Evaluation of genotoxic potential of avarol, avarone, and its methoxy and methylamino derivatives in prokaryotic and eukaryotic test models

<p>In this study, mutagenic and genotoxic potential of anti-tumor compounds avarol, avarone, and its derivatives 3′-methoxyavarone, 4′-(methylamino)avarone and 3′-(methylamino)avarone was evaluated and compared to cytostatics commonly used in chemotherapy (5-fluorouracil, etoposid, and cisplatin). Mutagenic potential of selected hydroquinone and quinones was assessed in prokaryotic model by the SOS/<i>umuC</i> assay in <i>Salmonella typhimurium</i> TA1535/pSK1002. Genotoxic potential was also assessed in eukaryotic models using comet assay in human fetal lung cell line (MRC-5), human adenocarcinoma epithelial cell line (A549), and in human peripheral blood cells (HPBC). The results indicated that avarol and avarone do not exert mutagenic/genotoxic potential. Among the studied avarone derivatives, mutagenic potential was detected by SOS/<i>umuC</i> test for 3′-(methylamino)avarone, but only after metabolic activation. The results of comet assay indicated that 3′-methoxyavarone and 3′-(methylamino)avarone have a significant impact on the level of DNA damage in the MRC-5 cell line. Genotoxic potential was not observed in A549 cells or HPBC probably due to a different uptake rate for the compounds and lower in metabolism rate within these cells.</p

Evaluation of Genotoxic Pressure along the Sava River

<div><p>In this study we have performed a comprehensive genotoxicological survey along the 900 rkm of the Sava River. In total, 12 sites were chosen in compliance with the goals of GLOBAQUA project dealing with the effects of multiple stressors on biodiversity and functioning of aquatic ecosystems. The genotoxic potential was assessed using a complex battery of bioassays performed in prokaryotes and aquatic eukaryotes (freshwater fish). Battery comprised evaluation of mutagenicity by SOS/<i>umuC</i> test in <i>Salmonella typhimurium</i> TA1535/pSK1002. The level of DNA damage as a biomarker of exposure (comet assay) and biomarker of effect (micronucleus assay) and the level of oxidative stress as well (Fpg—modified comet assay) was studied in blood cells of bleak and spirlin (<i>Alburnus alburnus/Alburnoides bipunctatus</i> respectively). Result indicated differential sensitivity of applied bioassays in detection of genotoxic pressure. The standard and Fpg—modified comet assay showed higher potential in differentiation of the sites based on genotoxic potential in comparison with micronucleus assay and SOS/<i>umuC</i> test. Our data represent snapshot of the current status of the river which indicates the presence of genotoxic potential along the river which can be traced to the deterioration of quality of the Sava River by communal and industrial wastewaters. The major highlight of the study is that we have provided complex set of data obtained from a single source (homogeneity of analyses for all samples).</p></div

Overview of the parameters analyzed within the Sava survey 2015.

<p>Overview of the parameters analyzed within the Sava survey 2015.</p

Cell viability, frequency of hedgehogs and frequency of micronuclei in fish blood samples (mean ± SE).

<p>Cell viability, frequency of hedgehogs and frequency of micronuclei in fish blood samples (mean ± SE).</p

Fpg—modified comet assay, tail intensity values for buffer and Fpg—exposed slides and net contribution of 8-oxoG sites (mean ± SE); different letters denote significant differences among studied sites (p < 0.05).

<p>Fpg—modified comet assay, tail intensity values for buffer and Fpg—exposed slides and net contribution of 8-oxoG sites (mean ± SE); different letters denote significant differences among studied sites (p < 0.05).</p

Cryopreservation effects on the cell viability and DNA damage level.

<p>*statistical significance in comparison with fresh sample (p < 0.05).</p

Integrated Biomarker Response (IBR) ranking of the studied sites.

<p>Integrated Biomarker Response (IBR) ranking of the studied sites.</p

SOS induction rate (mean ± SE) in SOS/<i>umuC</i> assay.

<p>Red line represents threshold induction value (1.5); PC—positive controls: 4NQO (0.5 μg/mL) in experiments without metabolic activation and benzo(a)pyrene (10 μg/mL) in experiments with metabolic activation.</p

The average element concentrations in whole body composite (wbc) samples of <i>A</i>.<i>alburnus</i> and <i>A</i>. <i>bipunctatus</i> (marked with *) in mgkg^-1 ww, and MPI values for each sampling station; different letters denote significant differences among studied sites (p < 0.05).

<p>The average element concentrations in whole body composite (wbc) samples of <i>A</i>.<i>alburnus</i> and <i>A</i>. <i>bipunctatus</i> (marked with *) in mgkg^-1 ww, and MPI values for each sampling station; different letters denote significant differences among studied sites (p < 0.05).</p

Sampling sites along the Sava River.

<p>Sampling sites along the Sava River.</p