Florence Anna Zeman

Comparative potency approach based on H2AX assay for estimating the genotoxicity of polycyclic aromatic hydrocarbons

Polycyclic Aromatic Hydrocarbons (PAHs) constitute a family of over one hundred compounds and can generally be found in complex mixtures. PAHs metabolites cause DNA damage which can lead to the development of carcinogenesis. Toxicity assessment of PAH complex mixtures is currently expressed in terms of toxic equivalents, based on Toxicity Equivalent Factors (TEFs). However, the definition of new TEFs for a large number of PAH could overcome some limitations of the current method and improve cancer risk assessment. The current investigation aimed at deriving the relative potency factors of PAHs, based on their genotoxic effect measured in vitro and analyzed with mathematical models. For this purpose, we used a new genotoxic assay (γH2AX) with two human cell lines (HepG2 and LS-174T) to analyze the genotoxic properties of 13 selected PAHs at low doses after 24h treatment. The dose-response for genotoxic effects was modeled with a Hill model; equivalency between PAHs at low dose was assessed by applying constraints to the model parameters. In the two cell lines tested, we observed a clear dose-response for genotoxic effects for 11 tested compounds. LS-174T was on average ten times more sensitive than HepG2 towards PAHs regarding genotoxicity. We developed new TEFs, which we named Genotoxic Equivalent Factor (GEF). Calculated GEF for the tested PAHs were generally higher than the TEF usually used. Our study proposed a new in vitro based method for the establishment of relevant TEFs for PAHs to improve cancer risk assessment.

Increased effects of internal alpha irradiation in Daphnia magna after chronic exposure over three successive generations

A 70-day experiment was performed with Daphnia magna exposed to waterborne Am-241 on a range of concentrations (from 0.4 to 40 Bq ml(-1)) in order to test chronic effects of internal alpha irradiation on respiration, somatic growth and reproduction over three successive generations. Changes in Am-241 concentrations were followed in the water and in daphnid tissues, eggs and cuticles. Corresponding average dose rates of 0.3, 1.5 and 15 mGy h(-1) were estimated. This study confirmed that oxygen consumption increased significantly in the first generation (F0) after 6 days of exposure to a dose rate >or=1.5 mGy h(-1). Consequences were limited to a reduction in body length (5%) and dry mass of females (16%) and eggs (8%) after 23 days of exposure, while mortality and fecundity remained unaffected. New cohorts were started with neonates of broods 1 and 5, to examine potential consequences of the reduced mass of offspring for subsequent exposed generations. Results strongly contrasted with those observed in F0. At the highest dose rate, an early mortality of 38-90% affected juveniles while survivors showed delayed reproduction and reduced fecundity in F1 and F2. At 0.3 and 1.5 mGy h(-1), mortality ranged from 31 to 38% of daphnids depending on dose rate, but was observed only in generation F1 started with neonates of the brood 1. Reproduction was affected through a reduction in the proportion of breeding females, occurring in the first offspring generation at 1.5 mGy h(-1) (to 62% of total daphnids) and in the second generation at 0.3 mGy h(-1) (to 69% of total daphnids). Oxygen consumption remained significantly higher at dose rates >or=0.3 mGy h(-1) than in the control in almost every generation. Body size and mass continued decreasing in relation to dose rate, with a significant reduction in mass ranging from 15% at 0.3 mGy h(-1) to 27% at 15 mGy h(-1) in the second offspring generation.

Biology-Based Modeling To Analyze Uranium Toxicity Data on Daphnia magna in a Multigeneration Study

Recent studies have investigated chronic toxicity of waterborne depleted uranium on the life cycle and physiology of Daphnia magna. In particular, a reduction in food assimilation was observed. Our aims here were to examine whether this reduction could fully account for observed effects on both growth and reproduction, for three successive generations, and to investigate through microscope analyses whether this reduction resulted from direct damage to the intestinal epithelium. We analyzed data obtained by exposing Daphnia magna to uranium over three successive generations. We used energy-based models, which are both able to fit simultaneously growth and reproduction and are biologically relevant. Two possible modes of action were compared - decrease in food assimilation rate and increase in maintenance costs. In our models, effects were related either to internal concentration or to exposure concentration. The model that fitted the data best represented a decrease in food assimilation related to exposure concentration. Furthermore, observations of consequent histological damage to the intestinal epithelium, together with uranium precipitates in the epithelial cells, supported the assumption that uranium has direct effects on the digestive tract. We were able to model the data in all generations and showed that sensitivity increased from one generation to the next, in particular through a significant increase of the intensity of effect, once the threshold for appearance of effects was exceeded.

Variability of urinary concentrations of non-persistent chemicals in pregnant women and school-aged children

BACKGROUND Exposome studies are challenged by exposure misclassification for non-persistent chemicals, whose temporal variability contributes to bias in dose-response functions. OBJECTIVES We evaluated the variability of urinary concentrations of 24 non-persistent chemicals: 10 phthalate metabolites, 7 phenols, 6 organophosphate (OP) pesticide metabolites, and cotinine, between weeks from different pregnancy trimesters in pregnant women, and between days and between seasons in children. METHODS 154 pregnant women and 152 children from six European countries were enrolled in 2014-2015. Pregnant women provided three urine samples over a day (morning, midday, and night), for one week in the 2nd and 3rd pregnancy trimesters. Children provided two urines a day (morning and night), over two one-week periods, six months apart. We pooled all samples for a given subject that were collected within a week. In children, we also made four daily pools (combining morning and night voids) during the last four days of the first follow-up week. Pools were analyzed for all 24 metabolites of interest. We calculated intraclass-correlation coefficients (ICC) and estimated the number of pools needed to obtain an ICC above 0.80. RESULTS All phthalate metabolites and phenols were detected in >90% of pools whereas certain OP pesticide metabolites and cotinine were detected in <43% of pools. We observed fair (ICC = 0.40-0.59) to good (0.60-0.74) between-day reliability of the pools of two samples in children for all chemicals. Reliability was poor (<0.40) to fair between trimesters in pregnant women and between seasons in children. For most chemicals, three daily pools of two urines each (for weekly exposure windows) and four weekly pools of 15-20 urines each would be necessary to obtain an ICC above 0.80. CONCLUSIONS This quantification of the variability of biomarker measurements of many non-persistent chemicals during several time windows shows that for many of these compounds a few dozen samples are required to accurately assess exposure over periods encompassing several trimesters or months.