Review

For reprint orders, please contact reprints@future-science.com

Bioanalytical techniques for detecting

biomarkers of response to human asbestos
exposure

Asbestos exposure is known to cause lung cancer and mesothelioma and its health and Clementina Mesaros‡,1,2,3,
economic impacts have been well documented. The exceptionally long latency periods Andrew J Worth‡,1,2,3,,
of most asbestos-related diseases have hampered preventative and precautionary Nathaniel W Snyder2,4, Melpo
steps thus far. We aimed to summarize the state of knowledge on biomarkers of Christofidou-Solomidou1,2,5,
Anil Vachani1,2,5, Steven M
response to asbestos exposure. Asbestos is not present in human biological fluids;
Albelda1,2,5 & Ian A Blair*,1,2,3
rather it is inhaled and trapped in lung tissue. Biomarkers of response, which reflect 1
Penn SRP Center, Perelman School of
a change in biologic function in response to asbestos exposure, are analyzed. Several Medicine, University of Pennsylvania,
classes of molecules have been studied and evaluated for their potential utility as Philadelphia, PA 19104-6160, USA
biomarkers of asbestos exposure. These studies range from small molecule oxidative 2
Center of Excellence in Environmental
Toxicology, Perelman School of
stress biomarkers to proteins involved in immune responses.
Medicine,University of Pennsylvania,
Philadelphia, PA 19104-6160, USA
Asbestos background fibers for both MM and lung cancer [10] . 3
Department of Systems Pharmacology
Asbestos is a class of silicate materials with a Despite widespread knowledge of the hazards & Translational Therapeutics, Perelman
School of Medicine,University of
history of industrial uses due to its inert phys- of asbestos and bans on any use of asbestos
Pennsylvania, Philadelphia,
icochemical properties such as fire- and heat- in more than 50 countries, an estimated two PA 19104-6160, USA
resistance and tensile strength, which makes million tons of asbestos continue to be used 4
A.J. Drexel Autism Institute, Drexel
it an ideal material for many commonly used around the world each year. University, Philadelphia, PA 19104, USA
products in both the construction indus-
5
Department of Medicine, Perelman
School of Medicine, University of
try and the home [1] . There are two major Asbestos-related diseases
Pennsylvania, Philadelphia,
forms of asbestos: serpentine (chrysotile) It has been clearly established in multiple PA 19104-6160, USA
and amphibole (which includes crocido- models that asbestos fiber inhalation can *Author for correspondence:
lite amosite, tremolite, actinolite and antho- lead to neoplastic diseases such as MM and Tel.: +1 2155739885
phyllite)  [2–4] (Figure 1) . Though it was long lung cancer [7,8] , as well as pulmonary fibro- Fax: +1 2155739889
ianblair@mail.med.upenn.edu
thought to pose no risk to human health, sis. MM is a highly heterogeneous and highly ‡
Authors contributed equally
asbestos is now known to cause a variety of aggressive cancer of serosal surfaces, such as
adverse health effects and has been classi- the pleura and the peritoneum [11] . The rela-
fied as a human carcinogen [5,6] . It has been tive 5-year survival rate for mesothelioma
clearly established in both animal models compared with age-matched controls is
and human studies that asbestos fiber inhala- between 5 and 10%. Current therapies, with
tion can lead to neoplastic diseases such as the exception of surgery in very early disease
malignant mesothelioma (MM) and states, are not curative [12] . The most recent
lung cancer [7,8] , as well as pulmonary fibrosis statistics on MM mortality show that there
(i.e., asbestosis). The toxic forms of chryso- were 2704 deaths in the USA in 2005 with
tile and crocidolite asbestos fibers are consid- little evidence that rates are declining [13] .
ered to be longer than 5 μm, with a thick- Although asbestos use has been restricted
ness between 250 nm and 1 μm, and with an in many western countries, it is still mined
aspect ratio ≥ 3:1 as determined by transmis- in Russia, China, Canada, Brazil, Kazakh-
sion electron microscopy [9,10] . Fibers that are stan and Zimbabwe [13,14] . It has been esti-
>10 μm in length and <250 nm in thickness mated approximately 125 million people
part of
are more potent than shorter and thicker are exposed to asbestos globally within their

10.4155/BIO.15.53 © IA Blair et al. Bioanalysis (2015) 7(9), 1157–1173 ISSN 1757-6180 1157
Review  Mesaros, Worth, Snyder et al.
Serpentine
Chrysotile Crocidolite
(white) (blue)
Commercial
forms
Figure 1. Classification of asbestos fibers.
Modified with permission from [4] © BMJ Publishing Group Ltd. (1990).
place of work, often in third world countries that do
not have strictly regulated workplace safety controls.
Thus, there will likely be an increase in MM cases in
the third world, and in developed countries due to
occupational exposure (e.g. plumbers, pipefitters, insu-
lators, etc.) [15] . In addition, there are some nonoccu-
pational environmental exposures. For example, there
is an increased risk of asbestos-related diseases
(ARDs) from old mining sites such as Libby Mon-
tana  [16,17] or where asbestos-related industries once
operated such as the BoRit site in Ambler, PA, USA [18] .
Another major concern regarding environmental expo-
sures includes areas within Europe and Australia that
have large numbers of buildings where asbestos was
used in construction materials and are thus likely to
result in ongoing exposures [19–26] .
The economic impact of asbestos is large and
far reaching, from burdening health care systems
to extraordinary costs associated with removal of
Key terms
Serpentine asbestos: Curly class of fibers, chrysotile is the
only member of the serpentine class.
Amphibole asbestos: Needle-like class of fibers, where
crocidolite, amosite, tremolite, anthophyllite and actinolite
are members of the amphibole class. Crocidolite, often
known as blue asbestos, is considered to be one of the
most toxic forms of asbestos due to its high iron content.
Malignant mesothelioma: Rare form of cancer that
develops from cells of the mesothelium, the protective
lining that covers many of the internal organs of the
body. Mesothelioma is most commonly caused by
exposure to asbestos. The most common anatomical site
for mesothelioma is the pleura (the outer lining of the
lungs and internal chest wall), but it can also arise in the
peritoneum and the pericardium.
Asbestos-related diseases: Asbestos-related diseases
include: malignant mesothelioma, lung cancer, pleural
plaques, asbestosis and diffuse pleural thickening.
Oxidative stress: Reflects an imbalance between the
formation of reactive oxygen species, reactive nitrogen
species, or electrophilic reactive intermediates and a
biological system’s ability to readily detoxify the reactive
species and electrophilic intermediates.
1158 Bioanalysis (2015) 7(9)
Amphiboles
Amosite Tremolite/ Anthophyllite
(brown) actinolite
environmental contaminants. One of the most chal-
lenging factors to address with ARDs is the considerable
latency period between initial exposure (Figure 2)  [27]
and subsequent adverse health effects, as most clini-
cal cases result from exposures decades (20–40 years)
beforehand. For this reason it is imperative that mark-
ers of asbestos exposure be developed to facilitate detec-
tion of at-risk individuals prior to the onset of disease.
Furthermore, individuals suspected of being exposed to
asbestos would be able to confirm they are not likely
to suffer from an ARD. Finally, a quantitative assay
capable of assessing asbestos exposure would facilitate
confirmation of the beneficial impacts resulting from
environmental remediation of asbestos sites.
Pleural changes resulting from asbestos
exposure
Biologically significant asbestos exposure is associated
with the presence of pleural changes (especially pleural
plaques) visualized on chest x-ray (CXR), or more com-
monly by computerized tomography (CT) scan [28,29] .
Pleural plaques are circumscribed areas of thickening
of parietal or diaphragmatic pleura composed of avas-
cular collagen connective tissue. Although they can
be seen occasionally after pleural infections or pleu-
ral hemorrhage, they are most commonly the result
of prior asbestos exposure, generally appearing some
20–40 years after first exposure (that is, after a ‘latent’
interval) and tend to calcify over time. Plaques tend
to grow slowly over time, but they do not cause symp-
toms. It has been estimated that approximately 5–15%
of those with occupational exposure to asbestos will
have noncalcified pleural plaques 20 years after this
exposure and about a third or more will have calcified
plaques after 30 years [28] . A recent study of over 5000
French asbestos-exposed workers that used more sensi-
tive CT screening found a pleural plaque prevalence
of 20%. After adjustments for asbestos exposure and
latency time, patients with plaques had a significantly
elevated risk of mesothelioma [30] . Studies evaluating
the association of pleural plaques and lung cancer risk
have also provided conflicting evidence [31] . This means
future science group

that there are significant limitations in using plaques as
biomarkers of asbestos exposure. Since plaques evolve
slowly over time and become large enough to see only
many years after initial exposures, their reported
prevalence increases with longer latency periods and
at older ages. Furthermore, only a small percentage of
subjects with pleural plaques develop mesothelioma.
Thus, asbestos exposure history and imaging are use-
ful but are not sensitive or specific enough to accurately
identify those at greatest risk for developing malignant
disease or asbestosis. This means that there is a real
need for more specific and sensitive biomarkers of
asbestos exposure that could permit personalization of
risk assessment.
Asbestos exposure & oxidative stress
The most highly cited studies of asbestos exposure
have tended to focus on biomarkers of oxidative
stress  [32,33] , which cannot distinguish asbestos
exposure from other causes of oxidative stress such as
from cigarette smoking [34,35] or from diseases such as
atherosclerosis  [36] . However, there is substantial evi-
dence that oxidative stress is involved in the etiology
of ARDs such as pulmonary fibrosis [37] and mesothe-
lioma  [38] . One factor driving MM carcinogenesis is
the DNA damage caused by asbestos fibers. The DNA
damage could potentially be caused directly through
mechanical interference of asbestos fibers with chro-
mosome segregation during mitosis [39] . However, it
is now largely believed that DNA damage is in fact
caused by the oxidation that arises through sustained
inflammation and subsequent production of reactive
oxygen species (ROS) (Figure 3) [40] .
Since only a small fraction of patients with signifi-
cant asbestos exposure develop MM, the disease is now
thought to progress through a combination of both
genetic and environmental factors. For example, the Sim-
ian virus 40 T antigen has been detected in many meso-
theliomas and has been implicated in the etiology of the
disease [41] . Furthermore, some studies have shown that
the cyclin-dependent kinase inhibitor 4a (P16INK4a)
and ARF tumor suppressor (p14ARF) genes are fre-
quently inactivated in mesothelioma and approximately
50% of MM cases contain missense or nonsense muta-
tions in the neurofibromin type 2 gene. Mice lacking
these genes are relatively normal but develop MM at an
accelerated rate when exposed to asbestos by intraperi-
toneal injection. In addition, within the last few years,
germ line and sporadic mutations of the tumor suppres-
sor, breast cancer susceptibility (BRCA)-1 associated
protein-1 (BAP-1) gene, which expresses a deubiquiti-
nating enzyme, appear to predispose patients to DNA
damage after environmental stress [2] . Inhaled asbestos
fibers work their way into the lung and ultimately reach
future science group
Biomarkers of asbestos exposure Review
2000
All asbestos fibers
Asbestos fibres (ff/ml)
Chrysotile fibers
1500 Amphiboles fibers
1000
500
0
0–10 11–20 > 20
Time since last exposure (years)
Figure 2. Concentrations of asbestos fibers per milliliter
of bronchoalveolar lavage fluid in the workplace-
exposed subjects. Data are subdivided on the basis of
time spent between last exposure and bronchoscopy
(geometric mean).
Reprinted with permission [27] © Oxford University
Press (2007).
the pleural surface (Figure 4) [42] where they are engulfed
by tissue phagocytes, primarily macrophages. This pro-
cess stimulates intracellular ROS production and acti-
vates the nuclear factor kappa-light-chain-enhancer of
activated B cells (NF-κB) pathway, inducing the release
of numerous cytokines (Figure 3). Recent studies suggest
that asbestos also activates the nucleotide-binding oligo-
merization domain-like receptor family, pyrin domain
containing 3 (NLRP3) inflammasome thus promoting
the release of interleukin (IL)-1β and IL-18 [43] . Fur-
thermore, genetic studies indicate that polymorphisms
in genes within the NRLP3 family can both predispose
and protect against fibrosis following asbestos exposure.
Mice exposed to asbestos exhibit recruitment of acti-
vated macrophages to the mesothelium interacting with
asbestos fibers. Macrophages are also thought to directly
interact with and phagocytose asbestos. It is hypothe-
sized that macrophages and mesothelial cells exposed to
asbestos undergo frustrated phagocytosis of elongated
fibers; this process is thought to cause chronic produc-
tion of ROS and cytokines, which contribute to DNA
damage and transformation of mesothelial cells [44] .
The Carbone group has also suggested that high lev-
els of the high-mobility group box 1 (HMGB1) protein
are secreted after asbestos exposure and during the pro-
gression of MM. HMGB1 is a lysine-rich protein with
an acidic carboxy-terminal region containing multiple
aspartate and glutamate residues [45] . During tissue
necrosis, such as the hepatic necrosis induced by high
doses of acetaminophen, many of the lysine residues
are acetylated, which facilitates secretion from tissues
into the circulation [45] . A similar process after asbestos
exposure would provide a useful biomarker (Table 1) .
HMGB1 amplifies the inflammatory response in gen-
eral (by chemotaxing neutrophils and mast cells [46]),
www.future-science.com 1159
Review  Mesaros, Worth, Snyder et al.
Serum biomarkers
MDA
Isoprostanes
Modified lipids
Asbestos
Mesothelial cell membrane
Lipids ROS
Cytosol
NK-kB
8-oxo-dGuo NK-kB
Nucleus
Figure 3. The effect of asbestos fibers on mesothelial cells.
but also serves as an important protumor cytokine that
enhances the growth, survival and invasiveness of the
mesothelial cells [3] . MM tumor samples are associated
with chronic inflammation including macrophage
infiltration and inflammatory cytokine production. In
addition to the cytokines discussed above, the activated
macrophages contribute to tumorigenesis by forming
harmful ROS and reactive nitrogen species that can
in turn induce further DNA damage. This can lead
to genomic instability; by increasing tissue prolifera-
tion, and inducing tissue remodeling and angiogenesis-
promoting factors, as well as inducing extravasation of
tumor cells from the microenvironment [47] .
Biomarkers of response to asbestos
exposure
Biomarkers of exposure can be used to assess the
amount of a chemical that is present within the
body  [64] . However, asbestos is not present in readily
accessible biological fluids; it is inhaled and trapped
in lung tissue for long periods of time. Although the
asbestos fibers can appear in bronchoalveolar lavage
1160 Bioanalysis (2015) 7(9)
180 nm IL-1β
IL-18 TNF-α
5 µm
NLRP3
inflammasome
Transcription
TNF-α
(BAL) fluid, such measurements are more suitable
for a qualitative/categorical approach to exposure
assessment than a quantitative one [65] . This means
that it is necessary to analyze biomarkers of response
to asbestos rather than directly measuring asbestos
exposure. Biomarkers of biological response can,
in principle, provide more direct insight into the
potential for adverse health effects than biomarkers
of exposure. Before being implemented in a clini-
cal setting, response biomarkers must first be fully
characterized and validated in large sample sets.
Candidate biomarkers often lack diagnostic utility
because of poor sensitivity (false negatives) and/or
inadequate specificity (false positives) derived from
confounding exposures, nonspecific biomarkers or
individual variability. To circumvent some of these
issues, multiple biomarkers can be combined which
may result in a more complex but thorough analysis.
Additional issues may arise from variations in sample
preparation and analytical methodology throughout
sample collection, processing and analysis. For this
reason robust and reproducible analytical techniques
future science group

are critical to ensuring proper validation and utiliza-
tion of a biomarker. Once fully validated and char-
acterized, biomarkers can serve a critical role in char-
acterizing levels of exposure, which would otherwise
be unknown.
The primary route of exposure to asbestos is
through inhalation. The fibers are able to travel deep
into the respiratory system and penetrate pleural cells
within the lining of the lung. This penetration can
lead to innate immune responses and subsequent oxi-
dative stress (Figure 3) . Disrupted cellular processes
can ultimately lead to the generation of fibrotic tissue
and in some cases cancer. These pathogenic responses
to asbestos exposure afford the use of resulting bio-
chemical characteristics as biomarkers of exposure.
For example, oxidative stress leads to modified lipids,
proteins and DNA. These biochemical changes can be
confounded by smoking cigarettes, which is known
to induce oxidative stress, and these biomarkers are
higher in tobacco smokers than nonsmokers [35] . In
keeping with this concept, quantitative analysis of
documented markers of oxidative stress may be use-
ful for characterizing asbestos exposure across human
populations, but may lack specificity without includ-
ing other variables. In addition, several factors involved
in immune response, cell proliferation and generation
of fibrotic tissue also hold promise as useful biomark-
ers of asbestos exposure. The factors likely to impact
disease risk include common environmental variables
per se, functional polymorphisms in genes, [2,66] and
differential expression of genes that interact with such
variables [67,68] .
Here we provide a review of biomarkers of response
(including small molecules and proteins) that result
from exposure of human populations to asbestos. An
important point is that many studies have emphasized
the need for biomarkers of MM and other ARDs such
as lung cancer and pleural plaques [69,70] . However, in
this review we have focused primarily on biomarkers
of asbestos-exposed subgroups versus nonexposed indi-
viduals. Establishing biomarkers of response resulting
from asbestos exposure will enable better classifica-
tion of exposed populations, improving preventative
measures, and help characterize previously unknown
biochemical impacts of asbestos exposure.
DNA-adduct biomarkers
Urinary 8-oxo-7,8-dihydro-2’-deoxyguanosine (8-oxo-
dGuo), often incorrectly referred to as 8-hydroxy-2’-de-
oxyguansoine, is a widely used nonspecific biomarker
of oxidative stress. During oxidative stress, ROS can
cause oxidative damage to dGuo to form 8-oxo-dGuo
in DNA [71] and the trinucleotide pool. 8-oxo-dGuo
arises from human Mut T homolog 1 (hMTH1)-
future science group
Biomarkers of asbestos exposure Review
5 μl
Figure 4. Electron microscopy of an ashed lung
section after exposure to crocidolite and chrysotile.
The crocidolite fibers are at the upper left and the
chrysotile fibers are at the lower right.
Reprinted with permission from [42] © BMJ Publishing
Group Ltd. (1972).
mediated hydrolysis of 8-oxo-dGuo-triphosphate in
the trinucleotide pool followed by excretion in the
urine  [35] . Three base excision repair enzymes, human
MutY homolog (hMutY), hOGG and hOGG2 [72] , are
involved in the repair of 8-oxo-dGuo-derived lesions in
DNA. However, the trinucleotide pool is considered to
be the major source of urinary 8-oxo-dGuo. The gold
standard for quantification of urinary 8-oxo-dGuo [73]
involves the use of stable isotope dilution LC–MS
coupled with the addition of [13C]-dGuo to monitor
any artifactual formation.
There are several population studies that have exam-
ined levels of oxidative DNA base damage in workers
exposed to asbestos with a large number of subjects and
appropriate control groups (Table 1)  [48,49] . Unfortu-
nately, the LC-electrochemical detection (ED) meth-
odology employed in the studies was not of very high
quality because internal standards were not used and
the assays were not validated by more specific LC–MS
methodology [35] . Nevertheless, the older study, which
examined levels of urinary 8-oxo-dGuo in asbestos
workers, found a significant increase when compared
with unexposed controls [48] . The more recent study,
which was conducted in Japan [49] , analyzed urinary
8-oxo-dGuo in construction workers who had been
exposed to low levels of asbestos. The asbestos exposed
group, which consisted of 48 construction workers
with a history of suspected exposure to asbestos and
CXR evidence of irregular opacities or pleural plaques,
www.future-science.com 1161
Review  Mesaros, Worth, Snyder et al.

Table 1. Response biomarkers of asbestos exposure.

Asbestos biomarker
7,8-dihydro-8-oxo-2’-
deoxyguanosine (8-hydroxy-
2’-deoxyguanosine)
 
8-iso-isoprostane PGF2α
Soluble mesothelin-related
peptide or soluble mesothelin-   glycoproteins from proteolytic cleavage
related protein
 
Osteopontin
Fibulin-3
Fibronectin
 
Abbreviation Description Bioanalytical technique Ref.
8-oxo-dGuo Oxidative stress DNA base adduct LC-ED [48,49]
  biomarker arises in urine from hMTH1- Fluorometric OxyDNA kit [50–52]
mediated hydrolysis of 8-oxo-dGuo-
triphosphate. Alternatively, isolated
from DNA or analyzed intact in the DNA
of formalin-fixed cells 
8-iso-PGF2α Oxidative stress lipid biomarker formed Stable isotope dilution [53]
by ROS-mediated oxidation of esterified LC–MS
arachidonic acid appears as the free
form in plasma, urine, an EBC after
hydrolysis of esterified lipids
SMRP Mesothelin and SMRP are 40-kDa Mesomark ELISA kit [54]
of the 69-kDa mesothelin precursor
protein - both are analyzed Sandwich ELISA with [55]
  monoclonal antibodies
None An integrin-binding protein involved ELISA kit [56]
in tumorigenesis, progression and
metastasis
None Fibulin-3 is protein that belongs to ELISA kit [57]
a family of extracellular proteins
expressed in the basement membranes
of blood vessels
None Fibronectin is a glycoprotein involved in Immunochemical assay [58]
  the extracellular matrix structure that with laser nephelometer
plays a role in the generation of fibrotic detection
tissue 
ELISA [59]

High mobility group box 1 HMGB1 HMGB1 is a chromatin protein. The ELISA kit [60]
unmodified protein has a nuclear
location - lysine hyperacetylation causes
translocation of HMGB1 into the cytosol
Interleukin 6 and interleukin 8 IL-6 and IL-8 Members of a large family of cytokines Solid phase ELISA [61,62]
that promote the development,
differentiation and activation of
lymphocytes and play an important role
in the immune response
Regulated on activation RANTES A chemokine also known as chemokine Magnetic bead multiplex [63]
normal T cell expressed (C–C motif) ligand 5 (CCL5) immunoassay
protein
EBC: Exhaled breath condensate; SMRP: Soluble mesothelin-related peptide.

was compared with a control group of 41 unexposed
healthy controls. This revealed that 8-oxo-dGuo levels
in the asbestos exposed group were higher than those
in the control group, although the levels did not reach
statistical significance.
Additional studies have quantified 8-oxo-dGuo
in the DNA of white blood cells (WBC) or circu-
lating lymphocytes using a commercially available
fluorometric OxyDNA kit (Table 1) [50–52] . This method
for analyzing oxidative DNA damage is based upon
the direct binding of a fluorescent probe to 8-oxogua-
nine moieties in DNA for formalin-fixed cells. Fluores-
cence is then quantified using flow cytometry. It is also
necessary to remove all transition metal ions in buffers
during formalin fixing in order to prevent additional
artifactual DNA oxidation [71] . As no such precautions
were used, the reported levels of 8-oxo-dGuo must be
treated with some caution. The studies showed that
asbestos-exposed subjects had significantly elevated
8-oxo-dGuo levels in WBCs and circulating lympho-

1162 Bioanalysis (2015) 7(9) future science group


cytes from asbestos exposed populations compared
with unexposed controls [50–52] .
Another study conducted in a Chinese popula-
tion measured levels of 8-oxo-dGuo in the DNA of
peripheral blood leukocytes as a biomarker of asbes-
tos exposure in a population occupationally exposed
primarily to chrysotile asbestos [74] . Leukocyte DNA
was extracted from 5 ml samples of peripheral blood
and 8-oxo-dGuo levels were measured by LC after
hydrolysis of the DNA. DNA isolation for the analysis
8-oxo-dGuo requires the use of chaotropic methods in
order to prevent artifactual DNA oxidation [71] . It is
also necessary to remove all transition metal ions from
hydrolysis buffers in order to prevent additional arti-
factual DNA oxidation [71] . In this study, no attempt
was made to prevent DNA oxidation during isolation
and hydrolysis. Furthermore, the statistical power was
very limited with only 19 controls and ten asbestos-
exposed workers group-matched for age and sex. The
geometric mean of 8-oxo-dGuo levels showed no sig-
nificant difference between the control and asbestos
exposed groups [74] .
The largest studies to date were conducted in
Germany by Marczynski et al. using a longitudinal
design with annual measurements over 3 consecutive
years  [75,76] . In these studies, the ability of inhaled
asbestos fibers to induce the formation of 8-oxo-dGuo
9
Mean:
8
Median:
7
8-oxo-dGuo/105 dGuo
0
Controls
1994–97 1994/95
(n = 214) (n = 496)
median = 1.51 median = 2.49
mean ± SD = 1.52 ± 0.39 mean ± SD = 2.61 ± 0.91*
Figure 5. 8-oxo-dGuo levels in the white blood cell DNA of asbestos-exposed workers over a period of three
study years.
*p < 0.001, significantly different from nonasbestos-exposed controls.
Reprinted with permission from [75] © Elsevier (2000).
future science group
Biomarkers of asbestos exposure Review
in the DNA from WBCs of workers highly exposed
was evaluated with 636 workers in the exposed group
and 214 healthy unexposed individuals as the control
group. The asbestos-exposed workers had an average
exposure of around 19 years, so the levels of 8-oxo-
dGuo were measured at steady state in hydrolyzed
DNA using LC-ED. The study used WBCs isolated
from 9 ml of whole blood but again there was no con-
trol for artifactual oxidation during DNA isolation
and hydrolysis. The study reported 8-oxo-dGuo lev-
els in asbestos-exposed workers that were significantly
increased (p < 0.001) compared with that in the con-
trol group, with this significant difference identified in
all 3 years of the study. Asbestos-exposed individuals
displayed a mean value of 2.61 ±0.91 8-oxo-dGuo/105
dGuo (median 2.49; n = 496) in 1994–1995, 2.96
±1.10 8-oxo-dGuo/105 dGuo (median 2.76; n = 437)
in 1995–1996 and 2.55 ±0.56 8-oxo-dGuo/105 dGuo
(median 2.53; n = 447) in 1996–1997 (Figure 5)  [75] .
The mean levels in the control group were also
extremely high at 1.52 ±0.39 8-oxo-dGuo/105 dGuo
(median 1.51; n = 214) [75] , likely because there was
no inhibition of DNA oxidation during isolation and
hydrolysis. The studies indicated that levels of oxida-
tive stress were between 1.7- and 2.0-times the level
of oxidative damage relative to that found in control
samples in all 3 years of the study [75,76] ; however, the
Asbestos-exposed workers
1995/96 1996/97
(n = 437) (n = 447)
median = 2.76 median = 2.53
mean ± SD = 2.96 ± 1.10* mean ± SD = 2.55 ± 0.56*
www.future-science.com 1163
Review  Mesaros, Worth, Snyder et al.
data must be treated with caution. Typical levels of
8-oxo-dGuo in cells where artifactual DNA oxidation
is prevented are at least two orders of magnitude lower
at 1.0 8-oxo-dGuo/107 dGuo [71] . Therefore, oxidative
DNA damage could be higher in the WBCs of work-
ers highly exposed to asbestos fibers but more rigorous
assay methodology will be required for validation. If
these findings are confirmed it would indicate that pre-
ventive and therapeutic approaches using antioxidants
might be useful.
The most recent in vivo studies of 8-oxo-dGuo
surveyed subjects occupationally exposed to asbestos
in Italy [50,51] . The first study included 119 asbestos-
exposed workers from the shipbuilding industry,
and 54 aged matched controls unlikely to have been
exposed to asbestos based on their job history. The
second study examined 94 asbestos-exposed subjects
and 54 controls. The studies quantified 8-oxo-dGuo
in WBCs using the OxyDNA assay kit with no control
of artifactual DNA oxidation [50,51] . Levels of WBC
8-oxo-dGuo were significantly elevated in subjects
heavily exposed to asbestos but were identical with
subjects with MM [50] . The area under the receiver
operating characteristic curves (AUC) of 0.775 showed
that WBC 8-oxo-dGuo levels poorly differentiated
asbestos-exposed subjects from healthy controls [50] .
Isoprostanes as biomarkers
Arachidonic acid is a major fatty acyl component of the
lipidome that is present as the free fatty acid in plasma
as well as esterified in sterol lipids and at the sn-2 posi-
tion of glycerolipids and glycerophospholipids [77] .
IsoPs are oxidation products of arachidonic acid, which
were originally discovered as artifacts present in stored
plasma samples [78] . Subsequently, we showed that they
are also formed by ROS-mediated oxidation of esteri-
fied arachidonic acid, which appear in biofluids after
hydrolysis of esterified lipids [79] . 8-iso-isoprostane
PGF2α (8-iso-PGF2α also known as 8-epi-PGF2α and
iPF2α-III) is the most widely analyzed isoP, with rela-
tively few reports describing the detection of more than
one isoP in a single LC–MS analysis [80] . Each class of
isoP forms a specific product ion during LC–MS/MS
analysis  [81] , making it possible to readily differentiate
the four isoP classes [80] . IsoPs have been rigorously
validated as reliable biomarkers of oxidative stress in
two multilaboratory collaborative studies [82,83] .
Very few studies have assessed isoP levels in asbes-
tos exposed populations [53,84–87] . In one of the studies,
8-iso-PGF2α was analyzed by stable isotope dilution
LC–MS in exhaled breath condensate (EBC) from
healthy subjects who had been exposed to asbestos
(Table 1)  [53] . The limit of quantification for this
method was 5 pg/ml and the mean levels were approxi-
1164 Bioanalysis (2015) 7(9)
mately ten-times higher. The study evaluated 92 for-
mer asbestos workers with mean age 68.8 ±1.7 years
and mean duration of asbestos exposure of 24.1 ±2.0
years. The control group had 46 subjects with mean
age 65.2 ±3.3 years. The mean level of 8-iso-PGF2α was
higher in asbestos-exposed subjects (69.5 ±6.6 pg/ml;
p = 0.0001) compared with the control group, where
the concentration was 47.0 ±7.8 pg/ml. There was no
correlation with asbestos fiber exposure years. How-
ever, the results support the hypothesis that oxidative
stress will lead to increased 8-iso-PGF2α in EBC, and
there is a direct correlation between asbestos exposure
and oxidative stress. Therefore, the analysis of isoPs
in EBC is a promising noninvasive tool for assessing
asbestos exposure but may also reveal any condition
leading to oxidative stress. Thus, the measurements of
isoPs could be coupled with more specific biomarkers
of asbestos exposure. Strikingly, to date, there are no
other known small molecule biomarkers available to
distinguish between asbestos-exposed individuals and
healthy controls. This lack of studies highlights the
acute need for discovery of such biomarkers.
Protein biomarkers
Mesothelin, which was first identified on mesothelial
cells, is known to be overexpressed in several types of
cancer. The soluble form, which is known as soluble
mesothelin-related protein or soluble mesothelin-
related peptide (SMRP), has emerged as a potential
biomarker for the early detection of asbestos-induced
mesothelioma  [88,89] . Multiple enzyme-linked immu-
nosorbent assay (ELISA) kits have been developed to
analyze serum SMRP including the Mesomark assay,
which has been approved by the US FDA as a bio-
marker for mesothelioma (Table 1) [54] . A recent review
reported a meta analysis of 30 publications to deter-
mine the sensitivity and specificity of SMRP as a bio-
marker of mesothelioma [90] . Studies on occupational
exposure to asbestos in the Czech Republic revealed
distinguishing serum levels of SMRP in which exposed
subjects with benign disease had higher levels than nor-
mal subjects but lower levels than subjects with MM
as determined by CXRs [91] . An additional study from
Australia used a sandwich ELISA with two monoclo-
nal antibodies (OV569 and 4H3) comparing a non-
exposed control group of 28 controls with 40 asbestos
exposed (Table 1) [55] . This study showed increased lev-
els of SMRP in mesothelioma patients, but the small
sample size did not permit adequate statistical power.
There was no significant difference in the SMRP levels
between asbestos exposed and controls.
Similar ELISA-based studies utilizing plasma and
serum have shown elevation of SMRP levels in MM
subgroups with no statistical significance between
future science group

exposed individuals and unexposed controls [51,92–
94] . In contrast, analyses involving asbestos-exposed
workers revealed no association between total expo-
sure and serum SMRP levels [88,95] . Creaney et al. also
reported steady serum concentrations of mesothelin
in asbestos-exposed individuals over a time span of 4
years, suggesting their findings were not a result of
sampling time in relation to exposure occurrence. In
contrast to these reports, other studies found higher
serum levels of SMRP in asbestos-exposed individu-
als than unexposed controls [96–98] . One of these
studies  [96] also examined the correlation between
SMRP levels and frequency of micronuclei in blood.
Measurement of micronuclei frequency in peripheral
blood lymphocytes is extensively used to evaluate
the presence of DNA damage in humans exposed
to genotoxic agents and for the prediction of cancer
risk. The micronuclei frequency test is easier to con-
duct than the chromosomal aberration test. It uses
fluorescent in situ hybridization with probes targeted
to the centromere region. A statistically significant
positive correlation of the SMRP levels with the fre-
quency of mononuclei in the mononucleated lym-
phocytes was observed. Additional evidence indicates
serum SMRP levels may be useful for monitoring
the progression of diseases resulting from asbestos
exposure (Figure 6) [99] . Mean serum levels of SMRP
differed between the four groups that were exam-
ined with the mean level in healthy subjects being
lower than in subjects who were entitled to compen-
sation for their ARD (Figure 6) . The authors have
suggested that asbestos biomarkers could potentially
be used for monitoring disablement assessment in
ARDs  [99] . A potential issue with studies involving
SMRP quantification is the confounding effects of
sample storage, subject age, body mass index, glo-
merular filtration rate and smoking status within the
cohorts [89,100–101] .
Osteopontin is a glycoprotein that is overex-
pressed in lung cancer and several other types of can-
cer  [102] . It is an extracellular cell adhesion protein
that plays a key role in cytokine mediated immune
response. High levels of osteopontin are correlated
with tumor progression and metastasis. In a rat
model, osteopontin was upregulated in asbestos-
induced tumors [103] . Compellingly, osteopontin
levels in plasma or serum were able to differentiate
between healthy subjects exposed to asbestos and
mesothelioma patients [104] . Several human popula-
tion studies looked at serum levels of osteopontin in
asbestos-exposed subjects [56,92,95,104–106] . All studies
employed a commercially available ELISA kit, which
had a detection limit of 3 ng/ml (Table 1)  [56] . The
first population study [105] , which was conducted in
future science group
Biomarkers of asbestos exposure Review
Michigan, examined 69 asbestos-exposed subjects
and 45 healthy controls. The level of osteopontin was
elevated in the asbestos group (30 ±3 ng/ml) com-
pared with the unexposed group (20 ±4 ng/ml) but
was of borderline statistical significant (p = 0.06).
Creaney et al.  [56] examined an Australian cohort to
compare osteopontin levels in patients with different
lung disorders, including ten healthy controls exposed
to asbestos and ten nonexposed. Their findings agreed
with the Michigan study in that osteopontin was not
significantly different in the asbestos exposed popula-
tion. Furthermore, even if serum and plasma levels
of osteopontin were highly correlated with asbestos
exposure  [107] , osteopontin can be hydrolyzed by
thrombin and so the results might not be a true reflec-
tion of protein levels in the blood. A recent study con-
ducted in Turkey had 120 healthy controls and 123
subjects exposed to asbestos [92] . This was the first
study to examine a cohort of subjects exposed to nat-
urally occurring surface asbestos. The subjects lived
within 10 km of ophiolitic areas, which are known
sources of naturally occurring asbestos. The study
measured levels of mesothelin as well as osteopontin
and concluded that both proteins were related to the
5
4
SMRP (nmol/l)
3
2
1
0
Healthy exposed
plaques
Noncompensated
Compensated
Pleural
to asbestos
ARDs
ARDs
Disablement (%)
Figure 6. Serum soluble mesothelin-related peptide
concentrations in asbestos-exposed and nonexposed
subjects. Categories from left to right: exposed to
asbestos but apparently healthy; with pleural plaques;
with ARDs (includes asbestosis, DPT and asbestosis/
DPT) but not eligible for compensation due to their 0%
disablement; and with compensated ARDs due to their
10–100% disablement. Horizontal scale bars denote
mean concentrations. There was a significant difference
between the groups (analysis of variance, p < 0.0001).
ARD: Asbestos-related disease; DPT: Diffuse pleural
thickening; SMRP: Soluble mesothelin-related peptide.
Reproduced with permission from [99] © Elsevier (2012).
www.future-science.com 1165
Review  Mesaros, Worth, Snyder et al.
severity of the inflammation observed in individual
subjects. The difference in the levels of osteopontin
was significant (p < 0.05) in controls versus asbestos-
exposed individuals but was not useful to predict
malignant transformations (Figure 7) . Another recent
study  [95] analyzed mesothelin and osteopontin levels
using ELISA kits in a very large number of asbestos-
exposed workers (n = 1894) together with a smaller
number of unexposed controls (n = 102). The levels of
osteopontin were not significantly different between
the two groups. This study also found no correlation
between osteopontin and mesothelin levels with the
duration of the asbestos exposure.
Fibulins are a group of secreted glycoproteins that
act as connectors with components of the extracellular
matrix and thus play an important role in the devel-
opment of fibrotic tissue [108] . Fibulin-3 has recently
emerged as a potential plasma protein biomarker of
asbestos exposure with the capability of distinguish-
ing between exposed and disease states within multi-
ple cohorts using an ELISA kit (Table 1) [57] . Interest-
ingly, Pass et al. also found that in a parallel analysis
of matched samples fibulin-3 levels were lower in
serum than in plasma [57] . The authors suggest this
might be due to the presence of thrombin cleavage
sites within fibulin-3, which has implications for
other serum-based assays exploring the potential of
protein biomarkers. Furthermore, the same study
40
35
p < 0.001
30
Osteopontin (ng/ml)
25
20671
20
15
10
Mesothelioma Pleural plaque
n: 24
Figure 7. The median and quartile serum osteopontin levels. Samples were from mesothelioma patients, pleural
plaque patients, healthy subjects exposed to asbestos and a control nonexposed group.
Reprinted with permission from [92] © Springer (2013).
1166 Bioanalysis (2015) 7(9)
showed a poor correlation of fibulin-3 levels in
plasma and effusions from matched samples, bring-
ing to light a question of the validity of blood-based
assays for fibulin-3 quantification. Additional studies
have shown that fibulin-3 was not able to distinguish
between patients with MM and asbestosis because
serum levels were elevated in both groups [109] . How-
ever, it has been suggested that fibulin-3 is a better
prognostic biomarker for MM than a diagnos-
tic biomarker, because plasma fibulin-3 levels better
predicted survival in MM patients [110] . Additional
validation studies will be required to fully elucidate
the utility of fibulin-3 as a dependable biomarker of
asbestos exposure in human populations.
Fibronectin is a glycoprotein that is also involved in
the extracellular matrix structure and therefore plays
an important role in the generation of fibrotic tis-
sue. For this reason there has been interest in fibro-
nectin as a potential biomarker of asbestos exposure.
Unfortunately, relatively few human studies have
been performed to assess the utility of fibronectin for
this role. In vitro experiments utilizing lung fibro-
blasts have shown that expression of the fibronectin
gene increases in response to many of the cytokines
known to be released in response to asbestos fibers.
Bégin  et al. reported increased fibronectin levels in
BAL fluid (as determined immunochemically with a
laser nephelometer instrument) from sheep exposed
p < 0.001
p < 0.001
p < 0.05
p < 0.05 p < 0.05
8318
6448 7137
Healthy subjects
Control group
exposed to asbestosis
n: 279 n: 120
n: 123
future science group

to asbestos fibers [58] . This same study found that
humans exposed to asbestos who were suffering from
ARDs also had elevated BAL fluid levels of fibronectin.
Schwartz et al. performed a similar study involving 93
men who had been occupationally exposed to asbestos
and found increased BAL fluid levels of fibronectin (as
determined by ELISA) in patients with restricted lung
function (Table 1)  [59] . A similar finding was reported
the following year by the same group with a different
study population [111] .
The HMGB1 protein has a known regulatory role
in inflammatory immune responses and has received
attention as a potential novel therapeutic target in
MM  [112] . Normally located in the nucleus, HMGB1
is acetylated and translocates during cell necrosis due
to asbestos fibers into the cytosol and extracellular
space, where it binds to and activates proinflamma-
tory mediators. Specifically, activation of the NF-κB
signaling pathway and release of tumor necrosis fac-
tor-alpha (TNFα) are downstream consequences of
HMGB1 activity. Given the role it plays in inflamma-
tory processes, HMGB1 may hold promise as a bio-
marker of cell transformative processes and thus hold
utility as an indicator of asbestos exposure. Genetic
profiling of mesothelial cells exposed to asbestos
has shown upregulation of many genes targeted by
HMGB1 [112] . Furthermore, exposing mice to asbes-
tos increased serum levels of HMGB1, although
they were only maintained as long as the exposure
continued  [112] . One possible explanation for this
finding is that the mice were exposed to asbestos by
intraperitoneal injection, whereas most human expo-
sure are through inhalation, suggesting the route of
exposure may have influences on the long-term bio-
chemical impacts of asbestos fibers. One study found
elevated serum levels of HMGB1, using an ELISA
kit, in asbestos-exposed individuals compared with
both smoking and nonsmoking controls (Table 1) [60] ,
indicating serum HMGB1 could be exploited for
assessing asbestos exposure in human populations. In
agreement with this finding, serum levels of HMGB1
have also been reported to be elevated in MM patients
using an HMBG1 ELISA kit [113–115] .
TNFα can be produced by many different cell
types and is heavily involved in the regulation of
immune cells. Generated through an NF-κB depen-
dent pathway, TNFα is likely to increase in response
to oxidative insults (Figure 3) . Typically upregulated
after phagocytosis of fibers by macrophages, genera-
tion of TNFα may induce further binding of parti-
cles, thus exacerbating the effects of asbestos expo-
sure. It has also been proposed that TNFα plays an
important anticytotoxic role for human mesothelial
cells and therefore results in a higher abundance
future science group
Biomarkers of asbestos exposure Review
Key term
Prognostic biomarker: Indicator of the likely course of a
disease in an untreated individual.
of damaged cells that may go through a malignant
transformation  [116] . Although numerous cell-cul-
ture-based studies have confirmed the importance of
TNFα in responding to asbestos fibers, it remains to
be seen if TNFα levels hold promise as a marker of
asbestos exposure in human studies.
The large family of cytokines, the ILs, promote the
development, differentiation and activation of lym-
phocytes and therefore play an important role in the
immune response. As such, levels of several ILs have
been assessed as biomarkers of asbestos exposure.
In particular, increased IL-6 and IL-8 production
is associated with activation of the NRLP3 inflam-
masome in response to asbestos fibers [117] . Workers
occupationally exposed to asbestos have increased
serum levels of IL-6 and IL-8 when compared con-
trols in two studies using a solid phase ELISA test
(Table 1)  [61,62] . In each of these studies, workers
who were occupationally exposed to asbestos fibers
within factories were compared with unexposed con-
trols. Interestingly, each study employed two sets of
controls both within each factory and within each
town containing the factory. In all cases serum levels
of IL-6 and IL-8 were significantly elevated in the
asbestos exposed group over both sets of controls.
In agreement with this finding, other work found
serum levels of IL-6 to track with asbestos exposure
in a cohort of workers differentially exposed to asbes-
tos  [50] . This particular study involved 119 subjects
with a history of occupational exposure to asbestos
that were compared with 54 age-matched controls.
For IL-6 serum quantification the exposed popula-
tion was divided into three groups based on cumula-
tive asbestos exposure. The highest exposure group
was found to have statistically increased levels of IL-6
while there was no statistical significance between
the other two groups. In contrast to this finding, one
comparison of healthy controls to exposed individu-
als with asbestosis found no difference in serum IL-8
levels  [109] . It is noteworthy that IL polymorphisms
do not associate with risk factors for diseases resulting
from asbestos exposure [118] .
The chemokine, regulated on activation normal T
cell expressed and secreted (RANTES), also known
as chemokine (C–C motif ) ligand 5 (CCL5) has
recently emerged as a potential biomarker of asbestos
exposure. By studying Italian workers with a mean
of 25 years of asbestos exposure one group found a
significant elevation in serum levels of RANTES
as determined by a magnetic bead multiplex
www.future-science.com 1167
Review  Mesaros, Worth, Snyder et al.
immunoassay (Table 1)  [63] . Importantly, RANTES
levels were also able to distinguish between asbestos-
exposed and MM subgroups. However, it remains
to be seen whether this biomarker will continue to
be valid in studies involving larger sample sizes and
different ARDs.
Currently available biomarkers of response to asbes-
tos exposure represent a good starting point for devel-
oping a specific and sensitive biomarker panel. The
availability of such a panel would make it possible to
identify individuals who have been exposed to asbestos
and to distinguish them from subjects who are at risk
for progression to mesothelioma. Availability of such a
panel would have a significant impact on the health of
asbestos-exposed individuals by providing early warn-
ing for monitoring the potential occurrence of ARDs.
It would also provide significant evidence to regulatory
authorities such as the US FDA that their remediation
efforts have been successful, particularly when indi-
vidual monitoring can be conducted on a regular basis.
Conclusion & future perspective
The detrimental health effects resulting from asbestos
exposure are not likely to subside in the near future.
Despite being banned in the USA, asbestos is still being
used in other parts of the world with much less strin-
gent controls on possible exposure of the workers [13] .
In addition, the often decades-long latency periods of
ARDs means that they will continue to manifest in the
USA for years to come [7] . As such, a reliable biomarker
panel capable of assessing levels of asbestos exposure
would provide a useful clinical tool in screening, diag-
nosis, prevention, and alleviating concerns about possi-
ble exposure and ensuring effective removal of asbestos
from the environment. The majority of response bio-
marker studies thus far have focused on small molecule
biomarkers of oxidative stress, signaling factors for cell-
mediated and humoral immune responses, and growth
factors generated in response to inhalation of asbestos
fibers. These studies have shed light on important fac-
tors involved with the pathogenesis of diseases resulting
from asbestos exposure as well as guided future efforts
for developing an effective biomarker panel for asbestos
exposure. However, considering the lack of specificity
for many of the biomarkers, it is likely that assessment
of new classes of compounds in response to asbestos
exposure will result in a synergistic effect on biomarker
performance.
LC–MS-based methodology coupled with careful
exclusion of artifact formation [35,71,80] could provide
more definitive information on the role of oxidative
DNA and lipid damage in response to asbestos expo-
sure. One approach that could be applied to discovery
of biomarkers of response resulting from asbestos expo-
1168 Bioanalysis (2015) 7(9)
sure involves the implementation of untargeted serum
metabolomics using ultraperformance LC coupled with
high-resolution MS [119] . This could potentially lead to
the discovery of a comprehensive panel of metabolomic
biomarkers of response that would be capable of iden-
tifying individuals exposed to asbestos. The biomarker
panel would also be useful for monitoring populations
to ensure environmental sources of asbestos exposure
have been adequately remediated. Stable isotope dilu-
tion LC–high-resolution MS could also be employed [73]
to distinguish the multiple SMRPs that are present in
serum (Table 1) as well as for defining the role of lysine
acetylation in the secretion HMBG1 protein (Table 1) .
The resulting assays could potentially improve specific-
ity in assessing biological responses to asbestos exposure
as well as improving the early detection of mesotheli-
oma. Finally, untargeted serum protein profiling, using
either Slow Off-rate Modified Aptamers as employed
for mesothelioma biomarker discovery [120] or LC–MS-
based proteomics methodology as we described recently
for biomarkers of preterm birth [121] , could be employed
to discover additional protein biomarkers of asbestos
exposure. Linking more specific response biomarkers
with existing radiographic markers could have a signifi-
cant impact on our ability to diagnose and treat ARDs.
Financial & competing interests disclosure
This work was supported by National Institutes of Health grants
P42ES023720, P30ES013508 and T32ES019851. C Mesaros is
a Senior Research Investigator in Systems Pharmacology and
Translational Therapeutics, A Worth is a Graduate Student in
Pharmacology, M Christofidou-Solomidou is a Research Asso-
ciate Professor of Medicine, A Vachani is an Assistant Professor
of Medicine, SA Albelda is the William Maul Measey Professor
of Medicine, IA Blair is the A N Richards Professor of Systems
Pharmacology and Translational Therapeutics and Director of
the Penn SRP Center. They are all employees of the University
of Pennsylvania. N Snyder is an Assistant Professor in the A J
Drexel Autism Institute at Drexel University. Studies described
in the article were not influenced by the authors’ current ap-
pointments, although research in many of the areas is currently
on-going. The authors have no other relevant affiliations or
financial involvement with any organization or entity with a
financial interest in or financial conflict with the subject mat-
ter or materials discussed in the manuscript apart from those
disclosed.
No writing assistance was utilized in the production of this
manuscript.
Open Access
This work is licensed under the Creative Commons Attribu-
tion-NonCommercial 3.0 Unported License. To view a copy
of this license, visit http://creativecommons.org/licenses/by-
nc-nd/3.0/
future science group
 Biomarkers of asbestos exposure Review

Executive summary
• Asbestos exposure is now known to cause lung cancer and mesothelioma.
• Despite being banned in the US, asbestos is still used industrially in other parts of the world.
• The detrimental health impacts resulting from asbestos exposure are not likely to subside in the near future.
• The often decades-long latency periods of ARDs means health impacts will result for years to come.
• A validated biomarker panel capable of assessing levels of asbestos exposure would provide a great service in
preventative care.
• Asbestos is not present in human biological fluids; this means that it is necessary to analyze biomarkers of
effect, which reflect a change in biologic function in response to asbestos exposure.
• Current biomarkers of effect, which include small molecule markers of oxidative stress and proteins involved
in immune responses, are not specific for asbestos exposure
• A validated panel of biomarkers of effect of asbestos exposure would alleviate concerns about exposure as
well as identify subjects at risk for asbestos-related disease.

References 11 Montjoy C, Parker J, Petsonk L, Luis T, Fallon K.

Papers of special note have been highlighted as either:
• of interest •• of considerable interest
1 Huncharek M. Asbestos and cancer: epidemiological and
public health controversies. Cancer Invest. 12(2), 214–222
(1994).
2 Testa JR, Cheung M, Pei J, Below JE, Tan Y, Sementino E
et al. Germline BAP1 mutations predispose to malignant
mesothelioma. Nat. Genet. 43(10), 1022–1025 (2011).
•• Discussion on pre-disposition of patients to DNA damage
and genetic mutations after environmental stress.
Mesothelioma review. W. V. Med. J. 105(3), 13–16 (2009).
12 Sterman DH, Albelda SM. Advances in the diagnosis,
evaluation, and management of malignant pleural
mesothelioma. Respirology 10(3), 266–283 (2005).
13 Linton A, Vardy J, Clarke S, van ZN. The ticking time-bomb
of asbestos: its insidious role in the development of malignant
mesothelioma. Crit. Rev. Oncol. Hematol. 84(2), 200–212
(2012).
14 Frank AL, Joshi TK. The global spread of asbestos. Ann.
Glob. Health 80(4), 257–262 (2014).

3 Carbone M, Yang H. Molecular pathways: targeting 15 Stayner L, Welch LS, Lemen R. The worldwide pandemic
mechanisms of asbestos and erionite carcinogenesis in of asbestos-related diseases. Annu. Rev. Public Health 34,
mesothelioma. Clin. Cancer Res. 18(3), 598–604 (2012). 205–216 (2013).

•• Excellent description mechanism of asbestos-induced •• Excellent overview of issues related to worldwide asbestos
oxidative stress. exposure.

4 Gibbs AR. Role of asbestos and other fibres in the
development of diffuse malignant mesothelioma. Thorax
45(9), 649–654 (1990).
5 International Agency for Research on Cancer. Arsenic,
metals, fibres, and dusts. IARC Monogr. Eval. Carcinog. Risks
Hum. 100(Pt C), 11–465 (2012).
16 Peipins LA, Lewin M, Campolucci S et al. Radiographic
abnormalities and exposure to asbestos-contaminated
vermiculite in the community of Libby, Montana, USA.
Environ. Health Perspect. 111(14), 1753–1759 (2003).
17 Noonan CW. Exposure matrix development for the Libby
cohort. Inhal. Toxicol. 18(12), 963–967 (2006).

• In-depth description of risks associated with asbestos 18 BoRit asbestos superfund site community advisory group.
www.boritcag.org 
exposure.
19 Rushton L, Hutchings SJ, Fortunato L et al. Occupational
6 Cogliano VJ, Baan R, Straif K et al. Preventable exposures
cancer burden in Great Britain. Br. J. Cancer 107(Suppl. 1),
associated with human cancers. J. Natl Cancer Inst. 103(24),
S3–S7 (2012).
1827–1839 (2011).
20 Paglietti F, Malinconico S, Di Molfetta V, Giangrasso M.
7 Carbone M, Ly BH, Dodson RF et al. Malignant
Guidelines for asbestos remediation at Italian superfund sites.
mesothelioma: facts, myths, and hypotheses. J. Cell. Physiol.
J. Environ. Sci. Health C. Environ. Carcinog. Ecotoxicol. Rev.
227(1), 44–58 (2012).
30(3), 253–286 (2012).
8 Neri M, Ugolini D, Boccia S et al. Chemoprevention of
21 Szeszenia-Dabrowska N, Swiatkowska B, Szubert Z,
asbestos-linked cancers: a systematic review. Anticancer Res.
Wilczynska U. Asbestos in Poland: occupational health
32(3), 1005–1013 (2012).
problems. Int. J. Occup. Med. Environ. Health 24(2),
9 Berman DW, Crump KS. Update of potency factors for 142–152 (2011).
asbestos-related lung cancer and mesothelioma. Crit. Rev.
22 Offermans NS, Vermeulen R, Burdorf A et al. Occupational
Toxicol. 38(Suppl. 1), 1–47 (2008).
asbestos exposure and risk of pleural mesothelioma, lung
10 Stayner L, Kuempel E, Gilbert S, Hein M, Dement J. An cancer, and laryngeal cancer in the prospective Netherlands
epidemiological study of the role of chrysotile asbestos cohort study. J. Occup. Environ. Med. 56(1), 6–19 (2014).
fibre dimensions in determining respiratory disease risk in
23 Kameda T, Takahashi K, Kim R et al. Asbestos: use, bans
exposed workers. Occup. Environ. Med. 65(9), 613–619
and disease burden in Europe. Bull. World Health Organ.
(2008).
92(11), 790–797 (2014).

future science group www.future-science.com 1169

Review  Mesaros, Worth, Snyder et al.
24 MacTiernan A, Fritschi L, Slevin T, Jalleh G, Donovan
R, Heyworth J. Public perceptions of cancer risk factors: a
Western Australian study. Health Promot. J. Austr. 25(2),
90–96 (2014).
25 Reid A, de Klerk NH, Magnani C et al. Mesothelioma risk
after 40 years since first exposure to asbestos: a pooled analysis.
Thorax 69(9), 843–850 (2014).
26 Van den Borre L, Deboosere P. Asbestos in Belgium: an
underestimated health risk. The evolution of mesothelioma
mortality rates (1969–2009). Int. J. Occup. Environ. Health
20(2), 134–140 (2014).
27 Sartorelli P, Scancarello G, Romeo R et al. Asbestos exposure
assessment by mineralogical analysis of bronchoalveolar lavage
fluid. J. Occup. Environ. Med. 43(10), 872–881 (2001).
28 The Industrial Injuries Advisory Council. Pleural Plaques,
Postion Paper 23, 1–60 (2009).
• Comprehensive report on asbestos-induced pleural plaques in
the UK.
29 American Thoracic Society. Diagnosis and initial management
of nonmalignant diseases related to asbestos. Am. J. Respir.
Crit. Care Med. 170(6), 691–715 (2004).
•• Excellent overview of diagnosis of asbestos-related diseases
(ARDs).
30 Pairon JC, Laurent F, Rinaldo M et al. Pleural plaques and
the risk of pleural mesothelioma. J. Natl Cancer Inst. 105(4),
293–301 (2013).
• Very informative paper describing the relationship between
asbestos exposure, pleural plaques, and mesothelioma
31 Cullen MR, Barnett MJ, Balmes JR et al. Predictors of lung
cancer among asbestos-exposed men in the {beta}-carotene
and retinol efficacy trial. Am. J. Epidemiol. 161(3), 260–270
(2005).
32 Pilger A, Rudiger HW. 8-Hydroxy-2’-deoxyguanosine as a
marker of oxidative DNA damage related to occupational and
environmental exposures. Int. Arch. Occup. Environ. Health
80(1), 1–15 (2006).
• Discussion of 8-oxo-dGuo as a biomarker of oxidative stress.
33 Valavanidis A, Fiotakis K, Vlachogianni T. Airborne
particulate matter and human health: toxicological assessment
and importance of size and composition of particles for
oxidative damage and carcinogenic mechanisms. J. Environ.
Sci. Health C. Environ. Carcinog. Ecotoxicol. Rev. 26(4),
339–362 (2008).
34 Valavanidis A, Vlachogianni T, Fiotakis C. 8-hydroxy-2’-
deoxyguanosine (8-OHdG): A critical biomarker of oxidative
stress and carcinogenesis. J. Environ. Sci. Health C. Environ.
Carcinog. Ecotoxicol. Rev. 27(2), 120–139 (2009).
35 Mesaros C, Arora JS, Wholer A, Vachani A, Blair IA. 8-Oxo-
2’-deoxyguanosine as a biomarker of tobacco-smoking-induced
oxidative stress. Free Radic. Biol. Med. 53(3), 610–617 (2012).
•• Rigorous bioanalytical method for the analysis of urinary
8-oxo-dGuo.
36 Serban C, Dragan S. The relationship between inflammatory
and oxidative stress biomarkers, atherosclerosis and
rheumatic diseases. Curr. Pharm. Des. 20(4), 585–600
(2014).
1170 Bioanalysis (2015) 7(9)
37 Cheresh P, Morales-Nebreda L, Kim SJ et al. Asbestos-
induced pulmonary fibrosis is augmented in 8-oxoguanine
DNA glycosylase knockout mice. Am. J. Respir. Cell Mol.
Biol. 52(1), 25–36 (2015).
38 Sezgi C, Taylan M, Sen HS et al. Oxidative status and acute
phase reactants in patients with environmental asbestos
exposure and mesothelioma. ScientificWorldJournal 902748
doi: 10.1155/2014/902748 (2014).
39 Lamote K, Nackaerts K, van Meerbeeck JP. Strengths,
weaknesses, and opportunities of diagnostic breathomics
in pleural mesothelioma-a hypothesis. Cancer Epidemiol.
Biomarkers Prev. 23(6), 898–908 (2014).
40 Mossman BT, Shukla A, Heintz NH, Verschraegen CF,
Thomas A, Hassan R. New insights into understanding the
mechanisms, pathogenesis, and management of malignant
mesotheliomas. Am. J. Pathol. 182(4), 1065–1077 (2013).
41 Procopio A, Marinacci R, Marinetti MR et al. SV40
expression in human neoplastic and non-neoplastic tissues:
perspectives on diagnosis, prognosis and therapy of human
malignant mesothelioma. Dev. Biol. Stand. 94, 361–367
(1998).
42 Pooley FD. Electron microscope characteristics of inhaled
chrysotile asbestos fibre. Br. J. Ind. Med. 29(2), 146–153
(1972).
43 Hillegass JM, Miller JM, Macpherson MB et al. Asbestos and
erionite prime and activate the NLRP3 inflammasome that
stimulates autocrine cytokine release in human mesothelial
cells. Part. Fibre Toxicol. 10, 39 (2013).
44 Ramos-Nino ME, Testa JR, Altomare DA et al. Cellular and
molecular parameters of mesothelioma. J. Cell. Biochem.
98(4), 723–734 (2006).
45 Antoine DJ, Williams DP, Kipar A et al. High-mobility
group box-1 protein and keratin-18, circulating serum
proteins informative of acetaminophen-induced necrosis and
apoptosis in vivo. Toxicol. Sci. 112(2), 521–531 (2009).
•• Description of how acetylation of HMGB1 results in
secretion of the protein.
46 Hold GL, El-Omar EM. Genetic aspects of inflammation
and cancer. Biochem. J. 410(2), 225–235 (2008).
47 Colotta F, Allavena P, Sica A, Garlanda C, Mantovani
A. Cancer-related inflammation, the seventh hallmark of
cancer: links to genetic instability. Carcinogenesis 30(7),
1073–1081 (2009).
48 Tagesson C, Chabiuk D, Axelson O, Baranski B, Palus J,
Wyszynska K. Increased urinary excretion of the oxidative
DNA adduct, 8-hydroxydeoxyguanosine, as a possible early
indicator of occupational cancer hazards in the asbestos,
rubber, and azo-dye industries. Pol. J. Occup. Med. Environ.
Health 6(4), 357–368 (1993).
49 Yoshida R, Ogawa Y, Shioji I et al. Urinary 8-oxo-7,
8-dihydro-2’-deoxyguanosine and biopyrrins levels among
construction workers with asbestos exposure history. Ind.
Health 39(2), 186–188 (2001).
50 Amati M, Tomasetti M, Mariotti L, Tarquini LM, Valentino
M, Santarelli L. Assessment of biomarkers in asbestos-
exposed workers as indicators of cancer risk. Mutat. Res.
655(1–2), 52–58 (2008).
future science group

51 Amati M, Tomasetti M, Scartozzi M et al. Profiling
tumor-associated markers for early detection of malignant
mesothelioma: an epidemiologic study. Cancer Epidemiol.
Biomarkers Prev. 17(1), 163–170 (2008).
52 Tomasetti M, Amati M, Nocchi L et al. Asbestos exposure
affects poly(ADP-ribose) polymerase-1 activity: role in
asbestos-induced carcinogenesis. Mutagenesis 26(5), 585–591
(2011).
53 Pelclova D, Fenclova Z, Kacer P, Kuzma M, Navratil T,
Lebedova J. Increased 8-isoprostane, a marker of oxidative
stress in exhaled breath condensate in subjects with asbestos
exposure. Ind. Health 46(5), 484–489 (2008).
54 Beyer HL, Geschwindt RD, Glover CL et al. MESOMARK:
a potential test for malignant pleural mesothelioma. Clin.
Chem. 53(4), 666–672 (2007).
55 Robinson BW, Creaney J, Lake R et al. Mesothelin-family
proteins and diagnosis of mesothelioma. Lancet 362(9396),
1612–1616 (2003).
56 Creaney J, Yeoman D, Demelker Y et al. Comparison
of osteopontin, megakaryocyte potentiating factor, and
mesothelin proteins as markers in the serum of patients with
malignant mesothelioma. J. Thorac. Oncol. 3(8), 851–857
(2008).
57 Pass HI, Levin SM, Harbut MR et al. Fibulin-3 as a blood
and effusion biomarker for pleural mesothelioma. N. Engl.
J. Med. 367(15), 1417–1427 (2012).
•• Landmark paper describing the use of ELISA-based
bioanalytical methodology for analysis of Fibulin-3 as a
biomarker or ARDs.
58 Begin R, Martel M, Desmarais Y et al. Fibronectin and
procollagen 3 levels in bronchoalveolar lavage of asbestos-
exposed human subjects and sheep. Chest 89(2), 237–243
(1986).
59 Schwartz DA, Galvin JR, Frees KL et al. Clinical relevance
of cellular mediators of inflammation in workers exposed to
asbestos. Am. Rev. Respir. Dis. 148(1), 68–74 (1993).
60 Yang H, Rivera Z, Jube S et al. Programmed necrosis induced
by asbestos in human mesothelial cells causes high-mobility
group box 1 protein release and resultant inflammation. Proc.
Natl Acad. Sci. USA 107(28), 12611–12616 (2010).
• Excellent paper describing the potential utility of ELISA-
based bioanalytical methodology for analysis of HMGB1 as
a response biomarker of asbestos exposure.
61 Ilavska S, Jahnova E, Tulinska J et al. Immunological
monitoring in workers occupationally exposed to asbestos.
Toxicology 206(2), 299–308 (2005).
62 Tulinska J, Jahnova E, Dusinska M et al. Immunomodulatory
effects of mineral fibres in occupationally exposed workers.
Mutat. Res. 553(1–2), 111–124 (2004).
63 Comar M, Zanotta N, Bonotti A et al. Increased levels of
C-C chemokine RANTES in asbestos exposed workers and
in malignant mesothelioma patients from an hyperendemic
area. PLoS ONE 9(8), e104848 (2014).
• Interesting paper on the use ELISA-based bioanalytical
methodology for analysis of regulated on activation normal
T cell expressed and secreted (RANTES) as a biomarker of
asbestos exposure.
future science group
Biomarkers of asbestos exposure Review
64 US EPA. Pesticides: Science and Policy.
www.epa.gov
65 Sartorelli P, Romeo R, Scancarello G, Montomoli L,
Muzzupappa C, Barabesi L. Measurement of asbestos fibre
concentrations in fluid of repeated bro-choalveolar lavages of
exposed workers. Ann. Occup. Hyg. 51(5), 495–500 (2007).
66 Altomare DA, Menges CW, Pei J et al. Activated TNF-
alpha/NF-kappaB signaling via down-regulation of
Fas-associated factor 1 in asbestos-induced mesotheliomas
from Arf knockout mice. Proc. Natl Acad. Sci. USA 106(9),
3420–3425 (2009).
67 Gustafson AM, Soldi R, Anderlind C et al. Airway PI3K
pathway activation is an early and reversible event in lung
cancer development. Sci. Transl. Med. 2(26), 26ra25 (2010).
68 Hillegass JM, Shukla A, Lathrop SA et al. Inflammation
precedes the development of human malignant
mesotheliomas in a SCID mouse xenograft model. Ann. NY
Acad. Sci. 1203, 7–14 (2010).
69 Lotti M, Bergamo L, Murer B. Occupational toxicology of
asbestos-related malignancies. Clin. Toxicol. (Phila) 48(6),
485–496 (2010).
70 Myers R. Asbestos-related pleural disease. Curr. Opin. Pulm.
Med. 18(4), 377–381 (2012).
71 Mangal D, Vudathala D, Park JH, Lee SH, Penning TM,
Blair IA. Analysis of 7,8-dihydro-8-oxo-2’-deoxyguanosine
in cellular DNA during oxidative stress. Chem. Res. Toxicol.
22(5), 788–797 (2009).
•• Techniques for preventing oxidation of DNA in
bioanalytical methods for 8-oxo-dGuo.
72 Weiss JM, Goode EL, Ladiges WC, Ulrich CM.
Polymorphic variation in hOGG1 and risk of cancer: a
review of the functional and epidemiologic literature. Mol.
Carcinog. 42(3), 127–141 (2005).
73 Ciccimaro E, Blair IA. Stable-isotope dilution LC-MS for
quantitative biomarker analysis. Bioanalysis 2(2), 311–341
(2010).
•• Comprehensive overview describing the important role
that stable isotope analog internal standards can play in
bioanalytical methods for small molecule and protein
biomarkers.
74 Takahashi K, Pan G, Kasai H et al. Relationship between
Asbestos Exposures and 8-Hydroxydeoxyguanosine Levels in
Leukocytic DNA of Workers at a Chinese Asbestos-material
Plant. Int. J. Occup. Environ. Health 3(2), 111–119 (1997).
75 Marczynski B, Rozynek P, Kraus T, Schlosser S, Raithel HJ,
Baur X. Levels of 8-hydroxy-2’-deoxyguanosine in DNA of
white blood cells from workers highly exposed to asbestos in
Germany. Mutat. Res. 468(2), 195–202 (2000).
76 Marczynski B, Kraus T, Rozynek P, Raithel HJ, Baur X.
Association between 8-hydroxy-2’-deoxyguanosine levels in
DNA of workers highly exposed to asbestos and their clinical
data, occupational and non-occupational confounding
factors, and cancer. Mutat. Res. 468(2), 203–212 (2000).
77 Roberts LJ, Morrow JD. Measurement of F(2)-isoprostanes
as an index of oxidative stress in vivo. Free Radic. Biol. Med.
28(4), 505–513 (2000).
www.future-science.com 1171
Review  Mesaros, Worth, Snyder et al.
•• Excellent overview of isoprostanes as biomarkers of
oxidative stress.
78 Morrow JD, Harris TM, Roberts LJ. Noncyclooxygenase
oxidative formation of a series of novel prostaglandins:
analytical ramifications for measurement of eicosanoids.
Anal. Biochem. 184(1), 1–10 (1990).
• Original description of isoprostanes.
79 Morrow JD, Awad JA, Boss HJ, Blair IA, Roberts LJ. Non-
cyclooxygenase-derived prostanoids (F2-isoprostanes) are
formed in situ on phospholipids. Proc. Natl Acad. Sci. USA
89(22), 10721–10725 (1992).
• Original description of the formation of esterified
isoprostanes.
80 Mesaros C, Lee SH, Blair IA. Targeted quantitative analysis
of eicosanoid lipids in biological samples using liquid
chromatography-tandem mass spectrometry. J. Chromatogr.
B. Analyt. Technol. Biomed. Life Sci. 877(26), 2736–2745
(2009).
•• Overview of bioanalytical methods for oxidized lipids and
isoprostanes.
81 Li H, Lawson JA, Reilly M et al. Quantitative high
performance liquid chromatography/tandem mass
spectrometric analysis of the four classes of F(2)-isoprostanes
in human urine. Proc. Natl Acad. Sci. USA 96(23), 13381–
13386 (1999).
82 Kadiiska MB, Gladen BC, Baird DD et al. Biomarkers of
oxidative stress study III. Effects of the nonsteroidal anti-
inflammatory agents indomethacin and meclofenamic acid
on measurements of oxidative products of lipids in CCl4
poisoning. Free Radic. Biol. Med. 38(6), 711–718 (2005).
83 Kadiiska MB, Gladen BC, Baird DD et al. Biomarkers of
oxidative stress study II: are oxidation products of lipids,
proteins, and DNA markers of CCl4 poisoning? Free Radic.
Biol. Med. 38(6), 698–710 (2005).
• Bioanalytical validation of isoprostanes as oxidative stress
biomarkers.
84 Pelclova D, Fenclova Z, Syslova K et al. Oxidative stress
markers in exhaled breath condensate in lung fibroses are not
significantly affected by systemic diseases. Ind. Health 49(6),
746–754 (2011).
85 Lehtimaki L, Oksa P, Jarvenpaa R et al. Pulmonary
inflammation in asbestos-exposed subjects with borderline
parenchymal changes on HRCT. Respir. Med. 104(7),
1042–1049 (2010).
86 Lehtonen H, Oksa P, Lehtimaki L et al. Increased alveolar
nitric oxide concentration and high levels of leukotriene B(4)
and 8-isoprostane in exhaled breath condensate in patients
with asbestosis. Thorax 62(7), 602–607 (2007).
87 Chow S, Campbell C, Sandrini A, Thomas PS, Johnson
AR, Yates DH. Exhaled breath condensate biomarkers
in asbestos-related lung disorders. Respir. Med. 103(8),
1091–1097 (2009).
88 Creaney J, Olsen NJ, Brims F et al. Serum mesothelin
for early detection of asbestos-induced cancer malignant
mesothelioma. Cancer Epidemiol. Biomarkers Prev. 19(9),
2238–2246 (2010).
1172 Bioanalysis (2015) 7(9)
89 Park EK, Thomas PS, Creaney J, Johnson AR, Robinson
BW, Yates DH. Factors affecting soluble mesothelin related
protein levels in an asbestos-exposed population. Clin. Chem.
Lab. Med. 48(6), 869–874 (2010).
90 Cui A, Jin XG, Zhai K, Tong ZH, Shi HZ. Diagnostic values
of soluble mesothelin-related peptides for malignant pleural
mesothelioma: updated meta-analysis. BMJ Open 4(2),
e004145 (2014).
• Comprehensive review and meta-analysis of soluble
mesothelin-related protein (SMRP) biomarkers.
91 Jakubec P, Pelclova D, Smolkova P, Kolek V, Nakladalova
M. Significance of serum mesothelin in an asbestos-exposed
population in the Czech Republic. Biomed. Pap. Med
Fac. Univ Palacky. Olomouc. Czech Repub. doi: 10.5507/
bp.2014.015 (2014) (Epub ahead of print).
92 Bayram M, Dongel I, Akbas A, Benli I, Akkoyunlu ME,
Bakan ND. Serum biomarkers in patients with mesothelioma
and pleural plaques and healthy subjects exposed to naturally
occurring asbestos. Lung 192(1), 197–203 (2014).
93 Hollevoet K, Nackaerts K, Thimpont J et al. Diagnostic
performance of soluble mesothelin and megakaryocyte
potentiating factor in mesothelioma. Am. J. Respir. Crit. Care
Med. 181(6), 620–625 (2010).
94 Iwahori K, Osaki T, Serada S et al. Megakaryocyte
potentiating factor as a tumor marker of malignant pleural
mesothelioma: evaluation in comparison with mesothelin.
Lung Cancer 62(1), 45–54 (2008).
95 Felten MK, Khatab K, Knoll L, Schettgen T, Muller-
Berndorff H, Kraus T. Changes of mesothelin and
osteopontin levels over time in formerly asbestos-exposed
power industry workers. Int. Arch. Occup. Environ. Health
87(2), 195–204 (2014).
96 Marini V, Michelazzi L, Cioe A, Fucile C, Spigno F,
Robbiano L. Exposure to asbestos: correlation between
blood levels of mesothelin and frequency of micronuclei in
peripheral blood lymphocytes. Mutat. Res. 721(1), 114–117
(2011).
97 Rodriguez Portal JA, Rodriguez BE, Rodriguez RD et
al. Serum levels of soluble mesothelin-related peptides in
malignant and nonmalignant asbestos-related pleural disease:
relation with past asbestos exposure. Cancer Epidemiol.
Biomarkers Prev. 18(2), 646–650 (2009).
98 Pass HI, Wali A, Tang N et al. Soluble mesothelin-related
peptide level elevation in mesothelioma serum and pleural
effusions. Ann. Thorac. Surg. 85(1), 265–272 (2008).
99 Park EK, Yates DH, Creaney J, Thomas PS, Robinson BW,
Johnson AR. Association of biomarker levels with severity
of asbestos-related diseases. Saf. Health Work 3(1), 17–21
(2012).
100 Park EK, Wilson D, Yates DH. A predictive equation to
adjust for clinical variables in soluble mesothelin-related
protein (SMRP) levels. Clin. Chem. Lab. Med. 50(12),
2199–2204 (2012).
101 Weber DG, Johnen G, Taeger D, Weber A, Gross IM, Pesch
B et al. Assessment of confounding factors affecting the
tumor markers SMRP, CA125, and CYFRA21–1 in Serum.
Biomarker Insights 5, 1–8 (2010).
future science group

102 Denhardt DT, Chambers AF. Overcoming obstacles to
metastasis–defenses against host defenses: osteopontin
(OPN) as a shield against attack by cytotoxic host cells.
J. Cell. Biochem. 56(1), 48–51 (1994).
103 Sandhu H, Dehnen W, Roller M, Abel J, Unfried K. mRNA
expression patterns in different stages of asbestos-induced
carcinogenesis in rats. Carcinogenesis 21(5), 1023–1029
(2000).
104 Grigoriu BD, Scherpereel A, Devos P et al. Utility of
osteopontin and serum mesothelin in malignant pleural
mesothelioma diagnosis and prognosis assessment. Clin.
Cancer Res. 13(10), 2928–2935 (2007).
105 Pass HI, Lott D, Lonardo F et al. Asbestos exposure, pleural
mesothelioma, and serum osteopontin levels. N. Engl. J. Med.
353(15), 1564–1573 (2005).
106 Park EK, Yates DH, Creaney J, Thomas PS, Robinson BW,
Johnson AR. Association of biomarker levels with severity
of asbestos-related diseases. Saf. Health Work 3(1), 17–21
(2012).
107 Park EK, Thomas PS, Johnson AR, Yates DH. Osteopontin
levels in an asbestos-exposed population. Clin. Cancer Res.
15(4), 1362–1366 (2009).
108 Argraves WS, Greene LM, Cooley MA, Gallagher WM.
Fibulins: physiological and disease perspectives. EMBO Rep.
4(12), 1127–1131 (2003).
109 Corradi M, Goldoni M, Alinovi R et al. YKL-40 and
mesothelin in the blood of patients with malignant
mesothelioma, lung cancer and asbestosis. Anticancer Res.
33(12), 5517–5524 (2013).
110 Creaney J, Dick IM, Meniawy TM et al. Comparison
of fibulin-3 and mesothelin as markers in malignant
mesothelioma. Thorax 69(10), 895–902 (2014).
111 Schwartz DA, Davis CS, Merchant JA et al. Longitudinal
changes in lung function among asbestos-exposed workers.
Am. J. Respir. Crit. Care Med. 150(5 Pt 1), 1243–1249
(1994).
112 Qi F, Okimoto G, Jube S et al. Continuous exposure to
chrysotile asbestos can cause transformation of human
future science group
Biomarkers of asbestos exposure Review
mesothelial cells via HMGB1 and TNF-alpha signaling. Am.
J. Pathol. 183(5), 1654–1666 (2013).
113 Jube S, Rivera ZS, Bianchi ME et al. Cancer cell secretion of
the DAMP protein HMGB1 supports progression in malignant
mesothelioma. Cancer Res. 72(13), 3290–3301 (2012).
114 Tabata C, Kanemura S, Tabata R et al. Serum HMGB1 as a
diagnostic marker for malignant peritoneal mesothelioma.
J. Clin. Gastroenterol. 47(8), 684–688 (2013).
115 Tabata C, Shibata E, Tabata R et al. Serum HMGB1 as a
prognostic marker for malignant pleural mesothelioma. BMC
Cancer 13), 205–211 (2013).
116 Yang H, Bocchetta M, Kroczynska B et al. TNF-alpha
inhibits asbestos-induced cytotoxicity via a NF-kappaB-
dependent pathway, a possible mechanism for asbestos-induced
oncogenesis. Proc. Natl Acad. Sci. USA 103(27), 10397–10402
(2006).
117 Dostert C, Petrilli V, Van BR, Steele C, Mossman BT, Tschopp
J. Innate immune activation through Nalp3 inflammasome
sensing of asbestos and silica. Science 320(5876), 674–677
(2008).
• Overview of asbestos-mediated activation of the Nalp3
inflammasome.
118 Helmig S, Grossmann M, Wubbeling J, Schneider J.
Interleukin gene polymorphisms in pneumoconiosis. Int.
J. Mol. Med. 30(2), 401–408 (2012).
119 Snyder NW, Khezam M, Mesaros CA, Worth A, Blair IA.
Untargeted metabolomics from biological sources using
ultraperformance liquid chromatography-high resolution
mass spectrometry (UPLC-HRMS). J. Vis. Exp. (75), e50433
(2013).
120 Ostroff RM, Mehan MR, Stewart A et al. Early detection
of malignant pleural mesothelioma in asbestos-exposed
individuals with a noninvasive proteomics-based surveillance
tool. PLoS ONE 7(10), e46091 (2012).
121 Parry S, Zhang H, Biggio J et al. Maternal serum serpin B7 is
associated with early spontaneous preterm birth. Am.
J. Obstet. Gynecol. 211(6), 678, E1–E12 (2014).
www.future-science.com 1173