Mutation Research 748 (2012) 1–7 Contents lists available at SciVerse ScienceDirect Mutation Research/Genetic Toxicology and Environmental Mutagenesis journa l h omepage: www.elsevier.com/locate/gentox C om mun i ty a ddress: www.elsevier.com/locate/mutres Evaluation of the genotoxicity and cytotoxicity in the general population in Turkey by use of the cytokinesis-block micronucleus cytome assay Hamiyet Donmez-Altuntas ∗ , Nazmiye Bitgen Erciyes University, Medical Faculty, Department of Medical Biology, 38039 Kayseri, Turkey a r t i c l e i n f o Article history: Received 23 May 2011 Received in revised form 6 April 2012 Accepted 18 May 2012 Available online 12 June 2012 Keywords: Age Cytome assay Gender Healthy volunteers Micronucleus Smoking a b s t r a c t The cytokinesis-block micronucleus cytome (CBMN-Cyt) assay was originally developed as an ideal system for measuring DNA damage, cytostasis and cytotoxicity. The objective of the present study is to simultaneously evaluate the background levels of micronuclei (MN), nucleoplasmic bridges (NPBs), nuclear buds (NBUDs), cell death (necrosis or apoptosis) and nuclear division index (NDI) in the periph- eral blood lymphocytes of non-occupationally exposed, healthy subjects living in the city of Kayseri in Turkey. We used the CBMN-Cyt assay, taking into account factors – age, gender, and smoking habits – that might affect MN frequency and also other CBMN-Cyt assay parameters. Ninety-six healthy subjects (48 female and 48 male) were selected with ages varying between 21 and 60 years. The parameters, except for the number of binucleated (BN) cells with NPBs, showed no statistically significant difference between smokers and non-smokers. There were significant differences between female and male groups in MN frequency (higher in females) and in the number of NPBs (lower in females), while the other parameters were not significantly different between genders. The correlations between years of age and MN frequency, number of NPBs and the frequency of necrotic cells were statistically significant, while the correlations between the years of age and the other parameters were not. The results of the correlation analysis between years of smoking and MN frequency were positive, although no statistically significant correlation was found between the years of smoking and the other parameters. Among the smokers, no correlation was found either between the pack-years of smoking and the parameters assessed in this group. The results of the present study provide evidence of increasing MN frequency, number of NPBs and frequency of necrotic cells with increasing age in the peripheral blood lymphocytes of healthy individuals and influencing MN frequency and number of NPBs by gender. © 2012 Elsevier B.V. All rights reserved. 1. Introduction The cytokinesis-block micronucleus (CBMN) assay has been the most widely used method to study micronucleus (MN) frequency in human populations since 1985, when it was first described [1–5]. Recent developments now indicate that this method can also be used to evaluate chromosomal instability or damage status [presence of MN, nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs)], mitotic status (mononucleated, metaphase, anaphase, bi-nucleated and multi-nucleated cells) and viability status (necrosis, apoptosis) [6–9]. The idea of obtaining spontaneous or background MN values in peripheral blood lymphocytes of healthy individuals belonging to our own population occurred to us during the evaluation of MN frequencies of control individuals in our previous studies with the ∗ Corresponding author. Tel.: +90 352 4374937/23327; fax: +90 352 4375285/4374931. E-mail address: donmezh@erciyes.edu.tr (H. Donmez-Altuntas). CBMN assay. On the other hand, we are also following-up on the fact that the CBMN assay has evolved into the CBMN cytome (CBMN- Cyt) assay over the past 20 years [1–9]. The CBMN-Cyt method is now used to measure DNA damage (MN, NPBs and NBUDs), cytosta- sis (the proportion of mono-, bi- and multi-nucleated cells; nuclear division index, NDI) and cytotoxicity (necrotic and apoptotic cell ratio) [7,8]. We have already observed these cytological changes and abnormalities when scoring MN frequencies in bi-nucleated (BN) cells in our previous studies. Thus, our initial thought was necessarily focused on the evaluation of not only the MN frequen- cies, but also of other, new cytogenetic variables in the CBMN-Cyt assay. To our knowledge, there are many studies assessing spon- taneous and induced MN frequencies in the peripheral blood lymphocytes of healthy subjects by use of the CBMN assay. The effects of age, gender and smoking status on the frequency of MN have been evaluated as potential confounders in some of these studies. Many studies have shown that there is an increase in MN fre- quency with age [3,9–19], although others have not shown any 1383-5718/$ – see front matter © 2012 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.mrgentox.2012.05.013