Evaluation of genotoxic effects of lead in pottery-glaze workers using micronucleus assay, alkaline comet assay and DNA diffusion assay
- 462 Downloads
We investigated genotoxic effects of occupational exposure to lead acetate in pottery-glaze ceramic workers.
The study was carried out in 30 exposed workers and 30 matched controls, to whom several biochemical parameters—the blood lead (B-Pb; range: exposed, 41.68–404.77; controls, 12–52) and cadmium (B-Cd) level, the activity of delta-aminolevulinic acid dehydratase (ALAD), erythrocyte protoporphyrin (EP), the level of vitamin B12 and folate in serum—were measured. The genotoxic effects were evaluated by the alkaline comet assay, the DNA diffusion assay and micronucleus test in peripheral blood lymphocytes.
Subjects exposed to lead had significantly higher B-Pb level and, consequently, increased values of tail intensity (TI), frequency of apoptotic and necrotic cells, and frequency of micronuclei (MN). In contrast, their activity of ALAD, the level of vitamin B12 and folate in serum were significantly lower compared to controls. Poisson regression analysis showed a significant correlation of profession, duration of exposure, smoking, level of cadmium in blood, ALAD and EP with primary DNA damage. A majority of primary damage repairs in a short period after exposure to a genotoxic agent. In addition, the influence of gender and level of vitamin B12 and folate in serum MN frequency in exposed group was observed.
In this study, DNA diffusion and micronucleus test showed higher influence of tested parameters to DNA damage. The results indicate a need for concomitant use of at least two different biomarkers of exposure when estimating a genetic risk of lead exposure.
KeywordsCeramic workers Lead exposure Biological markers Alkaline comet assay Micronucleus assay
This investigation was supported by the Croatian Ministry of Science, Education and Sports as a part of the Projects No: 022-0222148-2137 and No: 022-0222411-2408. We thank Krešimir Nekić from Unit for analytical toxicology and mineral metabolism for performing ALAD and EP measurement.
Conflict of interest
None of authors had any personal or financial conflict of interest.
- Agency for toxic substances and disease registry (ATSDR) (1999) Toxicological profile for Lead. US Department of Health and Human Services, AtlantaGoogle Scholar
- Alessio L, Castoldi MR, Odone P, Franchini I (1981) Behaviour of indicators of exposure and effect after cessation of occupational exposure to lead. Br J Ind Med 38:262–267Google Scholar
- ARC (Seventh Annual report on carcinogens) (1994) National toxicology program. National Institute of environmental Health Sciences. Research Triangle Park, North CarolinaGoogle Scholar
- Barale R, Marrazzini A, Bacci E, Disibio A, Tessa A, Cocchi L, Scarcelli V, Lubrano V, Vassalle C, Landi S (1998) Sister chromatid exchange and micronucleus frequency in human lymphocytes of 1650 subjects in an Italian population: I. Contribution of methodological factors. Environ Mol Mutagen 31:218–227CrossRefGoogle Scholar
- Berlin A, Schaller H (1974) European method for determination of deltaamino-levulinic acid dehydratase activity in blood. Z Clin Chem Clin Biochim 2:389–390Google Scholar
- Bermúdez E, Stone K, Carter KM, Pryor WA (1994) Environmental tobacco smoke is just as damaging to DNA as mainstream smoke. Environ Health Perspect 102:870–874Google Scholar
- Bonassi S, Ceppi M, Fontana V, Merlo F (1997) Multiple regression analysis of cytogenetic human data. Mutat Res 313:69–80Google Scholar
- Bonassi S, Neri M, Lando C, Ceppi M, Lin Y, Chang WP, Holland N, Kirsch-Volders M, Wushou PC, Zeiger E, Fenech M (2003) The HUMN collaborative group, effect of smoking habit on the frequency of micronuclei in human lymphocytes: results from the human MicroNucleus project. Mutat Res 543:155–166CrossRefGoogle Scholar
- Calderón-Ezquerro C, Guerrero-Guerra C, Sansores-Martínez R, Calderón-Segura ME, Villalobos-Pietrini R, Amador-Muñoz O, Gómez-Arroyo S (2010) Genotoxicity in lymphocytes of smokers living in México City. Rev Int Contam Ambient 26:47–63Google Scholar
- Chislom JJ, Brown DH (1975) Micro-scale photofluorimetric determination of “free erythrocyte protoporphyrin” (protoporphyrin IX). Clin Chem 21:1669Google Scholar
- Fenech M, Bonassi S (2011) The effect of age, gender, diet and life style on DNA damage measured using micronucleus frequency in human peripheral blood lymphocytes. Mutagenesis 26(1):43–49Google Scholar
- Forni A, Camiaghi G, Sechi GC (1976) Initial occupational exposure to lead: chromosome and biochemical findings. Arch Environ Health 31:73–78Google Scholar
- Garcia-Leston J, Laffon B, Roma-Torres J, Teixeira JP, Costa C, Coelho P, Monteiro S, Mayan O, Valdiglesias V, Pasaro E, Mendez J (2009) Evaluation of genotoxic effects of occupational exposure to lead bay means of the comet assay. Abstract book of ICEM 2009, August 20–25, Firenze, Italy, pp 287–288Google Scholar
- Hartwig A (1994) Role of DNA repair inhibition in lead- and cadmium-induced genotoxicity: a review. Environ Health Perspect 102:45–50Google Scholar
- Hogstedt C, Hane M, Agrell A, Bodin L (1983) Neuropsychological test results and symptoms among workers with well-defined long-term exposure to lead. Br J Ind Med 40:99–105Google Scholar
- Kašuba V, Rozgaj R, Milić M, Želježić D, Kopjar N, Pizent A, Kljaković-Gašpić Z (2009) Evaluation of lead exposure in battery-manufacturing workers with focus on different biomarkers. J Appl Tox 30:321–328Google Scholar
- Kažimirova A, Barančokova M, Krajčovičova-Kudlačkova M, Volkovova K, Staruchova M, Valachovičova M, Paukova V, Blažiček P, Wsolova L, Dušinska M (2006) The relationship between micronuclei in human lymphocytes and selected micronutrients in vegetarians and non-vegetarians. Mutat Res 611:64–70CrossRefGoogle Scholar
- Manikantan P, Balachander V, Sasikala K (2010) DNA damage in workers occupationally exposed to lead, using comet assay. Int J Biol 2:103–110Google Scholar
- McKevith B (2004) Folate and folic acid. British Nutrition Foundation, www.nutrition.org.uk
- OSHA (2002) Employee Standard Summary—1910.1025 App B.U.S. Department of Labor, Occupational Health and Safety Administration. Available at: http://www.osha.gov/pls/oshaweb/ovaolisp.show_document?_table=STANDARDS&p_id=10032
- Popović M, McNeill FE, Chettle DR, Webber CE, Lee CV, Kaye WE (2005) Impact of occupational exposure on lead levels in women. Environ Med 113:478–484Google Scholar
- Sarto F, Stella M, Acqua A (1978) Cytogenetic studies in twenty workers occupationally exposed to lead (in Italian). Med Lav 69:120–180Google Scholar
- SAS Institute Inc (1999) SAS/STAT® user s guide, Version 8. SAS Institute Inc, CaryGoogle Scholar
- Seppäläinen AM, Hernberg S, Vesanto R, Kock B (1983) Early neurotoxic effects of occupational lead exposure: a prospective study. Neurotoxicology 4:181–192Google Scholar
- Singh NP (2005) Apoptosis assessment by the DNA diffusion assay. Chemosensitivity Vol.2: In vivo models. Imaging Mole Regulat 111:55–67Google Scholar
- Wagner C (1995) Biochemical role of folate in cellular metabolism. In: Bailey LB (ed) Folate in health and disease. Marcel Dekker, New York, pp 23–42Google Scholar
- Wetmur JG (1994) Influence of the common human delta-aminolevulinate dehydratase polymorphism on lead body burden. Environ Health Perspect 102(suppl. 3):215–219Google Scholar