV: Poster #IV-59

Inhibition of Rad18 by Arsenic

Lindsay Volk, University of New Mexico, Albuquerque, New Mexico Karen L Cooper, University of New Mexico, Albuquerque, New Mexico Xuejiao Dong, University of California, Riverside California Yinsheng Wang, University of California, Riverside California Laurie G Hudson, University of New Mexico, Albuquerque, New Mexico

Superfund Mandates: Advanced techniques for the detection, assessment, and evaluation of the effect on human health of hazardous substances

Superfund Center: University of New Mexico Superfund Research Center

Environmental exposure to arsenic enhances the carcinogenicity of ultraviolet radiation leading to increased risk of developing nonmelanoma skin cancers associated with increased cost, morbidity, and mortality. The impacts of arsenic on human health is a major concern for Southwestern United States where an estimated 16% of well water contains arsenic levels exceeding the United States Environmental Protection Agency maximum contaminant level of 10 ppb. The goal of this study is to elucidate mechanisms underlying the cocarcinogenic actions of arsenic to improve the health of the over 200 million individuals worldwide exposed to high levels of arsenic. Arsenite (AsIII) is known to bind and inhibit zinc finger domains within DNA repair proteins, resulting in the retention of DNA damage with coexposure to ultraviolet radiation. Accumulated DNA damage increases the burden on DNA damage tolerance mechanisms, the last line of defense against cell death or mutagenesis. Rad18 is an essential factor to DNA damage tolerance and DNA double-strand break repair and contains two functionally diverse zinc fingers which are structurally conducive to AsIII binding and disruption. Due to the increased propensity for mutagenesis with AsIII exposure, the central hypothesis of this study is AsIII disrupts the function of Rad18. Results have shown Rad18 zinc finger disruption by AsIII leading to zinc displacement and domain loss of function. In addition, findings demonstrate a multifaceted effect of AsIII on Rad18, including alterations in Rad18 cellular localization. By evaluating the impact of AsIII on Rad18, novel findings have been revealed supporting the link between environmental exposure to arsenic and the development of cancer. This information is pertinent to environmental policy decision making and the discovery of novel interventions.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

292 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-60

Inducible and conditional stimulation of adult hippocampal neurogenesis rescues mice from Cd-induced hippocampus-dependent learning and memory deficits

Hao Wang, University of Washington, Seattle Glen M. Able, University of Washington, Seattle

Superfund Mandates: N/A

Superfund Center: University of Washington Superfund Research Center

Cadmium (Cd) is a naturally occurring heavy metal that has been recognized as one of the most toxic environmental pollutants around the world. Cd is also suggested to be a neurotoxicant. However, the underlying mechanisms for its neurotoxicity are not fully understood. Our previous studies have demonstrated that Cd exposure can impair hippocampus-dependent learning and memory, as well as adult hippocampal neurogenesis in mice. To determine if these adverse effects of Cd can be mitigated by genetically and conditionally enhancing adult neurogenesis, we utilized the transgenic caMEK5 mouse strain we previously developed and characterized. This mouse strain enables us to genetically and conditionally stimulate adult neurogenesis by administering tamoxifen to induce expression of a constitutively active form of MEK5 (caMEK5) in adult neural progenitor cells, which can stimulate adult neurogenesis through the activation of the endogenous ERK5 MAP kinase pathway. The caMEK5 mice were exposed to Cd (0.6 mg/L) through drinking water for 38 weeks. Once memory impairment was confirmed, tamoxifen was administered to induce caMEK5 expression and to activate adult neurogenesis. Behavior tests were conducted at different time points to monitor hippocampus-dependent learning and memory. Upon completion of the behavior tests, brain tissues were collected for the cellular study of adult hippocampal neurogenesis. We found that Cd impaired hippocampus-dependent short-term spatial memory and contextual fear memory in caMEK5 mice. These deficits were rescued by the tamoxifen- induction of caMEK5 expression. In addition, Cd inhibition of adult hippocampal neurogenesis was also reversed. This rescue experiment provides evidence for a direct link between Cd-induced impairments of adult hippocampal neurogenesis and hippocampus-dependent memory.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 293 Poster Session IV: Poster #IV-61

Soluble epoxide hydrolase is an endogenous regulator of obesity-induced intestinal barrier dysfunction and bacterial translocation

Yuxin Wang, University of California, Davis Jun Yang, University of California, Davis Weicang Wang, University of California, Davis Katherine Z. Sanidad, University of Massachusetts, Amherst Maris A. Cinelli, Michigan State University Debin Wan, University of California, Davis Sung Hee Hwang, University of California-Davis Daeyoung Kim, University of Massachusetts, Amherst Kin Sing Stephen Lee, Michigan State University Bruce D. Hammock, University of California Davis Guodong Zhang, University of Massachusetts, Amherst

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California, Davis Superfund Research Center

Intestinal barrier dysfunction, which leads to translocation of bacteria or toxic bacterial products from the gut into the bloodstream and results in systemic inflammation, is a key pathogenic factor in many human diseases. However, the molecular mechanisms leading to intestinal barrier defects are not well understood, and there are currently no available therapeutic approaches to target intestinal barrier function. Here we show that soluble epoxide hydrolase (sEH) is an endogenous regulator of obesity- induced intestinal barrier dysfunction. We find that sEH is overexpressed in the colons of obese mice. In addition, pharmacologic inhibition or genetic ablation of sEH abolishes obesity-induced gut leakage, translocation of endotoxin lipopolysaccharide or bacteria, and bacterial invasion-induced adipose inflammation. Furthermore, systematic treatment with sEH-produced lipid metabolites, dihydroxyeicosatrienoic acids, induces bacterial translocation, and colonic inflammation in mice. The actions of sEH are mediated by gut bacteria-dependent mechanisms since inhibition or genetic ablation of sEH fails to attenuate obesity-induced gut leakage and adipose inflammation in mice lacking gut bacteria. Overall, these results support that sEH is a potential therapeutic target for obesity-induced intestinal barrier dysfunction, and that sEH inhibitors, which have been evaluated in human clinical trials targeting other human disorders, could be promising agents for prevention and/or treatment.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

294 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-62

Impact of Metals on Aromatic Amine N-Acetyltransferase Metabolism, Genotoxicity and Cytoxicity in Human Lung Cells

James T.F. Wise, University of Louisville, Louisville, Kentucky Raúl A. Salazar-González, University of Louisville, Louisville, Kentucky Mark A. Doll, University of Louisville, Louisville, Kentucky David W. Hein, University of Louisville, Louisville, Kentucky

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of Louisville Superfund Research Center

Humans are exposed to carcinogenic compounds via environmental and occupational exposures. Compounds of concern that may occur in mixtures together are aromatic amines (e.g. 2-aminofluorene [2-AF]) and heavy metals (arsenic, cadmium, hexavalent chromium, and nickel). Arylamine N- acetyltransferases 1 and 2 (NAT1/2) are key to the metabolism of aromatic amines and their genotoxicity. A previous report indicated cadmium reduces NAT1/2 activities. However, the implication of heavy metals on the metabolism of aromatic amines by NATs remains unknown and whether heavy metals alter the aromatic amines genotoxicity or cytotoxicity. This research seeks to assess whether heavy metals will alter the N-acetyltransferase metabolism of aromatic amines and their genotoxicity and cytotoxicity in immortalized human lung epithelial cells and Chinese hamster ovary cells (CHO) expressing NAT1 or NAT2. We will measure individual and combined effects of heavy metals and aromatic amines on NAT proteins, genotoxicity, and cytotoxicity. Our preliminary “in cell” western results show that exposure to Cd(II) decreased protein expression of NAT1/2 in CHO cells. Specifically, relative to the untreated control, 1.25, 2.5, and 5 uM Cd(II) NAT1 protein expression was 103, 64, and 35% and NAT2 protein expression was 61, 28, and 5%. We also observed Cd(II) reduced para-aminobenzoic acid and sulfamethazine N-acetylation by NAT1 and NAT2 respectfully. Using yeast extract expressing recombinant human NAT1 or NAT2, cadmium inhibited activity N-acetylation of 2-AF of NAT1 and NAT2. For 0.4 and 4 uM Cd(II) reduced NAT1 activity to 11 and 8% of control and NAT2 activity to 21 and 13% of control. We anticipate this observed reduction of NATs will alter the toxicity of an aromatic amine during co-exposures and have implications for understanding co-exposures.

Email: [email protected] Presenter Status: Trainee (pre-/post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 295 Poster Session IV: Poster #IV-63

The role of 7-dehydrocholesterol reductase (DHCR7) in alcoholic liver disease- induced hepatocellular carcinoma

Gen Yamamoto, University of California, San Diego Na Li, University of California, San Diego Xiao Liu, University of California, San Diego Li Liu, University of California, San Diego Jacopo Baglieri, University of California, San Diego David A. Brenner, University of California, San Diego Tatiana Kisseleva, University of California, San Diego

Superfund Mandates: Basic biological, chemical, and physical methods to reduce the amount and toxicity of hazardous substances

Superfund Center: University of California, San Diego Superfund Research Center

Background & Aim: We revealed that EtOH-fed IL-17RAΔHep mice devoid of IL-17RA in hepatocytes developed the fewest tumors, with reduced steatosis and fibrosis. Steatotic IL-17RA-deficient hepatocytes showed a striking defect in TNF/TNFR1-dependent caspase-2-SREBP1/2-DHCR7-mediated cholesterol synthesis. We aimed to reveal the role of DHCR7, the cholesterol producing enzyme, in the development of alcohol-induced hepatocellular carcinoma (HCC) in experimental mouse model. Methods: DHCR7-/- mice have developmental problems and some metabolic abnormalities leading to perinatal death. Therefore, we are using DHCR7+/- (which downregulate expression of DHCR7 protein by 50%) and wild type (WT) littermates (males, 2 weeks old) were injected i.p. with Diethylnitrosamine (DEN, 25mg/kg body weight) and at 3 months of age were subjected to isocaloric liquid ethanol (EtOH) feeding, or pair-fed, or chow fed for 18 weeks. Results: Development of HCC was reduced in DEN-injured and EtOH-fed DHCR7+/- mice. EtOH-fed DHCR7+/- mice were also protected from steatosis, fibrosis and inflammation, as shown by decreased staining for Sirius Red, Desmin, a-SMA and F4/80. However, hepatic or serum cholesterol level were not changed, even though hepatic triglyceride level was reduced in EtOH-fed DHCR7+/- mice. Serum level of vitamin D3 in EtOH-fed DHCR7+/- mice was much higher than that of EtOH-fed WT mice, since the conversion of cholesterol precursors into cholesterol was shifted toward vitamin D3 production. Conclusion: These findings suggested that loss-of-function of DHCR7 might provide a poor microenvironment for alcohol-induced HCC growth.

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

296 • NIEHS SRP 2020 Virtual Annual Meeting Program Poster Session IV: Poster #IV-64

Nearby Nature and Post-Traumatic Stress Disorder: Evidence from Hurricane- Affected Communities

Rui Zhu, Texas A&M University, College Station, TX Galen Newman, Texas A&M University, College Station, TX Dongying Li, Texas A&M University, College Station, TX

Superfund Mandates: None

Superfund Center: Texas A&M University Superfund Research Center

Exposure to a severe natural disaster such as a hurricane or flood can lead to mental health issues such as increased risks of stress, anxiety disorders, and post-traumatic stress disorder (PTSD). Exposure to nearby nature has consistently been demonstrated to help vulnerable or disadvantaged populations cope with stress. However, the effects of nearby nature on mitigating post-traumatic distress has not been discussed much. This study aims to understand the associations between nearby nature and levels of PTSD in Houston communities that were affected by Hurricane Harvey using a cross-sectional survey. We sampled 30 Houston neighborhoods that endured severe loss during Hurricane Harvey, and then employed an address-based sampling of households within the block groups in 2019. We had a total of 272 participants, among whom 62.3% were females, and 28.5% were Hispanic. Self-reported neighborhood green space was measured using an adapted NOS scale. PTSD was measured using the Posttraumatic Stress Disorder Checklist for DSM-5 (PCL-5) as a dichotomous outcome, and hurricane-related distress was measured using the subscales of the Impact of Event Scale (IES-R). Generalized linear mixed models were used to test the relationships. In the models, neighborhood- and individual-level confounding variables were controlled. Results showed significant associations between nearby nature and lower levels of PTSD, as well as the avoidance domain of hurricane-related distress. As the first study that links salutogenic environmental factors to post-hurricane mental disorders, this study contributes to the literature by identifying the mental health effects of green space in the aftermath of the most severe hurricane in recent years. The findings from this study can provide policy implications in establishing a holistic disaster planning that incorporates mental health promotion. This research was funded by grant from NIH (P42 ES027704).

Email: [email protected] Presenter Status: Trainee (pre- / post-doc)

NIEHS SRP 2020 Virtual Annual Meeting Program • 297 Attendees

Last Name First Name Email Other (if noted) Baylor College of Medicine Aagaard Kjersti [email protected] Alvarez Pedro [email protected] Rice University Bajomo Mary [email protected] Rice University Banerjee Sohini [email protected] Chu Chun [email protected] Coarfa Cristian [email protected] Couroucli Xanthi [email protected] The University of Texas Croisant Sharon [email protected] Medical Branch at Galveston Damaraju Aneel [email protected] Rice University Dias da Silva Priscilla [email protected] Rice University Dowst Heidi [email protected] Edwards Dean [email protected] Edwards Dean [email protected] The University of Texas Elferink Cornelis [email protected] Medical Branch at Galveston Finnell Richard [email protected] Gastelum Grady [email protected] Grimm Sandra [email protected] Halas Naomi [email protected] Rice University The University of Texas Hallberg Lance [email protected] Medical Branch at Galveston Hilsenbeck Susan [email protected] Kamal Abu Hena Mostafa [email protected] Kraushaar Daniel [email protected] Lingappan Krithika [email protected] Lupo Philip [email protected] Moorthy Bhagavatula [email protected] Neumann Oara [email protected] Rice University Nordlander Peter [email protected] Rice University Patel Ankit [email protected] Rice University Pedraza Emily [email protected] The University of Texas Prochaska John [email protected] Medical Branch at Galveston Putluri Nagireddy [email protected] Renard David [email protected] Rice University Robertson Matthew [email protected] The University of Texas Singleton Chantele [email protected] Medical Branch at Galveston Suter Melissa [email protected] Swanson Lauren [email protected] Symanski Elaine [email protected]

298 • NIEHS SRP 2020 Virtual Annual Meeting Program Attendees

Last Name First Name Email Other (if noted) Szafran Adam [email protected] Villanueva Ferdinand [email protected] Wensel Theodore [email protected] Xia Guobin [email protected] Boston University Foley Caredwen [email protected] Woods Hole Hahn Mark [email protected] Oceanographic Institution Heiger-Bernays Wendy [email protected] Labadorf Adam [email protected] Sherr David [email protected] Brown University Bilderback Marjorie [email protected] Carvalho Fernandes Deisy [email protected] Hurt Robert [email protected] Li Nan [email protected] Liu Yun [email protected] Martin Shannon [email protected] Plavicki Jess [email protected] Shukla Vidushi [email protected] Suuberg Eric [email protected] Columbia University Abuawad Ahlam [email protected] Bostick Benjamín [email protected] Connolly Craig [email protected] Gamble Mary [email protected] Graziano Joseph [email protected] LoIacono Nancy [email protected] Navas-Acien Ana [email protected] Nghiem Athena [email protected] Nigra Anne [email protected] Saxena Roheeni [email protected] Sobel Marisa [email protected] Spaur Maya [email protected] Dartmouth College Barrack Kaitlyn [email protected] Chen Celia [email protected] Disney Jane [email protected] MDI Biological Laboratory Gibson James [email protected] Goodale Britton [email protected] Guerinot Mary Lou [email protected] Hadlock Robyn [email protected]

NIEHS SRP 2020 Virtual Annual Meeting Program • 299 Attendees

Last Name First Name Email Other (if noted) Karagas Margaret [email protected] OToole George [email protected] Geisel School of Medicine Punshon Tracy [email protected] Rardin Laurie [email protected] Signes-Pastor Antonio [email protected] Taylor Vivien [email protected] Duke University Chernick Melissa [email protected] Clark Charlotte [email protected] Cohen Samuel [email protected] Crittenden Joshua [email protected] Di Giulio Richard [email protected] Gunsch Claudia [email protected] Ferguson Lee [email protected] Hawkey Andrew [email protected] Holloway Zade [email protected] Jasperse Lindsay [email protected] Kastleman Catherine [email protected] Leuthner Tess [email protected] Levin Edward [email protected] Luukinen Bryan [email protected] Meyer Joel [email protected] Ranasinghe Prabha [email protected] Shapiro Elizabeth [email protected] Sharp Alexis [email protected] Stapleton Heather [email protected] Varner Paige [email protected] Harvard School of Public Health Bidlack Felicitas [email protected] Forsyth Institute Goodman Julie [email protected] University of Colorado– James Katherine [email protected] Anschutz Medical Campus Korrick Susan [email protected] Lu Quan [email protected] University of Colorado– Macaluso Francesca [email protected] Anschutz Medical Campus Muhich Chris [email protected] Arizona State University Nwokonkwo Obinna [email protected] Arizona State University Park Hae-Ryung [email protected] Preston Emma [email protected] Von Stackelberg Katherine [email protected] Weisskopf Marc [email protected]

300 • NIEHS SRP 2020 Virtual Annual Meeting Program Attendees

Last Name First Name Email Other (if noted) Wirth Charlotte [email protected] Louisiana State University Ampiah Emmanuel [email protected] Aryal Ankit [email protected] Baconguis Liana [email protected] Billiot Gina [email protected] Chowdhury Pratiti Home [email protected] Coldiron Matt [email protected] Cormier Stephania [email protected] Dangal Prakash [email protected] Dugas Tammy [email protected] School of Veterinary Medicine Guan Xia [email protected] Hale H. [email protected] Harmon Ashlyn [email protected] Hasan Farhana [email protected] Irving Jennifer [email protected] Khachatryan Lavrent [email protected] Odera Matilda [email protected] North Carolina State University Richmond-Bryant Jennifer [email protected] North Carolina State University Sprunger Phillip [email protected] Massachusetts Institute of Technology Armijo Amanda [email protected] Beard Jessica [email protected] Ceriales Gabrielle [email protected] Corless Elliot [email protected] Corrigan Joshua [email protected] Croy Robert [email protected] Engelward Bevin [email protected] Essigmann John [email protected] Feng Haosheng [email protected] Furst Ariel [email protected] Hemond Harold [email protected] Hrdina Amy [email protected] Hu Irene [email protected] Kaushal Simran [email protected] Kelly Jamie [email protected] Kohale Ishwar [email protected] Levine Stuart [email protected] Lu Ruqiang [email protected] Moise Aimee [email protected] Owiti Norah [email protected]

NIEHS SRP 2020 Virtual Annual Meeting Program • 301 Attendees

Last Name First Name Email Other (if noted) Pribyl Lee [email protected] Reposa Kathy [email protected] Selin Noelle [email protected] Swager Timothy [email protected] Tat Amanda [email protected] Vandiver Kathy [email protected] White Forest [email protected] Michigan State University Blevins Lance [email protected] Cholico Giovan [email protected] Dean Rachel [email protected] Dodson Jack [email protected] Fling Russell [email protected] Hamm Joseph [email protected] Hashsham Syed [email protected] Jurgelewicz Amanda [email protected] Kaminski Norbert [email protected] Khan D.M. Isha Olive [email protected] Nault Rance [email protected] Orlowska Karina [email protected] Sallach J. Brett [email protected] Steidemann Michelle [email protected] Swagart Amy [email protected] Zylstra Gerben [email protected] Rutgers University North Carolina State University Alexander Nancy [email protected] Belcher Scott [email protected] Boatman Anna [email protected] Buchwalter David [email protected] DeWitt Jamie [email protected] East Carolina University Duckworth Owen [email protected] Farrell Matt [email protected] Foster Makayla [email protected] Guo Chuqi [email protected] Hopkins Zachary [email protected] Hoppin Jane [email protected] Jackson Thomas [email protected] Knappe Detlef [email protected] Kullman Seth [email protected] Mattingly Carolyn [email protected] May Katlyn [email protected] Peel Hannah [email protected]

302 • NIEHS SRP 2020 Virtual Annual Meeting Program Attendees

Last Name First Name Email Other (if noted) Phelps Drake [email protected] Planchart Tony [email protected] Polera Madi [email protected] Reif David [email protected] Starnes Hannah [email protected] Taylor Krystal [email protected] East Carolina University Yoder Jeff [email protected] Northeastern University Alshawabkeh Akram [email protected] Ashrap Pahriya [email protected] University of Michigan Brown Phil [email protected] Campbell Kyle [email protected] University of Michigan Cardona Cordero Nancy [email protected] University of Puerto Rico Cathey Amber [email protected] University of Michigan Compton Patrick [email protected] Downey Autum [email protected] West Virginia University Eaton Jarrod [email protected] University of Michigan Elkin Elana [email protected] University of Michigan Gal·n Rivera Chrystal [email protected] Gao Griffith [email protected] Harris Sean [email protected] University of Michigan Hernandez-Otero Nobel [email protected] Hojabri Shayan [email protected] Kaeli David [email protected] University of Puerto Rico– Lafarga Previdi Irene [email protected] Medical Sciences Campus Loch-Caruso Rita [email protected] University of Michigan Magrath Anne [email protected] McGrath Anne [email protected] Padilla Ingrid [email protected] University of Puerto Rico Riddell Jill [email protected] West Virginia University Santos Rivera Amailie [email protected] Sarrouf Stephanie [email protected] Sheahan Thomas [email protected] Slowinski Lura [email protected] Torres Zayas Héctor [email protected] Velez Vega Carmen [email protected] University of Puerto Rico Vesper Dorothy [email protected] West Virginia Univ Watkins Deborah [email protected] University of Michigan Xi Chuanwu [email protected] Zhao Yuwei [email protected] Zlatan Feric [email protected]

NIEHS SRP 2020 Virtual Annual Meeting Program • 303 Attendees

Last Name First Name Email Other (if noted) Oregon State University Adams Kaley [email protected] Anderson Kim [email protected] Barton Michael [email protected] Bonner Emily [email protected] Buermeyer Andrew [email protected] Caballero Clarisa [email protected] Colvin Victoria [email protected] Dasgupta Subham [email protected] Pacific Northwest Gaither Kari [email protected] National Laboratory Pacific Northwest Garcia Whitney [email protected] National Laboratory Ghetu Christine [email protected] Gosline Sara [email protected] Harper Stacey [email protected] Hoffman Peter [email protected] Huizenga Juliana [email protected] Kile Molly [email protected] Maier Monica [email protected] Marcus Craig [email protected] Morales Juan [email protected] Moran Ian [email protected] Rericha Yvonne [email protected] Rivera Brianna [email protected] Rohlman Diana [email protected] Schindler Jason [email protected] Scott Richard [email protected] Scotten Jessica [email protected] Semprini Lewis [email protected] Shankar Prarthana [email protected] Shepard Lisa [email protected] Smith Jordan [email protected] Tanguay Robyn [email protected] Tidwell Lane [email protected] Tilton Susan [email protected] Truong Lisa [email protected] Tucker Shawn [email protected] Waters Katrina [email protected] Wilson Lindsay [email protected] Texas A&M University Abbott Derek [email protected]

304 • NIEHS SRP 2020 Virtual Annual Meeting Program Attendees

Last Name First Name Email Other (if noted) Aly Noor [email protected] Baker Erin [email protected] Barteau Mark [email protected] Bera Gopal [email protected] Beykal Burcu [email protected] Bhandari Sharmila [email protected] Brinkmeyer-Langford Candice [email protected] Camargo Krisa [email protected] Chen Zunwei [email protected] Chiu Weihsueh [email protected] Clark Jessica [email protected] Cordova Alexandra [email protected] Dalaijamts Chimeddulam [email protected] Dodds James [email protected] Fawkes Leanne [email protected] Ford Lucie [email protected] Ganesh Hari [email protected] Grange Stacie [email protected] Han Arum [email protected] Hearon Sara [email protected] Horney Jennifer [email protected] University of Delaware Jang Suji [email protected] Johnson Natalie [email protected] Jun Mikyoung [email protected] Kaihatu James [email protected] Knap Anthony [email protected] Li Dongying [email protected] Lloyd Dillon [email protected] North Carolina State University Luo Yu-Syuan [email protected] Malecha Matthew [email protected] Mancini Michael [email protected] Baylor College of Medicine Mancini Maureen [email protected] Baylor College of Medicine McDonald Thomas [email protected] Mustapha Toriq [email protected] Moyer Haley [email protected] Newman Galen [email protected] Orr Asuka [email protected] Phillips Tim [email protected] Pistikopoulos Stratos [email protected] Rohde Arlean [email protected] Roman-Hubers Alina [email protected] Rusyn Ivan [email protected]

NIEHS SRP 2020 Virtual Annual Meeting Program • 305 Attendees

Last Name First Name Email Other (if noted) Safe Stephen [email protected] Safe Stephen [email protected] Sansom Garett [email protected] Stossi Fabio [email protected] Baylor College of Medicine Szafran Adam [email protected] Baylor College of Medicine Tamamis Phanourios [email protected] Tao Zhihan [email protected] Valdiviezo Alan [email protected] Wade Terry [email protected] Wang Meichen [email protected] Wright Fred [email protected] North Carolina State University Zhu Rui [email protected] University of Alabama–Birmingham Antony Veena [email protected] Agarwal Neelima [email protected] Athar Mohammad [email protected] Bae Sejong [email protected] Baskin Monica [email protected] Carter A. [email protected] Chen Nathan [email protected] Deshane Jessy [email protected] Kim Young-il [email protected] Larson-Casey Jennifer [email protected] Li Fu Jun [email protected] Lungu Claudiu [email protected] McMahan David [email protected] Mirov Sergey [email protected] Oh Jonghwa [email protected] Pillai Renjith [email protected] Shedd Jacob [email protected] Singh Pooja [email protected] Srivastava Ritesh [email protected] Stephens Crystal [email protected] Surolia Ranu [email protected] Thomas Vinoy [email protected] University of Arizona Babst-Kostecka Alicja [email protected] Bonsey Kendra [email protected] Brusseau Mark [email protected] Carini Paul [email protected] Chief Karletta [email protected] Ding Xinxin [email protected]

306 • NIEHS SRP 2020 Virtual Annual Meeting Program Attendees

Last Name First Name Email Other (if noted) Dodson Matthew [email protected] Estrella Rocio [email protected] Fontana Catherine [email protected] Kiela Pawel [email protected] Li Hui [email protected] Maier Raina [email protected] McKenna Anissa [email protected] Merchant Nirav [email protected] Moreno Denise [email protected] Murawska Kamila [email protected] Ooi Aikseng [email protected] Schmitt Harrison [email protected] Shakya Aryatara [email protected] Teres Kevin [email protected] Vicenti Anthony [email protected] Yung Michelle [email protected] Zaid Alghzawi Ma’in [email protected] Zhang Qing-yu [email protected] Zhang Donna [email protected] University of California–Berkeley Alvarez-Cohen Lisa [email protected] Bozack Anne [email protected] Cardenas Andres [email protected] Collender Phillip [email protected] Cuthbertson Amy [email protected] McHale Cliona [email protected] Morello-Frosch Rachel [email protected] Ocasio Daniel [email protected] Pace Clare [email protected] Rigutto Gabrielle [email protected] Sedlak David [email protected] Smith Martyn [email protected] Sun Mohan [email protected] University of California–Davis Atone Jogen [email protected] Black Gabrielle [email protected] Lawrence Livermore Buchholz Bruce [email protected] National Laboratory Chiamvimonvat Nipavan [email protected] Connolly Morgan [email protected] El-Moghazy Ahmed [email protected] Furlow David [email protected]

NIEHS SRP 2020 Virtual Annual Meeting Program • 307 Attendees

Last Name First Name Email Other (if noted) Gallegos Chantel [email protected] Gomes Aldrin [email protected] Haj Fawaz [email protected] Hammock Bruce [email protected] Hammond Charles [email protected] Hattaway Madison [email protected] Koike Shinichiro [email protected] Li Fangzhou [email protected] Li Dongyang [email protected] Li Wenting [email protected] Loge Frank [email protected] Matsumoto Naoki [email protected] McReynolds Cindy [email protected] Middleton Beth Rose [email protected] Morisseau Christophe [email protected] Qi Meng [email protected] Rocke David [email protected] Siegel Justin [email protected] Singh Nalin [email protected] Sirish Padmini [email protected] Solorzano Tamara [email protected] Sule Rasheed [email protected] Sun Yao-Hui [email protected] Sun Gang [email protected] Tachachartvanich Phum [email protected] Taha Ameer [email protected] Tancredi Daniel [email protected] Tang Peixin [email protected] Thai Phung [email protected] Wang Weicang [email protected] Wang Yuxin [email protected] Yang Jun [email protected] Young Thomas [email protected] University of California–San Diego Chen Shujuan [email protected] Feiock Michelle [email protected] Guo Joy [email protected] Jobe Timothy [email protected] University of Cologne Mennillo Elvira [email protected] Pezzoli Keith [email protected] Rio Jazmin [email protected] Schroeder Julian [email protected]

308 • NIEHS SRP 2020 Virtual Annual Meeting Program Attendees

Last Name First Name Email Other (if noted) Tukey Robert [email protected] Weber Andre [email protected] Yamamoto Gen [email protected] Yang Xiaojing [email protected] Zarabi Yasman [email protected] University of Iowa Andino Jessica [email protected] Ankrum James [email protected] Bako Christian [email protected] Bittle Maeve [email protected] Bullert Amanda [email protected] Doershuk Suzanne [email protected] Dong Qin [email protected] Duffel Michael [email protected] Ewald Jessica [email protected] Hornbuckle Keri [email protected] Jahnke Jacob [email protected] Janssen Brandi [email protected] Jones Michael [email protected] Keshei Moala [email protected] Klingelhutz Al [email protected] Lehmler Hans [email protected] Li Xueshu [email protected] Ludewig Gabriele [email protected] Mattes Tim [email protected] Paranjape Neha [email protected] Purdy David [email protected] Saktrakulkla Panithi [email protected] Simonsen Derek [email protected] Spak Scott [email protected] Wang Kai [email protected] Westra Brian [email protected] Zhang Duo [email protected] Zhang Chunyun [email protected] Zhang Qianjin (Marina) [email protected] University of Kentucky Bhattacharyya Dibakar [email protected] Deng Pan [email protected] Durham Jerika [email protected] Frazar Molly [email protected] Gutierrez Angela [email protected] Haynes Erin [email protected]

NIEHS SRP 2020 Virtual Annual Meeting Program • 309 Attendees

Last Name First Name Email Other (if noted) Hennig Bernhard [email protected] Hilt J. Zach [email protected] Ingram Britt [email protected] Klaus Victoria [email protected] Koempel Annie [email protected] Leniz Francisco [email protected] Li Ying [email protected] Mills Rollie [email protected] Moore Jennifer [email protected] Morris Andrew [email protected] Moseley Hunter [email protected] Mudd-Martin Gia [email protected] Pearson Kevin [email protected] Pennell Kelly [email protected] Powell Christian [email protected] Rice Brittany [email protected] Shah Rishabh [email protected] Tay Hong Cheng [email protected] University of Louisville Adhihetty Prasadi [email protected] Cave Matthew [email protected] Conklin Daniel [email protected] Dang Audrey [email protected] Fu Xiao-An [email protected] Gripshover Tyler [email protected] Halder Sujoy [email protected] Hart Joy [email protected] Heberle Lauren [email protected] Hein David [email protected] Lynch Jordan [email protected] Malovichko Marina [email protected] McClain Craig [email protected] McFall Samantha [email protected] McGraw Katlyn [email protected] Smith Ted [email protected] Srivastava Sanjay [email protected] Taylor Breandon [email protected] Toyoda Jennifer [email protected] Wahlang Banrida [email protected] Whitt Aaron [email protected] Winner Millie [email protected]

310 • NIEHS SRP 2020 Virtual Annual Meeting Program Attendees

Last Name First Name Email Other (if noted) Wise James [email protected] Wood Lindsey [email protected] Xie Zhenzhen [email protected] Young Jamie [email protected] University of New Mexico Southwest Research Adeky Sarah [email protected] & Information Center Anderson Tamara [email protected] Beene Daniel [email protected] Begay Jessica [email protected] Bolt Alicia [email protected] Brearley Adrian [email protected] Burchiel Scott [email protected] Campen Matt [email protected] Cerrato José [email protected] Cruz Nathan [email protected] Southwest Research Dan Rose [email protected] & Information Center Dashner-Titus Erica [email protected] De Pree Thomas [email protected] El Hayek Eliane [email protected] Cameron Agricultural Franklin Mae [email protected] Ad Hoc Committee Gonzales Melissa [email protected] Hudson Laurie [email protected] LaRosa-LoPresti Savannah [email protected] Lauer Fredine [email protected] Lewis Johnnye [email protected] Lin Yan [email protected] Luo Li [email protected] MacKenzie Debra [email protected] Medina Sebastian [email protected] Meza Isabel [email protected] Miller Curtis [email protected] Natoli Luna [email protected] Nozadi Sara [email protected] Roman Carolyn [email protected] Schilz Jodi [email protected] Shaikh Nabil [email protected] University of Illinois–Chicago Southwest Research Shuey Christopher [email protected] & Information Center

NIEHS SRP 2020 Virtual Annual Meeting Program • 311 Attendees

Last Name First Name Email Other (if noted) Southwest Research Swimmer Kyle [email protected] & Information Center Volk Lindsay [email protected] Wegele Ashley [email protected] Zychowski Katherine [email protected] University of North Carolina–Chapel Hill Armstrong Mikayla [email protected] Balogun Fatai [email protected] Bodnar Wanda [email protected] Bowers Christopher [email protected] Brown Eric [email protected] Coronell Orlando [email protected] Eaves Lauren [email protected] Fry Rebecca [email protected] George Andrew [email protected] Gleason Zamira [email protected] Gray Kathleen [email protected] Haine Dana [email protected] Lu Kun [email protected] Obeidy Chelsea [email protected] University of Oregon Pardo Manuel de Fernando [email protected] Villena Polizzotto Matthew [email protected] Rodgers Megan [email protected] Sethupathy Praveen [email protected] Cornell University Smeester Lisa [email protected] Smith Kriste [email protected] Styblo Miroslav [email protected] Venkatratnam Abhishek [email protected] Vickers Riley [email protected] Wu Jingyi [email protected] Cornell University Xenakis James [email protected] Yang Yifei [email protected] Yelton Sarah [email protected] Zou Fei [email protected] University of Rhode Island Agudelo Juliana [email protected] Blomberg Annelise [email protected] Harvard University (URI STEEP) Boving Tom [email protected] Dekker Harrison [email protected] Dunn Matt [email protected] Hrytsenko Yana [email protected]

312 • NIEHS SRP 2020 Virtual Annual Meeting Program Attendees

Last Name First Name Email Other (if noted) Lohmann Rainer [email protected] Lucht Wendy [email protected] Morales-McDevitt Maya [email protected] Neville Amber [email protected] Ryu Sangwoo [email protected] Schaider Laurel [email protected] Silent Spring Institute (URI STEEP) Shih Yu-Hsuan [email protected] Harvard University (STEEP) Slitt Angela [email protected] Snook Jarod [email protected] Swift Judith [email protected] Vinhateiro Nathan [email protected] Woodward Melissa [email protected] University of Washington Cole Toby [email protected] Faustman Elaine [email protected] Fung Samantha [email protected] Furlong Clement [email protected] Gallagher Evan [email protected] Garrick Jacqueline [email protected] Gawel James [email protected] Hayward Lisa [email protected] Marsillach Judit [email protected] Matsushita Megumi [email protected] Patridge Emily [email protected] Wang Hao [email protected] NIH/NIEHS Amoglebe Sara [email protected] Balshaw David [email protected] Bennett April [email protected] Bennett Michelle [email protected] Carlin Danielle [email protected] Drew Christie [email protected] Duncan Chris [email protected] Edwards Lisa [email protected] Greer Jenny [email protected] Heacock Michelle [email protected] Henry Heather [email protected] Holmgren Stephanie [email protected] Latoni Alfonso [email protected] Mastin James [email protected] Outen Sharmice [email protected] Suk William [email protected]

NIEHS SRP 2020 Virtual Annual Meeting Program • 313 Attendees

Last Name First Name Email Other (if noted) Thomas Laura [email protected] Trottier Brittany [email protected] Williams James [email protected] Woychik Rick [email protected] R25 Brisolara Kari [email protected] LSUHSC–NO Donald Mercante [email protected] LSUHSC–NO Raynor Peter [email protected] University of Minnesota RO1 University of Maryland– Damond Jada [email protected] Baltimore County Virginia Institute Prossner Kristen [email protected] of Marine Science Ranville james [email protected] Colorado School of Mines Virginia Institute Unger Michael [email protected] of Marine Science RO1/SBIR Campagna Shawn [email protected] University of Tennessee–Knoxville SBIR Ball Raymond [email protected] EnChem Engineering Inc. John Sanil [email protected] Lynntech Inc. Other Avakian Megan [email protected] MDB, Inc. Bakulski Kelly [email protected] University of Michigan Buschang Steve [email protected] Texas General Land Office Cordero Jose [email protected] University of Georgia Yurok Tribe Cosby Christine [email protected] Environmental Program Craft Elena [email protected] Environmental Defense Fund Eddington Susan [email protected] Ecocuriean Patlovich Scott [email protected] UTHealth–School of Public Health Rios Janelle [email protected] UTHealth–School of Public Health Rodriguez Natalie [email protected] MDB, Inc. Skalla Lesley [email protected] MDB, Inc Valasco Mali [email protected] MDB, Inc. Zygourakis Kyriacos [email protected] Rice University Texas Environmental Justice Advocacy Services (t.e.j.a.s.) Parras Ana [email protected] Parras Juan [email protected] Hildago Nalleli [email protected] Unaffiliated Baker Tracie [email protected] Wayne State University

314 • NIEHS SRP 2020 Virtual Annual Meeting Program Attendees

Last Name First Name Email Other (if noted) Baker Bridget [email protected] Wayne State University Bakulski Kelly [email protected] University of Michigan Blount-Pacheco Jessica [email protected] Wayne State University Briese Emily [email protected] Arizona State University Debarba Lucas [email protected] Wayne State University Ding Jiahui [email protected] Wayne State University Dittrich Timothy [email protected] Wayne State University Koshko Lisa [email protected] Wayne State University Maxwell Anthony [email protected] Wayne State University McNulty Reginald [email protected] University of California–Irvine University of Michigan– Meeker John [email protected] School of Public Health Miller Carol [email protected] Wayne State University Wayne State University– O’Leary Brendan [email protected] Engineering Papuga Shirley [email protected] Wayne State University Rudel Holly [email protected] Yale University Runge-Morris Melissa [email protected] Wayne State University Tummala Chandra mouli [email protected] Wayne State University Zimmerman Julie [email protected] Yale University US Environmental Protection Agency (EPA) Geller Andrew [email protected] Maurice Charles [email protected] Thomas Russell [email protected] Zaragoza Larry [email protected]

NIEHS SRP 2020 Virtual Annual Meeting Program • 315 NIEHS SRP 2020 Annual Meeting srp2020.org

superfund.tamu.edu

tamu.edu

www.niehs.nih.gov/research/supported/centers/srp/index.cfm