Details of test chemicals.

<p>Details of test chemicals.</p

The effect of Mixture 2 and its constituents on testosterone levels in H295R cells.

<p>H295R cells were incubated with chemicals or Mixture 2 in concentrations ranging from 0.04 to 30 µM for 48 h. The cell medium was next isolated and estradiol was measured by DELFIA. Data are mean ± SD. A <i>p</i>-value of less than 0.05 was considered significant, and in case of significance a sigmoidal curve fit (black line) was applied with a 95% confidence band (black dotted lines). Concentration addition (CA, dotted blue line), independent action (IA, dotted red line) and generalized concentration addition (GCA, green line) predictions were calculated and applied to the graph of the mixture data. The calculated contribution of each chemical is illustrated on the graph of the mixture data (abbreviated as “calculate” in the graph). This contribution is established by shifting the regression line of single chemical effects to the right along the x-axis by the reciprocal of its ratio in the mixture.</p

The effect of Mixture 2 and its constituents on progesterone levels in H295R cells.

<p>H295R cells were incubated with chemicals or Mixture 2 in concentrations ranging from 0.04 to 30 µM for 48 h. The cell medium was next isolated and progesterone was measured by DELFIA. Data are mean ± SD. A <i>p</i>-value of less than 0.05 was considered significant, and in case of significance a sigmoidal curve fit (black line) was applied with a 95% confidence band (black dotted lines). The calculated contribution of each chemical is illustrated on the graph of the mixture data (abbreviated as “calculate” in the graph). This contribution is established by shifting the regression line of single chemical effects to the right along the x-axis by the reciprocal of its ratio in the mixture.</p

The effect of Mixture 2 and its constituents on estradiol levels in H295R cells.

<p>H295R cells were incubated with chemicals or Mixture 2 in concentrations ranging from 0.04 to 30 µM for 48 h. The cell medium was next isolated and estradiol was measured by DELFIA. Data are mean ± SD. A <i>p</i>-value of less than 0.05 was considered significant, and in case of significance a sigmoidal curve fit (black line) was applied with a 95% confidence band (black dotted lines). The calculated contribution of each chemical is illustrated on the graph of the mixture data (abbreviated as “calculate” in the graph). This contribution is established by shifting the regression line of single chemical effects to the right along the x-axis by the reciprocal of its ratio in the mixture.</p

The effect of Mixture 1 and its constituents on testosterone levels in H295R cells.

<p>H295R cells were incubated with chemicals or Mixture 1 at concentrations ranging from 0.04 to 30 µM for 48 h. The cell medium was next isolated and testosterone was measured by DELFIA. Data are mean ± SD. A <i>p</i>-value of less than 0.05 was considered significant, and in case of significance a sigmoidal curve fit (black line) was applied with a 95% confidence band (black dotted lines). Concentration addition (CA, dotted blue line), independent action (IA, dotted red line) and generalized concentration addition (GCA, green line) predictions were calculated and applied to the graph of the mixture data. The calculated contribution of each chemical is illustrated on the graph of the mixture data (abbreviated as “calculate” in the graph). This contribution is established by shifting the regression line of single chemical effects to the right along the x-axis by the reciprocal of its ratio in the mixture.</p

The effect of Mixture 1 and its constituents on progesterone levels in H295R cells.

<p>H295R cells were incubated with chemicals or Mixture 1 at concentrations ranging from 0.04 to 30 µM for 48 h. The cell medium was next isolated and progesterone was measured by DELFIA. Data are mean ± SD. A <i>p</i>-value of less than 0.05 was considered significant, and in case of significance a sigmoidal curve fit (black line) was applied with a 95% confidence band (black dotted lines). The calculated contribution of each chemical is illustrated on the graph of the mixture data (abbreviated as “calculate” in the graph). This contribution is established by shifting the regression line of single chemical effects to the right along the x-axis by the reciprocal of its ratio in the mixture.</p

The effect of Mixture 1 on steroid hormone levels in H295R cells.

<p>Cells were incubated with Mixture 1 at concentrations ranging from 0.04 to 30 µM for 48 h. Hormones levels were measured by LC-MS/MS, except for estradiol that was measured by DELFIA. The figure shows the results of progesterone, 17-OH-progesterone, cortisol, dehydroepiandrosterone, androstenedione, estrone, testosterone and estradiol arranged according to appropriate steps in steroidogenesis. Data are mean ± SD expressed as per-cent of the control level. A <i>p</i>-value of less than 0.05 was considered significant, and in case of significance a sigmoidal curve fit (black line) was applied with a 95% confidence band (black dotted lines). Enzymes involved in steroidogenesis are illustrated by colour shaded boxes at appropriate steps. Abbreviations are as follows: CYP: Cytochrome P450, HSD: Hydroxysteroid dehydrogenase.</p

The effect of Mixture 1 and its constituents on estradiol levels in H295R cells.

<p>H295R cells were incubated with chemicals or Mixture 1 in concentrations ranging from 0.04 to 30 µM for 48 h. The cell medium was next isolated and estradiol was measured by DELFIA. Data are mean ± SD. A <i>p</i>-value of less than 0.05 was considered significant, and in case of significance a sigmoidal curve fit (black line) was applied with a 95% confidence band (black dotted lines). The calculated contribution of each chemical is illustrated on the graph of the mixture data (abbreviated as “calculate” in the graph). This contribution is established by shifting the regression line of single chemical effects to the right along the x-axis by the reciprocal of its ratio in the mixture.</p

Additional file 1: of A proposed framework for the systematic review and integrated assessment (SYRINA) of endocrine disrupting chemicals

A proposed framework for the systematic review and integrated assessment (SYRINA) of endocrine disrupting chemicals. (DOCX 124 kb