Genotoxic potential of selected cytostatic drugs in human and zebrafish cells

Due to their increasing use, the residues of anti-neoplastic drugs have become emerging pollutants in aquatic environments. Most of them directly or indirectly interfere with the cell’s genome, which classifies them into a group of particularly dangerous compounds. The aim of the present study was to conduct a comparative in vitro toxicological characterisation of three commonly used cytostatics with different mechanisms of action (5-fluorouracil [5-FU], cisplatin [CDDP] and etoposide [ET]) towards zebrafish liver (ZFL) cell line, human hepatoma (HepG2) cells and human peripheral blood lymphocytes (HPBLs). Cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and acridine orange/ethidium bromide staining. All three drugs induced time- and dose-dependent decreases in cell viability. The sensitivity of ZFL and HepG2 cells towards the cytotoxicity of 5-FU was comparable (half maximal inhibitory concentration (IC₅₀) 5.3 to 10.4 μg/mL). ZFL cells were more sensitive towards ET- (IC₅₀ 0.4 μg/mL) and HepG2 towards CDDP- (IC₅₀ 1.4 μg/mL) induced cytotoxicity. Genotoxicity was determined by comet assay and cytokinesis block micronucleus (CBMN) assay. ZFL cells were the most sensitive, and HPBLs were the least sensitive. In ZFL cells, induction of DNA strand breaks was a more sensitive genotoxicity endpoint than micronuclei (MNi) induction; the lowest effective concentration (LOEC) for DNA strand break induction was 0.001 μg/mL for ET, 0.01 μg/mL for 5-FU and 0.1 μg/mL for CDDP. In HepG2 cells, MNi induction was a more sensitive genotoxicity endpoint. The LOEC values were 0.01 μg/mL for ET, 0.1 μg/mL for 5-FU and 1 μg/mL for CDDP. The higher sensitivity of ZFL cells to cytostatic drugs raises the question of the impact of such compounds in aquatic ecosystem. Since little is known on the effect of such drugs on aquatic organisms, our results demonstrate that ZFL cells provide a relevant and sensitive tool to screen genotoxic potential of environmental pollutant in the frame of hazard assessment.     

The relation between the estimated dietary intake of PCDD/Fs and levels in blood in a Flemish population (50-65 years)

Dioxin-like activity was measured in the serum of 1425 Flemish men and women via the CALUX assay. The adults, aged between 50 and 65 years, participated in a large biomonitoring program, executed by the Flemish Center of Expertise for Environment and Health between 2002 and 2006. Within the context of this biomonitoring program also dietary intake of dioxin-like contaminants was assessed through a food frequency questionnaire. The relation between the estimated dietary intake and the dioxin-like activity in serum was evaluated using multivariate analyses: a logistic model was performed on the total population, while a linear regression analysis was done on the subsample with quantifiable dioxin activity levels in serum. Region, gender, age, BMI, smoking status, as well as dietary habits were entered in the model, with dioxin level as an outcome estimate. Both the logistic and linear model confirmed the contribution of dietary intake to the dioxin activity measured in serum. Also BMI and region were found to be associated with dioxin activity levels.     

Monitoring of firefighters exposure to smoke during fire experiments in Portugal

Forest fires represent a serious threat to public security in Europe due to the large burned area. Moreover, smoke pollution due to forest fire events is an important public health issue for the communities directly affected, and particularly for the personnel involved in firefighting operations. Aiming to contribute to the scientific knowledge concerning firefighters exposure to forest fires smoke, data of individual exposure to carbon monoxide, nitrogen dioxide, volatile organic compounds, and particulate matter were obtained during experimental field fires for a group of 10 firefighters equipped with portable “in continuum” measuring devices. Measured values are very high exceeding the Occupational Exposure Standard limits, in particular for peak limit thresholds. These are the first measurements and analysis of firefighter's individual exposure to toxic gases and particles in fire smoke experiments in Europe. However, they already indicate that urgent measures to avoid these levels of exposure are needed.     

Use of sensitive methods for detection of DNA damage on human lymphocytes exposed to p,p′-DDT: Comet assay and new criteria for scoring micronucleus test

Wide distribution, stability and long persistence in the environment of dichlorodiphenyltrichloroethane (DDT), probably the best-known and most useful insecticide in the world, imposes the need for further examination of the effect of this chemical on human health and especially on the human genome. In this study, peripheral blood human lymphocytes from a healthy donor were exposed to 0.025 mg/L concentration of p,p′-DDT at different time periods (1, 2, 24 and 48 h). For the assessment of genotoxic effect, the new criteria for scoring micronucleus test and alkaline comet assay were used. Both methods showed that p,p′-DDT induces DNA damage in low concentration used in this research. Results of micronucleus test showed a statistically significant (p < 0.05) genotoxic effect of p,p′-DDT on human lymphocytes compared with corresponding control and a different exposure time. A comet assay also showed increased DNA damage caused in p,p′-DDT-exposed human lymphocytes than in corresponding control cells for the tail length. Results obtained by measuring the level of DNA migration and incidence of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) indicate the sensitivity of these tests and their application in detection of primary genome damage after long-term exposure to establish the effect of p,p′-DDT on human genome.     

Copper-tolerant yeasts: Raman spectroscopy in determination of bioaccumulation mechanism

Environmental Science and Pollution Research - Modern, efficient, and cost-effective approach to remediation of heavy metal-contaminated soil is based on the application of microorganisms. In this...     

Concentration of ascorbic acid and antioxidant response in early life stages of Engraulis ringens and zooplankton during the spawning seasons of 2006–2009 off central Chile

This study reports changes in ascorbic acid (AA) in anchoveta eggs, copepods and zooplankton during the 2006, 2007 and 2009 main spawning seasons in the coastal area of the central Humboldt Current System, Chile. Anchoveta eggs, copepods and total zooplankton community shared a seasonal variation and an increasing trend in AA concentration from winter through spring which was associated with the spring diatom bloom. The lineal relationship observed between AA concentration in anchoveta eggs, chlorophyll a and Sea Surface Temperature (SST) suggests that the increase in phytoplankton abundance could also increase the amount of AA in the spawning female anchoveta incorporated through tissue, thus increasing the concentration in their eggs. Ascorbic acid concentrations in copepods presented size (weight) dependence. Small copepods (e.g. Acartia, Oithona) had AA concentrations two orders of magnitude higher than the heavier weight class copepods (e.g. Calanus, Rhincalanus). Results of the determination of glutathione and the antioxidant potential showed a similar trend in interannual variations, suggesting that cold SST conditions observed in the 2007 spawning season could increase the consumption of antioxidants in early stages. Potential connections between AA concentration in the food web on anchoveta reproduction and egg hatching and embryo malformations are discussed.     

Fourth WHO-coordinated survey of human milk for persistent organic pollutants (POPs): Belgian results

Persistent organic pollutants (POPs) are chemicals that accumulate in the food chain and are toxic to humans and wildlife. The fourth World Health Organization (WHO) survey on POP levels in human milk (2006-2009) aims to provide baseline and trend information on human exposure to POPs. So far Belgium participated in all three previous rounds (1988, 1992, 2001). Whereas the first three rounds focused on determination of dioxins and PCBs in pooled (mixed) samples, the fourth survey comprised the analyses of individual milk samples for nine "basic POPs" (chlorinated pesticides and indicator PCBs) and of pooled milk samples for "basic POPs", "advanced POPs" (dioxins and dioxin-like PCBs) and "optional POPs" (polybrominated diphenylethers [PBDEs], polybrominated dioxins and dibenzofurans [PBrDD/F], mixed halogenated dioxins and dibenzofurans [PXDD/F] and hexabromocyclododecane [HBCD]). For the Belgian participation human milk samples were collected during the summer of 2006 from 197 women between 18 and 30 years old distributed over all Belgian provinces. The individual samples were analyzed in a Belgian Laboratory for "basic" POPs. A pooled sample was made from 178 individual samples and analyzed by the WHO Reference Laboratory for the "basic, advanced and optional" POPs. The results indicate that most organochlorinated pesticides banned 25-30 years ago were below or around detection limits in Belgian human milk samples although DDE was still found at low levels in all samples. Over the last five years the levels of marker PCBs and PCDD/Fs in Belgian human milk decreased, respectively, by 58% and 39%. For some of the other emerging or older compounds recent international data are needed to allow comparison. This shows the importance of international studies as run by WHO.     

Synthesis of zirconia-immobilized copper chelates for catalytic decomposition of hydrogen peroxide and the oxidation of polycyclic aromatic hydrocarbons

Chelating sorbents with diethylenetriaminepenta(methylene-phosphonic acid) (DTPMPA) and ethylenediaminetetraacetic acid ligands immobilized on zirconia matrix were prepared and subsequently saturated with Cu(II). All the Cu chelates catalyzed decomposition of H(2)O(2) yielding highly reactive hydroxyl radicals. All of them were also able to catalyze degradation of polycyclic aromatic hydrocarbons (anthracene, benzo[a]pyrene and benzo[b]fluoranthene). The most effective DTPMPA-based catalysts G-32 and G-35 (10 mg ml(-1) with 100 mmol H(2)O(2)) caused almost complete decomposition of 15 ppm anthracene and benzo[a]pyrene during a five day catalytic cycle at 30 degrees C. Anthracene-1,4-dione was the main product of anthracene oxidation by all catalysts. The catalysts were active in several cycles without regeneration.