Mahmut Coşkun

Micronuclei, nucleoplasmic bridges, and nuclear buds induced in human lymphocytes by the fungicide signum and its active ingredients (boscalid and pyraclostrobin)

The aim of this study was to investigate the genotoxic and cytotoxic potential of the Signum fungicide and its active ingredients (boscalid and pyraclostrobin) on human peripheral blood lymphocytes using the cytokinesis‐block micronucleus (CBMN) assay. Micronuclei (MNi), nucleoplasmic bridges (NPBs), nuclear bud (NBUDs) formations, and the cytokinesis‐block proliferation index (CBPI) were evaluated in treated lymphocytes in Go (cells were treated and then kept in culture without stimulation for 24 h) and proliferation phases (cells were treated after 44 h culture in medium containing phytohemagglutinin). MN formation in lymphocytes treated in G0 statistically increased at doses of 2, 6, and 25 μg/mL signum; 0.5 and 2 μg/mL boscalid; and 0.5, 1.5, and 2 μg/mL pyraclostrobin; while NPB formation increased at a dose of 0.25 μg/mL pyraclostrobin. All concentrations of each fungicide did not statistically increase NBUD formation, while the cytotoxicity increased the dependent on concentration in lymphocytes treated in G0. Doses of 0.5, 1, 1.5, and 3 μg/mL signum; 0.5, 1, and 1.5 μg/mL boscalid; and 0.75 μg/mL pyraclostrobin statistically increased the MN formation in proliferating lymphocytes. NPB formation increased in proliferating lymphocytes at doses of 1, 1.5, 2, and 3 μg/mL signum and at a dose of 0.75 μg/mL pyraclostrobin. In addition, a dose of 0.75 μg/mL pyraclostrobin increased NBUD frequencies. Cytotoxicity increased with increasing concentrations of each fungicide. It is concluded that signum, boscalid, and pyraclostrobin may be genotoxic and cytotoxic in vitro human peripheral blood lymphocytes in consideration of each of the two protocols.

Frequencies of micronuclei (MNi), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) in farmers exposed to pesticides in Çanakkale, Turkey

This study aimed to determine the incidence of micronuclei (MNi), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) in peripheral blood lymphocytes due to direct exposure to pesticides among 46 farmers in Çanakkale, Turkey. 48 non-exposed individuals living in the same socioeconomic conditions were chosen as control. In addition, a cytokinesis-block proliferation index (CBPI) was calculated. MNi and NBUDs frequencies were significantly higher among the farmers (p < 0.05). Although the NPB frequency of the farmers was higher than the controls, there was no statistical difference. Multiple linear regression analysis showed that apart from gender, no significant effects of various confounding factors were observed. Regarding CBPI, data obtained for the controls were higher than that of the farmers; however, there was no statistically significant difference.

Radioactivity concentrations in topsoil samples from the Thrace region of Turkey and assessment of radiological hazard

The activity concentrations of gamma emitting radionuclides were measured in surface soil samples from the Thrace region. 232Th, 238U, 226Ra, 40K and 137Cs activities and physico-chemical parameters of the soil samples were determined in samples collected from 73 sampling stations. Radionuclide concentrations measured were compared with those found in the samples from other locations of Turkey and from different countries. The average activity concentrations of 232Th, 238U, 226Ra, 40K and 137Cs were found to be 24.71 ± 8.79, 22.30 ± 7.93, 32.09±12.44, 509.00±160.05, 32.74±29.24 Bq kg−1, respectively. The mean value of the annual effective dose equivalent from the outdoor terrestrial gamma radiation was calculated as 57.2 μ Sv.

Heavy metal deposition in moss samples from East and South Marmara Region, Turkey

A survey of atmospheric heavy metal deposition in the east and south Marmara region, Turkey was carried out in September 2004. For this purpose, moss samples (Hypnum cupressiforme) were collected in a systematic network of 125 sites. Concentrations of the elements (Al, B, Ba, Ca, Cd, Fe, Co, Cr, Cu, K, Li, Mn, Na, Ni, Pb, Sr, Mg, Ti, and Zn) in the moss were used as an indication of the level of air pollution in the region. Significant differences in heavy metal concentrations, especially for Pb, Cd, Cu, and Zn, were recorded in the moss samples collected around industrialized and heavily populated cities (Istanbul, Bursa, Bandırma, Kocaeli, Biga-Çan) and in an abandoned lead-mining area (Balıkesir-Balya). A map of the spatial distribution of each element in the region was plotted, and enrichment factors were calculated. VARIMAX principal component analysis was applied to the data obtained, and five different components were obtained. The results showed that Pb, Cu, Cd, and Zn derived from anthropogenic and industrial sources while other elements came mostly from natural sources.

Comparison of Epigeic Moss (Hypnum cupressiforme) and Lichen (Cladonia rangiformis) as Biomonitor Species of Atmospheric Metal Deposition

In the present work epigeic moss (Hypnum cupressiforme Hedw.) and epigeic lichen (Cladonia rangiformis Hoffm.) samples were collected simultaneously in the Thrace region, Turkey according to a regular sampling grid. Whereas the moss was found at all 68 sampling sites, the lichen could be collected only at 25 of the sites, presumably because lichens are more sensitive than mosses with respect to air pollution and climatic variations. All elements showed higher accumulation in the moss than in the lichen whereas element inter-correlations were generally higher in the lichen. All considered the moss was judged to be a better choice than the lichen for biomonitoring of atmospheric deposition of metals in this case, and it is argued that mosses may be generally more suited than lichens for this purpose.

Cross-border response of mosses to heavy metal atmospheric deposition in Southeastern Bulgaria and European Turkey

First cross-border atmospheric pollution of 11 heavy metals and toxic elements assessed by Hypnum cupressiforme was reported for a part of Southeastern Europe (Southeastern Bulgaria and European Turkey). Moss monitoring technique followed the main requirements of European Moss Survey. Moss samples were collected in April 2006 both in Bulgaria and Turkey. Concentration of Al, As, Cd, Cr, Cu, Fe, Ni, Pb, Sb, V, and Zn were determined by ICP-AES. Interlaboratory parallel calibration (exchanged four moss samples from each country), standard reference moss materials (M2 and M3) results ensured the study. ANOVA showed no differences between measured results in both laboratories at the 99% confidence level. Principle Component Analyze proved two factors: F1 group of Al, As, Cd, Cr, Fe, Ni, and V and F2 of Cu, Pb, and Zn as main atmospheric pollutants. Results obtained showed approximately Cu and Pb high concentrations around Istanbul and Burgas and Zn pollution in Istanbul district. Arsenic cross-border atmospheric pollution in the study area of Southeastern Europe was found.

Evaluation of background DNA damage in a Turkish population measured by means of the cytokinesis-block micronucleus cytome assay

This study aimed to determine the background incidence of micronuclei (MN), nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) in the peripheral blood lymphocytes of a population in the province of Çanakkale, Turkey. Seventy-one individuals living under similar socio-economic conditions, not exposed to an X-ray examination during the last six months, receiving no medical treatment and not involved in agricultural activities were evaluated. In addition to genetic end-points, the cytokinesis-block proliferation index (CBPI) was determined. MN and NBUD frequencies gradually increased with age. MN frequencies of the older group were ∼7 fold higher than those of the younger group. Multiple linear regression analysis showed that apart from age, no significant effects of various confounding factors on MN and NBUD were observed. NPB frequencies were affected by gender. In the present study, no statistically significant effect was found of smoking on the genetic end-points evaluated, which agrees with results from other studies.

Prevelance of Common YMDD Motif Mutations in Long Term Treated Chronic HBV Infections in a Turkish Population

In the current study we aimed to show the common YMDD motif mutations in viral polymerase gene in chronic hepatitis B patients during lamivudine and adefovir therapy. Forty-one serum samples obtained from chronic hepatitis B patients (24 male, 17 female; age range: 34-68 years) were included in the study. HBV-DNA was extracted from the peripheral blood of the patients using an extraction kit (Invisorb, Instant Spin DNA/ RNA Virus Mini Kit, Germany). A line probe assay and direct sequencing analyses (INNO-LIPA HBV DR v2; INNOGENETICS N.V, Ghent, Belgium) were applied to determine target mutations of the viral polymerase gene in positive HBV-DNA samples. A total of 41 mutations located in 21 different codons were detected in the current results. In 17 (41.5%) patients various point mutations were detected leading to lamivudin, adefovir and/ or combined drug resistance. Wild polymerase gene profiles were detected in 24 (58.5%) HBV positive patients of the current cohort. Eight of the 17 samples (19.5%) having rtM204V/I/A missense transition and/or transversion point mutations and resistance to lamivudin. Six of the the mutated samples (14.6%) having rtL180M missense transversion mutation and resistance to combined adefovir and lamivudin. Three of the mutated samples (7.5%) having rtG215H by the double base substituation and resistance to adefovir. Three of the mutated samples (7.5%) having codon rtL181W due to the missense transversion point mutations and showed resistance to combined adefovir and lamivudin. Unreported novel point mutations were detected in the different codons of polymerase gene region in the current HBV positive cohort fromTurkish population. The current results provide evidence that rtL180M and rtM204V/I/A mutations of HBV-DNA may be associated with a poor antiviral response and HBV chronicity during conventional therapy in Turkish patients.

In vitro genotoxic and cytotoxic effects of doxepin and escitalopram on human peripheral lymphocytes

Abstract Antidepressants are drugs used for the treatment of many psychiatric conditions including depression. There are findings suggesting that these drugs might have genotoxic, carcinogenic, and/or mutagenic effects. Therefore, the present in vitro study is intended to investigate potential genotoxic and cytotoxic effects of the antidepressants escitalopram (selective serotonin reuptake inhibitor) and doxepin (Tricyclic antidepressant) on human peripheral lymphocytes cytokinesis-block micronucleus (CBMN), sister chromatid exchange (SCE), and single cell gel electrophoresis (alkaline comet assay) were used for the purpose of the study. In the study, four different concentrations of both drugs (1, 2.5, 5, and 10 µg/mL) were administered to human peripheral lymphocytes for 24 h. The tested concentrations of both drugs were found to exhibit no cytotoxic and mitotic inhibitory effects. SCE increase caused by 5 and 10 µg/mL of escitalopram was found statistically significant, while no statistically significant increase was observed in DNA damage and micronucleus (MN) formation. Moreover, the increase caused by doxepin in MN formation was not found statistically significant. Besides, 10 µg/mL of doxepin was demonstrated to significantly increase arbitrary unit and SCE formation. These findings suggest that the investigated concentrations of escitalopram and doxepin were non-cytotoxic but potentially genotoxic at higher concentrations.

DNA damage and circulating cell free DNA in greenhouse workers exposed to pesticides

Pesticides widely used in agriculture and other applications have been linked to cancer and other diseases through several potential mechanisms. The goals of this study were to assess DNA damage in lymphocytes by the cytokinesis‐block micronucleus assay (CBMN), and to measure circulating cell free DNA (ccf‐DNA) in the blood of pesticide‐exposed greenhouse workers and matched controls living in the same area. CBMN was applied to peripheral blood lymphocyte samples taken at different times (spring and autumn) for each individual. We measured plasma ccf‐DNA levels using a Qubit® fluorometer. The results indicated that the MNL, BNMN, and NBUDs frequencies of pesticide‐exposed individuals were significantly higher than non‐exposed individuals. Apart from MNL, BNMN and CBPI, a season‐related effect was found for the NPB and NBUD frequencies. With MNL and BNMN as the dependent variables, multiple regression analysis showed that age and gender affected MN formation. The ccf‐DNA level in the pesticide‐exposed group was significantly higher than the control group. There was no seasonal variation regarding the free DNA amount. Ccf‐DNA in males was found to be higher than females. The MNL, BNMN, NPB, and CBPI did not correlate with the ccf‐DNA amount. It can be concluded that pesticide exposure can modulate DNA integrity via different mechanisms. Also, elevated levels of ccf‐DNA could be recommended as a biomarker of pesticide exposure. Environ. Mol. Mutagen. 59:161–169, 2018.