# Annex C. C ontrolled Human Exposure Studies

Table C-1. Cardiovascular effects.

Study Pollutant Exposure Findings

Reference: Barregard et al. (2006, 091381 )

Subjects: 13 healthy adults

Gender: 6 M/7 F

Age: 20-56 yr Wood smoke

Particle Size: Session 1: GMD 42 nm; Session 2: GMD 112 nm

Particle Number/Count: Session 1: 180,000/cm 3 ; Session 2: 95,000/cm 3

Concentration: Session 1: median: 279 μg/m 3 ; Session 2: median 243 μg/m 3 Subjects exposed in two groups for 4 h to filtered air, followed a wk later by a 4-h exposure to wood sm oke. Exposures conducted with two 25-mi n periods of light exercise. Other measured combustion products:

Session 1: NO 2 (0.08 ppm), CO (13 ppm), formaldehyde (114 μg/m 3 ), acetaldehyde (75 μg/m 3 ), benzene (30 μg/m 3 ), 1,3-butadiene (6.3 μg/m 3 );

Session 2: NO 2 (0.09 ppm), CO (9.1 ppm), formaldehyde (64 μg/m 3 ), acetaldehyde (40 μg/m 3 ), benzene (20 μg/m 3 ), 1,3-butadiene (3.9 μg/m 3 ).

Time to analysis : Immediately followingexposure as well as 3 and 20 h postexposure. Statistically significant increase in plasma factor VIII 20 h post wood smoke exposure relative to filtered air. The factor VIII/von Willebrand ratio in plasma was increased with wood smoke relative to filtered air at 0, 3, and 20 h post-exposure. Wood smoke exposure increased the urinary excretion of free 8-iso-prostaglandin2 α relative to clean air 20 h post-exposure (n = 9). These findings were more pronounced in session 1 than session 2 (similar mass concentration but higher number concentration in Session 1).

Reference: Beckett et al. (2005, 156261 )

Subjects: 12 healthy adults

Gender: 6 M/6 F

Age: 23-52 yr Ultrafine and fine zinc oxide

Particle Size: UF: <0.1 μm; Fine: 0.1-1.0 μm

Particle Number/Count: UF: 4.6 × 10 7 /cm 3 ; Fine: 1.9 × 10 5 /cm 3

Concentration: 500 μg/m 3 Subjects exposed via mouthpiece for 2 h during rest to filtered air, ultrafine, and fine zinc oxide in a randomized crossover study design. Exposures were separated by at least 3 wk.

Time to analysis: Immediately following exposure and 3, 6, 11, 23, and 24 h after
exposure. Exposure to ultrafine and fine zinc oxide did not affect HRV (time and frequency domain parameters) relative to clean air immediately following exposure, or at 3, 6, 11, and 23 h post-exposure. Exposure did not affect blood pressure through 24 h post-exposure. No effects of exposure to either fine or ultrafine zinc oxide observed on factor VII, von Willebrand factor (vWf), tissue plasminogen activator (t-PA), or fibrinogen. No effect of exposure observed on peripheral blood cell counts or levels of pro- inflammatory cytokines.

Reference: Blomberg et al. (2005, 191991 )

Subjects: 15 older adults (former smokers) with COPD

Age: 56-72 yr DE

Concentration: 300 μg/m 3 Subjects exposed for 1 h with intermittent exercise to DE and filt ered air in a randomized crossover study design.

Time to analysis: 6 and 24 h post-exposure. DE was not observed to af fect blood levels of C-reactive protein, fibrinogen, D-Dimer, prothrombin factor 1-2, or von Willebrand factor activity at 6 and 24 h post-exposure.

Reference: Brauner et al. (2007, 091152 )

Subjects: 29 healthy adults

Gender: 20 M/9 F

Age: 20-40 yr Urban traffic particles

Particle Number/Count: 6



# - 700nm: 10,067/cm 3

Concentration: PM 2.5 : 9.7 μg/m 3 ; PM 10-2.5 : 12.6 μg/m 3 Subjects exposed to ur ban traffic particles and filtered air for 24 h with and without two 90-min periods of lig ht exercise in a randomized crossover study design. Concentrations of NO X and NO were low and did not differ between filtered and unfiltered exposures. CO concentra tions were higher with filtered air (0.35 and 0.41 ppm), while O 3 concentrations were lower with filtered air (12.08 and 4.29 ppb).

Time to analysis: 6 and 24 h after the start of
exposure. An increase in DNA strand breaks and formamidopyrimidine-DNA glycosylase sites in peripheral blood mononuclear cells were observed after 6 and 24 h of exposure to urban particulates. The partic le concentration at the 57nm mode was shown to be the major contributor to these effects.

Note: Hyperlinks to the reference citations throughout this doc ument will take you to the NCEA HERO database (Health and Environmental Research Online) at http://epa.gov/hero . HERO is a database of scientific literature used by U.S. EPA in the process of developing science assessments such as the Integrated Science Assessments (ISA) and the Integrated Risk Information System (IRIS)

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### December 2009 C-1

### Study Pollutant Exposure Findings

Reference: Brauner et al. (2008, 156293 )

Subjects: 42 healthy older adults (21 couples)

Age: 60-75 yr Indoor air particles

Particle Number/Count: 10-700 nm: 10,016/cm 3

Concentration: Coarse: 9.4 μg/m 3 ; Fine: 12.6 μg/m 3 Exposures consisted of two 48 h periods in the home of each subject with or without the use of a HEPA filter (randomized crossover design). HEPA filters reduced coarse concentration from 9.4 to 4.6 μg/m 3 , and fine concentration from 12.6 to 4.7 μg/m 3

. Concentrations of NO 2 did not differ between the 2 sessions (20 ppb).

Time to analysis: After the completion of each 48 h session. The use of HEPA filter s significantly improved microvascular function (p = 0.04) after 48 h (reactive hyperemia- peripheral arterial tonometry). Microva scular function was assessed using a scoring system representing the extent of reac tive hyperemia. The reduction in PM concent ration through the use of HEPA filters did not significantly affect blood pressure following the 48-h exposures. Lowering PM concentration did not significantly affect inflammatory response markers in peripheral venous blood (IL-6, TNF- α

, C-reactive protein, plasma amyloid A).

Reference: Brauner et al. (2008, 191966 )

Subjects: 29 healthy adults

Gender: 20 M, 9 F

Age: M avg 27 yr, F avg 26 yr Urban traffic particles

Particle Number/Count: 11,600/cm 3

Concentration: PM 2.5 : 10.5 μg/m 3 ; PM 10-2.5 : 13.8 μg/m 3 Subjects exposed to ur ban traffic particles and filtered air for 24 h with and without two 90-min periods of lig ht exercise in a randomized crossover study design. Concentrations of NO X and NO were low and did not differ between filtered and unfiltered exposures. CO concentra tions were higher with filtered air, while O 3 concentrations were lower with filtered air.

Time to analysis: 6 and 24 h after the start of
exposure. Exposure to urban traffic particles was not observed to affect microvascular function (digital peripheral artery tone) at 6 or 24 h after the start of exposure. No difference in various blood markers of coagulation, inflammation, or protein oxidation (e.g., fibrinogen, platelet count, CRP, IL-6, TNF- α ) were demonstrated between particle and filtered air exposure.

Reference: Carlsten et al. (2007, 155714 )

Subjects: 13 healthy adults

Gender: 11 M/2 F

Age: 20-42 yr DE

2002 Cummins B-series diesel engine (6BT5.9G6, 5.9 L) operating at load

Concentration: Fine PM: 100, 200 μg/m 3 Subjects exposed for 2 h at rest to filtered air and each of the two DEPs concentrations in a randomized crossover study design. Exposures were separated by at least 2 wk. Other diesel emissions measured: NO 2 (10-35 ppb), CO (0.7-1.8 ppm).

Time to analysis: 3, 6, and 22 h after the start of exposure. No statistically significant changes in plasminogen activator inhibitor-1 (PAI-1), vWf, D-dimer, or platelet c ount observed 3, 6, or 22 h following exposure to DE relative to filtered air. Non-stat istically significant increases in D-dimer, vWf, and platelet count were observed at 6 h following the start of exposure (4 h pos t-exposure). No diesel-induced increase in C-reactive protein observed relative to filtered air in peripheral venous blood at 1 or 20 h post-exposure.

Reference: Carlsten et al. (2008, 156323 )

Subjects: 16 adults with metabolic syndrome

Gender: 10 M/6 F

Age: 25-48 yr DE

2002 Cummins B-series diesel engine (6BT5.9G6, 5.9 L)

Concentration: Fine PM: 100, 200 μg/m 3 Subjects exposed for 2 h at rest to filtered air and each of the two DE particle concentrations in a randomized crossover study design. Exposures were separated by at least 2 wk. Other diesel emissions measured: NO 2 (30 ppb), NO (1.69 ppm), CO (0.65 ppm).

Time to analysis: 3, 7, and 22 h after the start of exposure. At 5 h after the end of diesel exposure (fine particulate concentration 200 μg/m 3 ), the authors observed a signifi cant decrease in vWf in peripheral venous blood. No other changes in thrombotic markers (vWf, D-dimer, PAI-1) were observed at eit her concentration between 1 and 20 h post-exposure.

Reference: Danielsen et al. (2008, 156382 )

Subjects: 13 healthy adults

Gender: 6 M/7 F

Age: 20-56 yr Wood smoke

Particle Size:

Session 1: GMD 42 nm; Session 2: GMD 112 nm

Particle Number/Count: Session 1: 180,000/cm 3 ; Session 2: 95,000/cm 3

Concentration:

Session 1: median: 279 μg/m 3 ; Session 2: median 243 μg/m 3 Subjects exposed in two groups for 4 h to filtered air, followed a wk later by a 4-h exposure to wood sm oke. Exposures conducted with two 25-mi n periods of light exercise. Other measured combustion products:

Session 1: NO 2 (0.08 ppm), CO (13 ppm), formaldehyde (114 μg/m 3 ), acetaldehyde (75 μg/m 3 ), benzene (30 μg/m 3 ), 1,3-butadiene (6.3 μg/m 3 );

Session 2: NO 2 (0.09 ppm), CO (9.1 ppm), formaldehyde (64 μg/m 3 ), acetaldehyde (40 μg/m 3 ), benzene (20 μg/m 3 ), 1,3-butadiene (3.9 μg/m 3 ).

Time to analysis: 3 and 20 h post-exposure. Exposure to wood smoke increased the mRNA levels of hOGG1 in PBMCs relative to filtered air 20 h after exposure. DNA strand breaks were shown to decrease in PBMCs 20 h after wood smoke exposure.

Reference: Devlin et al. (2003, 087348 )

Subjects: 10 healthy older adults

Gender: 7 M/3 F

Age: Avg 66.9 yr Fine CAPs (Chapel Hill, NC)

Concentration: Mean: 40.5 μg/m 3 , Range: 21.2-80.3 μg/m 3 Exposures conducted for 2 h at rest to filtered air and CAPs in a randomized crossover study design.

Time to analysis: Immediately following exposure and 24 h post-exposure. CAPs exposure resulted in statistically significant reductions (p <0.05) in time domain (PNN50) and frequency domain (HF power) parameters relative to clean air immediately following exposure. Thes e relative decreases were still apparent 24 h after exposure (p <0.08).



### December 2009 C-2

### Study Pollutant Exposure Findings

Reference: Fakhri et al. (2009, 191914 )

Subjects: 50 adults (40 healthy, 10 asthmatic)

Gender: 24 M/26 F

Age: 19-48 yr Fine CAPs (Toronto)

Concentration: 127 ± 62 μg/m 3 with and without co-exposure to O 3 (114 ± ppb) Exposures conducted through a facemask which covered the subject’s nose and mouth. Subjects were exposed to CAPs, O 3 , CAPs + O 3 and filtered air for 2 h at rest in a randomized crossover study design.

Time to analysis: Every 30 min during exposure, with the fi nal measurement made immediately prior to the end of the exposure. Exposure to CAPs or O 3 , alone or in combination, resulted in no significant changes in HRV or blood pressure relative to filtered air. However, a negative concentration response relationship was reported between CAPs concentration with co-exposure to O 3 and SDNN, rMSSD, HF power and LF power (statistically significant for LF power). Diastolic blood pressure was observed to increase with exposure to CAPs + O 3 , but not with either pollutant alone. There was no difference in response between asthmatics and healthy subjects.

Reference: Frampton et al. (2006, 088665 )

Subjects: 16 asthmatic adults, 40 healthy adults

Gender: Asthmatics: 8 M/8 F, Healthy: 20 M/20 F

Age: 18-40 yr Ultrafine EC

Particle Size: CMD ~25 nm

Particle Number/Count: 10 μg/m 3 : ~2.0 × 10 6 /cm 3 ; 25 μg/m 3 : ~7.0 × 10 6 /cm 3 ; 50 μg/m 3 : ~10.8 × 10 6 /cm 3

Concentration: 10, 25, and 50 μg/m 3 Study conducted using a randomized crossover design with 2-h exposures. Asthmatics (n = 16) exposed to filtered air and 10 μg/m 3 . 12 healthy adults exposed to filtered air and 10 μg/m 3 at rest; 12 healthy adults exposed to filtered air, 10 and 25 μg/m 3 with intermittent exerci se; 16 healthy adults exposed to filtered air and 50 μg/m 3 with intermittent exerci se. Exposures were conducted via mouthpiece.

Time to analysis: Immediately following exposure as well as 3. 5, 21, and 45 h postexposure. No effect of ultrafin e particle exposure on leukocyte counts or leuk ocyte expression of adhesion molecules observed in healthy subjects exposed at rest to 10 μg/m 3 . Among healthy adults exposed to ultrafine carbon during exercise, monocyte expression of adhesion molecules CD54 and CD18 decreased relative to filtered air immediately following exposure. An ultrafine particle-induced decrease in PMN expression of CD18 was also observed 0-21 h post-exposure. Expression of CD11b on monocytes and eosinophils was reduced following exposure to ultrafine particles in exercising asthmatics 0-21 h post-exposure. A decrease in total leukocyte count was observed following ultraf ine particle exposure in exercising healthy and asthmatic subjects.

Reference: Gong et al. (2004, 087964 )

Subjects: 13 older adults with COPD, 6 healthy older adults

Gender: COPD: 5 M/8 F, Healthy: 2 M/4 F

Age: COPD: avg 68 yr, Healthy: avg 73 yr Fine CAPs (Los Angeles)

Particle Size: 85% of mass between 0.1 and 2.5 μm

Concentration: Mean: 194 μg/m 3 , Range: 135-229 μg/m 3 Exposures to CAPs and filtered air (randomized crossover) for 2 h with intermittent light ex ercise (four 15-min periods). Exposures were separated by at least 2 wk.

Time to analysis: Immediately followingexposure as well as 4 and 22 h postexposure. SDNN shown to decr ease following CAPs exposure relative to filtered air in healthy older adults (4-22 h post-exposure). No CAPs-induced changes in HRV were observed in older adults with COPD. Ectopic heart beats were observed to increas e slightly with CAPs relative to filtered air among healthy subjects, but decreased among subjects with COPD. Exposure to CAPs did not affect platelet or white blood cell count, or levels of fibrinogen, vWF, or factor VII.

Reference: Gong et al. (2004, 055628 )

Subjects: 12 adult asthmatics, 4 healthy adults

Gender: Asthmatics: 4 M/8 F, Healthy: 2 M/2 F

Age: Asthmatics: avg 38 yr, Healthy: avg 32 yr Coarse CAPs (Los Angeles)

Particle Size: 80% of mass between 2.5 and 10 μm, 20% of mass <2.5 μm

Concentration: Mean: 157 μg/m 3 , Range: 56-218 μg/m 3 Exposures to CAPs and filtered air (randomized crossover) for 2 h with intermittent light ex ercise (four 15-min periods). Exposures were separated by at least 2 wk.

Time to analysis: Immediately followingexposure as well as 4 and 22 h postexposure. SDNN shown to decr ease following CAPs exposure relative to filtered air in healthy adults (4-22 h post-ex posure). Decrease in PNN50 also observed in healthy adults at 4 h post-exposure. No CAPs -induced decreases in HRV demonstrated in asthmatics.

Reference: Gong et al. (2008, 156483 )

Subjects: 14 adult asthmatics, 17 healthy adults

Gender: Asthmatics: 9 M/5 F, Healthy: 5 M/12 F

Age: Asthmatics: 34 ± 12 yr, Healthy: 24 ± 8 yr Ultrafine CAPs (Los Angeles)

Particle Number/Count: 145,000/cm 3 , Range 39,000-312,000/cm 3

Concentration: Mean100 μg/m 3 , Range13-277 μg/m 3 Subjects exposed for 2 h during intermittent exercise (15-min periods) to both CAPs and filtered air in random order. The first 7 subjects underwent w hole body exposure, while the remaining subjects were exposed through a facemask. Facemask exposures had higher particle counts but lower particle mass than whole body exposures. Exposures were separated by at least 2 wk.

Time to analysis: Immediately followingexposure as well as 4 and 22 h postexposure. Relative to filtered air, exposure to ultrafine CAPs resulted in a transient decrease in LF power 4 h post-exposure. This effect of CAPs on HRV was not influenced by health status. CAPs exposure was not observed to affect any other measures of HRV, blood pressure, or blood markers of inflamma tion or coagulation. There were no differences in response observed between facemask and whole body
exposures.



### December 2009 C-3

### Study Pollutant Exposure Findings

Reference: Graff et al. (2009, 191981 )

Subjects: 14 healthy adults

Gender: 8 M/6 F

Age: 20-34 yr Coarse CAPs (Chapel Hill, NC)

Concentration: 89 ± 49.5 μg/m 3 (estimated inhaled dose ≈ 67% of measured particle mass) Subjects exposed for 2 h with intermittent exercise (15-min periods ) to coarse CAPs and filtered air in a randomized crossover design. Exposures were separated by at least 2 mos.

Time to analysis: 0-1 and 20 h postexposure. At 20 h post-exposure, tPA was observed to decrease by 32.9% fr om baseline (preexposure) per 10 μg/m 3 increase in CAPs concentration (p = 0.01). D-dimer concentration decreased 11.3% per 10 μg/m 3

, a change of marginal statistical significance (p = 0.07). No other coarse CAPs-induced changes in blood biomarkers of coagulation (e.g., vWF, factor VI I, plasminogen, fibrinogen, or PAI-1) or inflammation (e.g., CRP) were observed. At 20 h post-e xposure, overall HRV (SDNN) was shown to decrease by 14.4% relative to pre-expos ure measurements per 10 μg/m 3 increase in CAPs concentration. No other changes in HRV were observed following exposure to coarse CAPs.

Reference: Huang et al. (2003, 087377 )

Subjects: 38 healthy adults

Gender: 36 M/2 F

Age: Avg 26.2 ± 0.7 yr Fine CAPs (Chapel Hill, NC)

Concentration: 23.1-311.1 μg/m 3 Subjects exposed to CAPs (n = 30) or filtered air (n = 8) for 2 h with intermittent exercise (subjects did not serve as their own controls). Component data of CAPs was available for 37 of the 38 subjects.

Time to analysis: 18 h after exposure. The increase in blood fibrinogen following exposure to fine CAPs r eported by Ghio et al. (2000, 012140 ) was shown to be associated with copper, zinc, and vanadium content in the CAPs.

Reference: Larsson et al. (2007, 091375 )

Subjects: 16 healthy adults

Gender: 10 M/6 F

Age: 19-59 yr Traffic particles (road tunnel)

Particle Size: PM 2.5 , PM 10 ; PM 2.5 mass constituted ~36% of PM 10 mass

Particle Number/Count: 20-1,000 nm: 1.1 × 10 5 /cm 3

, < 100 nm: 0.85 × 10 5 /cm 3

Concentration: PM 2.5 - 46-81 μg/m 3 ; PM 10 - 130-206 μg/m 3 Exposures were conducted for 2 h with intermittent exercise in a room adjacent to a busy road tunnel. Study used a randomized crossover design with each subject also exposed to normal air (control). Exposures were separated by 3-10 wk. No exposures to filtered air were conducted. Other traffic emissions measured: NO (874 μg/m 3 ), NO 2 (230 μg/m 3 ), CO (5.8 μg/m 3 reported, likely 5.8 mg/m 3 ).

Time to analysis: 14 h post-exposure. No change in plasma leve ls of fibrinogen or PAI-1 observed 14 h post-exposure.

Reference: Lucking et al. (2008, 191993 )

Subjects: 20 healthy adults

Gender: M

Age: 21-44 yr DE

Protocol 1 (n=8): idling Deutz diesel engine (F3M2011, 2.2 L, 500 rpm) using gas oil

Protocol 2 (n=12): idling Volvo diesel engine (TD45, 4.5 L, 4 cylinders, 680 rpm) using Gasoil E10

Particle Number/Count: Protocol 1: 1.2 × 10 6 /cm 3 ; Protocol 2: 1.26 × 10 6 /cm 3

Concentration: Protocol 1: 348 μg/m 3 , Protocol 2: 330 μg/m 3 In both protocols, ex posures were conducted with intermittent exerci se (15-min periods) to DE and filtered air in a randomized crossover design with exposures se parated by at least one wk.

Protocol 1 (n=8): Ex posures conducted for 2 h. Other diesel emissions measured: NO X (0.58 ppm), NO 2 (0.23 ppm), NO (0.36 ppm), CO (3.54 ppm), total hydrocarbon (2.8 μg/m 3 ).

Time to analysis: 6 h post-exposure.

Protocol 2 (n=12): Exposures conducted for 1h. Other diesel emissions measured: NO X (2.78 ppm), NO 2 (0.62 ppm), NO (2.15 ppm), CO (3.08 ppm), total hydrocarbon (1.58 μg/m 3 ).

Time to analysis: 2 and 6 h post-exposure. Thrombus formation wa s observed to increase with diesel 2 and 6 h post-exposure using anex vivo perfusion chamber. Both plateletneutrophil and platelet -monocyte aggregates increased relative to filtered air 2 h following exposure to diesel (onl y evaluated in Protocol 2). Plasma concentrati ons of soluble CD40L were also observed to increase with diesel. Exposure to diesel was not shown to affect total leukocyte, monocyte, or platelet counts.

Reference: Lund et al. (2009, 180257 )

Subjects: 10 healthy adults

Gender: 4 M/6 F

Age: 18-40 yr DE

Idling Cummins diesel engine (5.9 L) using commercial No. 2 fuel

Particle Size: MMAD 0.10 μm

Concentration: 100 μg/m 3 Subjects exposed for 2 h with intermittent exercise (15-min periods) to DE and filtered air in a randomized crossover study design. Other diesel emissions measured: NO X (4.7 ppm), NO 2 (0.8 ppm), CO (2.8 ppm), total hydrocarbons (2.4 ppm).

Time to analysis: 30 min and 24 h postexposure. Exposure to diesel resulted in an increase in MMP-9 plasma concent ration and activity as well as an increase in endothelin-1 plasmaconcentration at both 30 min and 24 h postexposure.



### December 2009 C-4

### Study Pollutant Exposure Findings

Reference: Lundback et al. (2009, 191967 )

Subjects: 12 healthy adults

Gender: M

Age: 21-30 yr DE

Idling Volvo diesel engine (TD45, 4.5 L, 4 cylinders, 680 rpm) using Gasoil E10

Particle Number/Count: 1.26 × 10 6 /cm 3

Concentration: 330 μg/m 3 Subjects exposed for 1 h with intermittent exercise (15-min periods) to DE and filtered air in a randomized crossover study design. Exposures were separated by at least one wk. Other diesel emissions measured: NO X (2.78 ppm), NO 2 (0.62 ppm), NO (2.15 ppm), CO (3.08 ppm), total hydrocarbon (1.58 μg/m 3 ).

Time to analysis: 10, 20, 30, and 40 min post-exposure. Diesel-induced increase in arterial stiffness (increases in augmentation pressure and augmentation index, as well as decrease in time to wave reflection) observed at 10 and 20 min post-exposure using radial artery pulse wave analysis. No effect of diesel observed on carotid-femoral pulse wave velocity which was assessed 40 min post-exposure, but not at earlier time points. No ef fect of diesel observed on blood pressure 10-30 min post-exposure.

Reference: Mills et al. (2005, 095757 )

Subjects: 30 healthy adults

Gender: M

Age: 20-38 yr DE

Idling 1991 Volvo diesel engine (TD45, 4.5 L, 4 cylinders, 680 rpm)

Particle Number/Count: 1.2 × 10 6 /cm 3

Concentration: 300 μg/m 3 Subjects exposed for 1 h with intermittent exercise (15-min periods) to DE and filtered air in a randomized crossover study design. Exposures were separated by two wk. Other diesel emissions measured: NO 2 (1.6 ppm), NO (4.5 ppm), CO (7.5 ppm), total hydrocarbon (4.3 ppm), formaldehyde (0.26 μg/m 3 ).

Time to analysis: 2-4 h post-exposure for 15 subject; 6-8 h post-expos ure for the other 15 subjects. Forearm blood flow increase (induced by bradykinin, acetylcholine, and sodium nitroprusside) was attenuated by DE 2 and 6 h post-exposure. A 6 mmHg increase in diastolic blood pressure (p = 0.08) 2 h following exposure to DE was obs erved relative to filtered air control. Br adykinin-induced release of t-PA was attenuated by diesel exposure 6 h post-exposure. DE did not affect the release of t-PA 2 h post-exposure. No diesel-induced changes in serum IL-6 or TNF- α observed 6 h post-exposure.

Reference: Mills et al. (2007, 091206 )

Subjects: 20 older adults with prior myocardial infarction

Gender: M

Age: 60 ± 1 yr DE

Idling 1991 Volvo diesel engine (TD45, 4.5 L, 4 cylinders, 680 rpm) using low sulfur gas-oil E10

Particle Size: Median particle diameter 54 nm, Range 20-120 nm

Particle Number/Count: 1.26 × 10 6 /cm 3

Concentration: 300 μg/m 3 Subjects exposed for 1 h with intermittent exercise (15-min periods) to DE and filtered air in a randomized crossover study design. Exposures were separated by at least two wk. Other diesel emissions measured: NO X (4.45 ppm), NO 2 (1.01 ppm), NO (3.45 ppm), CO (2.9 ppm), total hydrocarbon (2.8 ppm).

Time to analysis: During exposure and 6-8 h post-exposure. A greater increase in exercise induced ST-segment depression and ischemic burden was observed during ex posure to DE than clean air. No diesel-induced effects on vasomotor dysfunction observed 6 h post-exposure. Bradykini n-induced release of t-PA was attenuated by diesel exposure relative to filtered air 6 h post-exposure. Effect of diesel on t-PA release was not evaluated at earlier times post-exposure. No diesel-induced changes in blood leukocyte counts or serum C-reactive protein 6 h post-exposure.

Reference: Mills et al. (2008, 156766 )

Subjects: 12 adults with coronary heart disease, 12 healthy adults

Gender: M

Age: CHD: 59 ± 2 yr, Healthy: 54 ± 2 yr Fine CAPs (Edinburgh, Scotland, UK)

Particle Size: Mean 1.23 μm

Particle Number/Count: 99,400/cm 3

Concentration: 190 ± 37 μg/m 3 Exposures conducted for 2 h with intermittent exercise. Subjects exposed to CAPs and filtered air using a randomized crossover design with exposures se parated by at least 2 wk.

Time to analysis: 2, 6-8, and 24 h postexposure. CAPs exposure had no significant effect on vascular function in healthy adults or adults with coronary heart disease 6-8 h post-exposure (i.e., no change in forearm blood flow as assessed using venous occlusion plethysm ography). The authors attributed this lack of response to a low concentration of combus tion-derived particles. Small increase in blood platelet and monocyte concentration observed following CAPs exposure. Exposure to CAPs did not affect serum CRP concentration or total leukocyte or neutrophil count.

Reference: Peretz et al. (2007, 156853 )

Subjects: 5 healthy adults

Gender: M

Age: 20-31 yr DE

2002 Cummins B-series diesel engine (6BT5.9G6, 5.9 L); operating at 75% of rated capacity

Concentration: Fine PM 50, 100, 200 μg/m 3 Subjects exposed for 2 h at rest to filtered air and each of the three DE particle concentrations in a randomized crossover study design. Exposures were separated by at least 2 wk. Other diesel emissions measured, 200 μg/m 3 exposure: NO 2 (23 ppb), NO (1.75 ppm), CO (1.58 ppm).

Time to analysis: 6 and 22 h after the start of
exposure. PBMC expression of 10 genes involved in the inflammatory response were observed to be significantly affected by exposure to DE at the highest concentration tested (8 upregulated, 2 downregulated) 6 h after the start of exposure. The expression of 4 genes (1 upregulated, 3 downregulated) associated with the inflammatory response showed significant changes 22 h after diesel exposure. PBMC expression of 5 genes involved in the oxidative stress pathways showed significant changes at 6 h after the start of diesel exposure at the highest concentration tested (4 upregulated, 1 downregulated). 7 genes involved in the oxidative stress pathwa ys showed significant changes at 22 h following exposure (4 upregulated, 3 downregulated).



### December 2009 C-5

### Study Pollutant Exposure Findings

Reference: Peretz et al. (2008, 156854 )

Subjects: 17 adults with metabolic syndrome, 10 healthy adults

Gender: Metabolic syndrome: 11 M/6 F, Healthy: 8 M/2 F

Age: Metabolic syndrome: 20-48 yr, Healthy: 20-42 yr DE

2002 Cummins B-series diesel engine (6BT5.9G6, 5.9 L) using No. 2 undyed on-highway fuel; operating at 75% of rated capacity

Particle Size: Median particle diameter 1.04 μm

Concentration: Fine PM 100, 200 μg/m 3 Subjects exposed for 2 h at rest to both concentrations of DE as well as filtered air in a randomized crossover design. Exposures were separated by at l east 2 wk. Other diesel emissions measured, 100 μg/m 3 exposure: NO 2 (16.5 ppb), NO (0.96 ppm), CO (0.51 ppm); 200 μg/m 3 exposure: NO 2 (24.7 ppb), NO (1.54 ppm), CO (0.89 ppm).

Time to analysis: Immediately following exposure (within 30 mi n post-exposure) and 3 h from the start of exposure. Exposure to 200 μg/m 3 elicited a statistically significant decrease in br achial artery diameter relative to filtered air immediately following exposure. A smaller decreas e in brachial artery diameter was also observed following exposure to DE at 100 μg/m 3 . Plasma levels of endothelin-1 were observed to increase following DE exposure (200 μg/m 3 ). The observed effects were more pronounced in healthy subjects than in subjects with metabolic syndrome. DE did not affect endothelium-dependent flow-mediated dilatation. No effect of DE on blood pressure was demonstrated immediately following
exposure.

Reference: Peretz et al. (2008, 156855 )

Subjects: 13 adults with metabolic syndrome, 3 healthy adults

Gender: Metabolic syndrome: 8 M/5 F, Healthy: 3 M/0 F

Age: M etabolic syndrome: 31-48 yr, Healthy: 24-39 yr DE

2002 Cummins B-series diesel engine (6BT5.9G6, 5.9 L) using No. 2 undyed on-highway fuel; operating at 75% of rated capacity

Concentration: Fine PM 100, 200 μg/m 3 Subjects exposed for 2 h at rest to both concentrations of DE as well as filtered air in a randomized crossover design. Exposures were separated by at l east 2 wk. Other diesel emissions measured, 100 μg/m 3 exposure: NO 2 (20.6 ppb), NO (0.95 ppm), CO (0.47 ppm); 200 μg/m 3 exposure: NO 2 (28.3 ppb), NO (1.63 ppm), CO (0.74 ppm).

Time to analysis: 1, 3, 6, and 22 h from the start of exposure. Exposure to 200 μg/m 3 increased HF power and decreased the LF/HF ratio 1h post-exposure; how ever, this effect was not consistent across subjec ts. No effect of DE was observed at later time points. Subjects with metabolic syndrome did not experience greater changes in HRV than healthy subjects.

Reference: Power et al. (2008, 191982 )

Subjects: 5 adults with mildto-moderate allergic asthma

Gender: 1 M/4 F

Age: 28-51 yr Carbon and ammonium nitrate particles

Concentration:

With co-exposure to 0.2ppm O 3 : 255 μg/m 3

, Without co-exposure to 0.2ppm O 3 : 313 μg/m 3 Subjects exposed for 4 h with intermittent exercise (30-min periods) to filtered air, particles, and particles + O 3 in a crossover study design. Exposures were separated by at least 3 wk.

Time to analysis: 3 h 40 min from the start of
exposure. Time and frequency domain HRV parameters were not affected by particle exposure relative to filtered air. However, exposure to particles with O 3 resulted in a significant decrease in SDNN as well as changes to both high and low frequency power normalized to the difference between total and very low frequency power.

Reference: Routledge et al. (2006, 088674 )

Subjects: 20 older adults with coronary artery disease, 20 healthy older adults

Gender: CAD: 17 M/3 F, Healthy: 10 M/10 F

Age: CAD: 52-74 yr, Healthy: 56-75 yr Ultrafine carbon

Particle Size: <10-300 nm; mode at 20-30 nm

Concentration: Ultrafine carbon: 50 μg/m 3 ; SO 2 : 200 ppb Exposures conducted (head dome system) to filtered air, ultrafine carbon, SO 2 , and ultrafine carbon + SO 2 for 1 h at rest using a randomized crossover study design.

Time to analysis: Immediately followingexposure as well as 3 and 23 h postexposure. No PM-induced changes in HRV observed among subjects with cor onary artery disease. Among healthy subjects, small increase in HRV (RR, SDNN, rMSSD, and LF power) demonstrated immediately post-carbon exposure. Relative to filtered air control, exposure to ultrafine carbon did not significantly affect blood pressure in healthy adults or adults with coronary artery disease 0-3 h post-exposure. Ex posure to ultrafine carbon, either alone or with SO 2 , did not affect plasma levels of fibrinogen or D-dimer at 3 or 23 h post-exposure. Ex posure to ultrafine carbon did not affect peripheral blood leukocyte count or C-reac tive protein levels 3 or 23 h post-exposure.

Reference: Rundell and Caviston (2008, 191986 )

Subjects: 15 healthy college athletes

Gender: M

Age: Avg 19.5 yr Gasoline emissions

2.5 hp gasoline engine running 10 s each min during exposure and in the min prior to exposure

Particle Size: PM1.0

Particle Number/Count: Trial 1: 336,730 ± 149,206/cm 3 ;

Trial 2: 396,200 ± 82,564/cm 3 Subjects were exposed twice to both clean air and dilute gasoline exhaust during 6-min periods of maximal exercise on a cycle ergometer. Clean air exposures occurred first and were separated by 3 days. Gasoline exhaust exposures were also separated by 3 days, with the first occurring 7 days after the second clean air exposure. Other emissions measured: CO (6.3 ± 3.4 ppm).

Time to analysis: 6 min There was no difference in total work done (kJ) between the clean air exposures or between the clean air exposures and the first exposure to gasoline exhaust. However, the second gasoline exhaust expos ure was demonstrated to significantly decrease work accumulated over the 6min exercise period compared with either of the other ex posure conditions (p < 0.01).



### December 2009 C-6

### Study Pollutant Exposure Findings

Reference: Samet et al. (2007, 156940 )

Subjects: Ultrafine CAPs: 20 healthy adults, Coarse CAPs: 14 healthy adults

Gender: Ultrafine CAPs: 11 M/9 F, Coarse CAPs: 8 M/6 F

Age: Ultrafine CAPs: 18-35 yr, Coarse CAPs: 18-35 yr CAPs (Chapel Hill, NC)

Particle Size: Ultrafine 0.049 ± 0.009 μm; Coarse 3.59 ± 0.58 μm

Concentration: Ultrafine 47.0 ± 20.2 μg/m 3 ; Coarse 89.0 ± 49.5 μg/m 3 Preliminary report comparing effects of controlled exposures to coarse, fine, and ultrafine CAPs among healthy adults (3 separate studies). A randomized crossover design was used in evaluating effects of coarse CAPs (n=14) and ultrafine CAPs (n=20) relative to filtered air following 2-h exposures with intermi ttent exercise. Results compared with previous study of controlled exposure to fine C APs (Chapel Hill, NC) where subjects did not serve as their own controls (Ghio et al., 2000, 012140 ).

Time to analysis: 0-20 h post-exposure. Statistically signific ant decrease in SDNN observed 20 h following exposure to coarse CAPs relative to filter ed air. Subjects in the high ultrafine CAPs group experienced a decrease in SDNN based on an analysis of 24 h ambulatory Holter monitoring relative to filtered air. Fine CAPs did not significantly affect HRV. Increased levels of D-dimer observed 18 h following exposure to ultrafine CAPs. No CAPs-induc ed changes in plasma factor VII, plasminogen, fibrinogen, PAI-1, vWf, or t-PA. No CAPs-induced changes in C-reactive protein le vels were observed.

Reference: Samet et al. (2009, 191913 )

Subjects: 19 healthy adults

Gender: 10 M/9 F

Age: 18-35 yr Ultrafine CAPs (Chapel Hill, NC)

Particle Size: < 0.16 μm

Particle Number/Count: 120,662 ± 48,232 particles/cm 3

Concentration: 49.8 ± 20 μg/m 3 Subjects exposed for 2 h with intermittent 15 periods of exercise to UF CAPs and filtered air using a randomized crossover study design.

Time to analysis: Immediately following exposure and 1 and 18 h post-exposure. UF CAPs exposure resulted in an increase in plasma concentrations of D-dimer both immediately following exposure (20.6% increase per 10 5 particles/cm 3 ) as well as 18 h post-exposure (18.2% increase per 10 5 particles/cm 3 ). Plasma concentration of PAI1 also increased with UF CAPs, although this increase was not statisti cally significant (24% increase, p = 0.1). No UF CAPs-induced changes observed in plas ma concentrations of tPA, vWF, CRP, fibrinogen, plasminogen, or Factor VII. HF and LF power were both observed to increase wi th UF CAPs exposure relative to filtered ai r at 18 h post-exposure (41.8% and 36%, respectively, per 10 5 particles/cm 3 increase in UF CAPs). UF CAPs expressed as mass concentration was not observed have a statistica lly significant effect in HF total power. UF CAPs was not observed to affect time domain measures of HRV over 24 h. The QT interval was shown to decrease both immediately following and at 18 h post exposure (not statis tically significant immediately following exposure).

Reference: Shah et al. (2008, 156970 )

Subjects: 16 healthy adults

Age: 26.9 ± 6.9 yr Ultrafine EC

Particle Number/Count: 10.8 ± 1.7 × 10 6 /cm 3

Concentration: 50 μg/m 3 Exposures conducted via mouthpiece for 2 h with intermittent exerci se to filtered air and ultrafine carbon in a randomized crossover study design.

Time to analysis: Immediately following exposure as well as 3. 5, 21, and 45 h postexposure. Exposure to ultrafine carbon attenuated peak forearm blood flow after ischemia relative to filtered air 3.5 h post-ex posure. Venous nitrate levels were significantly lower at 21 h following exposure to UF carbon compared with filtered air exposure. PM expos ure was not observed to affect blood pressure relative to filtered air at times 0-45 h post-exposure.

Reference: Tornqvist et al. (2007, 091279 )

Subjects: 15 healthy adults

Gender: M

Age: 18-38 yr DE

Idling 1991 Volvo diesel engine (TD45, 4.5 L, 4 cylinders, 680 rpm)

Concentration: 300 μg/m 3 Subjects exposed for 1h with intermittent exercise (15-min periods) to DE and filtered air in a randomized crossover study design. Exposures were separated by at least two wk. Other diesel emissions measured: NO X (4.44 ppm), NO 2 (0.82 ppm), NO (3.62 ppm), total hydrocarbon (2.21 ppm).

Time to analysis: 24 h post-exposure. DE was observed to significantly attenuate endothelium-dependent vasodilation 24 h post-exposure. Endothelium-independent vasodilation was not affected by diesel exposure. Exposure to DE did not affect blood pressure relative to filtered air 24 h after exposure. DE signific antly increased plasma levels of IL-6 and TNF- α 24 h following exposure. Exposure to diesel resulted in an increase in total antioxidant capacity of plasma relative to filtered air 24 h post-exposure.

Reference: Urch et al. (2004, 055629 )

Subjects: 24 healthy adults

Gender: 14 M/10 F

Age: 35 ± 10 yr Fine CAPs (Toronto)

Concentration: 150 μg/m 3 (range 101-257 μg/m 3 ) with 120 ppb O 3 Exposures conducted through a facemask which covered the subject’s nose and mouth. Subjects were exposed to CAPs + O 3 and filtered air for 2 h at rest in a randomized crossover study design. Exposures were separated by at least 2 days.

Time to analysis: Immediately following
exposure. CAPs + O 3 exposure resulted in a significant decrease in brachial artery diameter immediately post-exposure (Brook et al., 2002, 024987 ), which was demonstrated to be associated with both the organic and EC fractions of the CAPs.



### December 2009 C-7

### Study Pollutant Exposure Findings

Reference: Urch et al. (2005, 081080 )

Subjects: 23 healthy adults

Gender: 13 M/10 F

Age: 32 ± 10 yr Fine CAPs (Toronto);

Concentration: 150 μg/m 3 (range 102-214 μg/m 3 ) with 120 ppb O 3 Exposures conducted through a facemask which covered the subject’s nose and mouth. Subjects were exposed to CAPs + O 3 and filtered air for 2 h at rest in a randomized crossover study design.

Time to analysis: Every 30 min during exposure, with the fi nal measurement made immediately prior to the end of the exposure. An increase in diastolic blood pressure of 6 mmHg was observed at the end of CAPs + O 3 exposure, which was statistically different from the change in blood pressure experienced during exposure to filter ed air (1 mmHg). This effect was associated with the organic fraction of PM 2.5

.

Reference: Zareba et al. (2009, 190101 )

Subjects: 24 healthy adults

Gender: 12 M/12 F

Age: 18-40 yr Ultrafine EC

Particle Size: Count median diameter 25 nm

Particle Number/Count: 2×10 6 /cm 3 (10 μg/m 3 ), 7×10 6 /cm 3 (25 μg/m 3 )

Concentration: 10 μg/m 3 ; 25 μg/m 3 Protocol 1 (n=12, 6 M/6 F): Subjects exposed to 10 μg/m 3 UF carbon and filtered air for 2 h at rest in a randomized crossover design. Exposures were separated by at least 2 wk.

Protocol 2 (n=12, 6 M/6 F): Subjects exposed to 10 μg/m 3 , 25 μg/m 3 , and filtered air for 2 h with intermittent exerci se (15-min periods) in a restricted randomized crossover design (all subjects exposed to 10 μg/m3 before 25 μg/m3). Exposures were separated by at least 2 wk.

Time to analysis (both protocols): Immediately following exposure and 3.5 and 21 h post-exposure. Exposure to 10 μg/m 3 at rest resulted in no change in HRV frequency domain parameters relative to filtered air exposure. Time domain parameters were observed to increase slightly with UF carbon exposure (10 μg/m 3 at rest), however, only the increase in rMSSD was statistically significant (p = 0.032). Some trends toward less short ening of QT interval, increase in ST segment, and increase in variability of repolarization (variability of T wave complexity) were observed with exposure to 10 μg/m 3 at rest, none of which were statistically significant.

In Protocol 2, ex posure to 10 μg/m 3 UF carbon was observed to slightly increase HRV time domain parameters as was demonstrated in Protocol 1. However, this was not observed at the higher concentration (25 μg/m 3 ). As with exposure at rest, exposure to UF carbon during exercise was observed to affect repolarization (reducti on in QT duration and increase in T-wave amplitude), although this effect was not statistically significant.



### December 2009 C-8

# Table C-2. Respiratory effects.

Reference Pollutant Exposure Findings

Reference: Alexis et al. (2006, 154323 )

Subjects: 9 healthy adults

Gender: 3 M/6 F

Age: 18-35 yr Coarse fraction particles (Chapel Hill, NC)

Heat-treated (biologically inactive) and non-heated particles

Particle Size: MMAD 5 μm

Concentration: 0.65 mg per subject Subjects were administered heat-treated PM 10-2.5 , non-heated PM 10-2.5 , and 0.9% saline (control) via nebulization in a randomized crossover study design. Exposures were separated by at least 1 wk.

Time to analysis: 2-3 h post-inhalation. Both heat-treated and non-heated coarse PM were observed to increase neutrophil counts in induced sputum 2-3 h post-inhalation. Biologically active PM (non-heated) induced an increase expression of macrophage TNF- α mRNA, eotaxin, and immune surface phenotypes on macrophages (mCD14, CD11b/CR3, and HLA-DR).

Reference: Barregard et al. (2008, 155675 )

Subjects: 13 healthy adults

Gender: 6 M/7 F

Age: 20-56 yr Wood smoke

Particle Size:

Session 1: geometric mean diameter 42 nm, Session 2: geometric mean diameter 112

nm

Particle Number/Count: Session 1: 180,000/cm 3 ; Session 2: 95,000/cm 3

Concentration: Session 1: median 279 μg/m 3 ; Session 2: median 243 μg/m 3 Subjects exposed in two groups for 4 h to filtered air, followed a wk later by a 4-h exposure to wood smoke. Exposures conducted with two 25-min periods of light exercise. Other measured combustion products:

Session 1: NO 2 (0.08 ppm), CO (13 ppm), formaldehyde (114 μg/m 3 ), acetaldehyde (75 μg/m 3 ), benzene (30 μg/m 3 ), 1,3-butadiene (6.3 μg/m 3 );

Session 2: NO 2 (0.09 ppm), CO (9.1 ppm), formaldehyde (64 μg/m 3 ), acetaldehyde (40 μg/m 3 ), benzene (20 μg/m 3 ), 1,3-butadiene (3.9 μg/m 3 ).

Time to analysis: Immediately following exposure as well as 3 and 20 h post-exposure. Relative to filtered air, exposure to wood smoke was observed to increase levels of eNO 3 h post-exposure. Serum Clara cell protein increased 20 h after wood smoke exposure. Wood smoke was observed to increase levels of malondialdehyde in breath condensate immediately after as well as 20 h post-exposure. Effects of wood smoke on eNO and malondialdehyde levels were similar between the two sessions of wood smoke exposure. However, serum Clara cell protein was significantly increased with wood smoke in session 1 (higher particle count) but not in session 2.

Reference: Bastain et al. (2003, 098690 )

Subjects: 18 nonsmoking adults with positive allergy skin test to short ragweed

Gender: 7 M/11 F

Age: 18-38 yr DEP

Isuzu diesel engine, 4 cylinder, 4JB1

Concentration: 0.3 mg in 200 μl saline Subjects underwent nasal provocation challenge (intranasal spray) with allergen and either DEP or placebo (saline) in a randomized crossover study design. Challenges were separated by 30 days. This protocol was then repeated 30 days after the last exposure.

Time to analysis: 24 h post-exposure and 4 and 8 days after exposure. DEP significantly increased allergic responses to short ragweed. Relative to allergen + placebo, allergen + DEP increased allergen specific IgE 4days following exposure, and increased IL-4 1 day post-exposure. The enhancement of allergic response with DEP was observed to be reproducible within subjects.

Reference: Beckett et al. (2005, 156261 )

Subjects: 12 healthy adults

Gender: 6 M/6 F

Age: 23-52 yr Ultrafine and fine zinc oxide

Particle Size: UF: <0.1 μm; Fine: 0.1-1.0 μm

Particle Number/Count: UF: 4.6 × 10 7 /cm 3 ; Fine: 1.9 × 10 5 /cm 3

Concentration: 500 μg/m 3 Subjects exposed via mouthpiece for 2 h during rest to filtered air, ultrafine, and fine zinc oxide in a randomized crossover study design. Exposures were separated by at least 3 wk.

Time to analysis: 11 and 24 h after exposure. No changes observed in neutrophil count in induced sputum. No PM (zinc oxide)-induced changes in respiratory symptoms observed 0-24 h post-exposure.

Reference: Behndig et al. (2006, 088286 )

Subjects: 15 healthy adults

Gender: 8 M/7 F

Age: 21-27 yr DE

Idling 1991 Volvo diesel engine (TD45, 4.5 L, 4 cylinders, 680 rpm)

Particle Size: PM 10 ; majority of PM mass made up of particles < 1 μm in diameter

Concentration: 100 μg/m 3 Exposures conducted for 2 h with intermittent exercise to both DE and filtered air in a randomized crossover design. Exposures were separated by at least 3 wk. Other diesel emissions measured: NO X (1.8 ppm), NO 2 (0.4 ppm), NO (1.3 ppm), CO (10.4 ppm), total hydrocarbons (1.3 ppm).

Time to analysis: 18 h post-exposure. Exposure to DE increased neutrophil and mast cell numbers in bronchial mucosa at 18 h post-exposure. Neutrophils, IL-8, and myeloperoxidase observed to increase in bronchial lavage fluid following exposure relative to filtered air. No inflammatory response observed in the alveolar compartment. Exposure to DE increased urate and reduced glutathione bronchoalveolar lavage at 18 h post-exposure.

Reference: Blomberg et al. (2005, 191991 )

Subjects: 15 older adults (former smokers) with COPD

Age: 56-72 yr DE

Concentration: 300 μg/m 3 Subjects exposed for 1 h with intermittent exercise to DE and filtered air in a randomized crossover study design.

Time to analysis: 6 and 24 h post-exposure. DE was not observed to affect levels of Clara cellprotein in peripheral blood at 6 and 24 h postexposure.



### December 2009 C-9

### Reference Pollutant Exposure Findings

Reference: Bosson et al. (2007, 156286 )

Subjects: 16 healthy adults

Gender: 7 M/9 F

Age: 20-28 yr DE

Idling Volvo diesel engine

Concentration: PM 300 μg/m 3 followed by exposure to filtered air or 0.2 ppm O 3 Subjects exposed to DE for 1 h followed 5 h later by a 2-h exposure to either filtered air or O 3 (0.2 ppm) using a randomized crossover study design. All exposures were conducted with subjects engaged in intermittent exercise.

Time to analysis: 18 h after second exposure (filtered air or O 3 ). The percentage of neutrophils and concentration of myeloperoxidase in induced sputum (18 h post-O 3 /air exposure) was significantly higher following diesel + O 3 than diesel + air.

Reference: Bosson et al. (2008, 196659 )

Subjects: 14 healthy adults

Gender: 9 M/5 F

Age: 21-29 yr DE

Idling 1991 Volvo diesel engine (TD45, 4.5 L, 4 cylinders)

Concentration: PM 300 μg/m 3 or filtered air followed by exposure to 0.2 ppm O 3 Subjects exposed to DE or filtered air for 1h followed 5 h later by a 2-h exposure to O 3 (0.2 ppm) using a randomized crossover study design. All exposures were conducted with subjects engaged in intermittent exercise. Other diesel emissions measured: NO 2 (0.51 ppm), NO (1.65 ppm), total hydrocarbons (1.18 ppm).

Time to analysis: 24 h after the start of the initial
exposure. Neutrophil and macrophage numbers in bronchial wash were significantly increased 16 h following O 3 exposure when preceded by exposure to diesel, compared to O 3 exposure preceded by exposure to filtered air.

Reference: Brauner et al. (2009, 190244 )

Subjects: 29 healthy adults

Gender: 20 M, 9 F

Age: M avg 27 yr, F avg 26 yr Urban traffic particles

Particle Size: PM 2.5 , PM 10-2.5
Particle Number/Count: 6- 700 nm: 10,067/cm 3

Concentration: PM 2.5 : 9.7 μg/m 3 , PM 10-2.5 : 12.6 μg/m 3 Subjects exposed to urban traffic particles and filtered air for 24 h with and without two 90-min periods of light exercise in a randomized crossover study design. Concentrations of NO X and NO were low and did not differ between filtered and unfiltered exposures. CO concentrations were higher with filtered air (0.35 and 0.41 ppm), while O 3 concentrations were lower with filtered air (12.08 and 4.29 ppb).

Time to analysis: 2.5, 6, and 24 h after the start of exposure. Epithelial membrane integrity and blood-gas barrier permeability, assessed using pulmonary clearance of 99mTc-labeled diethylenetriamine pentaacetic acid (DTPA), was observed to increase with exercise, but was not affected by exposure to urban particles (2.5 h of exposure). Exposure to urban particles was not shown to affect plasma or urine concentration of Clara cell 16 protein at 6 and 24 h after the start of exposure. No relationship between exposure and pulmonary function was observed at 2.5 h.

Reference: Gilliland et al. (2004, 156471 )

Subjects: 19 adults with allergic rhinitis and positive skin test to ragweed, GSTM1 (14 null, 5 present); GSTT1 (9 null, 10 present); GSTP1 codon 105 variants (13 I/I, 6 I/V, 0 V/V)

Gender: 7 M/12 F

Age: 20-34 yr DEP

Isuzu diesel engine, 4 cylinder, 4JB1

Concentration: 0.3 mg DEP in 300 μL saline Subjects were challenged intranasally with allergen and placebo (saline) as well as allergen plus DEP in saline in a randomized crossover design. Challenges were separated by at least 6 wk.

Time to analysis: 10 min, 24 h, and 72 h postchallenge. Subjects who were GSTM1 null or homozygous for GSTP1 I105 wild-type allele experienced significantly greater increase in nasal IgE and histamine with diesel plus allergen compared to subjects with functional GSTM1 or who were heterozygous for GSTP1 I/V(105).

Reference: Gong et al. (2004, 087964 )

Subjects: 13 older adults with COPD, 6 healthy older adults

Gender: COPD: 5 M/8 F, Healthy: 2 M/4 F

Age: COPD: avg 68 yr, Healthy: avg 73 yr Fine CAPs (Los Angeles)

Particle Size: 85% of mass between 0.1 and 2.5 μm

Concentration: Mean: 194 μg/m 3 , Range: 135-229 μg/m 3 Exposures to CAPs and filtered air (randomized crossover) for 2 h with intermittent light exercise (four 15-min periods). Exposures were separated by at least 2 wk.

Time to analysis: Immediately following exposure as well as 4 and 22 h post-exposure. No CAPs-induced respiratory symptoms observed in healthy older adults or older adults with COPD at 0, 4, or 22 h post-exposure. Exposure to CAPs did not significantly affect FVC or FEV 1 . CAPs exposure caused a decrease in arterial oxygen saturation immediately following exposure which was more pronounced in healthy older adults than in older adults with COPD. Exposure to CAPs was not observed to affect the levels of white blood cells, columnar epithelial cells, IL-6, or IL-8 in induced sputum.

Reference: Gong et al. (2004, 055628 )

Subjects: 12 adult asthmatics, 4 healthy adults

Gender: Asthmatic: 4 M/8 F, Healthy: 2 M/2 F

Age: Asthmatic: avg 38 yr, Healthy: avg 32 yr Coarse CAPs (Los Angeles)

Particle Size: 80% of mass between 2.5 and 10 μm, 20% of mass <2.5 μm

Concentration: Mean: 157 μg/m 3 ; Range: 56-218 μg/m 3 Exposures to CAPs and filtered air (randomized crossover) for 2 h with intermittent light exercise (four 15-min periods). Exposures were separated by at least 2 wk.

Time to analysis: Immediately following exposure as well as 4 and 22 h post-exposure. No effect of CAPs exposure on spirometry or arterial oxygen saturation was observed 0, 4, or 22 h post-exposure. No respiratory symptoms reported 0-22 h post-exposure in either healthy orasthmatic adults. Sputum cell counts at 22 h postexposure did not differ between CAPs and filtered air.



### December 2009 C-10

### Reference Pollutant Exposure Findings

Reference: Gong et al. (2005, 087921 )

Subjects: 18 older adults with COPD, 6 healthy older adults

Gender: COPD: 9 M/9 F, Healthy: 2 M/4 F

Age: COPD: avg 72 yr, Healthy: avg 68 yr Fine CAPs (Los Angeles)

Concentration: CAPs: 200 μg/m 3 ; NO 2 : 0.4 ppm Each subject was exposed to CAPs, NO 2 , CAPs + NO 2 , and filtered air for 2 h with intermittent exercise. Exposure order was not fully counterbalanced as NO 2 exposures were conducted after the majority of the CAPs and filtered air exposures had been completed. Exposures were separated by at least 2 wk.

Time to analysis: Immediately following exposure as well as 4 and 22 h post-exposure. Exposure to CAPs was observed to decrease maximal mid-expiratory flow and arterial oxygen saturation relative to filtered air 4-22 h post-exposure. This response was more pronounced in healthy older adults than in older adults with COPD. Concomitant exposure to NO 2 did not enhance the response. No other respiratory responses (symptoms, spirometry, sputum cell counts) were affected by exposure to CAPs.

Reference: Gong et al. (2008, 156483 )

Subjects: 14 adult asthmatics, 17 healthy adults

Gender: Asthmatics: 9 M/5 F, Healthy: 5 M/12 F

Age: Asthmatics: 34 ± 12 yr, Healthy:24 ± 8 yr Ultrafine CAPs (Los Angeles)

Particle Number/Count: 145,000/cm 3 , Range 39,000- 312,000/cm 3

Concentration: Mean: 100 μg/m 3 , Range: 13-277 μg/m 3 Subjects exposed for 2 h during intermittent exercise (15-min periods) to both CAPs and filtered air in random order. The first 7 subjects underwent whole body exposure, while the remaining subjects were exposed through a facemask. Facemask exposures had higher particle counts but lower particle mass than whole body exposures. Exposures were separated by at least 2 wk.

Time to analysis: Immediately following exposure as well as 4 and 22 h post-exposure. No significant differences in respiratory symptoms observed between filtered air and ultrafine CAPs exposures. Individuals exposed to higher particle counts tended to experience greater symptoms with CAPs than with filtered air. An ultrafine CAPs-induced decrease in arterial oxygen saturation (0.5%) was observed at 0, 4, and 22 h post-exposure. A decrease in FEV 1 (2%) was also observed 22 h post-exposure relative to filtered air. Responses were not significantly different between healthy and asthmatic adults. CAPs exposure was not observed to affect total sputum cell counts or cytokine levels. There were no differences in response observed between facemask and whole body exposures.

Reference: Graff et al. (2009, 191981 )

Subjects: 14 healthy adults

Gender: 8 M/6 F

Age: 20-34 yr Coarse CAPs (Chapel Hill, NC)

Concentration: 89 ± 49.5 μg/m 3 (estimated inhaled dose ≈ 67% of measured particle mass) Subjects exposed for 2 h with intermittent exercise (15-min periods) to coarse CAPs and filtered air in a randomized crossover design. Exposures were separated by at least 2 mos.

Time to analysis: 0-1 and 20 h post-exposure. Pulmonary function (FVC, FEV 1 , and carbon monoxide diffusing capacity) was not affected by exposure to coarse CAPs either immediately following exposure or 20 h post-exposure. A significant increase in percent PMNs (10.7% increase per 10 μg/m 3 coarse CAPs) was observed in BAL fluid 20 h post-exposure. Percent monocytes in BL fluid were slightly decreased at 20 h post-exposure (2.0% decrease per 10 μg/m 3 CAPs; p = 0.05). Total protein in BAL fluid was also observed to decrease following CAPs exposure (1.8% decrease per 10 μg/m 3 CAPs). Markers of inflammation in BAL and BL fluids including IL-6, IL-8, and PGE2 were not affected by exposure to coarse CAPs.

Reference: Huang et al. (2003, 087377 )

Subjects: 38 healthy adults

Gender: 36 M/2 F

Age: Avg 26.2 ± 0.7 yr Fine CAPs (Chapel Hill, NC)

Concentration: 23.1-311.1 μg/m 3 Subjects exposed to CAPs (n = 30) or filtered air (n = 8) for 2 h with intermittent exercise (subjects did not serve as their own controls). Component data of CAPs was available for 37 of the 38 subjects.

Time to analysis: 18 h after exposure. The increase in bronchoalveolar lavage fluid neutrophils observed by Ghio et al. (2000, 012140 ) following exposure to fine CAPs was shown to be associated with iron, selenium, and sulfate content of the CAPs.

Reference: Kongerud et al. (2006, 156656 )

Subjects: 17 asthmatic adults, 46 healthy adults

Gender: Asthmatics6 M/11 F, Healthy- 24 M/22 F

Age: Asthmatics: avg 23 yr, Healthy: avg 26 yr DEP

NIST 1650, heavy duty diesel engine

Concentration: Untreated and treated with 0.1 ppm O 3 (48 h); 300 μg per nostril DEP (with and without O 3 pre-treatment) were intranasally instilled, usi ng the saline vehicle as control. Subjects did not serve as their own controls (not a crossover design).

Time to analysis: 4 and 96 h post-instillation. Exposure to DEP was not observed to alter markers of inflammation in nasal lavage fluid (e.g., cell counts, IL-8, IL-6) at 4 or 96 h post-instillation.

Reference: Larsson et al. (2007, 091375 )

Subjects: 16 healthy adults

Gender: 10 M/6 F

Age: 19-59 yr Traffic particles (road tunnel)

Particle Size: PM 2.5 , PM 10 ; PM 2.5 mass constituted ~36% of PM 10 mass

Particle Number/Count: 20- 1,000 nm: 1.1 × 10 5 /cm 3 , < 100 nm: 0.85 × 10 5 /cm 3

Concentration: PM 2.5 - 46-81 μg/m 3 ; PM 10 - 130-206 μg/m 3 Exposures were conducted for 2 h with intermittent exercise in a room adjacent to a busy road tunnel. Study used a randomized crossover design with each subject also exposed to normal air (control). Exposures were separated by 3-10wks. No exposures to filtered air were conducted. Other traffic emissions measured: NO (874 μg/m 3 ), NO 2 (230 μg/m 3 ), CO (5.8 μg/m 3 reported, likely 5.8 mg/m 3 ).

Time to analysis: 14 h post-exposure. An increase in bronchoalveolar lavage fluid cell number, lymphocytes, and alveolar macrophages were observed 14 h after road tunnel exposure relative to control. Traffic particulate exposure was not shown to effect cytokine or adhesion molecule expression in bronchial tissues. Respiratory symptoms were reported to increase during exposure to road tunnel air relative to pre-exposure symptom ratings. Exposure to road tunnel air was not shown to affect lung function.



### December 2009 C-11

### Reference Pollutant Exposure Findings

Reference: Mudway et al. (2004, 180208 )

Subjects: 25 healthy adults

Gender: 16 M/9 F

Age: 19-42 yr DE

Idling 1991 Volvo diesel engine (TD45, 4.5 L, 4 cylinders, 680 rpm)

Concentration: PM 10 100 μg/m 3 Subjects exposed to DE and filtered air for 2 h with intermittent exercise (15-min periods) in a randomized crossover design. Exposures were separated by at least 3 wk. Other diesel emissions measured: NO 2 (0.2 ppm), NO (0.6 ppm), CO (1.7 ppm), total hydrocarbons (1.4 ppm), formaldehyde (43.5 μg/m 3 ).

Time to analysis: 1 h after the start of exposure,immediately following exposure, and 6 h postexposure. DE caused mild throat irritation in some subjects and a significant increase in airways resistance (Raw) during or immediately following exposure. No changes in FEV 1 or FVC were observed following exposure to diesel. Neutrophil numbers in the bronchial airways tended to increase following exposure to DE; however, this increase was highly variable between subjects and did not reach statistical significance. Exposure to DE did not affect levels of SOD or malondialdehyde in the airways. An increase in levels of ascorbate and GSH in nasal lavage fluid was observed 6 h following exposure to DE.

Reference: Pietropaoli et al. (2004, 156025 )

Subjects: 16 asthmatic adults, 40 healthy adults

Gender: Asthmatic: 8 M/8 F, Healthy: 20 M/20 F

Age: 18-40 yr Ultrafine EC

Particle Size: CMD ~25 nm

Particle Number/Count: 10 μg/m 3 : ~2.0 × 10 6 /cm 3 ; 25 μg/m 3 : ~7.0 × 10 6 /cm 3 ; 50 μg/m 3 : ~10.8 × 10 6 /cm 3

Concentration: 10, 25, and 50 μg/m 3 Study conducted using a randomized crossover design with 2-h exposures. Asthmatics (n = 16) exposed to filtered air and 10 μg/m 3 . 12 healthy adults exposed to filtered air and 10 μg/m 3 at rest; 12 healthy adults exposed to filtered air, 10 and 25 μg/m 3 with intermittent exercise; 16 healthy adults exposed to filtered air and 50 μg/m 3 with intermittent exercise. Exposures were conducted via mouthpiece.

Time to analysis: Immediately followingexposure as well as 3.5, 21, and 45 h postexposure. No PM-induced changes in eNO or cell counts, IL-6, or IL-8 in induced sputum were observed in any of the protocols 21 h following exposure. Ultrafine carbon was not observed to increase respiratory symptoms in any of the study protocols. Healthy adults experienced an ultrafine PM-induced reduction in maximal mid-expiratory flow and CO diffusing capacity relative to filtered air 21 h following exposure.

Reference: Pourazar et al. (2005, 088305 )

Subjects: 15 healthy adults

Gender: 11 M/4 F

Age: 21-28 yr DE

Idling Volvo diesel engine

Particle Number/Count: 4.3 × 10 6 /cm 3

Concentration: PM 10 300 μg/m 3 Subjects exposed to DE and filtered air for 1 h with intermittent exercise (randomized crossover study design). Other diesel emissions measured: NO 2 (1.6 ppm), NO (4.5 ppm), CO (7.5 ppm), total hydrocarbons (4.3 ppm), formaldehyde (0.26 mg/m 3 ).

Time to analysis: 6 h post-exposure. Exposure to DE significantly increased nuclear translocation of NF- κ B, AP-1, phosphorylated p38, and phosphorylated JNK in bronchial epithelium 6 h post-exposure.

Reference: Pourazar et al. (2008, 156884 )

Subjects: 15 healthy adults

Gender: 11 M/4 F

Age: 21-28 yr DE

Idling Volvo diesel engine

Particle Number/Count: 4.3 × 10 6 /cm 3

Concentration: PM 10 300 μg/m 3 Subjects exposed to DE and filtered air for 1 h with intermittent exercise (randomized crossover study design). Other diesel emissions measured: NO 2 (1.6 ppm), NO (4.5 ppm), CO (7.5 ppm), total hydrocarbons (4.3 ppm), formaldehyde (0.26 mg/m 3 ).

Time to analysis: 6 h post-exposure. Exposure to DE observed to enhance epidermal growth factor receptor (EGFR) expression in bronchial epithelium 6 h post-exposure.

Reference: Riechelmann et al. (2004, 180120 )

Subjects: 30 healthy adults

Gender: 11 M/19 F

Age: 22-32 yr Urban dust

NIST SRM 1649a

Concentration: 150, 500 μg/m 3 Subjects exposed to both concentrations of urban dust (nose only exposure system) as well as filtered air for 3h at rest in a randomized crossover design. Exposures were separated by at least 1 wk.

Time to analysis: 30 min, 8 h, and 24 h postexposure. An increase in nasal secretion (nasal cytology) of IL-6 and IL-8 were observed 24 h after exposure to 500 μg/m 3 urban dust.

Reference: Samet et al. (2007, 156940 )

Subjects: Ultrafine CAPs: 20 healthy adults, Coarse CAPs: 14 healthy adults

Gender: Ultrafine CAPs: 11 M/9 F, Coarse CAPs: 8 M/6 F

Age: 18-35 yr CAPs (Chapel Hill, NC)

Particle Size: Ultrafine: 0.049 ± 0.009 μm, Coarse: 3.59 ± 0.58 μm

Concentration: Ultrafine: 47.0 ± 20.2 μg/m 3 , Coarse: 89.0 ± 49.5 μg/m 3 Preliminary report comparing effects of controlled exposures to coarse, fine, and ultrafine CAPs among healthy adults (3 separate studies). A randomized crossover design was used in evaluating effects of coarse CAPs (n=14) and ultrafine CAPs (n=20) relative to filtered air following of 2-h exposures with intermittent exercise. Results compared with previous study of controlled exposure to fine CAPs (Chapel Hill, NC) where subjects did not serve as their own controls (Ghio et al., 2000, 012140 )

Time to analysis: 0-20 h post-exposure. As was shown with fine CAPs, exposure to coarse CAPs increased the percentage of neutrophils in bronchoalveolar lavage fluid 20 h following exposure. Unlike fine CAPs, coarse CAPs did not increase the percent of monocytes in bronchoalveolar lavage fluid. Ultrafine CAPs were not shown to affect any markers of pulmonary inflammation in bronchoalveolar lavage fluid 18 h after exposure. No CAPs-induced changes in lung function were observed.

Reference: Samet et al. (2009, 191913 )

Subjects: 19 healthy adults

Gender: 10 M/9 F

Age: 18-35 yr Ultrafine CAPs (Chapel Hill, NC)

Particle Size: < 0.16 μm

Particle Number/Count: 120,662 ± 48,232 particles/cm 3

Concentration: 49.8 ± 20 μg/m 3 Subjects exposed for 2 h with intermittent 15 periods of exercise to UF CAPs and filtered air using a randomized crossover study design.

Time to analysis: Immediately following exposure and 1 and 18 h post-exposure. No effect of UF CAPs observed on pulmonary function immediately following exposure or 18 h post-exposure. IL-8 in BAL fluid was observed to increase with UF CAPs 18 h post-exposure. UF CAPs was not shown to alter any other inflammatory markers in BAL fluid.



### December 2009 C-12

### Reference Pollutant Exposure Findings

Reference: Schaumann et al. (2004, 087966 )

Subjects: 12 healthy adults

Gender: 4 M/8 F

Age: Avg 27 ± 2.5 yr Fine PM

Collected (filter) from industrialized and non-industrialized areas in Germany

Concentration: 100 μg per subject Bronchoscopic instillation of particles collected from both areas was conducted in contralateral lung segments for each subject.

Time to analysis: 24 h post-instillation. Particles collected from the industrialized area (transition metal-rich) increased the percentage of monocytes and oxidant radical generation in bronchoalveolar lavage fluid 24 h after exposure compared with PM containing less metal.

Reference: Stenfors et al. (2004, 157009 )

Subjects: 15 asthmatic adults, 25 healthy adults

Gender: Asthmatic: 10 M/5 F, Healthy: 16 M/9 F

Age: Asthmatic: 22-52 yr, Healthy:19-42 yr DE

Volvo diesel engine

Concentration: PM 10 108 μg/m 3 Subjects were exposed for 2 h with intermittent exercise to DE and filtered air using a randomized crossover study design. Other diesel emissions measured: NO 2 (0.7 ppm).

Time to analysis: 1 h after the start of exposure,immediately following exposure, and 6 h postexposure. DE was observed to increase neutrophilia and IL-8 in bronchial lavage fluid among healthy subjects 6 h after exposure. Among asthmatic subjects, exposure to DE did not cause anincrease in inflammatory markers. No dieselinduced change in pulmonary function was observed during or immediately following
exposure.

Reference: Tunnicliffe et al. (2003, 088744 )

Subjects: 12 asthmatic adults, 12 healthy adults

Gender: Asthmatics: 7 M/5 F, Healthy: 5 M/7 F

Age: Asthmatics: avg 35.7 yr, Healthy: avg 34.5 yr Aerosols of ammonium bisulfate and sulfuric acid

Particle Size: MMD 0.3 μm

Concentration: 200, 2,000 μg/m 3 Subjects were exposed for 1 h at rest to ammonium bisulfate (low and high concentrations), sulfuric acid (low and high concentrations) and filtered air in a randomized crossover design. Exposures were separated by at least 2 wk and were conducted using a head dome exposure system.

Time to analysis: Immediately following exposure as well as 5.5-6 h post-exposure. Neither ammonium bisulfate nor aerosolized sulfuric acid were observed to affect lung function or respiratory systems following exposures to 200 or 2,000 μg/m 3 among healthy or asthmatic adults. Exposures to ammonium bisulfate at both concentrations resulted in a significant increase in eNO in the asthmatic subjects.



# Table C- 3. Central nervous system effects.

Reference Pollutant Exposure Findings

Reference: Cruts et al. (2008, 156374 )

Subjects: 10 healthy adults

Gender: M

Age: 18-39 yr DE

Idling Volvo diesel engine (TD45, 4.5 L, 4 cylinders, 680 rpm)

Particle Number/Count: 1.2 × 10 6 /c m 3

Concentration: 300 μg/ m 3 Subjects were exposed to DE and filtered air for 1 h at rest in a randomized crossover study design. Exposures were separated by 2-4 days. Other diesel emissions measured: NO 2 (1.6 ppm), NO (4.5 ppm), CO (7.5 ppm), total hydrocarbons (4.3 ppm).

Time to analysis: From onset of exposure until 1 h post-exposure. Exposure to DE was observed to significantly increase the median power frequency (MPF) in the frontal cortex during exposure, as well as in the hour following the completion of the
exposure.



### December 2009 C-13

# Annex C References

Alexis NE; Lay JC; Zeman K; Bennett WE; Peden DB; Soukup JM; Devlin RB; Becker S (2006). Biological material on inhaled coarse fraction particulate matter activates airway phagocytes in vivo in healthy volunteers. J Allergy Clin Immunol, 117: 1396-1403. 154323

Barregard L; Sallsten G; Andersson L; Almstrand AC; Gustafson P; Andersson M; Olin AC (2008). Experimental exposure to wood smoke: effects on airway inflammation and oxidative stress. Occup Environ Med, 65: 319-324. 155675

Barregard L; Sallsten G; Gustafson P; Andersson L; Johanss on L; Basu S; Stigendal L (2006). Experimental exposure to wood-smoke particles in health humans: effects on markers of inflammation, coagulation, and lipid peroxidation. Inhal Toxicol, 18: 845-853. 091381

Bastain TM; Gilliland FD; Li Y-F; Saxon A; Diaz-Sanchez D (2003). Intraindividual reproducibility of nasal allergic responses to diesel exhaust particles indicates a susceptible phenotype. Clin Immunol, 109: 130-136. 098690

Beckett WS; Chalupa DF; Pauly-Brown A; Speers DM; Stewart JC; Frampton MW; Utell MJ; Huang LS; Cox C; Zareba W; Oberdorster G (2005). Compar ing inhaled ultrafine versus fine zinc oxi de particles in healthy adults - A human inhalation study. Am J Respir Crit Care Med, 171: 1129-1135. 156261

Behndig AF; Mudway IS; Brown JL; Stenfors N Helleday R Duggan ST; Wilson SJ; Boman C Cassee FR; Frew AJ; Kelly FJ; Sandstrom T Blomberg A (2006). Airway antioxidant and inflammatory responses to diesel exhaust exposure in healthy humans. Eur Respir J, 27: 359-365. 088286

Blomberg A; Tornqvist H; Desmyter L; Deneys V; Hermans C (2005). Exposure to diesel exhaust nanoparticles does not induce blood hypercoagulability in an at-risk population. J Thromb Haemost, 3: 2103-2105. 191991

Bosson J; Barath S; Pourazar J; Behndig AF; Sandstrom T; Blomberg A; Adelroth E (2008). Diesel exhaust exposure enhances the ozone-induced airway inflammation in healthy humans. Eur Respir J, 31: 1234-40. 196659

Bosson J; Pourazar J; Forsberg B; Adelroth E; Sandstrom T; Blomberg A (2007). Ozone enhances the airway inflammation initiated by diesel exhaust. Respir Med, 101: 1140-1146. 156286

Brauner EV; Forchhammer L; Moller P; Barregard L; Gunnarsen L; Afshari A; Wahlin P; Glasius M; Dragsted LO; Basu S; Raaschou-Nielsen O; Loft S (2008). Indoor particles affect vascular function in the aged: an air filtration-based intervention study. Am J Respir Crit Care Med, 177: 419-425. 156293

Brauner EV; Forchhammer L; Moller P; Simonsen J; Glasius M; Wahlin P; Raaschou-Nielsen O; Loft S (2007). Exposure to ultrafine particles from ambient air and oxidative st ress-induced DNA damage. Environ Health Perspect, 115:
1177-82. 091152

Brauner EV; Mortensen J; Moller P; Bernard A; Vinzents P; Wahlin P; Glasius M; Loft S (2009). Effects of ambient air particulate exposure on blood-gas barrier permeabili ty and lung function. Inhal Toxicol, 21: 38-47. 190244

Bräuner EV; Møller P; Barregard L; Dragsted LO; Glasius M; Wåhlin P; Vinzents P; Raaschou-Nielsen O; Loft S (2008). Exposure to ambient concentrations of particulate air pollution does not influence vascular function or inflammatory pathways in young healthy individuals. Part Fibre Toxicol, 5: 13. 191966

Brook RD; Brook JR; Urch B; Vincent R; Rajagopalan S; Silv erman F (2002). Inhalation of fine particulate air pollution and ozone causes acute arterial vasoconstriction in healthy adults. Circulation, 105: 1534-1536. 024987

Carlsten C; Kaufman JD; Trenga CA; Allen J; Peretz A; Sullivan JH (2008). Thrombotic markers in metabolic syndrome subjects exposed to diesel exhaust. Inhal Toxicol, 20: 917-921. 156323

Carlsten C; Kaufman Joel D; Peretz A; Trenga Carol A; Sheppard L; Sullivan Jeffrey H (2007). Coagulation markers in healthy human subjects exposed to diesel exhaust. Thromb Res Suppl, 120: 849-855. 155714

Cruts B; van Etten L; Tornqvist H; Blomberg A; Sandstrom T; Mills NL; Borm PJ (2008) . Exposure to diesel exhaust induces changes in EEG in human volunt eers. Part Fibre Toxicol, 5: 4. 156374

Note: Hyperlinks to the reference citations throughout this doc ument will take you to the NCEA HERO database (Health and Environmental Research Online) at http://epa.gov/hero . HERO is a database of scientific literature used by U.S. EPA in the process of developing science assessments such as the Integrated Science Assessments (ISA) and the Integrated Risk Information System (IRIS)

.



### December 2009 C-14

### Danielsen PH; Brauner EV; Barregard L; Sallsten G; Wall in M; Olinski R; Rozalski R; Moller P; Loft S (2008). Oxidatively damaged DNA and its repair after experiment al exposure to wood smoke in healthy humans. Mutat Res Fund Mol Mech Mutagen, 642: 37-42. 156382

Devlin RB; Ghio AJ; Kehrl H; Sanders G; Cascio W (2003). El derly humans exposed to concentrated air pollution particles
have decreased heart rate variab ility. Eur Respir J, 40: 76S-80S. 087348

Fakhri AA; Ilic LM; Wellenius GA; Urch B; Silverman F; Gold DR; Mittleman MA (2009). Autonomic effects of controlled fine particulate exposure in young healthy adults: Effect modification by ozone. Environ Health Perspect, 117: 1287-1292. 191914

Frampton MW; Stewart JC; Oberdorster G; Morrow PE; Chalupa D; Pietropaoli AP; Frasier LM; Speers DM; Cox C; Huang LS; Utell MJ (2006). Inhalation of ultrafine pa rticles alters blood leukocyte expression of adhesion molecules in humans. Environ Health Perspect, 114: 51-58. 088665

Ghio AJ; Kim C; Devlin RB (2000). Concentrated ambient ai r particles induce mild pulmonary inflammation in healthy human volunteers. Am J Respir Crit Care Med, 162: 981-988. 012140

Gilliland FD; Li YF; Saxon A; Diaz-Sanchez D (2004). Effect of glutathione-S-transferase M1 and P1 genotypes on xenobiotic enhancement of allergic responses: randomised , placebo-controlled crossover study. Lancet, 363: 119-
125. 156471

Gong H Jr; Linn WS; Clark KW; Anderson KR; Geller MD; Siout as C (2005). Respiratory responses to exposures with fine particulates and nitrogen dioxide in the elderl y with and without COPD. Inhal Toxicol, 17: 123-132. 087921

Gong H Jr; Linn WS; Clark KW; Anderson KR; Sioutas C; Al exis NE; Cascio WE; Devlin RB (2008). Exposures of healthy and asthmatic volunteers to concentrated ambient ul trafine particles in Los Angeles. Inhal Toxicol, 20: 533-
545. 156483

Gong H Jr; Linn WS; Terrell SL; Anderson KR; Clark KW; Sioutas C; Cascio WE; Alexis N; Devlin RB (2004). Exposures of elderly volunteers with and without chronic obstructive pulmonary disease (COPD) to concentrated ambient fine particulate pollution. Inha l Toxicol, 16: 731-744. 087964

Gong H Jr; Linn WS; Terrell SL; Clark KW; Geller MD; Anders on KR; Cascio WE; Sioutas C (2004). Altered heart-rate variability in asthmatic and healthy volunteers exposed to c oncentrated ambient coarse particles. Inhal Toxicol, 16:
335-343. 055628

Graff D; Cascio W; Rappold A; Zhou H; Huang Y; Devlin R ( 2009). Exposure to concentrated coarse air pollution particles causes mild cardiopulmonary effects in healthy young adults . Environ Health Perspect, 117: 1089-1094. 191981

Huang Y-CT; Ghio AJ; Stonehuerner J; McGee J; Carter JD; Grambow SC; Devlin RB (2003). The role of soluble components in ambient fine particles-induced changes in human lungs and blood. Inhal Toxicol, 15: 327-342.
087377

Kongerud J; Madden MC; Hazucha M; Peden D (2006). Nasal re sponses in asthmatic and nonasthmatic subjects following exposure to diesel exhaust partic les. Inhal Toxicol, 18: 589-594. 156656

Larsson B-M; Sehistedt M; Grunewald J; Skold CM; Lundin A; Blomberg A; Sandstrom T; Eklund A; Svartengren M (2007). Road tunnel air pollution induces bronchoalveolar infl ammation in healthy subjects. Eur Respir J, 29: 699-
705. 091375

Lucking A; Lundback M; Mills N; Faratian D; Barath S; Pourazar J; Cassee F; Donaldson K; Boon N; Badimon J; Sandstorm T; Blomberg A; Newby D (2008). Diesel exhaus t inhalation increases thrombus formation in man. Eur Heart J, 29: 3043-3051. 191993

Lund AK; Lucero J; Lucas S; Madden MC; McDonald JD; Seagrave JC; Knuckles TL; Campen MJ (2009). Vehicular emissions induce vascular MMP-9 expression and activity associated with endothelin-1 mediated pathways. Arterioscler Thromb Vasc Biol, 29: 511-517. 180257

Lundbäck M; Mills NL; Lucking A; Barath S; Donaldson K; Newby DE; Sandström T; Blomberg A (2009). Experimental exposure to diesel exhaust increases arterial st iffness in man. Part Fibre Toxicol, 6: 7. 191967

Mills NL; Robinson SD; Fokkens PH; Leseman DL; Miller MR; Anderson D; Freney EJ; Heal MR; Donovan RJ; Blomberg A; Sandstrom T; MacNee W; Boon NA; Dona ldson K; Newby DE; Cassee FR (2008). Exposure to concentrated ambient particles does not affect vascular f unction in patients with cor onary heart disease. Environ Health Perspect, 116: 709-715. 156766

December 2009 C-15

### Mills NL; Törnqvist H; Gonzalez MC; Vink E; Robinson SD; Soderberg S; Boon NA; Donaldson K; Sandstrom T; Blomberg A; Newby DE (2007). Ischemic and thrombotic ef fects of dilute diesel-exhaust inhalation in men with coronary heart disease. N Engl J Med, 357: 1075-1082. 091206

Mills NL; Tornqvist H; Robinson SD; Gonzalez M; Darnley K; MacNee W; Boon NA; Donaldson K; Blomberg A; Sandstrom T; Newby DE (2005). Diesel exhaust inhalati on causes vascular dysfunction and impaired endogenous fibrinolysis. Circulation, 112: 3930-3936. 095757

Mudway IS; Stenfors N; Duggan ST; Roxborough H; Zielinski H; Marklund SL; Blomberg A; Frew AJ; Sandstrom T; Kelly FJ (2004). An in vitro and in vivo investigation of the effects of diesel exhaust on human airway lining fluid antioxidants. Arch Biochem Biophys, 423: 200-212. 180208

Peretz A; Kaufman JD; Trenga CA; Allen J; Carlsten C; Aule t MR; Adar SD; Sullivan JH (2008). Effects of diesel exhaust inhalation on heart rate variability in human volunteers. Envi ron Res, 107: 178-184. 156855

Peretz A; Peck EC; Bammler TK; Beyer RP; Sullivan JH; Trenga CA; Srinouanprachnah S; Farin FM; Kaufman JD (2007). Diesel exhaust inhalation and assessment of peripheral blood mononuclear cell gene transcription effects: an exploratory study of healthy human vol unteers. Inhal Toxicol, 19: 1107-1119. 156853

Peretz A; Sullivan JH; Leotta DF; Trenga CA; Sands FN; Allen J; Carlsten C; Wilkinson CW; Gill EA; Kaufman JD (2008). Diesel exhaust inhalation elicits acute vasoconstriction in vivo. Environ Health Perspect, 116: 937-942.
156854

Pietropaoli AP; Frampton MW; Hyde RW; Morrow PE; Ober dorster G; Cox C; Speers DM; Frasier LM; Chalupa DC; Huang LS; Utell MJ (2004). Pulmonary function, diffusi ng capacity, and inflammation in healthy and asthmatic subjects exposed to ultrafine part icles. Inhal Toxicol, 16: 59-72. 156025

Pourazar J; Blomberg A; Kelly FJ; Davies DE; Wilson SJ; Holgate ST; Sandstrom T (2008) . Diesel exhaust increases EGFR and phosphorylated C-terminal Tyr 1173 in the br onchial epithelium. Part Fibre Toxicol, 5: 8. 156884

Pourazar J; Mudway IS; Samet JM; Helleday R; Blomberg A; Wilson SJ; Frew AJ; Kelly FJ ; Sandstrom T (2005). Diesel exhaust activates redox-sensitive transcription factors a nd kinases in human airways. Am J Physiol, 289: L724-
L730. 088305

Power K; Balmes J; Solomon C (2008). Controlled exposure to combined particles and ozone decreases heart rate variability. J Occup Environ Med, 50: 1253-1260. 191982

Riechelmann H; Rettinger G; Lautebach S; Schmittinger S; Deut schle T (2004). Short-term e xposure to urban dust alters the mediator release of human nasal mucosa. J Occup Environ Med, 46: 316-322. 180120

Routledge HC; Manney S; Harrison RM; Ayres JG; Townend JN (2006). Effect of inhaled sulphur dioxide and carbon particles on heart rate variability and markers of infla mmation and coagulation in human subjects. Heart, 92: 220-

227. 088674

Rundell KW; Caviston R (2008). Ultrafine and fine particulate matter inhalation decreases exercise performance in healthy subjects. J Strength Cond Res, 22: 2-5. 191986

Samet JM; Graff D; Berntsen J; Ghio AJ; Huang YC; Dev lin RB (2007). A comparison of studies on the effects of controlled exposure to fine, coarse and ultrafine ambient particulate matter from a single location. Inhal Toxicol, 19 Suppl 1: 29-32. 156940

Samet JM; Rappold A; Graff D; Cascio WE; Berntsen JH; Huang YC; Herbst M; Bassett M; Montilla T; Hazucha MJ; Bromberg PA; Devlin RB (2009). Concentrated ambient u ltrafine particle exposure induces cardiac changes in young healthy volunteers. Am J Respir Crit Care Med, 179: 1034-1042. 191913

Schaumann F; Borm PJA; Herbrich A; Knoch J; Pitz M; Schins RPF; Luettig B; Hohlfeld JM; Heinrich J; Krug N (2004). Metal-rich ambient particles (particulate matter 2.5) cause airway inflammation in healthy subjects. Am J Respir Crit Care Med, 170: 898-903. 087966

Shah AP; Pietropaoli AP; Frasier LM; Speers DM; Chalupa DC; Delehanty JM; Huang LS; Utell MJ; Frampton MW (2008). Effect of inhaled carbon ultr afine particles on reactive hyperemia in healthy human subjects. Environ Health Perspect, 116: 375-380. 156970

Stenfors N; Nordenhall C; Salvi SS; Mudway I; Soderberg M; Blomberg A; Helleday R; Levin JO; Holgate ST; Kelly FJ; Frew AJ; Sandstrom T (2004). Different airway inflammato ry responses in asthmatic and healthy humans exposed to diesel. Eur Respir J, 23: 82-86. 157009

December 2009 C-16

### December 2009 C-17 Tornqvist H; Mills NL; Gonzalez M; Miller MR; Robinson SD; Megson IL; MacNee W; Donaldson K; Soderberg S; Newby DE; Sandstrom T; Blomberg A (2007). Persistent endothelial dysfuncti on in humans after diesel exhaust inhalation. Am J Respir Crit Care Med, 176: 395-400. 091279

Tunnicliffe WS; Harrison RM; Kelly FJ; Dunster C; Ayres JG (2003). The effect of sulphurous air pollutant exposures on symptoms, lung function, exhaled nitric oxide, and nasal ep ithelial lining fluid antioxidant concentrations in normal and asthmatic adults. Occup Environ Med, 60: 1-7. 088744

Urch B; Brook JR; Wasserstein D; Brook RD; Rajagopalan S; Corey P; Silverman F (2004). Relative contributions of
PM2.5 chemical constituents to acute arterial vaso constriction in humans. I nhal Toxicol, 16: 345-352. 055629

Urch B; Silverman F; Corey P; Brook JR; Lukic KZ; Rajagopa lan S; Brook RD (2005). Acute blood pressure responses in healthy adults during controlled air pollution ex posures. Environ Health Perspect, 113: 1052-1055. 081080

Zareba W; Couderc JP; Oberdörster G; Chalupa D; Cox C; Huang LS; Peters A; Utell MJ; Frampton MW (2009). ECG parameters and exposure to carbon ultrafine particles in young healthy subjects. Inhal Toxicol, 21: 223-233. 190101