? VLIV ZNEČIŠTĚNÉHO OVZDUŠÍ NA ZDRAVOTNÍ STAV POPULACE Radim J. Šrám Ústav experimentální mediciny AV ČR Praha ARNIKA, Praha, 27. 11. 2008
VÝVOJ ZNEČIŠTĚNÍ OVZDUŠÍ V ČR
Air particles deposition in the airways > 10 mm < 10 mm (PM10) < 2.5 mm (PM2.5)
Prague Castle Old Town Square PRAGUE Prague-Smichov Prague- Libus STATIONARY MONITORING
7 6 Mont hly average concent rat ion of B[a]P Smichov Libus B[a]P ng/ m 3 5 4 3 2 1 0 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7 8 9 1011 12 1 2 3 4 5 6 7 8 9 10 1112 2004-2006
Roční průměry PM2.5 Praha - Smíchov a Libuš 2001-2006 32 30 28 Smíchov Libuš µg/m 3 26 24 22 20 18 16 2001 2002 2003 2004 2005 2006 rok
B[a]P - 2006 Concentrations [ng/m 3 ] 0.4 LAT > 0.4-0.6 (LAT, UAT > > 0.6-1.0 (UAT, LV > > 1.2-2.0 (LV,2 > > 2.0 > 2 39.7% 35.4% 15.9% 6.6% 2.4% R.J.Sram 2008
KONCENTRACE B[a]P 7 Ostrava-Poruba Praha-Smíchov 6 5 koncentrace [ng/m 3 ] 4 3 2 1 2.8 0 1 2 3 4 5 6 7 8 9 10 11 12 1 2 3 4 5 6 7 8 9 10 11 12 R. J. Sram 2007 2004 2005
BIOMARKERS
LIDSKÉ BIOMARKERY Biomonitorování Biomarkery Biomarkery expozice účinku Pre-maligní Diagnóza rakoviny Časná Pozdní léze 100% Prevence Léčba doba (roky) Ú m r t í R. J. Šrám 2006 R.J. Albertini 1998
E X P O S U R E HUMAN BIOMARKERS Genetic/Carcinogenic Risks Internal Dose Metabolic Biologically Effective Dose Genotype/Fenotype Early Genotoxic Effects DNA Repair Capacity BIOMARKERS OF SUSCEPTIBILITY Altered Structure Function Immune Functions C A N C E R Environ Health Perspect 74: 3-9, 1987
PREGNANCY OUTCOME EXPOSURE PAHs Cotinine DNA adducts HPRT BIRTH SMOKING ETS Internal dose Biologically Effective Dose Early Genotoxic Effects WEIGHT Metabolic Genotype/Phenotype DNA Repair Capacity
Autoradiographs of thin layer chromatograms with DNA adduct pattern of: DNA isolated from lymphocytes of subject sampled in January 2004 (1st sampling period) Water blank Positive control (DNA isolated from the lung of rats intraperitoneally treated with 100 mgb[a]p/kg b.w.)
CYTOGENETIC ANALYSIS FISH analysis t(ab);t(ab);t(ba) Three translocations between chromosome 1 and unpainted chromosomes
MICRONUCLEI
Reactive oxygen species Endogenous sources Exogenous sources O 2 - SOD H 2 O 2 Catalase GPx H 2 O + O 2 OH H 2 O Lipid peroxidation DNA oxidation Protein oxidation
Pathway-1 Pathway-2 Driven by: surface reactivity organic compounds OC-L PM OC-H PM Surface Reactivity ROS Oxidative Effect ROS NFkB API Nrf2 ROS Phase I (Cyp1a1 ) Bioactivation Biotransformation Phase II (Gst, Ngo1 ) Phase III Conjugation Elimination Proinflammatory response
BIOMARKERS AIR POLLUTION
Human studies and biomarkers of exposure, effect and susceptibility PM2.5 Stationary monitoring DNA adducts by 32 P-postlabeling c-pahs Personal monitoring Stationary monitoring VOC Personal monitoring Stationary monitoring Cotinine Chromosomal aberrations conventional, FISH, micronuclei Oxidative damage 8-oxodG, 15-F2T-isoP, proteins, SCGE Triglycerids, Total, HDL and LDL cholesterol Genetic polymorphisms Vitamins A, C. E, folic acid Gene expression
c-pahs CONCENTRATIONS PERSONAL MONITORING (median and range) Group N Age (years) B[a]P ng/m 3 carcpau ng/m 3 EXPOSED 53 31.6 ± 7.2 1.6 (0.3-8.7) 9.7 (3.1-58.2) Smokers 19 32.9 ± 7.0 1.6 (0.3-7.5) 10.8 (3.1-43.6) Nonsmokers 34 30.9 ± 7.3 1.5 (0.3-8.7) 8.7 (3.1-58.2) CONTROLS 52 29.6 ± 9.1 0.8 (0.3-2.8) 5.8 (3.1-19.3) Smokers 7 37.6 ± 14.2 0.3 (0.3-1.4) 3.3 (3.1-8.2) Nonsmokers 45 28.3 ± 7.6 0.9 (0.3-2.8) 6.1 (3.1-19.3)
DNA Adducts/10 8 Nucleotides 0.30 0.25 0.20 0.15 0.10 0.05 LIKE B[a]P-DNA ADDUCT "Like" B[a]P-DNA Adduct CON (N=52) EXP Cases (N=53) 0.00 All SM NS All SM NS EXP vs CON: P<0.01 Smokers vs Nonsmokers within groups: P<0.01
IMPACT OF ENVIRONMENTAL POLUTION TO DNA ADDUCTS 6 DNA adducts = 1,042 + benzo[a]pyren * 0,077 (p=0,0003) 5 DNA adducts [10 8 nucleotides] 4 3 2 1 0 0 1 2 3 4 5 6 7 8 benzo[a]pyren [ugm -3 ]
CYTOGENETIC ANALYSIS FISH 6.0 5.0 4.0 * P < 0.05 ** P < 0.05 EXP ** ** CON * 3.0 2.0 1.0 0 * * * F G /100 AB.C. NCJ B/1000 t rcp
IMPACT OF ENVIRONMENTAL POLUTION TO CHROMOSOMAL ABERRATIONS - FISH
SCGE repair capacities of DNA damage Policemen Controls 120 Repaired DNA damage [%] 110 100 90 80 p<0.005 p<0.04 70 whole group nonsmokers smokers
8-oxodG in urine of bus drivers and controls Urinary 8-oxodG levels in winter 2005, summer 2006 and winter 2006 14.00 12.00 p<0.01 p=0.05 p<0.001 8-oxodG (nmol/mmol creat.) 10.00 8.00 6.00 4.00 2.00 0.00 BUS1 CON1 BUS2 CON2 BUS3 CON3
Correlation between oxidative stress markers and other factors 15-F 2t -IsoP 8-oxodG carbonyl B[a]P cpahs PM2.5 PM10 15-F 2t -IsoP R P - - 0.212 <0.001 0.083 0.165 0.172 p<0.01 0.180 p<0.01 0.168 p<0.01 0.116 p<0.05 8-oxodG R P 0.212 <0.001 - - 0.057 0.338 0.009 0.874 0.009 0.874 0.212 <0.001 0.307 <0.001 carbonyl R P 0.083 0.165 0.057 0.338 - - -0.371 <0.001-0.379 <0.001-0.404 <0.001-0.030 0.613
R. J. Sram 2007 VLIV OVZDUŠÍ NA RESPIRAČNÍ ONEMOCNĚNÍ U DĚTÍ
Bronchitidy RR, 95% CI s, akutní expozice PAU, multivariátní analýza 2.00 1.80 0-24 months 2-4.5 years 1.60 1.40 1.20 1.00 0.80 Single day no lag 1 day lag 2 day lag 3 day lag 4 day lag 5 day lag 6 day lag 3 day prior average 7 day prior average Single day no lag 1 day lag 2 day lag 3 day lag 4 day lag 5 day lag 6 day lag 3 day prior average 7 day prior average
Bronchitidy RR, 95% CI s, akutní expozice PM2.5, multivariátní analýza 2.00 1.80 0-24 months 2-4.5 years 1.60 1.40 1.20 1.00 0.80 Single day no lag 1 day lag 2 day lag 3 day lag 4 day lag 5 day lag 6 day lag 3 day prior average 7 day prior average Single day no lag 1 day lag 2 day lag 3 day lag 4 day lag 5 day lag 6 day lag 3 day prior average 7 day prior average
Význam první léta života dítěte jsou kritickou periodou dýchacího ústrojí a imunity
VLIV OVZDUŠÍ NA 8-oxodG V MOČI DĚTÍ
Vztah 8-oxodG a kotininu 50 8-oxodG (nmol/mmol kreat.) 40 30 20 10 0 1 10 100 1000 kotinin (µg/mmol kreat.)
50 40 30 20 10 0 Vztah 8-oxodG a věku dítěte 5 6 7 8 9 10 11 Věk 8-oxodG (nmol/mmol kreat.)
Multivariate model of effect of PM2.5 on 8-oxodG levels (linear regression) Region All Pollutant Intercept PM2.5 Child age Cotinine (above/below 20 ng/mg) Allergic rhinitis Period (days before sampling 3-day (-4;-6) 0.16-2.74 3.20 1.34 33.56 <0.05 <0.0001 0.001 0.41 7-day (-1;-7) R p R p 0.16-2.74 3.23 1.28 33.94 0.08 <0.001 0.005 <0.005 Teplice Intercept PM2.5 Child age Cotinine (above/below 20 ng/mg) Allergic rhinitis 28.57 0.24-2.51 4.41 7.59 <0.01 <0.001 <0.005 0.01 29.81 0.23-2.56 4.321 7.64 <0.05 <0.001 0.005 0.01 Prachatice Intercept PM2.5 Child age Allergic rhinitis 43.71-0.13-3.07-3.03 0.30 <0.001 0.06 45.22-0.20-3.09-3.02 0.21 <0.001 0.06
Multivariate model of effect of B[a]P on 8-oxodG levels (linear regression) Region All Pollutant Intercept B[a]P Child age Cotinine (above/below 20 ng/mg) Allergic rhinitis Period (days before sampling 3-day (-4;-6) 34.01-1.44-2.65 3.04 1.19 0.03 <0. 001 <0.005 0.47 3-day (-1;-7) R p R p 34.94 1.20-2.74 3.49 1.27 0.08 <0.001 <0.01 0.55 Teplice Intercept B[a]P Child age Cotinine (above/below 20 ng/mg) Allergic rhinitis 31.09 1.73-2.54 4.42 7.76 <0.05 <0.001 <0.005 <0.01 31.30 2.13-2.71 4.58 8.46 <0.05 <0.001 0.01 <0.05 Prachatice Intercept B[a]P Child age Allergic rhinitis 40.25-0.02-3.05-1.62 0.98 <0.001 0.22 42.18-0.99-2.97-4.18 0.36 <0.001 0.05
8-oxodG V MOČI DĚTÍ zvýšen při expozici pasivnímu kouření snižuje se s věkem dětí je ovlivněn etnicitou a vzděláním matky zvýšen expozicí PM2.5 a PM10 7 dní před odběrem zvýšen expozicí k-pau 1-3 a 7-9 dní před odběrem zvýšen u dětí s alergickou rhinitidou 13/14
VLIV OVZDUŠÍ NA VÝSLEDKY TĚHOTENSTVÍ
IUGR by PM10 during 1994-1998 in TEPLICE 3,0 Low = 1 Med; NS High; NS 3,0 2,5 Med; P<0.04 High; P<0.0005 2,5 AOR with 95% CL 2,0 1,5 1.41 2.10 AOR 2,0 1,5 AOR with 95%CL 1,0 1,0 0,5 0,5 0,0 1st 2nd 3rd 4th 5th 6th 7th 8th 9th Months of gestation 0,0
CARCINOGENIC PAHs & IUGR IN TEPLICE 4.0 4.0 3.5 Low CI Medium ; NS 3.5 CI High ; NS 3.0 CI Medium ; P<0.05 3.0 AORs with 95% CI 2.5 2.0 1.5 1.0 2.16 1.67 CI High ; P<0.005 AORs 2.5 2.0 1.5 1.0 AORs with 95% CI 0.5 0.5 0.0 1st 2nd 3rd 4th 5th 6th 7th 8th 9th Months of gestation 0.0 R. J. Sram 2007
OVLIVNĚNÍ VÝVOJE DĚTÍ VLIV FAKTORŮ ŽP A ŽIVOTNÍHO STYLU: porodní hmotnost genotyp nemocnost dýchací funkce neuropsychické funkce R. J. Sram 2007
AIR POLLUTION IMPACT ON SPERM
100 SCSA - Acridine Orange Stained DNA Native DNA Stainability (green fluorescence) 0 100 Fragmented DNA (red fluorescence)
Air pollutions Prague 2007 (ng/m 3 ) 12.0 10.0 8.0 February May 6.0 4.0 2.0 0 PAH cpah B[a]P eqb[a]p
Sperm analysis February May Morphology (%) Vitality (%) Motility (%) Concentration (mil/ml) Volume (ml) 180 160 140 120 100 80 60 40 20 0
Policemen patrolling the streets in Prague centre with heavy traffic The level of air pollution will be assessed on the basis of information from two source: -data from stationary measuring stations AIM Prague -for 48 h using personal sampling devices (URG Corp, USA) det DFI (%) 25 20 15 10 5 February May high DFI (%) 16 12 8 4 February HDS (%) 20 15 10 5 May February P 0.001 P 0.001 P 0.001 May N=46 ddfi < 15% Feb 30 May 42 ddfi 15 30% Feb 16 May 4 ddfi >30% Feb 2 May 2 HDS >15% Feb 10 May 4
AIR POLLUTION RISK ASSESSMENT
ACCORDING TO MOLECULAR EPIDEMIOLOGY STUDIES concentrations > 1 ng B[a]P/m 3 in polluted air RISK FOR HUMAN HEALTH
NOVÉ POZNATKY VLIV ENVIRONMENTÁLNÍ EXPOZICE k-pau RIZIKO GENOTOXICITY VZTAH MEZI EXPOZICÍ k-pau, DNA adukty a chromosomovými aberacemi (FISH), snížením DNA reparace VÝZNAM OXIDAČNÍHO POŠKOZENÍ PRO NEMOCNOST DĚTÍ
VÝZNAM OXIDAČNÍHO POŠKOZENÍ U DĚTÍ zvyšuje výskyt respiračních onemocnění (bronchitidy, astma) ovlivňuje imunitu U DOSPĚLÝCH zvyšuje výskyt nádorových onemocnění aterosklerosy diabetes URYCHLUJE PROCES STÁRNUTÍ
POŠKOZENÍ GENOMU POČÁTEK NEMOCI NUTNOST PREVENCE!
ACKNOWLEDGEMENT B. J. M. I. A. Z. Z. P. P. A. O. I. V. J. Binkova Dejmek Dostal Chvatalova Milcova Novakova Lnenickova Rossner Rossner, Jr. Rossnerova Sevastyanova Solansky Svecova Topinka J. M. I. J. J. L. Z. P.B. A. E. Rubes Vozdova Benes Novak Nozicka Holy Pokorna Farmer Cebulska-Wasilewska Schallerova
ACKNOWLEDGEMENT Supported by the Czech Ministry of Environment VaV IC/5/6/04, SL/5/160/05, SP/1b3/50/07 AV CR IQS500390506