By Erin Madeen, Ph.D. candidate and Project 1 Trainee

Erin Madeen working at the Lawrence Livermore National Laboratory.
Erin Madeen working at the Lawrence Livermore National Laboratory.

Using new technology at Lawrence Livermore National Laboratory (LLNL), Oregon State University researchers are able to perform a controlled study of the human metabolism of environmental contaminate PAHs for the first time.

The Williams Laboratory has studied PAHs (polycyclic aromatic hydrocarbons) for over a decade, traditionally relying on animal and in vitro models of metabolism and toxicity. PAHs are produced by the burning of carbon-containing materials, for example forest fires, charcoal grilling, and engine combustion. After production, PAHs cling to foods such as vegetables, cereal grains, or smoked meats. Some of these compounds cause cancer at high doses in animal models.

As a graduate student in the Williams Lab, one of my projects is to relate PAH data to human health.  With our partners at LLNL, a sensitive tool known as an AMS (accelerator mass spectrometer) is used to detect very small doses of PAHs in urine or blood plasma.  We gave a model PAH called DBC [Dibenzo (def,p) chrysene] to human volunteers in doses less than what can be found in a charbroiled burger. This research has not been possible until now because of potential toxicity risks.  Traditional non-AMS methods need a larger dose of DBC which could pose too high of a risk to study participants.

With the support of LLNL staff and the OSU Superfund Research Program, I received a K.C. Donnelly Externship Supplement through the NIEHS Superfund Research Program.  This award supported my travel to LLNL for this project. My experience at LLNL greatly solidified my understanding of and appreciation for AMS. Maintaining and continuously developing unique instrumentation, such as AMS, requires a highly specialized, dedicated, and flexible team.  The environment of a national laboratory is different from that of university research.  Most notably this difference is in the concentration of specialists in a particular field and the team approach to problem solving. It was humbling to observe the amount of time, resources, and effort that the LLNL AMS staff dedicated to training and to progress on our DBC project. This externship allowed me to experience being part of the AMS team and to process my own samples, providing valuable insight that will help guide further work on our projects.

Accelerator Mass Spectrometry (AMS) is an instrument traditionally used for carbon dating. It has been modified to detect stable isotopes in biological samples. The AMS at LLNL is unique because it is able to use liquid samples.  The liquid biological samples are separated according to the changes the body makes to DBC, known as DBC metabolites.  The carbon isotope added to the DBC chemical structure was used to identify several different metabolites in human urine and plasma.  This project is ongoing as we continue to develop a profile of the human metabolism of DBC over time.

Related journal publications:

From OSU Superfund Research Program

From LLNL

 

Our Center is multi-investigator, multi-disciplinary and multi-institutional. In partnership with Pacific Northwest National Laboratories (PNNL), and other stakeholders and collaborators, we are developing new technologies to identify and quantitate known and novel polycyclic aromatic hydrocarbons (PAHs) found at many of the nation’s Superfund sites and assess the risk they pose for human health.

Women@Energy: Dr. Katrina Waters  Photo credit: energy.gov
Women@Energy: Dr. Katrina Waters
Photo credit: energy.gov

The research projects in our Center collect large amounts of molecular and chemical data. This data includes measuring PAH mixtures in environmental samples, determining toxicity of PAH mixtures, and the mechanism(s) of action for these toxic endpoints.

Our Biostatistics and Modeling Core, lead by Dr. Katrina Waters, greatly enhances our Center by providing expert statistical and bioinformatics data analysis support and software solutions for data management and interpretation.

Katrina Waters recently became the Deputy Director for the Biological Sciences Division at the Pacific Northwest National Lab (PNNL). Her expertise is in computational biology, and she works collaboratively with all of the research projects and co-authors with them.

This multidisciplinary training of toxicology students and fellows at OSU and PNNL is a unique strength of our program. Our SRP Trainees have benefited greatly from the PNNL partnership.  Students have gone to the lab in Richland, WA to be trained in Bioinformatics, Statistics and Study Design. More training workshops are being scheduled for this summer and fall.

Waters presented at SOT’s FutureTox II: In Vitro Data and In Silico Models for Predictive Toxicology on January 16, 2014. Her talk was entitled Computational Tools for Integration of High Throughout Screening (HTS) Data. She utilized examples from the collaboration with Robyn Tanguay and his zebrafish assay for toxicity testing (Project 3).

Susan Tilton
Susan Tilton works with Dr. Katrina Waters and the OSU SRP Biostatistics and Modeling Core Group

Dr. Susan Tilton, also from PNNL,  presented at FutureTox as well. The title of her presentation was ‘Pathway-based prediction of tumor outcome for environmental PAH mixtures’.  In this study, they developed a mechanism-based approach for prediction of tumor outcome after dermal exposure to PAHs and environmental PAH mixtures.  Their model was successfully utilized to distinguish early regulatory events during initiation linked to tumor outcome and shows the utility of short-term initiation studies in predicting the carcinogenic potential of PAHs and PAH mixtures.

“Dr. Waters and her group have proven to be of great value in not just the interpretation of extremely large and complicated data sets, but also in the “front-end” study design, which results in enrichment of the subsequent data obtained.”
Dr. David Williams, OSU SRP Center Director

This year the EPA Partners in Technical Assistance Program (PTAP) Pilot has launched the first project with a school located near the Black Butte Mine Superfund Site in rural Cottage Grove, Oregon.

“The overall objective of PTAP is to expand opportunities for cooperation between EPA and colleges, universities or nonprofits with the shared goal of assessing and addressing the unmet technical assistance needs of impacted communities. Through PTAP, colleges, universities, and nonprofit organizations cooperate with EPA and voluntarily commit to assist communities with their unaddressed technical assistance needs. At this time, PTAP is in the pilot phase, working with NIEHS Superfund Research Program grantees as PTAP pilot partners. Following this pilot phase, the intention is to expand this project so that any interested colleges, universities or nonprofits may also join the PTAP.”

OSU Superfund Research Program has begun a partnership with EPA through this Pilot to help them expand upon their community outreach capabilities surrounding the Black Butte site.

On December 18, 2013, we met with Laurie Briggs, the Principal of the London School, because she had a strong desire to give her students and their families’ science and environmental health knowledge. About 100 rural K – 8th grade students go to London school.

Our visit included getting to know one another, listening to the needs of the school, and a school tour. We were impressed with the beauty and organization. The school built and maintains a 1/4-acre organic garden, and has a trail to a river flowing behind the property.  72% of the students qualify for free/reduced lunch, and delicious healthy meals are cooked on site.

For this project, we plan to:

1) Maintain communication through monthly meetings, and share notes and project milestones on our web site. [Our next meeting is January 30th, 2014 at OSU.]

2) Address community and educational needs.

  • Create a hands-on, project-based integrated curriculum related to the science of the Superfund site and mercury contamination that can serve as a model for other rural, small schools.
  • Discuss ways to educate the students and community and expand and build a sustainable partnership.

3) Provide training opportunities for SRP Trainees wanting outreach experience.

4) Help students understand career opportunities in environmental and life sciences.

 

 

Project Team from left Diana Rohlman (OSU SRP CEC), Alanna Conley (EPA, Region 10), Dan Sudakin (OSU SRP RTC), Laura Briggs (London School Principle), Naomi Hirsch (SRP RTC OSU). Not pictured: Corey Fisher (OSU SRP CEC), Melissa Dreyfus (EPA Headquarters Superfund Community Involvement Program), Kira Lynch, (EPA Region 10, Science and Tech Liaison), and Richard Muza (Region 10 - Black Butte Mine, Project Manager)
The Project Team from left Diana Rohlman (OSU SRP CEC), Alanna Conley (EPA, Region 10), Dan Sudakin (OSU SRP RTC), Laura Briggs (London School Principal), Naomi Hirsch (OSU SRP RTC). Not pictured: Corey Fisher and Molly Kile (OSU SRP CEC), Melissa Dreyfus (EPA Headquarters Superfund Community Involvement Program), Kira Lynch, (EPA Region 10, Science and Tech Liaison), and Richard Muza (Region 10 – Black Butte Mine, Project Manager)

 

 

 

 

grilled meat

CORVALLIS, Ore. – Researchers at Oregon State University have discovered novel compounds produced by certain types of chemical reactions – such as those found in vehicle exhaust or grilling meat – that are hundreds of times more mutagenic than their parent compounds which are known carcinogens.

These compounds were not previously known to exist, and raise additional concerns about the health impacts of heavily-polluted urban air or dietary exposure. It’s not yet been determined in what level the compounds might be present, and no health standards now exist for them.

The findings were published in December in Environmental Science and Technology, a professional journal.

The compounds were identified in laboratory experiments that mimic the type of conditions which might be found from the combustion and exhaust in cars and trucks, or the grilling of meat over a flame.

“Some of the compounds that we’ve discovered are far more mutagenic than we previously understood, and may exist in the environment as a result of heavy air pollution from vehicles or some types of food preparation,” said Staci Simonich, a professor of chemistry and toxicology in the OSU College of Agricultural Sciences.

Dr. Staci Simonich, Project 5 Leader with the OSU Superfund Research Program
Dr. Staci Simonich, Project 5 Leader with the OSU Superfund Research Program

“We don’t know at this point what levels may be present, and will explore that in continued research,” she said.

The parent compounds involved in this research are polycyclic aromatic hydrocarbons, or PAHs, formed naturally as the result of almost any type of combustion, from a wood stove to an automobile engine, cigarette or a coal-fired power plant. Many PAHs, such as benzopyrene, are known to be carcinogenic, believed to be more of a health concern that has been appreciated in the past, and are the subject of extensive research at OSU and elsewhere around the world.

The PAHs can become even more of a problem when they chemically interact with nitrogen to become “nitrated,” or NPAHs, scientists say. The newly-discovered compounds are NPAHs that were unknown to this point.

This study found that the direct mutagenicity of the NPAHs with one nitrogen group can increase 6 to 432 times more than the parent compound. NPAHs based on two nitrogen groups can be 272 to 467 times more mutagenic. Mutagens are chemicals that can cause DNA damage in cells that in turn can cause cancer.

For technical reasons based on how the mutagenic assays are conducted, the researchers said these numbers may actually understate the increase in toxicity – it could be even higher.

These discoveries are an outgrowth of research on PAHs that was done by Simonich at the Beijing Summer Olympic Games in 2008, when extensive studies of urban air quality were conducted, in part, based on concerns about impacts on athletes and visitors to the games.

Beijing, like some other cities in Asia, has significant problems with air quality, and may be 10-50 times more polluted than some major urban areas in the U.S. with air concerns, such as the Los Angeles basin.

An agency of the World Health Organization announced last fall that it now considers outdoor air pollution, especially particulate matter, to be carcinogenic, and cause other health problems as well. PAHs are one of the types of pollutants found on particulate matter in air pollution that are of special concern.

Concerns about the heavy levels of air pollution from some Asian cities are sufficient that Simonich is doing monitoring on Oregon’s Mount Bachelor, a 9,065-foot mountain in the central Oregon Cascade Range. Researchers want to determine what levels of air pollution may be found there after traveling thousands of miles across the Pacific Ocean.

This work was supported by the National Institute of Environmental Health Sciences (NIEHS) and the National Science Foundation (NSF). It’s also an outgrowth of the Superfund Research Program at OSU, funded by the NIEHS, that focuses efforts on PAH pollution. Researchers from the OSU College of Science, the University of California-Riverside, Texas A&M University, and Peking University collaborated on the study.

[Credit: Oregon State University Press Release]

See video from KVAL news

Learn more about PAHs from the Superfund Research Program web site.

 

Using Integrated Problem-based Curriculum

Lisa Troy, an 8th grade science teacher at The Sage School in Foxboro, Massachusetts chose the NIEHS-funded Hydroville Curriculum as a way to give her students a real-world problem to solve, teach them collaboration and teamwork skills, and expand their understanding of “doing” science.  She was also very interested in environmental issues and once worked as an environmental consultant on EPA’s Superfund/RCRA Hotline.

Teacher Lisa Troy shares instructions for the team building activity (toxic popcorn).
Teacher Lisa Troy shares instructions for the team building activity (Toxic Popcorn). Photo credit: The Sage School

In the Hydroville Pesticide Scenario, students work in teams to examine and clean-up a large accidental spill of metam sodium near a river.  In this scenario students take on roles of an environmental chemist, environmental toxicologist, soil scientist, and mechanical engineer. It creates a valuable experience to learn about these careers and how they work together to solve problems.

I was very pleased with how involved my students were in their roles.  Since they were responsible for their own area of expertise, they took ownership of the skills and information that they learned.  The students also enjoyed fitting their solution into the constraints of a budget, as well as considering stakeholders’ varying viewpoints.  Their parents attended the presentations and took on roles as stakeholders when asking questions.  Their presence increased the feeling of a real town meeting, and it was fun to see the students dressed up! ~ Lisa Troy

Students divide up into expert groups of Soil Scientist, Environmental Toxicologist, Mechanical Engineer and Analytic Chemist.
Students do a number of background activities to learn about the science needed to solve the problem. Topics include reading labels, toxicity testing, analyzing pumps, soil texture and permeability, and decision analysis. Photo credit: The Sage School

Communicating with a Scientist

The students were learning about toxicity, LD50, and NOEL (No Observable Effect Level) through a seed germination lab.  Lisa Troy had read about Dr. Tanguay in the recent YALEe360 article, and she shared his research with the students. The students were very excited to speak to a “real” scientist who is engaged in meaningful work and making a difference.  A highlight for the students was when they Skyped with Dr. Robyn Tanguay.

Students were especially interested to learn how zebrafish are being used as models of human response to chemicals in research all over the world.  They shared a long list of questions with Dr. Tanguay in preparation for the Skype event.

The students were intrigued by the idea that, through research such as Dr. Tanguay’s, chemical manufacturers will know much more about the effects of individual chemicals and the possible synergistic effects of mixing chemicals. They were reassured to learn of the human treatment of the fish, as well.

Not only was Dr. Tanguay’s  interview incredibly valuable, it taught my students an important lesson about research: that you can contact scientists and experts in their fields and obtain information directly from the source.  Science is not just in a textbook. ~Lisa Troy

To increase career connections, Lisa Troy asked the parents, teachers, and administrators to identify any skills that were important to them in their work or life experience from a list she generated of all the skills the students learned or used during the course of Hydroville. They checked nearly all of the skills!

As the year progresses and we study other topics, I will continue to reinforce the concepts and skills the students acquired during Hydroville and know that they will be well prepared for the future. ~Lisa Troy

[This post was written in collaboration with Lisa Troy. We truly appreciate her sharing her experience with us. If you are an educator and want more information or have a story to share, please contact us.]