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488 Int Arch Occup Environ Health (2008) 81:487–493
higher pulmonary tissue Cr levels than controls (Tsuneta
et al. 1980), whereas Cr levels in occupationally unexposed
lung cancer patients and patients with other lung diseases
are lower and variable.
Exhaled breath condensate (EBC), which is obtained by
cooling the air exhaled under conditions of spontaneous
breathing, has recently been proposed as a promising biological
Xuid to monitor lung pathobiology. Various biomarkers
of inXammation and oxidative stress have been
identiWed and assayed (Rahman and Kelly 2003; Horvath
et al. 2005) and, over the last year, it has been demonstrated
that traces of pneumotoxic transition elements from
tobacco smoke or polluted environments, or involved in
oxidative stress as part of the redox system, can be found in
EBC (Mutti et al. 2006). Furthermore, a number of transition
metals have been measured in pulmonary tissue
(Adachi et al. 1991; Gerhardsson and Nordberg 1993;
Kubala-Kukus et al. 1999), and Cr has been related to biomarkers
of oxidative stress in the EBC of exposed chrome
plating workers (Caglieri et al. 2006). These Wndings suggest
that the measurement of EBC Cr seems to be a promising
method for deWning target organ Cr levels without
using invasive means.
The aim of this study was to measure Cr in EBC and in
urine in non-occupationally exposed non-small cell lung
cancer (NSCLC) patients before and after surgical intervention.
Moreover, we also took the advantage of this opportunity
to investigate the correlation between Cr levels in EBC
and pulmonary tissue.
Materials and methods
Subjects
We enrolled 20 patients, who underwent tumour resection
because of histological evidence of NSCLC at the University
of Parma’s Department of Thoracic Surgery. Tumour
size and node involvement were assessed using the International
Union Against Cancer TNM staging system, and all
of the patients were classiWed as having stage Ia, Ib or IIa
lung cancer. None of the patients received radiation or chemotherapy
before surgery. In particular, stage I (a or b) is
characterised by the absence of regional lymph node metastasis
(N0) and distant metastasis (M0) and stage IIa
(N1M0) is characterised by the presence of ipsilateral peribronchial
and/or ipsilateral hilar nodal metastases or intrapulmonary
nodes involved by direct extension of the
primary tumour, and by the absence of distant metastasis.
The asymptomatic group of control were 20 non-smokers,
not occupationally exposed to Cr, without any pulmonary
symptoms or a history of pulmonary disease, and with
normal lung spirometry results.
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Table 1 shows the characteristics of the study subjects.
In particular, 19 of the 20 NSCLC patients were smokers or
ex-smokers, all of whom had a history of at least 20 years
smoking of at least 15 cigarettes/day.
Study design
Before enrolment, the subjects completed a short questionnaire
concerning their current and previous medical history.
The EBC and urine samples of the NSCLC patients were
assessed immediately before and 2–4 days after tumour
resection, when patients were able to perform the manoeuvre.
Urine samples were obtained from only 15 patients, as
the other Wve presented with haematuria. Portions of cancerous
and non-cancerous lung tissue pieces were collected
during surgery, and immediately stored at ¡80°C until
analysis. The control EBC and urine samples were collected
during a normal working day in our laboratory.
All of the subjects gave their written informed consent to
the procedures, which were approved by our local ethics
committee. The biological material was sampled as laid
down in the Declaration of Helsinki.
EBC collection
EBC was collected using a TURBO-DECCS condenser
(Italchill, Parma, Italy) as previously described (Caglieri
et al. 2006; Goldoni et al. 2005, Mutti et al. 2006); the
selected condensation temperature was ¡5°C.
The subjects were asked to breathe tidally through the
mouthpiece for 10 min. They were instructed to form a complete
seal around the mouthpiece with their mouth, which
had to be kept dry by periodically swallowing excess saliva.
They were also asked to rinse their mouth with drinking
water thoroughly before, and every 5 min during the test. The
NSCLC patients had no diYculty in performing the manoeuvre
even during 2–4 days after tumour resection.
The EBC volume was measured using a calibrated
pipette, with salivary contamination being excluded by
Table 1 Demographic characteristics of studied groups
NSCLC Controls
Subjects (no.) 20 20
Age (median, years) 68.5 62.5
Sex (male/female) 17/3 14/6
Current smokers 10 0
Ex-smokers 9 0
Never smokers 1 20
The ex-smokers had stopped smoking for at least 1 year; otherwise
they were considered to be current smokers. All of the ex-smokers had
a 20–50-year history of smoking at least 15 cigarettes/day, like the current
smokers