Sensitive Biomarkers to Detect Biochemical Changes Indicating Multiple Stresses Including Chemically Induced Stresses

EPA Grant Number: R825433C043
Subproject: this is subproject number 043 , established and managed by the Center Director under grant R825433
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).

Center: EERC - Center for Ecological Health Research (Cal Davis)
Center Director: Rolston, Dennis E.
Title: Sensitive Biomarkers to Detect Biochemical Changes Indicating Multiple Stresses Including Chemically Induced Stresses
Investigators: Matsumura, Fumio
Institution: University of California - Davis
EPA Project Officer: Packard, Benjamin H
Project Period: October 1, 1996 through September 30, 2000
RFA: Exploratory Environmental Research Centers (1992) RFA Text |  Recipients Lists
Research Category: Center for Ecological Health Research , Targeted Research


This project seeks to study the combined effects of environmental pollutants and other biotic and abiotic stressors on sensitive biochemical systems to determine whether some of those changes could be used as reliable biomarkers to assess the end results of multiple stresses.


The investigators have recently discovered that protein kinase C (PKC), particularly the epsilon isoform, is a very sensitive biomarker, responding to even very subtle effects of chemicals. There are two sets of data indicating that PKC is a good marker. In one study (just published, Kiribuchi et al., see attached), investigators were able to show that this protein kinase in young rice plants is altered by winds, drought, fungus infection, the lack of sunlight, and high temperature in addition to herbicides, fungicides and cupric ions. The response of this enzyme to fungus infection was particularly interesting in view of its relationship to the plant defense signal transduction mechanism. It has been well established that pathogenic fungi give rise to the elicitor response in which plants are stimulated phenols, quinones and ether defensive substances, however, it has not been known that PKC is involved in this pathway. In the second set of data, we found heptachlor epoxide at low concentrations (0.1 - 1M) causes downregulation of PKC epsilon without well-known isoforms such as a and ß in cultured Hepa 1 cells from mouse liver hepatoma; such an action of heptachlor epoxide has not been reported before. Heptachlor epoxide is a very stable pesticidal environmental pollutant and is known to be the most potent congener of technical chlordane. PKC enzymes are known to play many vital roles in most of the organisms so far studied. Our findings indicate that the affected organisms are relying on the PKCs to respond and probably in some cases counteract toxic stresses they are subjected to.

Investigators are now moving into the area of endocrine disruptors. They are developing an MCF-7 cell foci forming assay method. This cell line is human breast cancer cells in origin. It responds to DDT, dioxin, HCH and other know disruptors by increasing foci (i.e. pile-ups of transformed, preneoplastic cells). In addition, we have begun an investigation of the possible role of c-Neu tyrosine kinase on the expression of estrogenicity of chlorinated pesticides. They have found that this kinase is activated by all of the above pollutants at very low concentrations.

Expected Results:

Investigators will test the possibility of developing the above foci formation assay on MCF-7 cells as a standard estrogenic biomarker to screen many chemicals. To achieve that goal they must define the cultural conditions, methods of application, concentration dependency, choice of standard chemicals and the nature of the truly estrogenic endpoint. Also we plan to study the basic mechanism by which these chemicals affect MCF-7 cells to evoke an estrogenic response. The main question we raise is: Why do these chemicals, which are reportedly weak estrogen receptor agonists, seem to evoke an estrogenic response at much lower concentrations than anticipated?

Supplemental Keywords:

Watershed, bioindicator, biomarker, endocrine disruptors, multiple stresses, protein kinase, pesticides, runoff, ecosystem stress., RFA, Health, Scientific Discipline, INTERNATIONAL COOPERATION, Water, ECOSYSTEMS, Ecosystem Protection/Environmental Exposure & Risk, Aquatic Ecosystems & Estuarine Research, Water & Watershed, Environmental Chemistry, Ecosystem/Assessment/Indicators, Ecosystem Protection, Endocrine Disruptors - Environmental Exposure & Risk, Aquatic Ecosystem, endocrine disruptors, Monitoring/Modeling, Ecological Effects - Environmental Exposure & Risk, Biochemistry, Environmental Monitoring, Ecological Monitoring, Ecology and Ecosystems, Endocrine Disruptors - Human Health, Watersheds, Ecological Indicators, bioindicator, nutrient dynamics, watershed development, pesticide exposure, immunoassay, wetlands, ecosystem monitoring, watershed management, fish habitat, biomarkers, accelerator mass spectrometry, anthropogenic effects, agricultural watershed, protein kinase, endocrine disrupting chemicals, pesticides, runoff, aquatic habitat, Clear Lake, watershed land use, watershed modeling, ecological assessment, hydrology, integrated watershed model, aquatic ecosystems, environmental stress, lake ecosysyems, water quality, watershed sustainablility, ecological risk, ecosystem health, ecosystem stress, agrochemicals, ecology assessment models, environmental stress indicators, water management options, ecosystem response, land use

Progress and Final Reports:

  • 1997
  • 1998
  • 1999 Progress Report
  • Final Report

  • Main Center Abstract and Reports:

    R825433    EERC - Center for Ecological Health Research (Cal Davis)

    Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
    R825433C001 Potential for Long-Term Degradation of Wetland Water Quality Due to Natural Discharge of Polluted Groundwater
    R825433C002 Sacramento River Watershed
    R825433C003 Endocrine Disruption in Fish and Birds
    R825433C004 Biomarkers of Exposure and Deleterious Effect: A Laboratory and Field Investigation
    R825433C005 Fish Developmental Toxicity/Recruitment
    R825433C006 Resolving Multiple Stressors by Biochemical Indicator Patterns and their Linkages to Adverse Effects on Benthic Invertebrate Patterns
    R825433C007 Environmental Chemistry of Bioavailability in Sediments and Water Column
    R825433C008 Reproduction of Birds and mammals in a terrestrial-aquatic interface
    R825433C009 Modeling Ecosystems Under Combined Stress
    R825433C010 Mercury Uptake by Fish
    R825433C011 Clear Lake Watershed
    R825433C012 The Role of Fishes as Transporters of Mercury
    R825433C013 Wetlands Restoration
    R825433C014 Wildlife Bioaccumulation and Effects
    R825433C015 Microbiology of Mercury Methylation in Sediments
    R825433C016 Hg and Fe Biogeochemistry
    R825433C017 Water Motions and Material Transport
    R825433C018 Economic Impacts of Multiple Stresses
    R825433C019 The History of Anthropogenic Effects
    R825433C020 Wetland Restoration
    R825433C021 Sierra Nevada Watershed Project
    R825433C022 Regional Transport of Air Pollutants and Exposure of Sierra Nevada Forests to Ozone
    R825433C023 Biomarkers of Ozone Damage to Sierra Nevada Vegetation
    R825433C024 Effects of Air Pollution on Water Quality: Emission of MTBE and Other Pollutants From Motorized Watercraft
    R825433C025 Regional Movement of Toxics
    R825433C026 Effect of Photochemical Reactions in Fog Drops and Aerosol Particles on the Fate of Atmospheric Chemicals in the Central Valley
    R825433C027 Source Load Modeling for Sediment in Mountainous Watersheds
    R825433C028 Stress of Increased Sediment Loading on Lake and Stream Function
    R825433C029 Watershed Response to Natural and Anthropogenic Stress: Lake Tahoe Nutrient Budget
    R825433C030 Mercury Distribution and Cycling in Sierra Nevada Waterbodies
    R825433C031 Pre-contact Forest Structure
    R825433C032 Identification and distribution of pest complexes in relation to late seral/old growth forest structure in the Lake Tahoe watershed
    R825433C033 Subalpine Marsh Plant Communities as Early Indicators of Ecosystem Stress
    R825433C034 Regional Hydrogeology and Contaminant Transport in a Sierra Nevada Ecosystem
    R825433C035 Border Rivers Watershed
    R825433C036 Toxicity Studies
    R825433C037 Watershed Assessment
    R825433C038 Microbiological Processes in Sediments
    R825433C039 Analytical and Biomarkers Core
    R825433C040 Organic Analysis
    R825433C041 Inorganic Analysis
    R825433C042 Immunoassay and Serum Markers
    R825433C043 Sensitive Biomarkers to Detect Biochemical Changes Indicating Multiple Stresses Including Chemically Induced Stresses
    R825433C044 Molecular, Cellular and Animal Biomarkers of Exposure and Effect
    R825433C045 Microbial Community Assays
    R825433C046 Cumulative and Integrative Biochemical Indicators
    R825433C047 Mercury and Iron Biogeochemistry
    R825433C048 Transport and Fate Core
    R825433C049 Role of Hydrogeologic Processes in Alpine Ecosystem Health
    R825433C050 Regional Hydrologic Modeling With Emphasis on Watershed-Scale Environmental Stresses
    R825433C051 Development of Pollutant Fate and Transport Models for Use in Terrestrial Ecosystem Exposure Assessment
    R825433C052 Pesticide Transport in Subsurface and Surface Water Systems
    R825433C053 Currents in Clear Lake
    R825433C054 Data Integration and Decision Support Core
    R825433C055 Spatial Patterns and Biodiversity
    R825433C056 Modeling Transport in Aquatic Systems
    R825433C057 Spatial and Temporal Trends in Water Quality
    R825433C058 Time Series Analysis and Modeling Ecological Risk
    R825433C059 WWW/Outreach
    R825433C060 Economic Effects of Multiple Stresses
    R825433C061 Effects of Nutrients on Algal Growth
    R825433C062 Nutrient Loading
    R825433C063 Subalpine Wetlands as Early Indicators of Ecosystem Stress
    R825433C064 Chlorinated Hydrocarbons
    R825433C065 Sierra Ozone Studies
    R825433C066 Assessment of Multiple Stresses on Soil Microbial Communities
    R825433C067 Terrestrial - Agriculture
    R825433C069 Molecular Epidemiology Core
    R825433C070 Serum Markers of Environmental Stress
    R825433C071 Development of Sensitive Biomarkers Based on Chemically Induced Changes in Expressions of Oncogenes
    R825433C072 Molecular Monitoring of Microbial Populations
    R825433C073 Aquatic - Rivers and Estuaries
    R825433C074 Border Rivers - Toxicity Studies