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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
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Air particles deposition in the airways
> 10 mm < 10 mm (PM10) < 2.5 mm (PM2.5)
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Prague Castle
Old Town Square
PRAGUE
Prague- Libus
Prague-Smichov
STATIONARY MONITORING
Mont hly average concent rat ion of B[a]P 7 Smichov
Libus
B[a]P ng/ m3
6 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 10 11 12 1 2 3 4 5 6 7 8 9 10 11 12
2004-2006
Roční průměry PM2.5 Praha - Smíchov a Libuš 2001- 2006 Smíchov
Libuš
32 30
µg/ m 3
28 26 24 22 20 18 16 2001
2002
2003 rok
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2004
2005
2006
B[a]P - 2006 Concentrations [ng/m3] ≤ 0.4 ≤ LAT > 0.4 - 0.6 (LAT, UAT > > 0.6 - 1.0 (UAT, LV > > 1.2 - 2.0 (LV,2 > > 2.0 >2
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39.7% 35.4% 15.9% 6.6% 2.4%
KONCENTRACE B[a]P Ostrava-Poruba
Praha-Smíchov
7
6
koncentrace [ng/m3]
5
4
3
2.8
2
1
0 1
2
3
4
5
6
7
2004
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8
9
10
11
12
1
2
3
4
5
6
7
2005
8
9
10
11
12
BIOMARKERS
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LIDSKÉ BIOMARKERY Biomonitorování Biomarkery expozice
Biomarkery účinku
Pre-maligní léze
Léčba
Diagnóza rakoviny Časná
Pozdní
100%
Ú m r t í
Prevence doba (roky) R. J. Šrám 2006
R.J. Albertini 1998
HUMAN BIOMARKERS Genetic/Carcinogenic Risks E X P O S U R E R. J. Sram 2008
Internal Dose
Biologically Effective Dose
Metabolic Genotype/Fenotype
Early Genotoxic Effects
Altered Structure Function
DNA Repair Capacity
Immune Functions
BIOMARKERS OF SUSCEPTIBILITY
C A N C E R
Environ Health Perspect 74: 3-9, 1987
PREGNANCY OUTCOME
EXPOSURE
Cotinine
DNA adducts
PAHs
HPRT BIRTH WEIGHT
SMOKING ETS
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Internal dose
Biologically Effective Dose
Early Genotoxic Effects
Metabolic
DNA Repair
Genotype/Phenotype
Capacity
Autoradiographs of thin layer chromatograms with DNA adduct pattern of:
DNA isolated from lymphocytes of subject sampled in January 2004 (1st sampling period)
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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) R. J. Sram 2008
Three translocations between chromosome 1 and unpainted chromosomes
MICRONUCLEI
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Reactive oxygen species Endogenous sources
Exogenous sources
O2
SOD
H2O2
OH•
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Lipid peroxidation
DNA oxidation
Catalase GP x
H2O + O2
H2O
Protein oxidation
Pathway-1
Pathway-2 organic compounds
OC-L PM
OC-H PM
Driven by: surface reactivity
Phase I (Cyp1a1 )
ROS
Surface Reactivity
ROS
ROS
Oxidative Effect
NFkB
Nrf2
API
Bioactivation Biotransformation
Phase II (Gst, Ngo1…)
Conjugation Proinflammatory response R. J. Sram 2008
Phase
III
Elimination
BIOMARKERS AIR POLLUTION
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Human studies and biomarkers of exposure, effect and susceptibility PM2.5
Stationary monitoring
c-PAHs Personal monitoring Stationary monitoring
DNA adducts by 32P-postlabeling Chromosomal aberrations conventional, FISH, micronuclei
VOC
Personal monitoring Stationary monitoring
Cotinine
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
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c-PAHs CONCENTRATIONS PERSONAL MONITORING (median and range)
Group
N
Age
B[a]P ng/m3
carcPAU ng/m3
(years) 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)
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„LIKE“"Like" B[a]P-DNA ADDUCT B[a]P-DNA Adduct DNA Adducts/108 Nucleotides
0.30
EXPCases (N=53)
CON (N=52)
0.25 0.20 0.15 0.10 0.05 0.00
All
SM
NS
All
SM
NS
EXP vs CON: P<0.01 Smokers vs Nonsmokers within groups: P<0.01
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IMPACT OF ENVIRONMENTAL POLUTION TO DNA ADDUCTS 6 DNA adducts = 1,042 + benzo[a]pyren * 0,077 (p=0,0003)
DNA adducts [108 nucleotides]
5
4
3
2
1
0
0
1
2
3
4
benzo[a]pyren [ugm-3 ]
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5
6
7
8
CYTOGENETIC ANALYSIS FISH EXP 6.0 5.0
* P < 0.05 ** P < 0.05
**
CON
** *
4.0 3.0 2.0
*
1.0 0
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* *
FG/100
AB.C.
NCJ
B/1000
t
rcp
IMPACT OF ENVIRONMENTAL POLUTION TO CHROMOSOMAL ABERRATIONS - FISH
SCGE repair capacities of DNA damage Policemen
Controls
Repaired DNA damage [%]
120 110 100
p<0.005
p<0.04
whole group
nonsmokers
90 80 70
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smokers
8-oxodG in urine of bus drivers and controls Urinary 8-oxodG levels in winter 2005, summer 2006 and winter 2006 14.00 p=0.05
p<0.01
p<0.001
8-oxodG (nmol/mmol creat.)
12.00 10.00 8.00 6.00 4.00 2.00 0.00 BUS1
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CON1
BUS2
CON2
BUS3
CON3
Correlation between oxidative stress markers and other factors 15-F2t-IsoP 8-oxodG carbonyl
B[a]P
cPAHs
PM2.5
PM10
15-F2t-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
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VLIV OVZDUŠÍ NA RESPIRAČNÍ ONEMOCNĚNÍ U DĚTÍ
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R. J. Sram 2008 3 day prior average 7 day prior average
6 day lag
5 day lag
4 day lag
3 day lag
0-24 months
2 day lag
1 day lag
Single day no lag
3 day prior average 7 day prior average
6 day lag
5 day lag
4 day lag
3 day lag
2 day lag
1 day lag
Single day no lag
Bronchitidy RR, 95% CI’s, akutní expozice PAU, multivariátní analýza
2.00
2-4.5 years
1.80
1.60
1.40
1.20
1.00
0.80
R. J. Sram 2008 3 day prior average 7 day prior average
6 day lag
5 day lag
4 day lag
3 day lag
0-24 months
2 day lag
1 day lag
Single day no lag
3 day prior average 7 day prior average
6 day lag
5 day lag
4 day lag
3 day lag
2 day lag
1 day lag
Single day no lag
Bronchitidy RR, 95% CI’s, akutní expozice PM2.5, multivariátní analýza
2.00
2-4.5 years
1.80
1.60
1.40
1.20
1.00
0.80
Význam první léta života dítěte jsou kritickou periodou dýchacího ústrojí a imunity
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VLIV OVZDUŠÍ NA 8-oxodG V MOČI DĚTÍ
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Vztah 8-oxodG a kotininu
8-oxodG (nmol/mmol kreat.)
50
40
30
20
10
0 1
10
100
kotinin (µg/mmol kreat.)
1000
Vztah 8-oxodG a věku dítěte 8-oxodG (nmol/mmol kreat.)
50 40 30 20 10 0 5
6
7
8 Věk
9
10
11
Multivariate model of effect of PM2.5 on 8-oxodG levels (linear regression)
Period (days before sampling Region All
Pollutant Intercept PM2.5 Child age Cotinine (above/below 20 ng/mg) Allergic rhinitis
3-day (-4;-6)
7-day (-1;-7)
R
p
R
p
0.16 -2.74 3.20 1.34
33.56 <0.05 <0.0001 0.001 0.41
0.16 -2.74 3.23 1.28
33.94 0.08 <0.001 0.005 <0.005
<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
0.30 <0.001 0.06
45.22 -0.20 -3.09 -3.02
0.21 <0.001 0.06
Teplice
Intercept PM2.5 Child age Cotinine (above/below 20 ng/mg) Allergic rhinitis
28.57 0.24 -2.51 4.41 7.59
Prachatice
Intercept PM2.5 Child age Allergic rhinitis
43.71 -0.13 -3.07 -3.03
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Multivariate model of effect of B[a]P on 8-oxodG levels (linear regression)
Period (days before sampling Region
Pollutant
3-day (-4;-6) R
All
Teplice
Prachatice
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Intercept B[a]P Child age Cotinine (above/below 20 ng/mg) Allergic rhinitis
34.01 -1.44 -2.65 3.04 1.19
Intercept B[a]P Child age Cotinine (above/below 20 ng/mg) Allergic rhinitis
31.09 1.73 -2.54 4.42 7.76
Intercept B[a]P Child age Allergic rhinitis
40.25 -0.02 -3.05 -1.62
p
3-day (-1;-7) R
p
0.03 <0. 001 <0.005 0.47
34.94 1.20 -2.74 3.49 1.27
0.08 <0.001 <0.01 0.55
<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
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
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IUGR by PM10 during 1994 - 1998 in TEPLICE Low = 1 Med; NS
3,0
3,0
High; NS
Med; P<0.04 2,5
2,5
High; P<0.0005
2.10
2,0
2,0
1,5
1,5
1.41 1,0
1,0
0,5
0,5
0,0
0,0
1st
2nd
3rd
4th
5th
6th
Months of gestation
7th
8th
9th
AOR with 95%CL
AOR with 95% CL
AOR
CARCINOGENIC PAHs & IUGR IN TEPLICE 4.0
4.0 Low CI Medium ; NS
3.5
3.5
CI High ; NS
3.0
CI Medium ; P<0.05 CI High ; P<0.005
2.5
2.5
AORs
2.16
2.0
2.0
1.67
1.5
1.5
1.0
1.0
0.5
0.5
0.0
0.0 1st
2nd
3rd
4th
5th
6th
Months of gestation R. J. Sram 2007
7th
8th
9th
AORs with 95% CI
AORs with 95% CI
3.0
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
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SCSA® - Acridine Orange Stained DNA
Native DNA Stainability (green fluorescence)
100
0 R. J. Sram 2008
Fragmented DNA (red fluorescence)
100
Air pollutions Prague 2007 (ng/m3) 12.0
February
May
B[a]P
eqB[a]P
10.0 8.0 6.0 4.0 2.0 0 PAH
cPAH
Sperm analysis 180
February
May
160 140 120 100 80 60 40
e m u l ) Vo (ml
tio a tr l) n c e il/m n Co (m
n
ty it li ) o M (%
it y l ta ) i V (%
M
0
or p (% hol ) og y
20
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)
16
15 10
20
12
HDS (%)
20
high DFI (%)
det DFI (%)
25
8
10
4
5 February May
February
P≤0.001
N=46 R. J. Sram 2008
15
May
5
P≤0.001 dDFI dDFI dDFI HDS
< 15% 15 – 30% >30% >15%
Feb 30 Feb 16 Feb 2 Feb 10
February May
P≤0.001 May 42 May 4 May 2 May 4
AIR POLLUTION RISK ASSESSMENT
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ACCORDING TO MOLECULAR EPIDEMIOLOGY STUDIES
concentrations > 1 ng B[a]P/m3 in polluted air
RISK FOR HUMAN HEALTH R. J. Sram 2008
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
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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 ! R. J. Sram 2008
ACKNOWLEDGEMENT B. J. M. I. A. Z. Z. P. P. A. O. I. V. J. R. J. Sram 2008
Binkova Dejmek Dostal Chvatalova Milcova Novakova Lnenickova Rossner Rossner, Jr. Rossnerova Sevastyanova Solansky Svecova Topinka
J. Rubes M. Vozdova I. Benes J. Novak J. Nozicka L. Holy Z. Pokorna P.B. Farmer A. Cebulska-Wasilewska E. 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
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