Home     Overview    Project tasks     Partners     Results     Jobs     Events      Links    Publications

Methodology

In order to identify substances as endocrine disrupters with possible pro- and anti-estrogenic, - antiandrogenic, -progestational,-glucocorticoid or thyroid hormone action the methods used by the consortium are:

- genomic and proteomic array assays

- recombinant cell lines with reporter genes

- cell lines with defined receptor populations and biological endpoints

For determination of acute and chronic effects in various organs of developing and steroid-deprived rats or mice, measurements of gene and protein expressions are made. ED concentrations in body extracts, organs and the blood of experimental animals and humans are performed.

Choice of substances

ED's, that will be studied due to their high production volume and exposure to human are:

substances that pollute the environment and are taken up through air, dermal contact or food chain:

- 4-nonylphenol

- dibutylphtalate

- bisphenol A

substances that are administered but unwillingly incorporated (cosmetics and UV absorbers):

- 4-methylbenzylidene camphor (4MBC)

- octylmethoxycinnamate (OMC)

- Benzophenone D

substances, that are deliberately and willingly administered to the human (through beer and wine):

- Resveratrol

- 8-Prenylnaringenin

- Silybinin

- a synthetic flavonoid for comparison with plant derived products

The effects of these ED's will first be tested in gene-targeted cell systems (reporter gene transfected immortalised cells and organ specific reporter cells). Those substances, which address other than ER or AR signals will then be subjected to animal experiments.

Workpackages

WP1     Programming, configuration and maintenance of a web-site, a data base and a tissue bank

WP2     Cell biology with gene transfected or organ specific reporter cells

WP3     Effects of ED's in human cells of various origin

WP4     In vitro  studies with hypothalamic fragments: effects on neurotransmitters and -peptides

WP5     Effects of ED's in developing rats

WP6     Effects of ED's in gonadectomized male and female rats

WP7     Influence of ED's in adrenalectomized male and female rats

WP8     Effects of ED's in thyroidectomized male and female rats

WP9     Effects of topical application of ED's in hairless rats

WP10   Experiments in ERa and ERß knock-out mice

WP11   Experiments in TRa and TRß knock-out mice

WP12   Experiments with AR knock-out mice

WP13   Experiments with reporter gene mice

WP14   Morphological analysis of gene and protein expression in cells and tissue of animal and human origin

WP15   Clinical studies with ED's produced for human application or consumption

WP16   Endocrine disrupter cluster activities

Expected outcomes

• Risk assessment through in vitro  effects of ED's in cell experiments with special reference to the steroid receptor type involved in these effects and whether these effects may be organ specific

• Risk assessment through in vivo effects of ED's in animal experiments with special reference to multi-organic effects and the resulting risk assessments thereof

• Risk assessment through in vivo effects in the human of ED produced for human use and the resulting risk assessments thereof

• Establishment of a focus for European Endocrine disrupter research