Graphical summary of the correlation between X and Y for the first two components.

<p>The correlation between X and Y (w*c) is represented by the loading plot. The PLS-DA model used was constructed on OSC-filtered and Pareto scaled data (N = 83; R2 = 90.7% and Q2 = 0.53), from the ¹H NMR urinary metabolic profiles from 83 pregnant women differently exposed to pesticides.</p

PLS-DA score plot from the ¹H NMR urinary metabolic profiles from 83 pregnant women.

<p>The score plot is the projection of the observations onto the first two latent variables. The PLS-DA model, constructed on OSC-filtered and Pareto-scaled data, includes 4 latent variables (N = 83; R2 = 90.7% and Q2 = 0.53). Three groups according to the percentage of the surface of land dedicated to cereal crops in the town of residence in early pregnancy: purple: group 0: 0–17%, green: group 1: >17–25%; orange: group 2: >25%.</p

17-hydroxyprogesterone (17-OHP) production is increased whereas androstenedione production is decreased by exposure of the fetal testes to MEHP.

<p>Effects of 10 µM MEHP on 17-OHP and androstenedione secretion by fetal rat testes cultured for 72 h, beginning at GD14.5. We determined 17-OHP and androstenedione concentrations with specific RIAs. Values are means +/−SEM of 8 (17-OHP) and 7 (androstenedione) testes from fetuses of 2 different litters in 2 independent experiments. The numbers indicate the percentage decrease or increase relative to the corresponding control. <i>* p<0.05 and ** p<0.01</i> in <i>Wilcoxon Mann-Whitney tests comparing treated and control testes</i>.</p

Flowchart of the metabolomics analysis.

<p>Flowchart of the metabolomics analysis.</p

Genes involved in testicular development and functions affected in fetal testis explants exposed to MEHP.

<p>Fold-change values expressed relative to time-matched untreated controls, with reciprocal transformation from expression ratios. NS: no significant change in gene expressions. Values in italics indicate a down-regulation and values in bold indicate an upregulation.</p><p>*p value<0.01 and</p><p>**p<0.001.</p

Genotoxicity of PhIP in Apc^+/+ and Apc^Min/+ cell lines.

<p>Apc^+/+ and Apc^Min/+ cells were treated with the indicated concentrations of PhIP for 24 h. (A) Genotoxicity was evaluated by HPLC-MS/MS quantification of dG-C8-PhIP in DNA. (B) Genotoxicity was evaluated with γHAX ICW assay. Bars represent the average of at least three independent experiments with SEM. Statistically significant increase in H2AX phosphorylation compared with DMSO control using Student’s test; *, <i>p</i><0.05; **, <i>p</i><0.01. Statistically significant difference between Apc^+/+ and Apc^Min/+ cells using Student’s test; b, <i>p</i><0.05; a, <i>p</i><0.01.</p

Production of B(<i>a</i>)P metabolites (supernatants and cell extracts) on Apc^+/+ and Apc^Min/+ cell lines after 24 h incubations with 3 µM [¹⁴C]-B(<i>a</i>)P.

<p>nd : not detected.</p><p>Values are expressed as a percentage of the total radioactivity and are means ± SD. Significant differences between Apc^+/+ and Apc^Min/+ cells were determined using Student’s test (**<0.01).</p

Synergic genotoxicity of B(<i>a</i>)P and PhIP in Apc^+/+ and Apc^Min/+ cell lines.

<p>Apc^+/+ and Apc^Min/+ cells were treated with the indicated concentrations of BaP and PhIP for 24 h. (A) Genotoxicity was evaluated by quantification by HPLC-MS/MS quantification of B(<i>a</i>)P- and PhIP-derived DNA adducts. (B) Genotoxicity was evaluated with γHAX ICW assay. Bars represent the average of at least three independent experiments with SEM. Statistically significant increase in H2AX phosphorylation compared with DMSO control using Student’s test; **, <i>p</i><0.01. Statistically difference between single and combined treatment using Student’s test; ns, not significant; a, <i>p</i><0.01.</p

Western blot analysis of P450c17 and Cytochrome b5 protein levels after MEHP exposure.

<p>A) Representative Western blot of protein extracts (20 µg) from a pool of 8 control testes or testes exposed to 10 µM MEHP. Blots were incubated with anti-p450c17, anti-Cytochrome b5 and anti-GAPDH antibodies, to control for equal loading. The apparent molecular masses (kDa) are indicated on the left. B) Quantification of band intensity on three blots, carried out with Quantity One software (Biorad). Data are expressed in arbitrary units relative to the corresponding control. The number indicates the percentage decrease relative to control. * <i>p<0.05 in Wilcoxon Mann-Whitney tests comparing treated and control testes. </i><i>NS: not significant</i>.</p

Suggested mechanisms of action of complex and low-dose pesticide mixtures.

<p>These suggestions are based on the modification of ¹H NMR urinary metabolic profile of pregnant women.</p