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Border Air Quality Strategy Toxicology of Urban Particulate Matter:
In Vitro and In Vivo Bioassays
Border Air Quality Strategy Toxicology of Urban Particulate Matter:
In Vitro and In Vivo Bioassays
Health CanadaSanté Canada
Dalibor Breznan MSc Renaud Vincent PhDDalibor Breznan MSc Renaud Vincent PhD
Inhalation Toxicology and Aerobiology Section Safe Environments Programme
Healthy Environments and Consumer Safety Branch
Inhalation Toxicology and Aerobiology Section Safe Environments Programme
Healthy Environments and Consumer Safety Branch
Second BAQS Investigator Workshop Detroit, 21 October 2005
Second BAQS Investigator Workshop Detroit, 21 October 2005
Health CanadaSanté CanadaHealth CanadaSanté Canada
Tit
Low Levels of Air Pollution Cause Health EffectsIncrease of +30 µg/m3 PM2.5 in southern Ontario associated the next day with +4% cardiac and +3% respiratory hospital admissions(Burnett et al, Am J Epidemiol 142:15-22, 1995)
Low Levels of Air Pollution Cause Health EffectsIncrease of +30 µg/m3 PM2.5 in southern Ontario associated the next day with +4% cardiac and +3% respiratory hospital admissions(Burnett et al, Am J Epidemiol 142:15-22, 1995)
Non-thresholdNon-threshold
ThresholdThreshold
Clean airClean airPolluted airPolluted air
Toxic airToxic air
SAFESAFE DANGERDANGER??
Health CanadaSanté Canada
Exposure characterization
Analytical chemistry
SourcesSources
SourcesSourcesSources AMBIENT
Sources INDOOR
Exposure assessment
Population and health studies
Risk assessment
Community outreach
Risk communication
Mechanisms
Biomarkers
Agents
Susceptible subgroups
Polymorphism
Pollutant interactions
Acute effects
Chronic effects
In uteroeffects
Inheritable effects
Molecular epidemiology
Samples collection
Biomathematics
In vivo bioassays
In vitro bioassays
Descriptors of potency
Inhalation exposures
Experimental animals
Human volunteers
Biomarkers of exposure and effects
Host factors of susceptibility
Current BAQSCurrent BAQS ToxicologyToxicology Burning IssuesBurning Issues
Exposure characterization
Analytical chemistry
Exposure characterization
Analytical chemistry
SourcesSources
SourcesSourcesSources AMBIENT
Sources INDOORSources
Sources
SourcesSourcesSources AMBIENT
Sources INDOOR
Exposure assessment
Population and health studies
Exposure assessment
Population and health studies
Exposure assessment
Population and health studies
Risk assessment
Community outreach
Risk communication
Mechanisms
Biomarkers
Agents
Susceptible subgroups
Polymorphism
Pollutant interactions
Acute effects
Chronic effects
In uteroeffects
Inheritable effects
Molecular epidemiology
Samples collection
Biomathematics
In vivo bioassays
In vitro bioassays
Descriptors of potency
Samples collection
Biomathematics
In vivo bioassays
In vitro bioassays
Descriptors of potency
Inhalation exposures
Experimental animals
Human volunteers
Biomarkers of exposure and effects
Host factors of susceptibility
Inhalation exposures
Experimental animals
Human volunteers
Biomarkers of exposure and effects
Host factors of susceptibility
Current BAQSCurrent BAQS ToxicologyToxicology Burning IssuesBurning Issues
Health CanadaSanté Canada
Animals
Inhalation exposures
Humans
Biomarkers of exposure and effects
Sample collection
In vitro bioassays
In vivo bioassays
Chemical Physical
Fractionation
Reconstitution
Biomathematics
Ambient Particles
Sources: Automotive
Diesel, GasolineIndustrial
Energy, Smelters, Incinerators
Model validation
Risk assessment
Integration of toxicity data to risk assessment and risk management
Integration of source-specific knowledge-base to risk management and health protection
Population and health
studies
A B
C D
E
BAQS Project StrategyBAQS Project StrategyBAQS Project Strategy
Health CanadaSanté Canada
Human inhalation
Animal inhalation
In vivo bioassays
In vitro bioassays
Risk assessment
Inte
grat
ion
©ITAB Health Canada 2005©ITAB Health Canada 2005
Human inhalation
Animal inhalation
In vivo bioassays
In vitro bioassays
Risk assessment
Human inhalation
Animal inhalation
In vivo bioassays
In vitro bioassays
Risk assessment
Inte
grat
ion
©ITAB Health Canada 2005©ITAB Health Canada 2005
Strengths andStrengths and Limitations ofLimitations of Assessment ApproachesAssessment Approaches
DirectDirect
HighHigh
MediumMedium
LowLow
Health Health relevancerelevance
HighHighNoneNoneHumanHumanInhalationInhalation
MediumMediumLowLowAnimal Animal InhalationInhalation
LowLowMediumMediumIn vivoIn vivobioassaysbioassays
Very lowVery lowVery highVery highIn vitroIn vitrobioassaysbioassays
TechnicalTechnicalchallengechallengeThroughputThroughput
Health CanadaSanté Canada
Xenometrix PM2.5
0
1
2
3
4
5
Fold
Indu
ctio
n
CYPGST
XREHMTIIA
HSP70GADD153
GADD45p53RE
NFkBRE0 µg
100 µg
200 µg
Ann Arbor
0
1
2
3
4
5
Fold
Indu
ctio
n
CYP
XRE
HSP70
GADD45
NFkBRE0 µg
100 µg
200 µg
Vermillion
0
1
2
3
4
5
Fold
Indu
ctio
n
CYPGST
XREHMTIIA
HSP70GADD153
GADD45p53RE
NFkBRE0 µg
100 µg
200 µg
Tiverton
0
1
2
3
4
5
Fold
Indu
ctio
n
CYPGST
XREHMTIIA
HSP70GADD153
GADD45p53RE
NFkBRE0 µg
100 µg
200 µg
Egbert
Vincent et al, Fund Appl Toxicol 39, 18-32, 1997Vincent et al, Fund Appl Toxicol 39, 18-32, 1997
Health CanadaSanté Canada
XRE (βi - βv)0.0 0.1 0.2 0.3 0.4 0.5
CY
P1A
1 (β
i - β
v)
0.00
0.05
0.10
0.15
0.20
0.25
0.30 1648
1649
EHCVER
DOR
ANA
TIV
A
EGB
TiO2
SAN R = 0.920p < 0.001
PAH (µg/g)0 10 20 30 40 50
CY
P1A
1 (β
i - β
v)
0.00
0.05
0.10
0.15
0.20
0.25
0.30B
SANTiO2
EGBDOR
EHC
ANA
TIV
VER
1649
1648
R = 0.863p = 0.001
XRE (βi - βv)0.0 0.1 0.2 0.3 0.4 0.5
CY
P1A
1 (β
i - β
v)
0.00
0.05
0.10
0.15
0.20
0.25
0.30 1648
1649
EHCVER
DOR
ANA
TIV
A
EGB
TiO2
SAN R = 0.920p < 0.001
PAH (µg/g)0 10 20 30 40 50
CY
P1A
1 (β
i - β
v)
0.00
0.05
0.10
0.15
0.20
0.25
0.30B
SANTiO2
EGBDOR
EHC
ANA
TIV
VER
1649
1648
R = 0.863p = 0.001
HMTIIA (βi - βv)0.0 0.1 0.2 0.3 0.4
HS
P70
(βi -
βv)
0.00
0.05
0.10
0.15
0.20
0.25
0.30
1648
1649
EHC
VER
EGB
DOR
SAN
ANA
TiO2
TIV
A
R = 0.880p < 0.001
Soluble Copper (µg/g)0 200 400 600 800 1000
HS
P70
(βi -
βv)
0.00
0.05
0.10
0.15
0.20
0.25
0.30
1648
1649
EHC
VER
EGB
DOR
SAN
ANA
TiO2
TIV
B
R = 0.955p < 0.001
HMTIIA (βi - βv)0.0 0.1 0.2 0.3 0.4
HS
P70
(βi -
βv)
0.00
0.05
0.10
0.15
0.20
0.25
0.30
1648
1649
EHC
VER
EGB
DOR
SAN
ANA
TiO2
TIV
A
R = 0.880p < 0.001
Soluble Copper (µg/g)0 200 400 600 800 1000
HS
P70
(βi -
βv)
0.00
0.05
0.10
0.15
0.20
0.25
0.30
1648
1649
EHC
VER
EGB
DOR
SAN
ANA
TiO2
TIV
B
R = 0.955p < 0.001
Potency
Vincent et al, Fund Appl Toxicol 39, 18-32, 1997Vincent et al, Fund Appl Toxicol 39, 18-32, 1997
Health CanadaSanté Canada
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
Sup
erna
tant
LD
H /
Tota
l Cel
l LD
Hra
te/m
in
0.0
0.5
1.0
1.5
2.0
2.5
3.0
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
Supe
rnat
ant L
DH
/ To
tal C
ell L
DH
Rat
e / m
in
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
LDH permeability assay(PM 24 h, n=3)
LDH permeability assay(PM 24 h, n=3)
A549
J774A.1
LDH
Health CanadaSanté Canada
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
Rel
ativ
e Lu
min
esce
nce
0.0
0.5
1.0
1.5
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
A549
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
Rel
ativ
e Lu
min
esce
nce
0.0
0.5
1.0
1.5
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
ATP metabolic assay(PM 24 h, n=3)
ATP metabolic assay(PM 24 h, n=3)
J774A.1
ATP
Health CanadaSanté Canada
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
Rel
ativ
e A
bsor
banc
e
0.0
0.3
0.6
0.9
1.2
1.5
1.8
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
0.0
10.0
20.0
40.0
80.0
160.
0
Rel
ativ
e Ab
sorb
ance
0.0
0.3
0.6
0.9
1.2
1.5
1.8
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
A549
BrdU proliferation assay(PM 24 h, n=3)
BrdU proliferation assay(PM 24 h, n=3)
J774A.1
BrdU
Health CanadaSanté Canada
Bioplex
TNF/WT
J774A.1 mouse macrophagesJ774A.1 mouse macrophages A549 human lung epithelial cellsA549 human lung epithelial cells
particulate matterparticulate matter
Bioplex cytokine / chemokine analysisBioplex cytokine / chemokine analysis
cell culture supernatantscell culture supernatants
Health CanadaSanté Canada
0.0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
0
2
4
6
8
10
12
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
A549 Human Lung Epithelial CellsA549 Human Lung Epithelial CellsGM-CSF
0.0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
0
300
600
900
1200
1500
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
0.0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
0
400
800
1200
1600
2000
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
IL-8
MCP-10.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
0
1
2
3
4
5
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
MIP-1ß0.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
0
500
1000
1500
2000
2500
3000
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
IL-6
Health CanadaSanté Canada
J774
Bioplex
J774A.1 Mouse Monocyte Cell LineJ774A.1 Mouse Monocyte Cell Line
0.0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
0
5
10
15
20
25
30
35
40
45
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
0.0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
00
3
6
9
12
15
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
0.0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
0
1000
2000
3000
4000
5000
6000
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
0.0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
00
5
10
15
20
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
0.0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
10.0
40.0
160.
0
0
5000
10000
15000
20000
25000
30000
35000
40000
EHC-93 EHC-98 DWR1SRM1648
Cristob.TiO2SRM1649
EHC-2000
GM-CSF
IL-1α
IL-6
IL-10
TNF-alpha
Health CanadaSanté Canada
0.0
0.0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
0
20
40
60
80
100
120
140
EHC-93 EHC-98 DWR1SRM1649
Cristob.TiO2SRM1650
EHC-2000control
sentinel
0.0
0.0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
0
20
40
60
80
100
120
EHC-93 EHC-98 DWR1SRM1649
Cristob.TiO2SRM1650
EHC-2000control
sentinel
Mouse In Vivo Bioassay (BALF)Mouse In Vivo Bioassay (BALF)GM-CSF
IL-100.
00.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
0
30
60
90
120
150
180
210
240
EHC-93 EHC-98 DWR1SRM1649
Cristob.TiO2SRM1650
EHC-2000control
sentinel
TNF-alpha
0.0
0.0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
0
3
6
9
12
15
EHC-93 EHC-98 DWR1SRM1649
Cristob.TiO2SRM1650
EHC-2000control
sentinel
IL-1α
0.0
0.0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
10.0
30.0
100.
0
0
30
60
90
120
150
180
EHC-93 EHC-98 DWR1SRM1649
Cristob.TiO2SRM1650
EHC-2000control
sentinel
IL-6
Health CanadaSanté Canada
VasoconstrictionHemodynamic changes VasoconstrictionHemodynamic changes
Endothelin-1Sustained higher plasma ET-1Endothelin-1Sustained higher plasma ET-1
ElectrophysiologyST-segment area depressionElectrophysiologyST-segment area depression
ParticlesParticles
Biological PlausibilityUrban particles cause elevation of circulating ET-1
Changes are sufficient to induce systemic vasoconstiction
May be compensated in healthy individuals
May not be compensated in individuals with heart diseases
Biological PlausibilityUrban particles cause elevation of circulating ET-1
Changes are sufficient to induce systemic vasoconstiction
May be compensated in healthy individuals
May not be compensated in individuals with heart diseases
Evidence in RatsCorrelate in time Maximal effects 36h post
Evidence in RatsCorrelate in time Maximal effects 36h post
VasoconstrictionHemodynamic changes VasoconstrictionHemodynamic changes
Endothelin-1Sustained higher plasma ET-1Endothelin-1Sustained higher plasma ET-1
ElectrophysiologyST-segment area depressionElectrophysiologyST-segment area depression
ParticlesParticles
VasoconstrictionHemodynamic changes VasoconstrictionHemodynamic changes
Endothelin-1Sustained higher plasma ET-1Endothelin-1Sustained higher plasma ET-1
ElectrophysiologyST-segment area depressionElectrophysiologyST-segment area depression
ParticlesParticlesParticlesParticles
Biological PlausibilityUrban particles cause elevation of circulating ET-1
Changes are sufficient to induce systemic vasoconstiction
May be compensated in healthy individuals
May not be compensated in individuals with heart diseases
Biological PlausibilityUrban particles cause elevation of circulating ET-1
Changes are sufficient to induce systemic vasoconstiction
May be compensated in healthy individuals
May not be compensated in individuals with heart diseases
Evidence in RatsCorrelate in time Maximal effects 36h post
Evidence in RatsCorrelate in time Maximal effects 36h post
Health CanadaSanté Canada
0 1 4 8 0 1 4 8 0 1 4 8 0 1 4 8 0 1 4 8
Rel
ativ
e ET
-1 s
ecre
tion
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4 * p<0.05** p=<0.001
CristobaliteTiO2EHC-93(total)
EHC-93(soluble)
EHC-93(insoluble)
****
*
***
***
****
**
A
0 1 4 8 0 1 4 8 0 1 4 8 0 1 4 8 0 1 4 8
Rel
ativ
e Bi
gET-
1 se
cret
ion
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4 * p<0.05** p=<0.001
CristobaliteTiO2EHC-93(total)
EHC-93(soluble)
EHC-93(insoluble)
**
**
**
**
**
****
**
**
**
B
0 1 4 8 0 1 4 8 0 1 4 8 0 1 4 8 0 1 4 8
Rel
ativ
e IL
-8 s
ecre
tion
0.0
0.5
1.0
1.5
2.0
2.5
3.0
CristobaliteTiO2EHC-93(total)
EHC-93(soluble)
EHC-93(insoluble)
***
*
**
* *
**
* p<0.05** p<0.001
*
**
*
C
0 1 4 8 0 1 4 8 0 1 4 8 0 1 4 8 0 1 4 8
rela
tive
VEG
F se
cret
ion
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
2.2
CristobaliteTiO2EHC-93(total)
EHC-93(soluble)
EHC-93(insoluble)
*
**
*
*
**
*
**
**
**
*
* p<0.05** p<0.001
**
D
ET-1 peptide bigET-1 peptide
IL-8 protein VEGF protein
A549 Human Lung Epithelial CellsA549 Human Lung Epithelial Cells
A
Chauhan et al, Cell Biol Toxicol 2005 (in press)Chauhan et al, Cell Biol Toxicol 2005 (in press)
Health CanadaSanté Canada
Human macrophages THP-1
Human macrophages THP-1
Human lung epithelial cells A549
Human lung epithelial cells A549
A549 + THP-1 Co-culture
A549 + THP-1 Co-culture
Human pulmonary artery endothelial cells
HPAE
Human pulmonary artery endothelial cells
HPAECysCys
Phe Phe Phe Phe TyrTyr
HisHis LeuLeu AspAsp IleIle IleIle TrpTrp
Ser Ser SerSerThrThr
ET-1ET-2ET-3
ET-1ET-2ET-3CysCys
CysCys CysCys
Ser Ser SerSerPhePhe
Leu Leu Trp Trp TyrTyrMetMet
Leu Leu LysLys
AspAsp
LysLys Glu Glu Glu Glu TyrTyr ValVal
TyrTyr
Ser Ser SerSerThrThr
N
C
7
65
43 2 1
8
10
9
11 1213
14 15 16 17 18 20 2119
CysCys
Phe Phe Phe Phe TyrTyr
HisHis LeuLeu AspAsp IleIle IleIle TrpTrp
Ser Ser SerSerThrThr
ET-1ET-2ET-3
ET-1ET-2ET-3CysCys
CysCys CysCys
Ser Ser SerSerPhePhe
Leu Leu Trp Trp TyrTyrMetMet
Leu Leu LysLys
AspAsp
LysLys Glu Glu Glu Glu TyrTyr ValVal
TyrTyr
Ser Ser SerSerThrThr
N
C
7
65
43 2 1
8
10
9
11 1213
14 15 16 17 18 20 2119
Bioplex protein liquid arrays
Bioplex protein liquid arrays
Endothelin isoforms
Endothelin isoforms
ProteomicsProteomics
In Vitro Cardiovascular ModelIn Vitro Cardiovascular Model
Health CanadaSanté Canada
Health CanadaSanté Canada
Examples protein spots
Health CanadaSanté Canada
Vascular sources
Inflammed atheroma
Hypertensive arteries
Aortic aneurysm
VascularVascular sourcessources
Inflammed atheromaInflammed atheroma
HypertensiveHypertensive arteriesarteries
Aortic aneurysmAortic aneurysm
Extravascular sources
Adipose tissue Lungs
Chronic infections (gingivitis, bronchitis, etc)
ExtravascularExtravascular sourcessources
AdiposeAdipose tissue tissue Lungs Lungs
ChronicChronic infections infections ((gingivitisgingivitis, , bronchitisbronchitis, , etcetc))
Primary inflammatory cytokines
Interleukin-1Tumor necrosis factor-"
etc.
Primary inflammatory cytokines
Interleukin-1Tumor necrosis factor-"
etc.
EndothelinsICAM-1sVCAM-1s-SelectinvWFtPAPAI-1
EndothelinsICAM-1sVCAM-1s-SelectinvWFtPAPAI-1
Acute phase proteins:CRPSAAFibrinogen
Acute phase proteins:CRPSAAFibrinogen
Serummarkers ofinflammationetc.
Serummarkers ofinflammationetc.
LiverLiver
Angiotensin IIAngiotensinAngiotensin IIII
Messenger cytokine
Interleukin-6
Messenger cytokine
Interleukin-6
Endothelial cellEndothelial cell
SelectinsSelectinsVCAM-1VCAM-1ICAM-1ICAM-1
CRP: C-reactive proteinN < 3 :g/mLR = 1000XStable
SAA: Serum amyloid A
CRP: C-reactive proteinN < 3 :g/mLR = 1000XStable
SAA: Serum amyloid A
Adapted from:Science 296:242-245 (2002)Adapted from:Science 296:242-245 (2002)
Health CanadaSanté Canada
-300s
-350s
-420s
-440s
-460s
Pre-exposure 2h 24h 32h 48hPost-exposure
-300s
-350s
-420s
-440s
-460s
Pre-exposure 2h 24h 32h 48hPost-exposure
Peptide 1400s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 1400s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 830s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 830s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 440s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 440s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 1400s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 1400s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 830s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 830s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 440s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Peptide 440s
Pre 32h24h 48h2h
Time post-exposure to urban particles
Health CanadaSanté Canada
Acknowledgements
Renaud Vincent Prem Kumarathasan
Dalibor BreznanVinita Chauhan
Marcelle PhaneufSubramanian Karthikeyan
Susantha MohottalagePatrick Goegan
Renaud Vincent Prem Kumarathasan
Dalibor BreznanVinita Chauhan
Marcelle PhaneufSubramanian Karthikeyan
Susantha MohottalagePatrick Goegan
Health CanadaBorder Air Quality Strategy
Health CanadaBorder Air Quality Strategy
Health CanadaHealth Canada
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