Olga Acampa

Determination of aldehydes in exhaled breath of patients with lung cancer by means of on-fiber-derivatisation SPME–GC/MS

A number of volatile organic compounds (VOCs) have been identified and used in preliminary clinical studies of the early diagnosis of lung cancer. The aim of this study was to evaluate the potential of aldehydes (known biomarkers of oxidative stress) in the diagnosis of patients with non-small cell lung cancer (NSCLC). We used an on-fiber-derivatisation SPME sampling technique coupled with GC/MS analysis to measure straight aldehydes C3-C9 in exhaled breath. Linearity was established over two orders of magnitude (range: 3.3-333.3×10(-12) M); the LOD and LOQ of all the aldehydes were respectively 1×10(-12) M and 3×10(-12) M. Accuracy was within 93% and precision calculated as % RSD was 7.2-15.1%. Aldehyde stability in a Bio-VOC(®) tube stored at +4°C was 10-17 h, but this became >10 days using a specific fiber storage device. Finally, exhaled aldehydes were measured in 38 asymptomatic non-smokers (controls) and 40 NSCLC patients. The levels of all of the aldehydes were increased in the NSCLC patients without any significant effect of smoking habits and little effect of age. The good discriminant power of the aldehyde pattern (90%) was confirmed by multivariate analysis. These results show that straight aldehydes may be promising biomarkers associated with NSCLC, and increase the sensitivity and specificity of previously identified VOC patterns.

The Effect of Inhaled Chromium on Different Exhaled Breath Condensate Biomarkers among Chrome-Plating Workers

Chromium is corrosive, cytotoxic, and carcinogenic for humans and can induce acute and chronic lung tissue toxicity. The aim of this study was to investigate Cr levels in exhaled breath condensate (EBC) of workers exposed to Cr(VI) and to assess their relationship with biochemical changes in the airways by analyzing EBC biomarkers of oxidative stress, namely, hydrogen peroxide (H2O2) and malondialdehyde (MDA). EBC samples were collected from 24 chrome-plating workers employed in a chrome-plating plant both before and after the Friday work shift and before the work shift on the following Monday. Cr-EBC levels increased from the beginning (5.3 μg/L) to the end of Friday (6.4 μg/L) but were considerably lower on Monday morning (2.8 μg/L). A similar trend was observed for H2O2-EBC levels (which increased from 0.36 μM to 0.59 μM on Friday and were 0.19 μM on Monday morning) and MDA-EBC levels (which increased from 8.2 nM to 9.7 nM on Friday and were 6.6 nM on Monday). Cr-EBC levels correlated with those of H2O2-EBC (r = 0.54, p < 0.01) and MDA-EBC (r = 0.59, p < 0.01), as well as with urinary Cr levels (r = 0.25, p < 0.05). The results of this study demonstrate that EBC is a suitable matrix that can be used to investigate both Cr levels and biomarkers of free radical production sampling the epithelial-lining fluid of workers exposed to Cr(VI).

Exhaled breath condensate as a suitable matrix to assess lung dose and effects in workers exposed to cobalt and tungsten.

The aim of the present study was to investigate whether exhaled breath condensate (EBC), a fluid formed by cooling exhaled air, can be used as a suitable matrix to assess target tissue dose and effects of inhaled cobalt and tungsten, using EBC malondialdehyde (MDA) as a biomarker of pulmonary oxidative stress. Thirty-three workers exposed to Co and W in workshops producing either diamond tools or hard-metal mechanical parts participated in this study. Two EBC and urinary samples were collected: one before and one at the end of the work shift. Controls were selected among nonexposed workers. Co, W, and MDA in EBC were analyzed with analytical methods based on mass spectrometric reference techniques. In the EBC from controls, Co was detectable at ultratrace levels, whereas W was undetectable. In exposed workers, EBC Co ranged from a few to several hundred nanomoles per liter. Corresponding W levels ranged from undetectable to several tens of nanomoles per liter. A parallel trend was observed for much higher urinary levels. Both Co and W in biological media were higher at the end of the work shift in comparison with preexposure values. In EBC, MDA levels were increased depending on Co concentration and were enhanced by coexposure to W. Such a correlation between EBC MDA and both Co and W levels was not observed with urinary concentration of either element. These results suggest the potential usefulness of EBC to complete and integrate biomonitoring and health surveillance procedures among workers exposed to mixtures of transition elements and hard metals.

Plasma and EBC microRNAs as early biomarkers of non-small-cell lung cancer

Abstract Lung cancer is a major cause of death in Western countries. Current screening methods are invasive and still lead to a high percentage of false positives. There is, therefore, a need to find biomarkers that increase the probability of detecting lung cancer early. MicroRNAs (miRNAs) are stable molecules in blood plasma and exhaled breath condensate (EBC). We quantified miRNA-21 and miRNA-486 expression from plasma and EBC samples from patients with a diagnosis of non-small-cell lung cancer (NSCLC) and controls. miRNA-21 was significantly higher in plasma and in EBC of the NSCLC patients and miRNA-486 was significantly lower. This difference indicates a significantly improved diagnostic value, and suggests that these miRNAs could be clinically used as a first-line screening test in high-risk subjects.

Development and set-up of a portable device to monitor airway exhalation and deposition of particulate matter.

The aim of this study was to assess and monitor airway exhalation and deposition of particulate matter (PM). After standardizing inspiratory/expiratory flow and volumes, a novel device was tested on a group of 20 volunteers and in a field study on workers exposed to cristobalite. Both male and female subjects showed a higher percentage of deposition in the 0.5 μm channel than in the 0.3 μm channel on a laser particle counter, but it was higher in the males because of their higher exhaled lung volumes. The device was tested on a wider range of particles (0.3–0.5–1.0–2.5 μm) in the cristobalite productive division. The device has low intrasubject variability and good reproducibility, with geometric mean of %CV < 5%. Such a measure can be used to assess individual susceptibility to PM, making repeated measures in different environments, and examining the persistence of particles in the airways after a period in polluted environments.

Metallic elements in exhaled breath condensate of patients with interstitial lung diseases

Epidemiological data support the hypothesis that environmental and occupational agents play an important role in the development of interstitial lung diseases such as idiopathic interstitial pneumonia (IIPs) and sarcoidosis. The aim of this study was to assess the elemental composition of exhaled breath condensate (EBC) in patients with interstitial lung diseases (ILDs) of unknown etiology and healthy subjects as an indirect evaluation of tissue burden, which could improve our understanding of the role of metals in the pathogenesis of ILDs. EBC was obtained from 33 healthy subjects, 22 patients with sarcoidosis, 15 patients with non-specific interstitial pneumonia (NSIP) and 19 with IIPs. Trace elements and toxic metals in the samples were measured by means of inductively coupled plasma-mass spectrometry. There are only small overall differences in the EBC levels of a number of metallic elements among patients with idiopathic pulmonary fibrosis (IPF), NSIP or sarcoidosis, and no pattern is capable of distinguishing them with a high degree of sensitivity and specificity. However, a pattern of pneumotoxic (Si, Ni) and essential elements (Zn, Se and Cu) with the addition of Co distinguished the patients with ILDs from healthy non-smokers with relatively high degrees of sensitivity (96.4%) and specificity (90.9%). Assessing the elemental composition of EBC in patients with different ILDs seems to provide useful information. The non-invasiveness of the EBC method makes it suitable for patients with pulmonary diseases, although further studies are required to confirm the usefulness of this approach and to better understand the underlying pathophysiological processes.

Inter- and intra-subject variability of kinetics of airway exhalation and deposition of particulate matter in indoor polluted environments

PM(2.5) generated by indoor combustion activities can contribute significantly to personal PM exposure. The aims of this study were: (1) to validate a device specifically designed to study the kinetics of particle exhalation and the percentage of airway particle deposition (%DEP) in polluted indoor environments (welding fumes, environmental tobacco smoke - ETS) and (2) to assess the intra- and inter-subject variability of the signal. The device was tested on 14 subjects exposed to welding fumes and 10 subjects exposed to environmental tobacco smoke (ETS), performing repeated measures at different environmental PM concentrations. The intra-subject variability of the signal for particles with diameter 0.3-1.0 μm showed a geometric mean of %CV always below 6%, despite the values of %DEP. In the welding fume study, the increase in airborne 0.5-1.0 μm PM concentrations between the consulting room and production department was explainable in terms of increased density due to the metallic composition of particles. The %DEP of 0.3-1.0 μm ETS particles decreased with airborne PM concentration due to the technical limits of a laser particle counter and the perturbation induced by the physical characteristics of ETS PM. However, also at those extreme conditions, the signal remained repeatable and the individual susceptibility to PM remained substantially unaltered. In conclusion, the versatility and portability of our device, together with the repeatability of the signal, confirmed that the kinetics of exhaled particles and %DEP could be routinely measured in polluted environments and used to define individual susceptibility to airborne particles.