1. A PILOT STUDY OF BIOMONITORING AMBIENT GENOTOXIC EXPOSURE OF CHILDREN L. Anna, B. Schoket, E. Győrffy, E. Erdei, P. Rudnai National Institute of Environmental Health, József Fodor National Centre for Public Health, Budapest, Hungary L. Anna, B. Schoket, E. Győrffy, E. Erdei, P. Rudnai National Institute of Environmental Health, József Fodor National Centre for Public Health, Budapest, Hungary

2. AIM OF THE STUDY To assess genotoxic exposure of children to air pollution To explore the suitability of aromatic DNA adducts as a biomarker of genotoxic exposure of children To assess genotoxic exposure of children to air pollution To explore the suitability of aromatic DNA adducts as a biomarker of genotoxic exposure of children

3. EPIDEMIOLOGICAL BACKGROUND OF THE STUDY Investigation of the respiratory health of 1800 children of 8 to 10 years of age (Nov-Dec 1998) Six towns in Hungary, three of those with elevated air pollution Exposure and health questionnaires filled in by the parents Investigation of the respiratory health of 1800 children of 8 to 10 years of age (Nov-Dec 1998) Six towns in Hungary, three of those with elevated air pollution Exposure and health questionnaires filled in by the parents

4. STUDY POPULATION FOR THE BIOMONITORING STUDY Exposure categories Polluted townsControl townsETS & TrafficETS onlyTraffic onlyNo ETS & No traffic Exposure categories Polluted townsControl townsETS & TrafficETS onlyTraffic onlyNo ETS & No traffic N = 105

5. METHODOLOGY Isolation of DNA from frozen whole blood (ca. 1.5 ml) Detection of aromatic DNA adducts by 32 P-postlabelling Radioactivity measurement by electronic autoradiograph Statistical analyses by Fisher’s exact test and Mann-Whitney U test Isolation of DNA from frozen whole blood (ca. 1.5 ml) Detection of aromatic DNA adducts by 32 P-postlabelling Radioactivity measurement by electronic autoradiograph Statistical analyses by Fisher’s exact test and Mann-Whitney U test

6. STRUCTURE OF BENZO[A]PYRENE DIOL- EPOXIDE-DNA ADDUCT

7. TYPICAL TWO-DIMENSIONAL DNA ADDUCT MAPS FROM CHILDREN’S BLOOD a ed cb f Blank

8. PERCENTAGE OF BLOOD SAMPLES WITH DNA ADDUCT SPOTS IN THE POLLUTED AND CONTROL TOWNS

9. PERCENTAGE OF BLOOD SAMPLES WITH DNA ADDUCT SPOTS POLLUTED TOWNS CONTROL TOWNS

10. ODDS RATIOS FOR DNA ADDUCT FORMATION DUE TO ELEVATED AIR POLLUTION (Main adduct spot)

11. INDIVIDUAL VARIATION IN AROMATIC DNA ADDUCT LEVELS IN BLOOD SAMPLES FROM CHILDREN Values for the main adduct spot Polluted towns Control towns P=0.02 for the Means P=0.02 for the Means

12. AROMATIC DNA ADDUCT LEVELS IN BLOOD SAMPLES FROM CHILDREN Polluted towns Control towns P=0.002 * * * * Values for the main spot (Mean ± SD)

13. CONCLUSIONS DNA adducts were detectable from frozen whole blood Detection limit was as low as 0.02-0.07 adducts in 10 8 nucleotides Two characteristic distinct adduct spots were related to the exposure Statistically significant difference was observed in DNA damage between the highest and lowest exposure categories Aromatic DNA adduct was a sensitive biomarker of genotoxic exposure of children to air pollution DNA adducts were detectable from frozen whole blood Detection limit was as low as 0.02-0.07 adducts in 10 8 nucleotides Two characteristic distinct adduct spots were related to the exposure Statistically significant difference was observed in DNA damage between the highest and lowest exposure categories Aromatic DNA adduct was a sensitive biomarker of genotoxic exposure of children to air pollution