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Cancer Epidemiology, Biomarkers & Prevention
Cancer Epidemiology, Biomarkers & Prevention
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Applicability of Induced Sputum for Molecular Dosimetry of Exposure to Inhalatory Carcinogens: 32P-Postlabeling of Lipophilic DNA Adducts in Smokers and Nonsmokers

Ahmad Besarati Nia, Lou M. Maas, Simone G. J. Van Breda, Daniëlle M. J. Curfs, Jos C. S. Kleinjans, Emiel F. M. Wouters and Frederik J. Van Schooten
Ahmad Besarati Nia
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Lou M. Maas
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Simone G. J. Van Breda
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Daniëlle M. J. Curfs
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Jos C. S. Kleinjans
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Emiel F. M. Wouters
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Frederik J. Van Schooten
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DOI:  Published April 2000
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    Fig. 1.

    Representative cell composition of induced sputum sample from a smoker [for visualization, three sections of the slide are assembled together by Image Processing and Analysis System (Quantimet 500, Leica, Cambridge, United Kingdom)]. A, BAM; B, neutrophil; C, bronchoepithelial cell; D, ciliated cell; E, lymphocyte; F, squamous cell. May Grünwald-Giemsa staining. Magnification, ×400. Bar, 30 μm.

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    Fig. 2.

    Representative chromatograms of the 32P-postlabeled DNA adducts in induced sputum of a smoker and a nonsmoker by the NP1 digestion method (A and B, respectively) and by the butanol extraction method (D and E, respectively; the radioactivity in the upper left-hand corner was also observed at the same location in standard DNA and therefore considered as background). C and F, BPDE-DNA adduct standards (1 adduct/107 nucleotides by NP1 and butanol enrichment methods, respectively).

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    Fig. 3.

    Levels of lipophilic DNA adducts in induced sputum of smokers and nonsmokers as determined by NP1 digestion and butanol extraction versions of the 32P-postlabeling assay. Lower and upper edges of the boxes, the 25th and the 75th percentiles, respectively. ○ and—— within the boxes, the means and the medians, respectively. Lower and upper bars, the 10th and the 90th percentiles, respectively. ○, individual values <10th or >90th percentiles.

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  • Table 1

    Characteristics of study population and summary of DNA adducts analysis

    SmokersNonsmokers
    No.2024
    Sex (male/female) 7/13 6/18
    Agea39 ± 1134 ± 8
    Cigarettes/daya17 ± 8 0
    Tar/day (mg)a345 ± 196 0
    Smoking yra20 ± 10  0
    Pack-yra18 ± 14 0
    Sputum induction (success/failure)18/2 21/3 
    Lipophilic DNA adducts in sputum (NP1 digestion method)a,b3.1 ± 1.4c0.6 ± 0.8
    Lipophilic DNA adducts in sputum (butanol extraction method)a,b4.6 ± 3.7d1.0 ± 1.9
    • a Results expressed as mean ± SD.

    • b Adducts/108 nucleotides as determined by 32P-postlabeling.

    • c Statistically significant compared to nonsmokers; P = 0.0007.

    • d Statistically significant compared to nonsmokers; P = 0.02.

  • Table 2

    Cellular characteristics and DNA yield of induced sputum

    Smokers (mean ± SD)Nonsmokers (mean ± SD)
    Total cell count (× 106)6.7 ± 6.04.7 ± 6.0
    Viability (%)80 ± 15a63 ± 17
    Squamous cells (%)17 ± 1316 ± 12
    Nonsquamous cells (%)83 ± 1384 ± 12
    BAMs (%)69 ± 24b92 ± 5
    Neutrophils (%)26 ± 26c4 ± 4
    Bronchoepithelial cells (%)5 ± 53 ± 4
    Lymphocytes (%)0.3 ± 0.50.3 ± 0.5
    Methachromatic cells (%)0.2 ± 0.60.2 ± 0.6
    DNA yield (μg)56 ± 5530 ± 35
    • a Statistically significant compared to nonsmokers; P = 0.01.

    • b Statistically significant compared to nonsmokers; P = 0.002.

    • c Statistically significant compared to nonsmokers; P = 0.008.

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April 2000
Volume 9, Issue 4
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Applicability of Induced Sputum for Molecular Dosimetry of Exposure to Inhalatory Carcinogens: 32P-Postlabeling of Lipophilic DNA Adducts in Smokers and Nonsmokers
Ahmad Besarati Nia, Lou M. Maas, Simone G. J. Van Breda, Daniëlle M. J. Curfs, Jos C. S. Kleinjans, Emiel F. M. Wouters and Frederik J. Van Schooten
Cancer Epidemiol Biomarkers Prev April 1 2000 (9) (4) 367-372;

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Applicability of Induced Sputum for Molecular Dosimetry of Exposure to Inhalatory Carcinogens: 32P-Postlabeling of Lipophilic DNA Adducts in Smokers and Nonsmokers
Ahmad Besarati Nia, Lou M. Maas, Simone G. J. Van Breda, Daniëlle M. J. Curfs, Jos C. S. Kleinjans, Emiel F. M. Wouters and Frederik J. Van Schooten
Cancer Epidemiol Biomarkers Prev April 1 2000 (9) (4) 367-372;
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