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1 Praeger Handbook of Environmental Health Robert H. Friis, Editor Volume III: Water Quality, Air Quality, and Solid Waste Disposal Chapter 8 Potential Health Effects of Exposure to Diesel Engine Exhausts Dr. Thomas W. Hesterberg, MA, PhD, MBA Director, Product Stewardship and Environmental Health Navistar 4201 Winfield Road Warrenville, IL 60555 Phone: 312-927-2697 Email: [email protected] Dr. William B. Bunn, MPH, MD, JD Vice President, Health, Safety, Security and Productivity Navistar 4201 Winfield Road Warrenville, IL 60555 Phone: 630-753-2301 Email: [email protected] Dr. Peter A. Valberg, MA, MS, PhD Gradient 20 University Road Cambridge, MA 02138-5756 Phone: 617-395-5570 Email: [email protected] Dr. Christopher M. Long, SM, ScD Gradient 20 University Road Cambridge, MA 02138-5756 Phone: 617-395-5532 Email: [email protected]

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Praeger Handbook of Environmental Health

Robert H. Friis, Editor

Volume III: Water Quality, Air Quality, and Solid Waste Disposal

Chapter 8

Potential Health Effects of Exposure to Diesel Engine Exhausts

Dr. Thomas W. Hesterberg, MA, PhD, MBA

Director, Product Stewardship and Environmental Health

Navistar

4201 Winfield Road

Warrenville, IL 60555

Phone: 312-927-2697

Email: [email protected]

Dr. William B. Bunn, MPH, MD, JD

Vice President, Health, Safety, Security and Productivity

Navistar

4201 Winfield Road

Warrenville, IL 60555

Phone: 630-753-2301

Email: [email protected]

Dr. Peter A. Valberg, MA, MS, PhD

Gradient

20 University Road

Cambridge, MA 02138-5756

Phone: 617-395-5570

Email: [email protected]

Dr. Christopher M. Long, SM, ScD

Gradient

20 University Road

Cambridge, MA 02138-5756

Phone: 617-395-5532

Email: [email protected]

2

Abstract

Diesel exhaust (DE) is a complex mixture of particulate matter, condensible vapors, and

gases, with a composition that can vary depending upon engine type, fuel type, and operating

conditions. Elevated exposure levels of DE have been linked with both cancer and non-cancer

health risks, primarily in studies of exhaust from diesel technology typical of the engines, fuels,

and lubricants in use prior to 1988 when diesel-engine emissions were not regulated in the

United States. Prompted by increasingly stringent emission standards implemented by the US

Environmental Protection Agency over the last two decades, major advances have occurred in

diesel technology that have resulted in quantitative and qualitative changes in diesel-engine

emissions, including dramatic reductions in emitted diesel exhaust particulate (DEP), nitrogen

oxides (NOx), and other components.

1.0 Diesel-Engine Exhaust: Introduction, Nature, and Human Exposure Potential

Diesel engines differ from spark-ignition, gasoline engines in that the fuel-air mixture is

ignited by the heat of compression. Diesel engines combust hydrocarbon fuels with oxygen to

produce mechanical energy, and thus, at the fundamental level, the products of combustion found

in "diesel exhaust" (DE) are gaseous carbon dioxide (CO2) and water (H2O). Because

combustion is generally incomplete, and because diesel fuel contains some level of impurities,

many other chemical compounds are present in DE, including a small quantity of airborne

particulate matter (PM), called "diesel exhaust particulate matter" (DEP).

3

At the outset, it should be pointed out that diesel-engine exhaust, its composition, and its

potential health effects have been the subject of frequent review. Among the major reviews is

the 2002 US EPA review entitled "Health Assessment Document for Diesel Exhaust," a 671-

page analysis (1). Additional, contemporary reviews include Hesterberg et al. 2006, 2009a,

2009b, 2010; Gamble, 2010; Institute of Medicine, 2005; Mauderly and Garshick, 2009; Pronk et

al. 2009; and Wichmann, 2006 (2-10). Older major reviews include the 1989 International

Agency for Research on Cancer (IARC) monograph (11), the 1995 report by the Health Effects

Institute (12), and the health-assessment documents developed by the California EPA (13).

In practice, DE is a complex mixture of particulate aerosol – DEP, condensible vapors,

and gases. DE composition varies widely depending upon engine type, fuel type, and operating

conditions. Data on the potential health effects of DE inhalation are available mostly for what

has been termed "traditional diesel exhaust," or TDE (2, 3, 14). TDE refers to the exhaust from

diesel technology typical of the engines, fuel injection systems, fuels, and lubricants in use prior

to 1988, when exhaust emissions from diesel were not regulated in the US.

Stimulated by progressively more stringent DE regulatory standards, major advances

have occurred in diesel technology over the past two decades that have resulted in quantitative

and qualitative changes in diesel-engine emissions, including dramatic reductions in emitted

DEP, nitrogen oxides (NOx), and other components. Present-day engines emit "new technology

diesel exhaust" (NTDE), where NTDE refers to the diesel exhaust from modern integrated

systems (engines, fuel injection systems, ultra-low-sulfur fuels, lubricants, and exhaust after-

treatment devices) manufactured and sold in the post-2006 era to meet the stringent US

4

Environmental Protection Agency (US EPA) emissions standards for heavy-duty highway diesel

engines. Both DE from post-2006 diesel engines, as well as DE from engines retrofitted with a

diesel particulate filter (DPF) and operated with ultra-low sulfur diesel fuel, are considered to be

representative of NTDE, providing that they can achieve the 2007 US EPA emissions standard

for DEP.

The US EPA-mandated changes in DEP and NOx emissions over time are illustrated in

Figure 1. In addition to > 90% reductions in DEP mass, the chemical composition of the DEP

and gases released from NTDE are qualitatively and quantitatively different from TDE (15, 16).

With NTDE, emissions of DEP and NOx are over 99% lower than emissions from engines prior

to regulations. Other aspects of NTDE are summarized elsewhere (15, 16). It is important to

note that most of the available health-effects research does not provide accurate information for

assessment of human health effects that might be associated with current and future exposures to

NTDE.

5

Figure 1. Emissions reductions due to US EPA standards for DEP from heavy-duty diesel

trucks (t) or urban buses (ub), relative to pre-1988 emissions. Panel A, DEP; Panel B,

NOx (data from (1); figures adapted by permission from Informa Healthcare: Critical

Reviews in Toxicology (2)).

US EPA standards for particulate emissions from heavy duty diesel trucks (t) or urban buses (ub),

calculated as grams particulate matter emitted per brake-horsepower-hour (g/b hp-hr) and adjusted

relative to pre-1988 unregulated engine emissions. From: US EPA Health Assessment Document for

Diesel Engine Exhaust, May 2002. Table 2-4, P. 2-16.

Part

icula

tes E

mis

sio

ns,

g/b

hp

-hr

(rela

tive t

o u

nre

gula

ted)

100%

60%

25%

10%5%

1%

Unregulated 1988 t 1991 t 1994 t 1996 ub 2007 t

Fig. 1. Reductions in Diesel Particulate Matter Emissions in the US

Traditional Diesel Transitional Diesel New Technology

Exhaust (TDE) Exhaust Diesel Exhaust (NTDE)

Panel A

NO

x (g

/bh

p-h

r)

15

6

5

4

2

0.2

1977 1990 1991 1998 2004 2010

US EPA standards for NOx emissions from heavy duty diesel engines, calculated as grams NOx

emitted per brake-horsepower-hour (g/b hp-hr). From: USEPA Health Assessment Document for

Diesel Engine Exhaust, May 2002, Table 2-4, p. 2-16.

Model Year

Fig. 2. Reductions in Diesel Nitrogen Oxide (NOx) Emissions in the US

Panel B

6

The majority of our review thus relates to historical and present-day exposures to TDE,

given that most of the available DE toxicological data are for studies of pre-2006 diesel engines

lacking modern aftertreatment systems. TDE is composed of a gas-and-vapor phase, which is

typically composed of about 98% carbon dioxide and water vapor, and also includes NOx, sulfur

dioxide, carbon monoxide, and methane plus non-methane volatile organic carbon compounds.

The particulate phase of TDE, i.e., DEP, from a traditional diesel engine running on high sulfur

fuel typically represents less than 1% of the mass of total diesel exhaust emissions (17).

Exposures to TDE are generally described by giving only the DEP concentration, which may

leave the erroneous impression that any observed health effects are due exclusively to the DEP

alone. In fact, several studies have shown that the gaseous components of engine exhausts may

be more important than the particulate components (18-20). In any case, DEP is often used as a

surrogate measure for all the components in the DE mixture.

A major problem in estimating DEP exposure is that most areas where diesel engines are

in use also include airborne particles from many other non-diesel sources, including carbon

compounds from, e.g., tobacco smoke, gasoline engine exhaust, tire-wear dust, combustion of

wood, paper and waste, meat-cooking fumes, hydrocarbon solvents, deposited and resuspended

pollen, and asphalt dust. The exhausts of engines (both gasoline- and diesel-powered) share

similar physical and chemical characteristics with each other and with airborne materials from

many other combustion sources. There is no known marker for distinguishing DEP from other

types of carbon-based dust. Thus, it has been difficult, if not impossible, to quantify the portion

of an individual's total airborne PM exposure that derives from vehicle exhaust generally, and

even more difficult to quantify the portion that is specifically related to DE. Indeed, DE and

7

gasoline engine exhaust exposures are generally present together and cannot be clearly separated

(21).

US EPA has collected data on environmental concentrations of DEP derived from

extrapolations of measured elemental carbon (EC) and other markers, and the agency has also

generated estimates of DEP levels using a variety of source apportionment strategies (1, 22).

The agency has estimated that U.S. long-term average urban and rural exposure concentrations

from on-road vehicles and stationary sources range between approximately 0.2 and 3.0 µg/m3.

Historically, DEP concentrations in occupational environments have been considerably higher

than the ambient levels, as illustrated in Table 1.

Table 1. Approximate DEP concentrations in ambient and occupational environments where

TDE is, or was, historically, present (9, 1, 23, 24).

Exposure Environment DEP (µg/m3)

Long-term Average Ambient Levels < 0.2 – 3.0

Truckers 7 – 10

Railroad workers 40 – 70

US surface miners 90 – 100

US underground coal miners 500 – 700

US underground metal/mineral

miners

800 – 1,000

For the general public, short-term DEP exposure levels can be considerably higher than

long term average exposure levels, and in fact, many human controlled-exposure studies use

8

DEP exposure levels in the range of 100 to 300 µg/m3. The range of typical DEP concentrations

for exposures to humans (both in the laboratory and ambient settings) is illustrated in Figure 2

(note logarithmic scale of DEP concentrations).

Figure 2: Examples of short-term and long-term DEP concentrations relevant to general public

exposure (figure adapted by permission from Informa Healthcare: Critical Reviews in

Toxicology (3)). The Fruin et al. (25) data (7 – 23 µg/m3) represent measurements made

inside vehicles during highway driving. The McCreanor et al. (26) data (EC

concentrations, 4 – 16 µg/m3) represent 2-hr averages during weekday periods along

London's heavily trafficked Oxford Street. (Note logarithmic scale of DEP

concentrations.)

DE

P C

on

ce

ntr

atio

n (

g/m

3)

0.1

1

10

100

1000

Short-term GeneralPopulation ExposureLevels

Typical Range of DEP Exposure Levels in RecentHuman Controlled Exposure Studies

Fruin et al. (25)

US EPA(1); Hesterberget al. (2)

US EPA(22)

Long-term GeneralPopulation ExposureLevels

McCreanor et al.(26)

2.0 Health Effects of Diesel-Engine Exhaust: Potential for Cancer Risk

In 1989, IARC (11) found limited evidence from studies of humans for carcinogenicity of

DE (evidence primarily derived from studies of railroad workers). The IARC panel noted in the

monograph that, in the epidemiologic studies they reviewed, there were no direct data for the

workers' DE exposures; historical exposures were either assumed based on job title or were

9

estimated from more recent exposure assessments (11, p. 132). The IARC finding of sufficient

evidence from animal studies for carcinogenicity of DE was based primarily on studies in which

rats (but not mice or hamsters) developed tumors after lifetime inhalation of very high

concentrations of DE ( ≥ 2,200 μg/m3 DEP). At the time of the 1989 IARC review, the Working

Group was not able to consider the strong evidence that developed subsequently regarding the

fact that the lung tumors in rats are due to a rat-specific lung-clearance-overload mechanism

triggered by long-term elevated exposures to PM (2).

The US EPA reviewed the science regarding health risks to humans from DE in 2002, in

their "Health Assessment Document for Diesel Exhaust." This document included an extensive

analysis of "Carcinogenicity of Diesel Exhaust" (Chapter 7), with a primary focus on a possible

link to lung cancer risk. The agency concluded that the weight of the evidence supports a

"likely" role for DE in the risk of lung cancer, but assessment of the DE database led to the

conclusion that neither the existing epidemiological studies nor the laboratory animal data were

adequate to quantitatively predict potential human health effects from exposure to DE, or to link

DE to increases in lung cancer (1, 2, 10). When evaluating the evidence for causal associations

between DE exposure and cancers to sites other than the lungs, the US EPA concluded: "Quite a

few studies have examined DE for other effects such as bladder cancer, leukemia,

gastrointestinal cancers, prostate cancer, etc. The evidence for these effects is inadequate" (1, pp.

7-80).

The Institute of Medicine of the National Academies also reviewed the science regarding

health risks to humans from fuels and fuel combustion products (7). Although the IOM did not

10

draw a conclusion specific to DE, it did offer the conclusion that: "there is sufficient evidence of

an association between exposure to combustion products and lung cancer."

Of the occupations listed in Table 1, truckers and railroad workers have been the focus of

most epidemiology studies regarding DE-exposed populations. A significant problem with the

data on truckers is that increases in lung cancer were seen in this occupation prior to dieselization

(See Table 9 in (2)). As discussed subsequently, studies of underground miners have generally

been negative for lung cancer risk. Consequently, U.S. railroad workers have formed the basis

of most studies regarding DE-exposure and its possible relationship to lung cancer risk. These

include case-control and retrospective cohort mortality studies reported by Garshick and

collaborators (27-30) and a reanalysis of the epidemiologic data by Crump (31). The railroad

epidemiologic studies were based on computerized work records maintained since 1959 by the

U.S. Railroad Retirement Board (RRB). By 1959, the transition from coal-powered to diesel-

powered locomotives was 95% complete in the U.S. However, one of the issues in the railroad

worker data is that the lung cancer mortality results do not show a clear trend with increasing

DEP concentration when the cohort is examined by job category. This is summarized in Table 2,

and additional details are available in Hesterberg et al. (2).

11

Table 2: Railroad-worker personal exposures, by job category, to estimated DEP a (n = number

of measurements) versus lung cancer mortality relative risk (n = number of lung cancer

deaths), based on U.S. lung cancer mortality rates (31, 32).

Exposure Lung cancer mortality

Job Category

n

Estimated DEP a

μg/m3, geometric

mean

n

Relative Risk (RR)

Unexposed

Clerks 59 17 315 b 0.98

b

Signal men 13 49

Exposed

Engineers/Firemen 128 39 – 73 298 1.23

Conductors/Brakemen 158 52 – 92

437 1.20 c

Hostlers 8 191

Shop Workers 174 114 – 134

318 1.08 a For each air sample, estimated fraction of cigarette smoke was subtracted from total

respirable particulate matter; adjusted data are presumed to reflect the concentration of

airborne DEP (32). b

Clerks and signal men combined. c

Conductors/brakemen and hostlers combined.

In the late 1990s, the Health Effects Institute (HEI) convened a "Diesel Epidemiology

Expert Panel" (33) to review the railroad worker data and the various analyses of these data. The

Panel was assisted by leading investigators who had conducted epidemiologic research on DE,

and by expert analysts who had conducted critical reviews of that research. The Panel noted that

lung cancer mortality for train workers/riders (engineers, conductors) was higher than for

clerks/signalmen (unexposed), with shop workers (highest DEP exposure) exhibiting

intermediate risks (See Table 2, above). However, the Panel's analyses also demonstrated that

within each category of worker, the risk of lung cancer decreased with increasing duration of

employment, and, further, that the decrease was statistically significant for clerks/signalmen and

train workers. HEI's Diesel Epidemiology Expert Panel concluded that the data: "are not

12

consistent with a steadily increasing association between cumulative diesel exposure and lung

cancer risk." The Panel noted that the negative exposure-response relationship could be due to a

number of possible reasons, including unmeasured confounding variables and/or "other sources

of pollution" that could have been responsible for the observed effect. The HEI Panel also found

that the conclusions initially reached by Garshick and co-workers (i.e., a positive exposure-

response relationship) resulted entirely from baseline differences in risk between the two job

categories (train workers versus clerks/signalmen). The Panel concluded: "These patterns are

not consistent with a monotonically increasing association between cumulative exposure to

diesel exhaust and lung cancer risk. If risk increased consistently with increasing exposure, a

positive trend with duration of employment would be expected for the exposed groups (including

train workers), even if exposure magnitudes were incorrect." That is, the weight of evidence

suggested that, in the railroad-worker studies, any occupational increase in lung cancer risk

among train workers was not due to DE exposures.

Laden et al. (34) published a report that used historical data on dieselization of individual

railroads and emission factors for locomotives in each railroad for an innovative exposure

intensity characterization. These investigators calculated annual railroad-specific probabilities of

worker exposure to DE, going back to 1945. As expected, RRs for lung cancer remained

elevated, but there was no evidence of an exposure-response relationship using these improved

measures of exposure intensity. Another innovative approach to RR-worker data is a case-

control analysis of lung-cancer mortality (1981-1982) by Lee et al. (35), using a new type of

Kaplan-Meier survival plot in which the time scale represents a measure of disease progression

rather than mortality over calendar time. The authors reported that workers who had been

13

engineers or brakemen had a better initial health status, but a steeper rate of health decline than

workers who had never been engineers or brakemen. This steeper rate of decline was attributed

by the authors to DE exposure, however, no quantitative measures of DE exposure were used

other than the "job categories" from the original study (28), which were shown to be problematic

in the HEI (33) analysis.

In a coal-dust analysis relevant to DEP, IARC reviewed studies of coal miners and

determined that evidence was inadequate to show any lung cancer risk from coal dust exposure;

IARC classified coal dust as Group 3, i.e., "cannot be classified as to its carcinogenicity to

humans" (36). This conclusion is important with regard to DEP for two reasons: (1) The studies

in the IARC review included coal miner cohorts of workers who historically inhaled some of the

highest occupational concentrations of total airborne particulates among all working

environments, and this potential "lung overlead" did not lead to lung cancer in humans as it does

in rats. (2) Hundreds of underground coal mines, particularly outside the US, had used diesel

equipment for many decades, exposing those coal miners to considerable DEP as well as to coal

dust. As illustrated in Table 1, and stated in US EPA's HAD for diesel exhaust, "The highest

occupational exposures to DEP are for workers in coal and non-coal mines using diesel-powered

equipment" (1, pp. 2-107). Yet, as IARC found, lung cancer did not appear to be elevated.

Studies of underground miners have been recently reviewed by Hesterberg and

colleagues (2), and the results indicate that between the high levels of DEP exposure and the

adequacy of latency periods, lung cancer risk, if any, should have been readily identified. That

is, the underground mining studies are potentially much more powerful than studies of other

14

occupational groups. However, while not definitive, the underground miner studies provide no

convincing evidence of a risk of lung cancer associated with DEP, and are generally consistent

with no increased risk. Thus, even with DEP exposures that were an order of magnitude higher

than those in either the trucking or railroad industries studies, the mining studies are, on balance,

consistent with no detectable increase in risk.

For more than 10 years now, a long-standing collaboration between the National Institute

of Occupational Safety and Health (NIOSH) and the National Cancer Institute (NCI) has been

evaluating mortality from lung cancer and other diseases among U.S. nonmetal miners in relation

to quantitatively measured exposure to DE, using a cohort and nested case control study (37).

NIOSH researchers (38, 39) just recently reported findings characterizing current and past levels

of surrogates of diesel exhaust, deriving historical estimates of exposure to respirable elemental

carbon (REC). These exposures are being used to investigate potential exposure-response

relationships with lung cancer, but as of this writing (late 2010), no epidemiologic results have

been published by NIOSH or NCI.

Laboratory studies of exposure to TDE, both in vivo and in vitro, have limited relevance

in assessing the carcinogenic potential of TDE in humans. Laboratory rats, when exposed for a

lifetime to high levels of TDE (> 2,200 μg/m3), develop excess lung tumors; however such lung

tumor increases are consistently observed in rats exposed to overload levels of other types of fine

particles (TiO2, talc, and carbon black). Other species (mice and hamsters) exposed at high DEP

levels do not show an excess of lung tumors, nor do rats exposed at lower DEP levels. In rats,

prolonged and elevated exposures to a variety of different particulates result in lung overload,

15

lung inflammation, lung epithelial cell proliferation, and eventually lung tumors. This

mechanism is not DEP-specific and did not occur in the rats at DEP exposure concentrations

below 600 μg/m3 (40), a concentration level that is 10-fold greater than TDE DEP levels to

which railroad and trucking industry workers might have been exposed, and 100-fold above

ambient concentrations. Thus, the tumorigenic effect of high levels of TDE in rats is considered

to be a non-specific particle effect that results from a unique rat-lung overload mechanism (14,

41, 42). In fact, if any occupation might have resulted in "lung overload" in humans,

underground coal miners might be the most likely in this regard. As described earlier, IARC did

not find evidence of lung cancer risk among coal-miner populations. Thus, the rat "lung

overload" phenomenon should not be considered relevant to humans exposed either to DEP in

occupational environments or to even lower ambient levels.

A final aspect of the laboratory-animal studies relates to the potential mutagenicity of

DEP organic extracts, and reviews of these data are available elsewhere (2, 8, 43, 44). Although

the organic compounds extracted from DEP using strong solvents like dichloromethane are

capable of damaging DNA in vitro, such damage has not been established to occur under the

conditions of the DEP inhalation bioassay. Moreover, DEP organic compounds are poorly

bioavailable (12), and the fact that lung tumors can be induced in rats exposed by lifetime

inhalation of non-DEP particles with virtually no adsorbed organics (e.g., carbon black, titanium

dioxide) supports the conclusion that PM overload per se, and not the organics bound to DEP,

are responsible for lung tumors in rats exposed to overloading levels of DEP.

16

Conclusions on Potential Cancer Risks of Inhaled DE

A number of careful analyses of the DE epidemiologic data have concluded that existing

epidemiological studies are unable to quantitatively link DE exposure levels to increases in lung

cancer (2, 6, 33, 45-48). Several factors that support this conclusion are: (1) the epidemiologic

database lacks consistency, with some studies showing a weak association between presumed

DE exposure and lung cancer and other studies showing no association; (2) there is an overall

lack of dose-response among DE-exposed populations, with truckers at low exposures showing

greater risks than underground miners at high DE exposures (49); (3) some studies show

negative exposure-response relationships, and for those with weak positive associations with DE

exposure, the result may be attributable to residual confounding (particularly by smoking); (4)

many of the DE epidemiologic studies suffer from inadequate latency periods; (5) given the

negative mutagenicity data on DEP per se, and a negative animal database (aside from lung-

overloaded rats), biological plausibility remains an issue; and (6) the epidemiological studies

lack adequate DEP exposure information, without which scientists cannot establish a quantitative

estimate of human cancer risk.

3.0 Health Effects of Diesel-Engine Exhaust: Potential for Non-Cancer Health Effects

Laboratory-Animal Data

Numerous non-cancer health endpoints for DE have been investigated in animal studies.

Compared with observational and experimental human studies, laboratory animal studies offer

several unique advantages, including the study of both healthy and diseased animals (e.g.,

17

spontaneously hypertensive rats, atherosclerotic mice, lung-compromised mice) over a range of

well-defined exposure levels and controlled study conditions. Importantly, laboratory studies

allow for the assessment of serious health impacts, including longer-duration, rare, and

permanent health effects as well as those that require invasive measurement methods (e.g., multi-

organ pathology), not possible in human studies. Through the use of elevated exposure levels,

large numbers of study animals, sensitive animal species, and animal models of disease,

laboratory animal studies can be designed to increase the likelihood of detecting adverse effects

from DE inhalation. In fact, sometimes specific genetic strains are used to discover possible

effects of DE on, say, resistance to infection (50) or endothelial function (51). However,

extrapolation of conclusions derived from animal species to humans can be highly uncertain

given that laboratory experiments are typically conducted at extremely high exposure levels,

experimental routes of administration may differ from physiological, real-life exposures (e.g.,

intratracheal instillation is not representative of real-life inhalation exposures), and there may be

large interspecies differences in sensitivity and mode of action.

The 2002 US EPA "Health Assessment Document for Diesel Engine Exhaust" (Diesel

HAD) examined evidence regarding potential non-cancer health effects of both long-term and

short-term exposure to DE and DEP. In terms of elevated, short-term exposures, the agency

stated that DE at sufficient levels can cause transient irritation, inflammatory symptoms, and

may exacerbate asthma symptoms. For long-term exposures, US EPA identified a DEP

reference concentration (RfC), representing a lifetime exposure concentration not expected to

cause adverse non-cancer health effects, of 5 μg/m3 (1). This RfC was based on exposure-

response data for inflammatory and histopathological changes in lungs of rats exposed to high

18

levels of DE over extended periods of time. However, as noted earlier, as cleaner NTDE engines

replace a substantial number of existing, traditional diesel engines, the applicability of the US

EPA RfC analysis will need to be reevaluated.

With regard to non-cancer effects of DEP exposure, Hesterberg et al. (3) provide a

critical evaluation of over 100 published articles on experimental research. It is important to

note that even recent studies continue to use DE emissions from a wide variety of old, traditional

diesel engines and fuels, and hence the results likely have limited relevance to the modern fleet

of vehicles emitting new technology diesel exhaust (NTDE). For example, out of all of the DE

studies, very few have used DE from an engine running on diesel fuel meeting the 2007 US EPA

standard, and most other studies have used engines or fuels no longer commercially available.

Also, animal studies were often carried out at elevated TDE concentrations, sometimes reaching

DEP levels as high as 20,000 to 100,000 μg/m3.

As reviewed by Hesterberg et al. (3), several large-scale experiments with animal species

inhaling DEP have been completed since the US EPA Diesel HAD (1). These data show that

highly elevated DEP doses (DEP concentration exposure duration) can lead to chronic lung

inflammation, but that at smaller doses, still much higher than typical ambient levels of DEP,

little has been reported in the way of adverse or irreversible effects. In fact, one group of

investigators suggested a no-observable-adverse-effect level (NOAEL) in the range between 200

to 1,000 μg/m3 DEP (52), and other investigators point out that rat exposure levels turn out to be

roughly equal to human exposure concentrations (HEC) (within a factor of 2) by taking into

account dose per lung (alveolar) surface area, relative lung deposition, and relative lung

19

clearance (53). Thus, more recent data, subsequent to US EPA setting the RfC at 5 μg/m3,

suggest that the 5 μg/m3 is quite conservative and health protective.

The McDonald et al. (17) laboratory animal study provides some of the few available

experimental data for a diesel exhaust mixture representative of NTDE. This study investigated

the relative toxicity of acute inhalation exposures (6 hrs per day over 7 days) for a baseline

uncontrolled, TDE emissions case (approximately 200 μg/m3 DEP) versus an emissions-

reduction, NTDE case (low sulfur fuel, catalyzed ceramic trap, near background levels for all

emissions but NOx) on a suite of sensitive measures of acute lung toxicity in mice, including

lung inflammation, RSV resistance, and oxidative stress. For the baseline TDE case, McDonald

et al. (17) observed statistically significant DE-induced effects for each class of responses, while

these effects were either nearly or completely eliminated for the NTDE case.

With its well-characterized exposure atmosphere and its assessment of a suite of sensitive

measures of acute lung toxicity, the McDonald et al. (17) study provides some of the strongest

experimental evidence of the reduced toxicity of NTDE. However, as a single study that

addressed only acute lung toxicity in a single animal species for a single engine and

aftertreatment configuration, there is clearly a need for more comprehensive toxicological

investigations of NTDE to confirm these findings of reduced NTDE toxicity for a broader range

of aftertreatment configurations and operating conditions and for other classes of health

endpoints (e.g., cardiovascular effects, allergenic effects). On this note, it is expected that the

comprehensive health effects components of the ongoing Advanced Collaborative Emissions

Study (ACES) will provide a wealth of information for assessing the carcinogenicity, subchronic,

20

and chronic non-cancer toxicity of NTDE. A core component of ACES is a chronic rat bioassay

where rats are exposed via inhalation for 24 or 30 months, with interim sacrifices at 1, 3, 12, and

24 months, to three dilutions of whole emissions from a 2007-compliant diesel engine with

advanced emission control technologies (and clean air controls). Health endpoints of interest

include not only carcinogenicity but also pulmonary function, pulmonary inflammation,

oxidative damage, lung cell proliferation, histopathological changes, and hematological effects.

Exposure of Human Volunteers to DE

Controlled-exposure studies with human volunteers have several major advantages,

including the direct study of human subjects; the use of well-defined, continuously measured

exposure concentrations and durations; the absence of confounding exposures to other

chemicals; and the use of sophisticated research equipment to simultaneously and precisely

measure a suite of physiological endpoints potentially relevant to health, including both subtle

biological responses and small functional decrements. Although exposures via nasal masks or

mouthpieces have been used in some studies, large state-of-the-art inhalation chambers (e.g., 25

to 100 m3 whole-body chambers with control of humidity, temperature, ventilation, etc.) are

currently used, wherein small numbers of volunteers receive whole-body exposures to one or

more pollutants while either exercising or at rest. Some controlled human exposure studies have

been performed using adults representative of potentially susceptible subpopulations, including

those with asthma, chronic obstructive pulmonary disease, and cardiovascular disease.

21

Studies on human volunteers also have limitations. Namely, (1) small groups of healthy

volunteers may not be representative of larger populations; (2) investigation of only acute effects

from short-duration exposures may not extrapolate to long-term exposures; and (3) use of

exposure protocols and pollutant concentrations expected to elicit only transient, clinically mild

health responses may not extrapolate to higher-level, worker exposures (54, 55). In addition,

there is generally no control of the recent exposure history of volunteers, which can be an

important modifier of study findings. To increase the likelihood of detecting effects among

small groups of healthy subjects, human exposure studies often use relatively high DE

concentrations, compared to actual real-world exposure levels, yet, of course, below those that

are suspected to increase the risk of long-term or irreversible health effects. Because of these

limitations, human controlled exposure studies may be underpowered to detect some health

responses that are of health significance.

Because of the mild nature of the responses measured (often with subjects reporting no

perceptible symptoms), the question arises as to whether the response is an "adverse health

effect." Although physiologic impairment or clinically significant decrements may reach

adverse levels, the American Thoracic Society (56) has pointed out that not all "effects"

measured in clinical-exposure studies reflect "adverse health" outcomes warranting regulatory,

preventive measures. Specifically, ATS (56) concluded that "not all changes in biomarkers

related to air pollution should be considered as indicative of an injury that represents an adverse

effect." Similarly, ATS (56) concluded that "a small, transient loss of lung function, by itself,

should not automatically be designated as adverse." ATS (56) further distinguished adverse

22

symptoms as those that are associated with "a diminished quality of life or with a change in

clinical status."

Since the preparation of the Diesel HAD, a number of new human controlled exposure

studies have been published, with several of these studies following up on previous studies of

lung and systemic inflammatory responses, and other recent studies addressing cardiovascular

health responses including cardiac, vasomotor, and fibrinolytic function (3). Studies of human

volunteers have used elevated concentrations of DE (about 100 to 300 μg/m3), markedly above

what would be experienced in typical non-occupational environments. An illustration of the

recent cardiovascular-related results that have been found in humans is given in Figure 3, where

the vertical axis gives the concentration of DEP, and the horizontal scale lists the type of studies

that have been done. Cardiovascular health responses have been a focus of a number of recent

DE human clinical studies, and as shown in Figure 3, these studies have generally reported

mixed findings with respect to various subclinical measures of acute cardiovascular health

responses. However, none of these studies have examined NTDE, the low emission technology

that is required for all diesel on-road vehicles starting in 2007. In fact, preliminary data are

available from two studies (57, 58) showing a reversal of effects on vasomotor function and

thrombus formation with addition of a commercially available retrofit particle trap to the test

heavy-duty diesel engine.

23

Figure 3: Graphical summary of key findings from recent controlled human exposure studies of

the cardiovascular-related health effects of inhaled DE (both particles and gases), with

Panel A addressing systemic inflammatory and fibrinolytic/thrombogenic responses and

Panel B showing effects on subclinical markers of cardiovascular disease (e.g., heart rate,

blood pressure, heart rate variability, etc.), effects on vascular function, and changes in

indicators of endothelial dysfunction. Green circles with – signs represent negative (null)

findings, red squares with + signs represent positive findings, and yellow triangles

represent equivocal findings.

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DE

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NegativeFindings

PositiveFindings

Inflammatory Cells/ Pro-InflammatoryMediators

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Fibrinolytic/Thrombogenic Response

Panel A

24

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NegativeFindings

PositiveFindings

Equivocal/MixedFindings

HR/BP Vascular Function,Arterial Compliance

+

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EKG/HRVIndicators of EndothelialDysfunction (e.g., VWF, ET-1)

Panel B

Overall, given the relatively small number of statistically significant DE-induced changes

observed in these recent human-exposure studies for some test parameters among a larger

number of non-significant findings, as well as some conflicting findings among studies (Figure

3), it remains difficult to interpret the broader health implications of what currently remains a

disparate body of findings from studies of small numbers of subjects and different study designs

and measurements. Although animal studies have provided some findings suggestive of

25

potential mechanisms (e.g., the Wong et al. (59) findings suggesting a neurokininergic

mechanism for DE-induced lung inflammation), our mechanistic understanding remains limited.

This is particularly true for low-level ambient DE concentrations, given that most animal studies

have been conducted at concentrations vastly higher than those used in controlled human

exposure studies. Broader interpretation of these human clinical data is limited by the fact that

all but a few of the available studies were conducted in the same exposure laboratory for freshly-

generated emissions from the same engine type under the same operating conditions (idling

Volvo TD45, model year 1991, 4.5-liter, 4 cylinder, 680 rpm, diesel engine). This places limits

on generalizing the study findings to emissions from other engines and operating conditions, and

to environmentally-aged diesel emissions. As summarized in the 2002 Diesel HAD (1) and Ris

(60), it is well-established that DE emissions vary both in chemical composition and particle size

distributions for different types and ages of diesel engines, and within engine types, depending

on operating conditions (fuel composition, load characteristics, lube oil composition, emission

control devices), with atmospheric aging also serving to modify DE properties.

The question regarding potential health effects of "aged" DE emissions might be best

addressed by studies such as that of McCreanor et al. (26), where both health effects and PM

levels were assessed for 2-hour weekday exposure periods between November 2003 and March

2005 along London's heavily-trafficked Oxford Street (Figure 2, EC concentrations, 4 – 16

µg/m3, median PM2.5 concentration 28 μg/m

3). The Oxford Street location was chosen to

represent a busy city street impacted by diesel-vehicle emissions, because only diesel-powered

buses and taxicabs are permitted on this heavily-trafficked street, while Hyde Park (EC

concentrations, 0.4 – 7 μg/m3, median PM2.5 concentration, 12 μg/m

3) is a traffic-free area of

26

London. Ultrafine-particle concentrations, in terms of 1,000's of particles per cm3, were 64 on

Oxford Street and 18 in Hyde Park.

The McCreanor et al. (26) study included 60 participants with mild to moderate asthma,

and each participant walked for 2 hours along a heavily trafficked street (Oxford Street) and, on

a separate occasion, through a nearby, traffic-free park (Hyde Park). The investigators observed

small, statistically significant, "asymptomatic but consistent" reductions in FEV1 (up to 6.1%)

and FVC (up to 5.4%) for the Oxford Street exposures compared to those for the Hyde Park

exposures. These mild, reversible lung function changes for the Oxford Street exposures

coincided with statistically significant increases in sputum myeloperoxidase levels, a biomarker

of neutrophilic inflammation, and airway acidification. However, between the two sites, there

were no statistically significant changes in FEF25-75%, mean FENO, sputum neutrophil counts,

interleukin-8, sputum eosinophil counts, or eosinophil cationic protein levels. Using pollutant-

specific, exposure-response analyses, the study investigators linked the observed health

responses with the significantly greater air pollutant exposure concentrations measured during

the Oxford Street exposure period (e.g., 5.7-fold higher EC concentrations and 3.5-fold greater

ultrafine particle count concentrations), noting that, in two-pollutant models, the effects of

ultrafine particles and elemental carbon retained statistical significance and were most consistent.

Although the study authors suggest elevated exposures to DE on Oxford Street may underlie

their study findings, they do acknowledge the possibility that other unmeasured pollutants

associated with roadside diesel-traffic exposure, namely road dust and engine or tire debris, may

instead be the causal agents. Likewise, noise, temperature (2°C hotter on Oxford Street), and

other stresses may have also played a role in the measured differences.

27

Although not specific to DE, a number of observational epidemiology studies have linked

cardiovascular and respiratory health effects to traffic-related air pollution (74-76). The HEI

Panel on the Health Effects of Traffic-Related Air Pollution (76) conducted one of the more

comprehensive reviews of the health effects literature related to traffic-related air pollution,

concluding that "the evidence is sufficient to support a causal relationship between exposure to

traffic-related air pollution and exacerbation of asthma." In addition, the HEI panel concluded

that there was "suggestive evidence of a causal relationship" with onset of childhood asthma,

nonasthma respiratory symptoms, impaired lung function, total and cardiovascular mortality, and

cardiovascular morbidity. Importantly, studies of traffic-related air pollution have generally

relied upon surrogate measures of exposure to traffic-related pollution, including both traffic-

pollutant surrogates (e.g., CO, NO2, EC) and proximity-based surrogates (e.g., distance of

residence to nearest road, and traffic density). As a result, the role of DE and DEP in these

associations cannot be clearly assessed due to the non-specificity of the pollutant surrogates, and

due to many potential confounding factors associated with surrogates based on proximity, such

as gasoline-engine exhaust, noise, fugitive road dust, socioeconomic status, and so forth (77).

Furthermore, as acknowledged in HEI (76), the available epidemiology studies are primarily for

past time periods and older vehicles, meaning that they are of uncertain, and possibly little,

relevance to today's modern engine technologies and fuels.

Conclusions on Potential Non-Cancer Health Effects of Inhaled DE

28

Human controlled-exposure studies provide evidence suggestive of lung inflammatory

effects and thrombogenic and ischemic effects of inhaled DE, albeit for older model diesel

engines and concentrations that are much higher (~300 μg/m3) than typical ambient or even

occupational levels. Recent animal studies provide insights on the potential mechanisms

underlying observed respiratory and cardiovascular health responses, but because of

unrealistically high DE concentrations, it is unclear whether the mechanisms elucidated in these

studies are relevant at lower DE exposure levels. A mechanism of action that allows reliable

prediction of adverse health impacts at DE exposure levels typical of the present-day ambient

and occupational environment has not emerged. Because of changing diesel-engine technology,

inhalation studies using realistic environmental and occupational exposures of new technology

diesel exhaust (NTDE) are of critical importance.

4.0 Conclusions

Much remains to be understood in terms of the constituents of TDE and NTDE that have

the greatest potential for eliciting health effects at adequately high exposure concentrations and

durations. There are important differences in mass emission rates, particle size distributions,

elemental carbon vs. organic carbon content, ratios of 3- and 4-ring polycyclic aromatic

hydrocarbons (PAHs), and other constituent concentrations, not only between TDE and NTDE,

but also among various types of engines within the two classes (15, 16). In fact, it cannot be

clearly stated whether DE is of greater or lesser toxicity than other combustion sources such as

gasoline-engine exhaust, wood smoke, fossil-fuel power plants, compressed-natural-gas engines,

and other combustion sources. These issues are crucial both for DEP and ambient PM, where

there is disagreement between observational epidemiology and toxicology as to the sufficient

29

dose and toxic role of chemical constituents and physical characteristics causing biologic

activity.

Although DE-exposed worker populations have been studied a great deal, both

inadequate estimates of DE exposure as well as poorly-understood exposure-response

relationships have prevented development of quantitative estimates of lung-cancer risk. In fact,

there does not appear to be an exposure-response relationship over different occupations that

have considerably different exposure potential for DE. A high priority is the reporting of results

from the longstanding NIOSH/NCI studies of US underground miners with historically elevated

DE exposures. These results need to be completed, analyzed, and reported in detail in the open

literature.

Emissions of DEP nanoparticles (i.e., with diameters < 100 nm) have received increased

attention in recent times, due to reports that nanoparticle emissions could be increased in NTDE,

as well as due to the intense interest in the toxicity of engineered nanoparticles. Nano-sized

particles, including individual nuclei mode particles and carbonaceous agglomerates, are well

known to dominate particle number concentrations of DEP (78). Hesterberg et al. (5) examined

findings from the DE human clinical studies to identify insights relevant to potential NP human

health hazards, concluding that the available DE human clinical data do not give evidence of a

unique toxicity for NPs (at least in the DEP context) as compared to other small particles. Given

that particle number and surface area concentrations are recognized as more relevant exposure

metrics for nanoparticle health effects than particle mass concentration, there is a real need for

DE health effect studies to quantify these exposure metrics (in addition to mass concentrations).

30

Recent studies of DE exposures to human volunteers have provided useful human

evidence of the potential respiratory and cardiovascular health hazards of short-term exposures to

elevated DE exposures, but there is a need to better understand the human clinical significance of

these findings and their relevance to lower, ambient levels of DE and a wider range of engine

types, operating conditions, and population subgroups. In addition, there is a need for more

comprehensive toxicological investigations of NTDE to confirm the toxicological differences

between NTDE and TDE indicated by the distinctions in their chemical and physical

characterization properties. Furthermore, with recent research indicating that other combustion

emissions (e.g., gasoline engine emissions) and ambient particle types can elicit similar

biological responses as DE, further study is also needed to understand how DE health risks

compare to those from other common air contaminants. With further study and improvements in

our understanding of not only DE hazard assessment, but also dose-response for realistic

inhalation exposures, it can be hoped that newly-collected data will stimulate continuing

progress toward a cleaner-burning, increasingly safer diesel engine technology.

31

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