Pollution of indoor environment, where people spend much of their time, comprises complex mixtures of compounds with vastly understudied hazard potential. This study examined several important specific toxic effects and pollutant levels (177 compounds) of indoor samples (air gas phase, PM10 and dust) from different microenvironments after two extractions with focus on their gas/particle/dust distribution and polarity. The endocrine disruptive (ED) potential was assessed by human cell-based in vitro bioassays addressing anti-/estrogenicity, anti-/androgenicity, aryl hydrocarbon, thyroid and peroxisome proliferator-activated receptor-mediated activities. Potential toxicity to respiratory tract tissue was assessed using human bronchial cell line. The toxicological analyses pointed out the relevance of both inhalation and ingestion exposure, with significant effects detected after exposure to extracts from all three studied matrices with distinct gas/particle distribution patterns. Chemical analyses document the high complexity of indoor pollutant mixtures with greatest levels of phthalates, their emerging alternatives, and PAHs in dust. Despite the detection of up to 108 chemicals, effects were explained only to low extent. This emphasizes data gaps regarding ED potencies of many detected abundant indoor contaminants, but also potential presence of other unidentified ED compounds. The omnipresent ED potentials in indoor environment rise concern regarding associated human health risk.
- Klíčová slova
- Bioassay, Dust, Endocrine disruption, Indoor air, Mixture,
- MeSH
- látky znečišťující životní prostředí * MeSH
- lidé MeSH
- polycyklické aromatické uhlovodíky * analýza MeSH
- prach analýza MeSH
- znečištění vzduchu ve vnitřním prostředí * analýza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- látky znečišťující životní prostředí * MeSH
- polycyklické aromatické uhlovodíky * MeSH
- prach MeSH
Zayandeh Rood river is the most important river in central Iran supplying water for a variety of uses including drinking water for approximately three million inhabitants. The study aimed to investigate the quality of water concerning the presence of pharmaceutical active compounds (PhACs) and hormonelike compounds, which have been only poorly studied in this region. Sampling was performed at seven sites along the river (from headwater sites to downstream drinking water source, corresponding drinking water, and treated wastewater) affected by wastewater effluents, specific drought conditions, and high river-water demand. The targeted and nontargeted chemical analyses and in vitro bioassays were used to evaluate the presence of PhACs and hormonelike compounds in river water. In the samples, 57 PhACs and estrogens were detected with LC-MS/MS with the most common and abundant compounds valsartan, carbamazepine, and caffeine present in the highest concentrations in the treated wastewater in the concentrations of 8.4, 19, and 140 μg/L, respectively. A battery of in vitro bioassays detected high estrogenicity, androgenicity, and AhR-mediated activity (viz., in treated wastewater) in the concentrations 24.2 ng/L, 62.2 ng/L, and 0.98 ng/L of 17β-estradiol, dihydrotestosterone and 2,3,7,8-TCDD equivalents, respectively. In surface water samples, estrogenicity was detected in the range of <0.42 (LOD) to 1.92 ng/L of 17β-estradiol equivalents, and the drinking water source contained 0.74 ng/L of 17β-estradiol equivalents. About 19% of the estrogenicity could be explained by target chemical analyses, and the remaining estrogenicity can be at least partially attributed to the potentiation effect of detected surfactant residues. Drinking water contained several PhACs and estrogens, but the overall assessment suggested minor human health risk according to the relevant effect-based trigger values. To our knowledge, this study provides some of the first comprehensive information on the levels of PhACs and hormones in Iranian waters.
- Klíčová slova
- Drugs, Endocrine disruptors, In vitro biotest, LC-MS/MS, Nontargeted screening,
- MeSH
- chemické látky znečišťující vodu * analýza MeSH
- chromatografie kapalinová MeSH
- estrogeny analýza MeSH
- léčivé přípravky * MeSH
- lidé MeSH
- odpadní voda MeSH
- pitná voda * MeSH
- tandemová hmotnostní spektrometrie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Írán MeSH
- Názvy látek
- chemické látky znečišťující vodu * MeSH
- estrogeny MeSH
- léčivé přípravky * MeSH
- odpadní voda MeSH
- pitná voda * MeSH
Estrogenic compounds are widely released to surface waters and may cause adverse effects to sensitive aquatic species. Three hormones, estrone, 17β-estradiol and 17α-ethinylestradiol, are of particular concern as they are bioactive at very low concentrations. Current analytical methods are not all sensitive enough for monitoring these substances in water and do not cover mixture effects. Bioassays could complement chemical analysis since they detect the overall effect of complex mixtures. Here, four chemical mixtures and two hormone mixtures were prepared and tested as reference materials together with two environmental water samples by eight laboratories employing nine in vitro and in vivo bioassays covering different steps involved in the estrogenic response. The reference materials included priority substances under the European Water Framework Directive, hormones and other emerging pollutants. Each substance in the mixture was present at its proposed safety limit concentration (EQS) in the European legislation. The in vitro bioassays detected the estrogenic effect of chemical mixtures even when 17β-estradiol was not present but differences in responsiveness were observed. LiBERA was the most responsive, followed by LYES. The additive effect of the hormones was captured by ERα-CALUX, MELN, LYES and LiBERA. Particularly, all in vitro bioassays detected the estrogenic effects in environmental water samples (EEQ values in the range of 0.75-304 × EQS), although the concentrations of hormones were below the limit of quantification in analytical measurements. The present study confirms the applicability of reference materials for estrogenic effects' detection through bioassays and indicates possible methodological drawbacks of some of them that may lead to false negative/positive outcomes. The observed difference in responsiveness among bioassays - based on mixture composition - is probably due to biological differences between them, suggesting that panels of bioassays with different characteristics should be applied according to specific environmental pollution conditions.
- Klíčová slova
- Bioassay, Chemical mixture, Endocrine disrupting compound (EDC), Environmental quality standard (EQS), Estrogenicity, Hormone mixture,
- MeSH
- biotest MeSH
- chemické látky znečišťující vodu * analýza toxicita MeSH
- endokrinní disruptory * analýza MeSH
- estrogeny analýza toxicita MeSH
- estron MeSH
- monitorování životního prostředí MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chemické látky znečišťující vodu * MeSH
- endokrinní disruptory * MeSH
- estrogeny MeSH
- estron MeSH
In the last decade, it has become evident that complex mixtures of cyanobacterial bioactive substances, simultaneously present in blooms, often exert adverse effects that are different from those of pure cyanotoxins, and awareness has been raised on the importance of studying complex mixtures and chemical interactions. We aimed to investigate cytotoxic and genotoxic effects of complex extracts from laboratory cultures of cyanobacterial species from different orders (Cylindrospermopsis raciborskii, Aphanizomenon gracile, Microcystis aeruginosa, M. viridis, M. ichtyoblabe, Planktothrix agardhii, Limnothrix redekei) and algae (Desmodesmus quadricauda), and examine possible relationships between the observed effects and toxin and retinoic acid (RA) content in the extracts. The cytotoxic and genotoxic effects of the extracts were studied in the human hepatocellular carcinoma HepG2 cell line, using the MTT assay, and the comet and cytokinesis-block micronucleus (cytome) assays, respectively. Liquid chromatography electrospray ionization mass spectrometry (LC/ESI-MS) was used to detect toxins (microcystins (MC-LR, MC-RR, MC-YR) and cylindrospermopsin) and RAs (ATRA and 9cis-RA) in the extracts. Six out of eight extracts were cytotoxic (0.04-2 mgDM/mL), and five induced DNA strand breaks at non-cytotoxic concentrations (0.2-2 mgDM/mL). The extracts with genotoxic activity also had the highest content of RAs and there was a linear association between RA content and genotoxicity, indicating their possible involvement; however further research is needed to identify and confirm the compounds involved and to elucidate possible genotoxic effects of RAs.
- Klíčová slova
- algae, chemical analysis, complex mixtures, cyanobacteria, cyanotoxins, cytotoxicity, extracts, genotoxicity, microcystins, retinoic acids,
- MeSH
- alkaloidy izolace a purifikace toxicita MeSH
- buňky Hep G2 MeSH
- Chlorophyta metabolismus MeSH
- chromatografie kapalinová MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- kometový test MeSH
- lidé MeSH
- mikrocystiny izolace a purifikace toxicita MeSH
- mikrojaderné testy MeSH
- mikrojádra chromozomálně defektní chemicky indukované MeSH
- poškození DNA * MeSH
- sinice metabolismus MeSH
- tandemová hmotnostní spektrometrie MeSH
- toxiny kmene Cyanobacteria MeSH
- tretinoin izolace a purifikace toxicita MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alkaloidy MeSH
- cylindrospermopsin MeSH Prohlížeč
- microcystin MeSH Prohlížeč
- mikrocystiny MeSH
- toxiny kmene Cyanobacteria MeSH
- tretinoin MeSH
We developed a simple and fast microplate assay for evaluation of the antimicrobial activity of electrospun nanofiber filtration membranes or similar porous materials for water treatment technologies. Resazurin (alamarBlue®) was used as an indicator of the amount of viable experimental microorganisms Gram-negative Escherichia coli, Gram-positive Enterococcus faecalis, and natural wastewater treatment plant effluent bacteria. A bacterial inoculum of concentration 1-3 × 105 CFU mL-1 was pipetted onto the surface of assessed both functionalized and respective control membranes and incubated in 12-well plates for 4 h at 37 °C. Kinetics of resazurin metabolization, i.e. its reduction to fluorescent resorufin, was evaluated fluorimetrically (λex520/λem590 nm). A number of viable bacteria on the membranes expressed as CFU mL-1 was calculated from the kinetic curves by using calibration curves that were constructed for both experimental bacterial species. Antimicrobial activities of the membranes were evaluated by either resazurin assay or modified ISO 20743 plate count assay. Results of both assays showed the significant antimicrobial activity of membranes functionalized with silver nanoparticles for both bacterial species and wastewater treatment plant effluent bacteria as well (log CFU reduction compared to control membrane > 4), while membranes containing specific quaternary ammonium salts were inefficient (log CFU reduction < 1). The suitability of resazurin microplate assay for testing nanofiber filtration membranes and analogous matrices has proven to be a faster and less demanding alternative to the traditionally used approach providing comparable results.
- Klíčová slova
- Antimicrobial activity, Electrospun nanofiber filtration membranes, Quaternary ammonium salts, Resazurin, Silver nanoparticles,
- Publikační typ
- časopisecké články MeSH
Detection of endocrine disrupting compounds in water and sediment samples has gained much importance since the evidence of their effects was reported in aquatic ecosystems in the 1990s. The aim of this review is to highlight the advances made in the field of in vitro analysis for the detection of hormonally active compounds with estrogenic, androgenic and progestogenic effects in water and sediment samples. In vitro assays have been developed from yeast, mammalian and in a few cases from fish cells. These assays are based either on the hormone-mediated proliferation of sensitive cell lines or on the hormone-mediated expression of reporter genes. In vitro assays in combination with various sample enrichment methods have been used with limits of detection as low as 0.0027 ng L-1 in water, and 0.0026 ng g-1 in sediments for estrogenicity, 0.1 ng L-1 in water, and 0.5 ng g-1 in sediments for androgenicity, and 5 ng L-1 in water for progestogenicity expressed as equivalent concentrations of standard reference compounds of 17β-estradiol, dihydrotestosterone and progesterone, respectively. The experimental results and limits of quantification, however, are influenced by the methods of sample collection, preparation, and individual laboratory practices.
- MeSH
- biotest metody MeSH
- chemické látky znečišťující vodu analýza toxicita MeSH
- endokrinní disruptory analýza toxicita MeSH
- geologické sedimenty analýza MeSH
- kultivované buňky MeSH
- lidé MeSH
- odběr biologického vzorku metody MeSH
- odpadní voda analýza MeSH
- pohlavní steroidní hormony analýza toxicita MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- chemické látky znečišťující vodu MeSH
- endokrinní disruptory MeSH
- odpadní voda MeSH
- pohlavní steroidní hormony MeSH
Yeast biosensors have become suitable tools for the screening and detection of environmental pollutants because of their various advantages compared to other sensing technologies. On the other hand, many limitations remain with regard to their optimal performance and applicability in several contexts, such as low-concentration samples and on-site testing. This review summarizes the current state of yeast biosensors, with special focus on screening and assessment of environmental contaminants, discusses both pros and cons, and suggests steps towards their further development and effective use in the environmental assessment.
- Klíčová slova
- environmental monitoring, on-site applicability, pollution, yeast biosensor,
- MeSH
- biosenzitivní techniky metody MeSH
- látky znečišťující životní prostředí analýza MeSH
- monitorování životního prostředí metody MeSH
- Saccharomyces cerevisiae metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- látky znečišťující životní prostředí MeSH
Recombinant yeast assays (RYAs) have been proved to be a suitable tool for the fast screening of compounds with endocrine disrupting activities. However, ready-to-use versions more accessible to less equipped laboratories and field studies are scarce and far from optimal throughputs. Here, we have applied freeze-drying technology to optimize RYA for the fast assessment of environmental compounds with estrogenic and androgenic potencies. The effects of different cryoprotectants, initial optical density and long-term storage were evaluated. The study included detailed characterization of sensitivity, robustness and reproducibility of the new ready-to-use versions, as well as comparison with the standard assays. Freeze-dried RYAs showed similar dose-responses curves to their homolog standard assays, with Lowest Observed Effect Concentration (LOEC) and Median effective Concentration (EC50) of 1 nM and 7.5 nM for testosterone, and 0.05 nM and 0.5 nM for 17β-estradiol, respectively. Freeze-dried cells stored at 4 °C retained maximum sensitivity up to 2 months, while cells stored at -18 °C showed no decrease in sensitivity throughout the study (10 months). This ready-to-use RYA is easily accessible and may be potentially used for on-site applications.
- Klíčová slova
- Androgen, Estrogen, Freeze-drying, Long-term, Ready-to-use, Recombinant yeast assay,
- MeSH
- alfa receptor estrogenů genetika MeSH
- androgenní receptory genetika MeSH
- androgeny farmakologie MeSH
- antagonisté estrogenového receptoru MeSH
- biosenzitivní techniky metody MeSH
- biotest metody MeSH
- endokrinní disruptory farmakologie MeSH
- estrogeny farmakologie MeSH
- kryoprotektivní látky farmakologie MeSH
- lyofilizace metody MeSH
- mikrobiální viabilita účinky léků MeSH
- monitorování životního prostředí metody MeSH
- reprodukovatelnost výsledků MeSH
- Saccharomyces cerevisiae účinky léků genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alfa receptor estrogenů MeSH
- androgenní receptory MeSH
- androgeny MeSH
- antagonisté estrogenového receptoru MeSH
- AR protein, human MeSH Prohlížeč
- endokrinní disruptory MeSH
- ESR1 protein, human MeSH Prohlížeč
- estrogeny MeSH
- kryoprotektivní látky MeSH
Recombinant yeast assays (RYAs) constitute a suitable tool for the environmental monitoring of compounds with endocrine disrupting activities, notably estrogenicity and androgenicity. Conventional procedures require yeast reconstitution from frozen stock, which usually takes several days and demands additional equipment. With the aim of applying such assays to field studies and making them more accessible to less well-equipped laboratories, we have optimized RYA by the immobilization of Saccharomyces cerevisiae cells in three different polymer matrices - gelatin, Bacto agar, and Yeast Extract Peptone Dextrose agar - to obtain a ready-to-use version for the fast assessment of estrogenic and androgenic potencies of compounds and environmental samples. Among the three matrices, gelatin showed the best results for both testosterone (androgen receptor yeast strain; AR-RYA) and 17β-estradiol (estrogen receptor yeast strain; ER-RYA). AR-RYA was characterized by a lowest observed effect concentration (LOEC), EC50 and induction factor (IF) of 1nM, 2.2nM and 51, respectively. The values characterizing ER-RYA were 0.4nM, 1.8nM, and 63, respectively. Gelatin immobilization retained yeast viability and sensitivity for more than 90d of storage at 4°C. The use of the immobilized yeast reduced the assay duration to only 3h without necessity of sterile conditions. Because immobilized RYA can be performed either in multiwell microplates or glass tubes, it allows multiple samples to be tested at once, and easy adaptation to existing portable devices for direct in-field applications.
- Klíčová slova
- Agar, Endocrine disruption, Environmental monitoring, Gelatin, Immobilization, Recombinant yeast assay,
- MeSH
- androgenní receptory MeSH
- biotest metody MeSH
- endokrinní disruptory analýza MeSH
- estradiol MeSH
- monitorování životního prostředí metody MeSH
- polymery * MeSH
- receptory pro estrogeny MeSH
- Saccharomyces cerevisiae metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- androgenní receptory MeSH
- AR protein, human MeSH Prohlížeč
- endokrinní disruptory MeSH
- estradiol MeSH
- polymery * MeSH
- receptory pro estrogeny MeSH
For long, natural organic matter (NOM) composed mainly of humic substances (HS) were regarded as inert in the ecosystems with respect to their possible chemical interaction with exposed organisms. However recently, NOM have been shown to elicit various adverse effects generally attributed to synthetic xenobiotics, including estrogenic effects translating into shifts of the gender ratios in populations. However, the anti/androgenic pathway was not yet evaluated. Here, we applied an anti/androgenic sensitive cell line MDA stably transfected with the firefly luciferase gene under transcriptional control of the androgen responsive element. With five out of twelve tested NOM preparations, particularly with soil and coal isolates, we identified a relatively high, concentration-dependent antiandrogenic effect. This appears to be the first study to show this endocrine disrupting pathway for a ubiquitous biogeochemical matrix, a potential activity which should not be neglected in forthcoming studies on synthetic endocrine disruptors in the environment.
- MeSH
- antagonisté androgenů toxicita MeSH
- chemické látky znečišťující vodu toxicita MeSH
- endokrinní disruptory toxicita MeSH
- huminové látky toxicita MeSH
- lidé MeSH
- luciferasy světlušek genetika metabolismus MeSH
- nádorové buněčné linie MeSH
- neutrální červeň chemie MeSH
- organické látky toxicita MeSH
- sladká voda MeSH
- transfekce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antagonisté androgenů MeSH
- chemické látky znečišťující vodu MeSH
- endokrinní disruptory MeSH
- huminové látky MeSH
- luciferasy světlušek MeSH
- neutrální červeň MeSH
- organické látky MeSH