Grantee Research Project Results

2004 Progress Report: Oxidation Stress Makers

EPA Grant Number: R827355C011
Subproject: this is subproject number 011 , established and managed by the Center Director under grant R827355
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: Center for Air, Climate, and Energy Solutions
Center Director: Robinson, Allen
Title: Oxidation Stress Makers
Investigators: Simpson, Chris , Kavanagh, Terrance J , Luchtel, Daniel L.
Institution: University of Washington
EPA Project Officer: Chung, Serena
Project Period: June 1, 1999 through May 31, 2004 (Extended to May 31, 2006)
Project Period Covered by this Report: June 1, 2003 through May 31, 2004
RFA: Airborne Particulate Matter (PM) Centers (1999) RFA Text | Recipients Lists
Research Category: Air Quality and Air Toxics , Particulate Matter , Air

Objective:

The objective of this research project is to assess oxidative stress in vivo in humans and animals exposed to diesel exhaust through the measurement of 3-nitrotyrosine and F₂-isoprostanes in biological samples.

Progress Summary:

The oxidative stress markers project is a new pilot-scale project that was established in Year 6 of the PM Center. The start of this project was delayed because of several factors described in the Future Activities section.

Future Activities:

The following work is to be completed during the no-cost extension period:

The project is relying on mouse tissue obtained from Glutamate-Cysteine Ligase Modifier Subunit (GCLm) knockout mice exposed to diesel exhaust in our exposure facility. Tissue from the mouse exposures is not yet available, as this project encountered unanticipated delays due to modification of the animal exposure protocols.
Synthesis of the ¹³C-labeled compounds for the nitrotyrosine assay proved more challenging than we had anticipated. We had to develop our own synthetic protocols as the yields from the literature methods were unacceptably low. The new protocols we developed proved to be successful, and we now have prepared substantial quantities of ¹³C₆-nitrotyrosine, ¹³C₆-3-nitro-4-hydroxyphenylacetate (NHPA), and ¹³C₈-4-hydroxyphenylacetate. These standards are an invaluable resource that will facilitate the accurate, artifact-free determination of nitrotyrosine in biological samples.
A new liquid chromatography/mass spectrometry (LC/MS) system (Agilent XCT Trap) was purchased to be used in part for the measurement of 3-nitrotyrosine and F₂-isoprostanes in biological samples. The purchase and installation of this instrument was delayed until January 2005 because of constraints placed by the University of Washington on the purchasing process for this instrument. Although we expect an initial learning curve as our analytical methods are adapted to the new instrument, the sensitivity and reliability of the new instrument are far superior to the LC/MS system we had been using previously, and the new instrument will greatly enhance the quality of the data generated in the current project.

As of April 1, 2005, we have acquired isotopically labeled standards for the F2-isoprostane assay, and we have completed synthesis, purification, and evaluation of the isotopically labeled standards for the nitrotyrosine assay. Optimization of the Agilent XCT Trap for analysis of nitrotyrsoine has been completed, and validation of our sample preparation protocols for measurement of free nitrotyrosine in urine and plasma, and protein-bound nitrotyrosine in plasma is complete. Blood and urine samples have been collected and archived from human volunteers exposed to diesel exhaust, and exposure of GCLm knockout mice to diesel exhaust is about to commence. We expect to complete the analysis of 3-nitrotyrosine and F2-isoprostanes in these archived samples, compile and interpret the results, and prepare manuscripts for publication.

Supplemental Keywords:

ambient particles, fine particles, combustion, health, exposure, biostatistics, susceptibility, human susceptibility, sensitive populations, air toxics, genetic susceptibility, indoor air, indoor air quality, indoor environment, tropospheric ozone, California, CA, polyaromatic hydrocarbons, PAHs, hydrocarbons, acute cardiovascular effects, aerosols, air pollutants, air pollution, air quality, airborne pollutants, airway disease, airway inflammation, allergen, ambient aerosol, ambient aerosol particles, ambient air, ambient air quality, ambient particle health effects, animal model, assessment of exposure, asthma, atmospheric aerosols, atmospheric chemistry, biological markers, biological response, cardiopulmonary response, cardiovascular disease, children, children’s vulnerability, combustion, combustion contaminants, combustion emissions, epidemiology, exposure, exposure and effects, exposure assessment, harmful environmental agents, hazardous air pollutants, health effects, health risks, human exposure, human health effects, human health risk, incineration, inhalation, lead, morbidity, mortality, mortality studies, particle exposure, particle transport, particulates, particulate matter, PM, risk assessment,, RFA, Health, Scientific Discipline, Air, particulate matter, Environmental Chemistry, Health Risk Assessment, air toxics, Risk Assessments, Biochemistry, automobile exhaust, air pollution, pariculate matter, particle exposure, diesel exhaust, human exposure, PM, airborne urban contaminants, human health risk

Relevant Websites:

http://depts.washington.edu/pmcenter/ Exit

Progress and Final Reports:

Original Abstract

Main Center Abstract and Reports:

R827355 Center for Air, Climate, and Energy Solutions

Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R827355C001 Epidemiologic Study of Particulate Matter and Cardiopulmonary Mortality
R827355C002 Health Effects
R827355C003 Personal PM Exposure Assessment
R827355C004 Characterization of Fine Particulate Matter
R827355C005 Mechanisms of Toxicity of Particulate Matter Using Transgenic Mouse Strains
R827355C006 Toxicology Project -- Controlled Exposure Facility
R827355C007 Health Effects Research Core
R827355C008 Exposure Core
R827355C009 Statistics and Data Core
R827355C010 Biomarker Core
R827355C011 Oxidation Stress Makers

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The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.