HPLC/fluorescence determination of anti-BPDE–DNA adducts in mononuclear white blood cells from PAH-exposed humans

Abstract

The aim of this study was to compare (⍨)-r-7,t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE)-DNA adduct levels in groups of humans subjected to various levels of polycyclic aromatic hydrocarbon (PAH) (benzo[a]pyrene) exposure. An HPLC/fluorescence method was applied to detect specifically anti-BPDE-DNA adducts in mononuclear white blood cells [lymphocyte plus monocyte fraction (LMF)] from humans exposed to PAHs. A total of 130 subjects comprised the sample population: 26 psoriatic patients (3 days after clinical coal tar treatment of the skin), 15 coke oven workers, 19 chimney sweeps, 36 aluminium anode plant workers and 34 non-occupationally PAH-exposed subjects (controls). PAH exposure was assessed in each group by means of the urinary excretion of 1-pyrenol (mean group levels: 1.2, 0.7, 0.3, 65.0 and 0.1 µmol/mol creatinine in coke oven workers, chimney sweeps, aluminium plant anode workers, psoriatic patients and non-occupationally PAH-exposed subjects, respectively). HPLC/fluorescence analysis of BPDE-DNA adducts showed that the percentage of subjects with adduct levels exceeding the 95 percentile control subject value (8.9 adducts/10 8 nucleotides) was significantly high in coke oven workers (46.7%) and chimney sweeps (21.0%) (χ 2 test, P < 0.01 and P < 0.05, respectively) but not in aluminium plant workers (11.1%) and psoriatic patients (0%). The increase in BPDE-DNA adduct levels in LMF (Ln values) was significantly related to chronic inhalatory and high PAH exposure (linear multiple regression analysis, F ⍧ 6.37, P < 0.01; t ⍧ 4.2, P < 0.001). Skin acute (or short-term) and high PAH exposure, charcoal-grilled meat consumption and smoking habit did not seem to influence BPDE-DNA adduct formation in LMF.

Integrating Field Analyses with Laboratory Exposures to Assess Ecosystems Health

“…One of the adducts co-migrated with the benzo [a]pyrene -7,8-diol-9,10-oxide-DNA adduct (Godschalk et al, 1998). Analysis of anti-benzo [a]pyrene -7,8-diol-9,10-oxide-DNA adduct levels by an HPLC/fluorescence method in a group of 26 psoriasis patients showed that the percentage of subjects with adduct levels that exceeded the 95th percentile of the control value was not significant (Pavanello et al, 1999a). The white blood cells of 23 psoriasis patients who were undergoing clinical coal-tar therapy were examined for benzo [a]pyrene-7,8-diol-9,10-oxide-DNA adducts by an enzyme-linked immunsorbent (ELISA) method.…”

“…The effect on lymphocyte micronuclei was more significant in pokeweed mitogen-stimulated cells, which may imply that T4 lymphocytes were preferentially damaged by the occupational exposure (Holmen et al, 1994). Analysis of benzo [a]pyrene -7,8-diol-9,10-oxide-DNA adduct levels in a group of 19 chimney sweeps showed that the percentage of subjects with adduct levels that exceeded the 95th percentile of the control value was significantly high in chimney sweeps (Pavanello et al, 1999a), and these higher levels were associated with the lack of GSTM1 activity (Pavanello et al, 1999b).…”

“…foundry workers Perera et al, 1988), coke-oven workers (Rojas et al, 1995;Pavanello et al, 1999a), cigarette smokers (Rojas et al, 1995;Lodovici et al, 1998), chimney sweeps (Pavanello et al, 1999a) and people exposed to mixtures in smoke emissions from coal combustion . Some bay-or fjord-region diol epoxides form DNA adducts in the human TP53 tumour-suppressor gene at sites that are hotspots for lung cancer .…”

“…In humans, the anti-benzo [a]pyrene -7,8-diol-9,10-oxide-DNA adducts have been detected in autopsy samples from human lungs of smokers and nonsmokers (Lodovici et al, 1998), in white blood cells of coke-oven workers (Rojas et al, 1995;Pavanello et al, 1999a), cigarette smokers (Rojas et al, 1995) and chimney sweeps (Pavanello et al, 1999a).…”

“…The anti-benzo [a]pyrene -7,8-diol-9,10-oxide-DNA adducts have been measured in populations exposed to benzo [a]pyrene in complex mixtures including coke-oven workers (Rojas et al, 1995;Pavanello et al, 1999a) and chimney sweeps (Pavanello et al, 1999a) (as mentioned above in (ii)).…”