Growth inhibition, oxidative stress and characterisation of mortality in green algae under the influence of beta-blockers and non-steroidal anti-inflammatory drugs.

Bisoprolol and ketoprofen are widely used pharmaceuticals in medical treatment hence these substances are occurring in wastewaters and in water environment. This research investigated the toxic effects of bisoprolol and ketoprofen on two microalgae taxa, Chlorella vulgaris and Desmodesmus armatus. The results showed that both drugs inhibited the growth of the species tested and induced a decrease in chlorophyll a content compared to controls. Ketoprofen turned out to be harmful to algae as the half maximal effective concentration (EC50) values (14 days) were 37.69 mg L-1 for C. vulgaris and 40.93 mg L-1 for D. armatus. On the other hand, for bisoprolol, the EC50 values were greater than the established NOEC, 100 mg L-1. Bisoprolol and ketoprofen induced oxidative stress in the tested microorganisms, as indicated by changes in the activities of antioxidant enzymes. Exposure to 100 mg L-1 of drugs significantly increased the activity of catalase, peroxidase and superoxide dismutase. Fluorescence microscopy showed that both medicaments changed the cells' morphology. There was atrophy of chlorophyll in the cells, moreover, dying multinuclear cells and cells without nuclei were observed. In addition, there were atrophic cells, namely cells that lacked nuclei and chlorophyll. Profile area analyses showed that bisoprolol and ketoprofen treated C. vulgaris cells were approximately 4 and 2 times greater compared to control ones. Our experimental findings highlight the ecotoxicological threats for aquatic primary producers from bisoprolol and ketoprofen and provide insight into the characteristics of their death.

Bisphenol A and its substitutes in the aquatic environment: Occurrence and toxicity assessment.

Bisphenol A is classified as a high production volume chemical commonly used in the manufacture of polycarbonate plastics, epoxy resins and thermal paper. The endocrine disrupting properties of this xenobiotic have led to the restriction and prohibition of its use in many consumer products. To date, many chemical compounds with a chemical structure similar to bisphenol A have been used in consumer products as its replacement. The ubiquitous occurrence of bisphenol A and its substitutes in the environment and their endocrine activity as well as adverse effects on aquatic organisms is a global concern, especially because many available literature reports show that many substitutes (e.g. bisphenol AF, bisphenol AP, bisphenol B, bisphenol C, bisphenol F, bisphenol G, bisphenol FL, tetrabromobisphenol A) exert adverse effects on aquatic organisms, similar to, or even stronger than bisphenol A. Therefore, the objective of this paper is to provide a comprehensive overview of the production, sources, occurrence and associated toxicity, as well as the endocrine activity of bisphenol A and its substitutes on aquatic species. The environmental levels and ecotoxicological data presented in this review allowed for a preliminary assessment and prediction of the risk of bisphenol A and its substitutes for aquatic organisms. Furthermore, the data collected in this paper highlight that several compounds applied in bisphenol A-free products are not safe alternatives and regulations regarding their use should be introduced.

Toxic effects of bisphenol A and its analogues on cyanobacteria Anabaena variabilis and Microcystis aeruginosa.

In the last decades, the use of bisphenol A has attracted global attention resulting from its actions as an endocrine disrupting compound. In this regard, various bisphenol analogues have been manufactured as a replacement for this compound in consumer products. As a result of the high production volumes, different bisphenol analogues are entered into the terrestrial and aquatic environment, which consequently leads to their increasing contamination and may pose serious risk to organisms. Nevertheless, only few studies have reported on the toxic effect of bisphenol analogues on phytoplankton. Therefore, in this study, the anticyanobacterial activity of six bisphenol analogues and their mixture were investigated for the first time. Bisphenol AF, bisphenol B and bisphenol C (14 d, EC50 12.88-54.87 mg L-1) exhibit more toxic effect to both tested species in comparison to bisphenol A (14 d, EC50 55.27-78.96 mg L-1). Moreover, data show that mixture of bisphenol analogues (14 d, EC50 32.32-60.88 mg L-1) exhibit toxic effect similar to or even stronger than that of bisphenol A. The toxic effect of bisphenol analogues, singly and in combination on the growth of both cyanobacteria species was arranged in the following order: bisphenol AF > bisphenol C> bisphenol B> bisphenol A> bisphenol E> bisphenol BP and bisphenol B> bisphenol AF > bisphenol C> bisphenol A> bisphenol E> bisphenol BP for Anabaena variabilis and Microcystis aeruginosa, respectively. This research aims to assure a basic understanding of the toxic effects of bisphenol analogues on cyanobacteria and provides a more comprehensive view on environmental risk assessment.

Molecularly imprinted polymer film grafted from porous silica for efficient enrichment of steroid hormones in water samples.

Steroid hormones as endocrine disrupting compounds can interfere with the functioning of hormonal systems of organisms and thus affect the health and reproduction of humans and wildlife. Unfortunately, these types of harmful endocrine disrupting compounds have been found in a variety of environmental samples at very low concentrations. Therefore, a simple, fast, and efficient method for enrichment of water samples is needed. A molecularly imprinted solid-phase extraction combined with high performance liquid chromatography coupled with diode array detection was developed for the determination of six steroid hormones, such as estrone, 17-ß-estradiol, estriol, 17-α-ethinylestradiol, progesterone, and testosterone in water samples. The recoveries obtained in the proposed method were in the range of 78.7-101.3%. Matrix effect below 20% suggests that the quantitative and qualitative results of the analysis were not significantly affected by the matrix. The results show that molecularly imprinted polymers based on spherical silica gel had the potential to be a highly innovative and selective sorbent. The proposed method was proved to be applicable for molecularly imprinted solid-phase extraction in selective and reliable extraction and enrichment of steroid hormones in environmental water samples.

Toxic effects of single animal hormones and their mixtures on the growth of Chlorella vulgaris and Scenedesmus armatus.

In their environments, aquatic organisms are simultaneously exposed to mixtures of several endocrine disrupting compounds, including hormones. However, most of the toxicity studies so far focused on effects of single contaminants. The available information on the potential toxicity of combined hormones on microalgae is extremely limited. For these reasons the aim of this study was to evaluate the individual and mixture effect of estrone (E1), ß-estradiol (E2), estriol (E3), 17-α-ethinylestradiol (EE2), progesterone (PRO), 5-pregnen-3ß-ol-20-one (PRE), levonorgestrel (LG) and testosterone (TST) on Chlorella vulgaris and Scenedesmus armatus. Green algae cells were exposed to different concentrations (0.1-100 mg L-1) of hormones for 14 days. Biomass in the form of dry weight and chlorophyll a was examined. The decreasing order of toxicity (based on EC50, 14d) to Chlorella vulgaris and Scenedesmus armatus was: EE2>PRO > E2>PRE > TST > E3>LG > E1 and EE2>PRO > TST > E2>PRE > LG > E1>E3, respectively. Chlorella vulgaris was more sensitive to the effects of hormones than Scenedesmus armatus. Although mixed hormones were more toxic to green algae than single hormones, in the ecosystem mixtures can pose higher ecological risk than single pollutants. Therefore, data on the toxicology of both single and mixed hormones is very valuable for assessment of the possibility of adverse ecological effects caused by these pollutants. Furthermore, these results suggest that environmental exposure to hormone mixtures may cause toxicity levels different to the sum of those of the single hormones and provides a basic understanding of their toxic effect on algae.

Cymantrenyl-Nucleobases: Synthesis, Anticancer, Antitrypanosomal and Antimicrobial Activity Studies.

The synthesis of four cymantrene-5-fluorouracil derivatives (1-4) and two cymantrene-adenine derivatives (5 and 6) is reported. All of the compounds were characterized by spectroscopic methods and the crystal structure of two derivatives (1 and 6), together with the previously described cymantrene-adenine compound C was determined by X-ray crystallography. While the compounds 1 and 6 crystallized in the triclinic P-1 space group, compound C crystallized in the monoclinic P21/m space group. The newly synthesized compounds 1-6 were tested together with the two previously described cymantrene derivatives B and C for their in vitro antiproliferative activity against seven cancer cell lines (MCF-7, MCF-7/DX, MDA-MB-231, SKOV-3, A549, HepG2m and U-87-MG), five bacterial strains Staphylococcus aureus (methicillin-sensitive, methicillin-resistant and vancomycin-intermediate strains), Staphylococcus epidermidis, and Escherichia coli, including clinical isolates of S. aureus and S. epidermidis, as well as against the protozoan parasite Trypanosoma brucei. The most cytotoxic compounds were derivatives 2 and C for A549 and SKOV-3 cancer cell lines, respectively, with 50% growth inhibition (IC50) values of about 7 µM. The anticancer activity of the cymantrene compounds was determined to be due to their ability to induce oxidative stress and to trigger apoptosis and autophagy in cancer cells. Three derivatives (1, 4 and 5) displayed promising antitrypanosomal activity, with GI50 values in the low micromolar range (3-4 µM). The introduction of the 5-fluorouracil moiety in 1 enhanced the trypanocidal activity when compared to the activity previously reported for the corresponding uracil derivative. The antibacterial activity of cymantrene compounds 1 and C was within the range of 8-64 µg/mL and seemed to be the result of induced cell shrinking.