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Pistacia lentiscus L. is one of the most popular medicinal plants attributed to its beneficial properties on human health. However, few toxicogenetic studies have been carried out. Therefore, the aim of this study was to examine the potential genotoxic/antigenotoxic and mutagenic/antimutagenic properties of oil, ethyl acetate and ethanolic extracts of P. lentiscus L. fruits using in vitro the Ames and Umu assays, as well as in vivo micronucleus (MN) test. Extracts did not exert any significant mutagenic/genotoxic effects but provided protection against standard mutagenic and genotoxic agents including 2 nitrofluorene (2-NF) at 2.5 and 5µg/ml; sodium azide at 5 and 10µg/ml; 3-methylcholanthrene (3-MC) at 25 and 50μg/ml; cyclophosphamide (CP) at 50 and 100μg/ml; 4-nitroquinoline 1-oxide (4-NQO) at 0.05µg/ml and 2-amino-anthracene (AA) at 0.2µg/ml. Further, cytotoxicity and selectivity were examined on human hepatocarcinoma (HepG2), and MCF-7 breast cancer cell lines as well as a human normal-like fibroblast cell line (TelCOFS02MA) using MTT assay. Among all extracts, PF1 (ethanolic) showed the most significant selectivity index (SI) (HepG2:11.98; MCF7:4.83), which led to further investigations using an animal model. Oral administration of PF1 (125–1000 mg/kg b.w.) significantly decreased the number of micronucleated cells in CP -initiated (50 mg/kg b.w.) mice, while the number of micronucleated reticulocytes (MNRET), micronucleated polychromatic erythrocytes (MNPCE) or mitotic index (MI) were not markedly affected. Further, PF1 significantly enhanced catalase (CAT) and superoxide dismutase (SOD) activities in the livers and kidneys of these animals. The obtained results indicated the beneficial properties of P. lentiscus L. fruits for use in therapy against harmful effects of genotoxic and mutagenic agents. However, while promising it should be noted that the obtained results are preliminary and need to be confirmed prior to therapeutic use.

Active packaging systems, interacting directly with the enclosed food, can delay or inhibit those phenomena responsible for food quality decay, contributing to the food shelf-life extension. In this work a vinyl resin-based coating containing free or loaded eugenol (EG) in Santa Barbara Amorphous (SBA)15 mesoporous silica nanoparticles is designed to coat flexible aluminium foils to obtain an antimicrobial material. Thermogravimetric analysis shows a good loading capacity of eugenol in SBA15 (48% wt/wt). SEM analysis shows a good dispersion of free EG in the hosting polymeric matrix, whereas some EG/SBA15 particles aggregations are observed in the material. Water contact angle highlights a higher hydrophobicity of the eugenol based-materials (>90°) compared to the pristine vinyl coating (85°). Electrochemical impedance spectroscopy highlights no corrosion phenomena of the VIN/5%(EG/SBA15EG) coating and corrosion phenomena of the VIN/5%EG coating after 7 days of exposure to lactic acid pH = 4. Finally, the two active coatings are studied to evaluate their antibacterial activity using the ISO 22196. Interestingly, results demonstrate that when eugenol is loaded in the SBA15 mesoporous silica nanoparticles the antimicrobial activity of the material significantly increases against both foodborne pathogens and food spoilage bacteria, achieving the highest microbial growth reduction on S. aureus (R = 3.62 log).

The ongoing COVID-19 pandemic is leading to an increase of the global production of plastics since the use of personal protective equipment (PPEs, i.e. gloves, gowns, masks, packaging items), has become mandatory to prevent the spread of the virus. Plastic breaks down into micro/nano particles due to physical or chemical or biological actions into environment. Due to small dimensions, ubiquitous and persistent nature, the plastic particles represent a significant threat to ecosystems and can entry into food chains. Among the plastic polymers used for PPEs, polystyrene is less studied regarding its eco-geno-toxicity. This study aims to investigate acute, chronic and subchronic effects of the microplastic polystyrene beads (PS-MP, size 1.0 μm) on three freshwater species, the alga Raphidocelis subcapitata, the rotifer Brachionus calyciflorus, the crustacean Ceriodaphnia dubia and the benthic ostracod Heterocypris incongruens. Furthermore, the potential genotoxicity and the ROS production due to the PS-MP were also determined in C. dubia. Results revealed that the acute effects occurred at concentrations of PS-MP in the order of dozens of mg/L in B. calyciflorus and C. dubia and hundreds of mg/L in H. incongruens. Regarding long-term toxicity, increasing chronic effects with EC50s in the order of units (C. dubia), hundreds (B. calyciflorus) and thousands (R. subcapitata) of μg/L were observed. Both for acute and chronic/sub chronic toxicity, daphnids were more sensitive to polystyrene than ostracods. Moreover, when C. dubia neonates were exposed to the PS-MP, alterations in genetic material as well as the production of ROS occurred, starting from concentrations in the order of units of μg/L, probably due to inflammatory responses. At last, the risk quotient (RQ) as a measure of risk posed by PS-MPs in freshwater environment, was calculated obtaining a value of 7.2, higher than the threshold value of 1.

Imidacloprid is a neonicotinoid systemic insecticide used worldwide. Despite its hazardous impact on non-target organisms, few studies have been conducted concerning the potential eco-genotoxic effects in invertebrates of surface waters where this pesticide is detected from units of ng/L to tens of μg/L. The aim of the present work was to determine the acute, the sub-chronic and the chronic toxicity of imidacloprid in producers and primary consumers of the freshwater trophic chain. The organisms under investigation were the green alga Raphidocelis subcapitata, the rotifer Brachionus calyciflorus, the cladoceran crustacean Ceriodaphnia dubia and the benthic ostracod Heterocypris incongruens. In addition, potential DNA damage and ROS production were evaluated in C. dubia. Furthermore, in accordance with European guidelines, toxicological risk assessment of imidacloprid was performed for all continents considering its global occurrence in surface waters. In addition, we assessed the genotoxicological risk and median inhibition of reproduction was observed at units of mg/L for rotifers and daphnids. Algae showed the lowest level of sensitivity to the pesticide with effective concentrations from units to hundreds of mg/L. DNA lesions were marked from 7μg/L with a significant increase in damage as concentrations increased. Chronic toxicity risk quotient values were generally below to a threshold value of 1, with no consequential environmental concern other than for the Canadian areas. On the contrary, the genotoxicological risk quotient values were found higher than the threshold value in all continents.

The antibacterial and antioxidant activities of three methoxyphenol phytometabolites, eugenol, capsaicin, and vanillin, were determined. The in vitro antimicrobial potential was tested on three common foodborne pathogens (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus) and three food spoilage bacteria (Shewanella putrefaciens, Brochothrix thermosphacta, and Lactobacillus plantarum). The antioxidant assays were carried out for studying the free radical scavenging capacity and the anti-lipoperoxidant activity. The results showed that eugenol and capsaicin were the most active against both pathogens and spoilage bacteria. S. aureus was one of the most affected strains (median concentration of growth inhibition: IC50 eugenol = 0.75 mM, IC50 capsaicin = 0.68 mM, IC50 vanillin = 1.38 mM). All phytochemicals slightly inhibited the growth of L. plantarum. Eugenol was the most active molecule in the antioxidant assays. Only in the oxygen radical absorbing capacity (ORAC) test did vanillin show an antioxidant activity comparable to eugenol (eugenol ORAC value = 2.12 ± 0.08, vanillin ORAC value = 1.81 ± 0.19). This study, comparing the antimicrobial and antioxidant activities of three guaiacol derivatives, enhances their use in future applications as food additives for contrasting both common pathogens and spoilage bacteria and for improving the shelf life of preserved food.

Wastewater irrigation of crops may be effective to avoid depletion (about 70%) of freshwater resources. However, the use of reclaimed waters containing persistent microcontaminants such as antineoplastic drugs is of high environmental concern. These active compounds may affect human health with potentially severe adverse effects. To better understand the impact on human health following irrigation of crops with reused contaminated waters, we exposed four edible plants, Brassica rapa, Lactuca sativa, Raphanus sativus, and Triticum durum, to two commonly used antitumoral drugs: 5-fluorouracil (5-FU), and Cisplatin (CDDP), using metabolomics as a potential functional genomics tool to combine with genotoxicity experiments. The metabolome of the treated and untreated plants was analysed to detect biochemical alterations associated to the exposure, and the potential genotoxic damage related to human exposure to the treated plants was evaluated using the comet assay in human lymphocytes, which are characterized by high sensitivity to genotoxic substances. The edible species were able to assimilate 5-FU and CDDP during the treatment, affecting the biochemical pathways of these plants with subsequent metabolome modifications. These metabolic alterations differed according to the specific species used for the test. Furthermore, all vegetables treated with two concentrations of the selected drugs (10 and 100 μg/L) caused significant (p < 0.0001) genotoxic damage in the cells of the immune system at a higher level than in the lymphocytes directly exposed to single antineoplastic drugs.

Capsaicin is a spicy capsaicinoid, produced as secondary metabolite by Capsicum fruits. This alkaloid has been used for years in folk medicine for its analgesic and antinflammatory properties although most data is referred to the raw fruit. In this study, the antiradical activity of the pure capsaicin has been studied using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays as well as its antiproliferative activity, using MTT assay, against two human tumour cell lines, the colorectal Caco-2 and the oesophageal OE19 cells. Furthermore, the antiproliferative activity observed on tumoral cells was compared with that of the human normal-like fibroblast cell line TelCOFS02MA. In addition, the apoptotic activity was evaluated using TUNEL assay. A higher radical scavenging activity was observed against ABTS radical cation than DPPH. Capsaicin showed also a higher cytotoxicity against cancer cells than normal-like cells with Selectivity index values greater than 2 at 72h. Capsaicin induced apoptosis especially in OE19 cell line.

In recent years, chronic degenerative diseases such as certain types of cancers, are becoming an evident issue. DNA damage has been for long recognized as a causal factor for cancer development because mutations or chromosomal aberrations affect oncogenes and tumor suppressor genes leading cells to malignant transformation and to the subsequent cancerous growth. Medicinal plants are often used for the prevention or treatment of various diseases with great scientific interest. Among the medicinal plants distributed in the Mediterranean region, Fraxinus angustifolia Vahl. has been used in traditional medicine for its remarkable curative properties. However, in spite of this popularity, little works have been performed on the activity so that further studies should be performed to investigate in depth the antimutagenic, antigenotoxic and antiproliferative activities of the plant. Thus, the present study was aimed to the evaluation of the potential antimutagenic, antigenotoxic and antiproliferative properties of leaves and stem bark extracts of this well-known tree. Antimutagenic activity was evaluated by Salmonella mutagenicity assay in Salmonella typhimurium TA98 and TA100 strains. The antigenotoxic potential was assessed by umu test in the strain of S. typhimurium TA1535/pSK1002. Antiproliferative activity was studied on human hepatoblastoma (HepG-2) and on breast adenocarcinoma (MCF-7) cell lines by MTT assay. Furthermore, the antiproliferative activity observed on cancer cells was compared with that on the human normal-like fibroblasts (TelCOFS02MA) and the selectivity index was calculated to understand if extracts were able to exert selective toxicity towards cancer cells. Moreover, phenolic compounds are plant substances with a large spectrum of biochemical activities with antioxidant, antimutagenic and anticarcinogenic effects. Based on the strong evidence of biological activities of phenolic compounds, the study was focused on the determination of total phenolics and flavonoids contents, and the phytochemical composition of the extracts assessed by LC/MS. The ethanol extracts of both leaves and stem barks showed significant from moderate to strong antimutagenic and antigenotoxic effects. In addition, selective cytotoxicity towards cancer cells was shown by ethanolic leaves extract and aqueous/chloroform leaves and stem bark extracts. The latter showed high levels of total phenolic contents among all the other extracts. Identified phenylethanoids (calceolariosides, verbascoside) and secoiridoids (oleuropein and ligstroside) could be responsible for the demonstrated broad spectrum of healthy properties.

Benzalkonium chloride (BAC) is a cationic surfactant commonly used as a disinfectant. Its ubiquitous nature is the result of high usage and frequent discharge into the environment and evidence of interaction with numerous contaminants, such as pharmaceutical active compound residues. Anticancer drugs, among these compounds, are able to exert eco-genotoxic effects at sub ng–µg/L. The purpose of this study was to assess the reproductive toxicity and the genotoxicity of 5-fluorouracil (5-FU), cisplatin (CDDP), etoposide (ET), and imatinib mesylate (IM)—binary mixtures combined with BAC in Ceriodaphnia dubia. The effects of the mixtures were assessed under the assumption of independent action in experiments that applied two effect levels. The type of interaction was not the same over the range of effect sizes. The combined action experiment on reproduction showed an antagonistic effect at higher effect levels for all binary combinations, except for BAC/IM, whereas independent action was observed in all mixtures at a low effect level. The results of binary combinations on genotoxicity showed antagonistic effects for BAC + ET and BAC + CDDP, whereas independence was expressed in BAC + IM and BAC + 5-FU. The antagonistic interactions still led to higher effects than those observed after single exposures at the same doses in most cases. The effects of mixtures of drugs should be taken into account for environmental risk assessment.

Metabolomics is the analysis of endogenous and exogenous low molecular mass metabolites within cells, tissues, or biofluids of an organism in response to an external stressor. In this chapter, we highlight the importance of the subdiscipline of environmental metabolomics, which investigates the interactions of organisms with environmental stressors such as biotic and abiotic factors, xenobiotics, temperature shifts, and chemical contaminants. Over the past decade, there has been increasing scientific interest in environmental metabolomics, most likely attributable to the comprehensive nature of nontargeted metabolomics. Hypotheses have therefore been developed on complex environmental stressor effects, especially those with unknown modes of action. The availability of a wide variety of model organisms such as freshwater organisms of the food chain has promoted the potential of metabolomics to detect stress from an extensive range of external factors. Furthermore, these dynamics may shift from individuals to populations, contemplating the traditional fields of the ecophysiology and ecology from instantaneous effects to those over evolutionary timescales. In this chapter, we provide an overview of analytical instrumentation, extraction methods, general experimental design, and the statistical methods generally used in environmental metabolomics. Despite the difficulty in understanding the consequences of environmental exposure due to inter- and intra-individual variability, we believe that environmental metabolomics may enrich our understanding of the responses of organisms to the numerous types of environmental stressors.

Antineoplastic drug residues released in the aquatic system represent a potential risk to exposed non-target organisms. Antineoplastic drugs are present in aquatic environments at lower concentration levels than in other therapeutic classes. In particular, antineoplastic drugs are known for their continuous release and subsequent exposure throughout the life span of aquatic organisms and will yield long-term toxicity rather than pose an immediate threat to the environment and human health. Furthermore, these drugs interfere directly or indirectly with DNA and their eco-toxicological effects, such as reproductive inhibition, may modify genetic material. In light of these observations, this chapter aims to investigate the acute and chronic effects of the most commonly used anticancer drugs in organisms of the freshwater trophic chain. The drugs investigated belong to the subgroup L01 of the Anatomical Therapeutic Chemical Classification System and have been chosen considering their occurrence, chemical structures, and modes of action.

Loratadine and desloratadine are second-generation antihistaminic drugs. Because of human administration, they are continuously released via excreta into wastewater treatment plants and occur in surface waters as residues and transformation products (TPs). Loratadine and desloratadine residues have been found at very low concentrations (ng/L) in the aquatic environment but their toxic effects are still not well known. Both drugs are light-sensitive even under environmentally simulated conditions and some of the photoproducts have been isolated and characterized. The aim of the present study was to investigate the acute and chronic ecotoxicity of loratadine, desloratadine and their light-induced transformation products in organisms of the aquatic trophic chain. Bioassays were performed in the alga Pseudokirchneriella subcapitata, the rotifer Brachionus calyciflorus and in two crustaceans, Thamnocephalus platyurus and Ceriodaphnia dubia. Loratadine exerted its acute and chronic toxicity especially on Ceriodaphnia dubia (LC50: 600 mu g/L, EC50: 28.14 mu g/L) while desloratadine showed similar acute toxicity among the organisms tested and it was the most chronically effective compound in Ceriodaphnia dubia and Pseudokirchneriella subcapitata. Generally, transformation products were less active in both acute and chronic assays.

Cannabidiol (CBD) and cannabidivarin (CBDV) are natural cannabinoids which are consumed in increasing amounts worldwide in cannabis extracts, as they prevent epilepsy, anxiety, and seizures. It was claimed that they may be useful in cancer therapy and have anti-inflammatory properties. Adverse long-term effects of these drugs (induction of cancer and infertility) which are related to damage of the genetic material have not been investigated. Therefore, we studied their DNA-damaging properties in human-derived cell lines under conditions which reflect the exposure of consumers. Both compounds induced DNA damage in single cell gel electrophoresis (SCGE) experiments in a human liver cell line (HepG2) and in buccal-derived cells (TR146) at low levels (≥ 0.2 µM). Results of micronucleus (MN) cytome assays showed that the damage leads to formation of MNi which reflect chromosomal aberrations and leads to nuclear buds and bridges which are a consequence of gene amplifications and dicentric chromosomes. Additional experiments indicate that these effects are caused by oxidative base damage and that liver enzymes (S9) increase the genotoxic activity of both compounds. Our findings show that low concentrations of CBD and CBDV cause damage of the genetic material in human-derived cells. Furthermore, earlier studies showed that they cause chromosomal aberrations and MN in bone marrow of mice. Fixation of damage of the DNA in the form of chromosomal damage is generally considered to be essential in the multistep process of malignancy, therefore the currently available data are indicative for potential carcinogenic properties of the cannabinoids. Electronic supplementary material The online version of this article (10.1007/s00204-018-2322-9) contains supplementary material, which is available to authorized users.

International audience; The photochemical behavior of etodolac was investigated under various irradiation conditions. Kinetic data were obtained after irradiation of 10− 4 M aqueous solutions by UVB, UVA and direct exposure to sunlight. The Xenon lamp irradiation was used in order to determine the photodegradation quantum yield under sun-simulated condition (ϕsun). The value was determined to be = 0.10 ± 0.01. In order to obtain photoproducts and for mechanistic purposes, experiments were carried out on more concentrated solutions by exposure to sunlight and to UVA and UVB lamps. The drug underwent photooxidative processes following an initial oxygen addition to the double bond of the five membered ring and was mainly converted into a spiro compound and a macrolactam. Ecotoxicity tests were performed on etodolac, its photostable spiro derivative and its sunlight irradiation mixture on two different aquatic trophic levels, plants (algae) and invertebrates (rotifers and crustaceans). Mutagenesis and genotoxicity were detected on bacterial strains. The results showed that only etodolac had long term effects on rotifers although at concentrations far from environmental detection values. A mutagenic and genotoxic potential was found for its derivative.

The eco-genotoxicity of six anti-neoplastic drugs, 5-fluorouracil, capecitabine, cisplatin, doxorubicin, etoposide, and imatinib, belonging to five classes of anatomical therapeutic classification (ATC), was studied applying the in vivo comet assay on cells from whole organisms of Daphnia magna and Ceriodaphnia dubia. For the first time, this test was performed in C. dubia. In addition, to have a wider genotoxic/mutagenic profile of the anticancer drugs selected, SOS chromotest and Salmonella mutagenicity assay were performed. The comet results showed that all drugs induced DNA damage, in both Cladocerans, with environmental concern; indeed Doxorubicin induced DNA damage in the order of tens of ng L(-1) in both crustaceans, as well as 5-flurouracil in C. dubia and cisplatin in D. magna. In the SOS Chromotest all drugs, except imatinib, were able to activate the repair system in Escherichia coli PQ37 while in the Salmonella mutagenicity assay, doxorubicin was the only drug able to cause direct and indirect frameshift and base-pair substitution mutations. Comet assay was the most sensitive tool of genotoxic exposure assessment, able to detect in vivo the adverse effects at concentration lower than those evaluated in vitro by bacterial assays.

Cyclophosphamide (CP) and Ifosfamide (IF) are two nitrogen mustard drugs widely prescribed in cancer therapy. They are continuously released via excreta into hospital and urban wastewaters reaching wastewater treatment plants. Although CP and IF, their metabolites and transformation products (TPs) residues have been found in the aquatic environment from few ng L−1 to tens of μg L−1, their environmental toxic effects are still not well known. The present study aimed to investigate the acute and chronic ecotoxicity of CP and IF and their commercially available human metabolites/TPs, i.e. carboxy-CP, Keto-CP and N-dechloroethyl-CP on different organisms of the aquatic trophic chain. The experiments were performed using the green alga Pseudokirchneriella subcapitata, the rotifer Brachionus calyciflorus and the crustaceans Thamnocephalus platyurus and Ceriodaphnia dubia. Moreover, to assess the treatment conditions in regards to parent compound removal and formation of new TPs, CP and IF were UV- irradiated for 6 h, 12 h, 24 h, 36 h and 48 h, followed by toxicity evaluation of treated samples by algae, rotifers and crustaceans. Between the parent compounds, IF resulted as more toxic drug under tested conditions, exerting both acute and chronic effects especially on C. dubia (LC50:196.4 mg L−1, EC50:15.84 mg L−1). Among the tested metabolites/TPs, only carboxy-CP inhibited the reproduction in the rotifer. However, LOEC and NOEC values were calculated for CP and IF for all organisms. In addition, despite a low degradation of CP (28%) and IF (36%) after 48 h UV-irradiation, statistically significant effect differences (p < 0.05) from not-irradiated and irradiated samples were observed in both acute and chronic assays, starting from 6 h UV-irradiation. Our results suggest that the toxic effects found in the aquatic organisms may be attributable to interactions between the parent compounds and their metabolites/TPs. The interactions among CP, IF and their metabolites/TPs may cause environmental toxic effects on aquatic organisms.

none 6 The growing use of cytostatic drugs is gaining relevance as an environmental concern. Environmental and distribution studies are increasing due to the development of accurate analytical methods, whereas ecotoxicological studies are still lacking. The aim of the present study was to investigate the acute and chronic toxicity of six cytostatics belonging to five classes of Anatomical Therapeutic Classification (ATC) (5-fluorouracil, capecitabine, cisplatin, doxorubicin, etoposide, and imatinib) on primary consumers of the aquatic chain (Daphnia magna, Ceriodaphnia dubia, Brachionus calyciflorus, and Thamnocephalus platyurus). Acute ecotoxicological effects occurred at concentrations in the order of mg L-136 , higher than those predicted in the environment, and the most acutely toxic drugs among those tested were cisplatin and doxorubicin for most aquatic organisms. For chronic toxicity, cisplatin and 5-fluorouracil were the most toxic drugs in all test organisms, inducing reproduction inhibition in 50% of crustaceans at a concentration on the order of μg L-140 . Rotifers were less susceptible to this class of pharmaceuticals. On the basis of chronic results, the low concentrations for an effect suggest a potential environmental risk of cytostatics. Thus, this study represents an important starting point for establishing the real environmental impact of these substances. Parrella A; Lavorgna M; Criscuolo E; Russo C; Fiumano V; Isidori M Parrella A; Lavorgna M; Criscuolo E; Russo C; Fiumano V; Isidori M

To increase the knowledge about the biological properties of tomatoes, the antioxidant and antiproliferative activities of extracts of nine cultivars of Lycopersicon esculentum, as well as their chemical composition, were studied. The highest antioxidant capacity has been revealed in San Marzano Cirio 3 and Pomodoro Giallo cultivars, both in hydrophilic and lipophilic fractions, while San Marzano, Corbarino di Corbara, and Pomodoro Giallo exhibited the best radical scavenger activity in methanolic fraction. As regards the antiproliferative activity, the median inhibition concentrations of the lipophilic extracts ranged from 272.3 (Pomodoro Giallo) to 364.4 (Corbarino di Corbara) mg dried extract L-1 on Hep-G2 and from 324.6 (San Marzano Cirio 3) to 455.4 (Nero di Sicilia) on Caco-2. The methanolic extracts were more active on Caco-2 than Hep-G2, while the hydrophilic extracts were not active. These biological properties could be ascribed to the identified carotenoids and phenolic acids as well as to a pull of minor compounds exerting their synergistic effect.

Cyclodextrins are natural macrocyclic oligosaccharides able to form inclusion complexes with a wide variety of guests, affecting their physicochemical and pharmaceutical properties. In order to obtain an improvement of the bioavailability and solubility of 5-fluorouracil, a pyrimidine analogue used as chemotherapeutic agent in the treatment of the colon, liver, and stomac cancers, the drug was complexed with alpha- and beta-cyclodextrin. The inclusion complexes were prepared in the solid state by kneading method and characterized by Fourier transform-infrared (FT-IR) spectroscopy and X-ray powder diffractometry. In solution, the 1:1 stoichiometry for all the inclusion complexes was established by the Job plot method and the binding constants were determined at different pHs by UV-VIS titration. Furthermore, the cytotoxic activity of 5-fluorouracil and its complexation products were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on MCF-7 (breast cancer cell line), Hep G2 (hepatocyte carcinoma cell line), Caco-2 (colon adenocarcinoma cell line), and A-549 (alveolar basal epithelial carcinoma cell line). The results showed that both inclusion complexes increased the 5-fluorouracil capability of inhibiting cell growth. In particular, 5-fluorouracil complexed with beta-cyclodextrin had the highest cytotoxic activity on MCF-7; with alpha-cyclodextrin the highest cytotoxic activity was observed on A-549. The IC50 values were equal to 31 and 73 µM at 72 h, respectively. Our results underline the possibility of using these inclusion complexes in pharmaceutical formulations for improving 5-fluorouracil therapeutic efficacy.

Anticancer drugs are continuously released into hospital and urban wastewaters, where they, most commonly, undergo conventional treatment in wastewater treatment plants (WWTPs). Wastewaters contain complex mixtures of substances including parent compounds, their metabolites and transformation products (TPs). In this study, samples of hospital effluents and WWTP influents and effluents from Slovenia and Spain were analyzed for twenty-two selected anticancer drugs, their metabolites and transformation products. Acute and chronic toxicity tests were performed on the crustacean Ceriodaphnia dubia, genotoxicity was determined with Tradescantia and Allium cepa micronucleus (MN) assays and in vitro comet assay in zebrafish (Danio rerio) liver cell line (ZFL cells). Sixty of the two hundred-twenty determinations revealed detectable levels of anticancer drug residues. Among the targeted compounds, platinum based were most frequently detected (90%). Furthermore, erlotinib was detected in 80%, cyclophosphamide and tamoxifen in 70% and methotrexate in 60% of the samples. Seven of ten samples were toxic to C. dubia after acute exposure, whereas after chronic exposure all samples reduced reproduction of C. dubia at high sample dilutions. Allium cepa proved insensitive to the potential genotoxicity of the tested samples, while in Tradescantia increased MN frequencies were induced by a hospital effluent and WWTP influents. In ZFL comet assay all but one sample induced a significant increase of DNA strand breaks. Correlations of chemotherapeutics or their TPs were detected for all bio-assays except for Allium cepa genotoxicity test, however for each test the highest correlations were found for different substances indicating differential sensitivities of the test organisms. (C) 2016 Elsevier Ltd. All rights reserved.

The antioxidant properties of nine different samples of freshly drawn, pasteurised and UHT milk were examined before and after fermentation with several lactobacilli, individually and in combination with the yeast Saccharomyces boulardii. The results showed no statistical differences for the content in proteins, fats and carbohydrates of the different milk. The antioxidant properties of the unfermented milk were positively affected by the thermal treatment. All samples of fermented milk had a greater antioxidant activity than unfermented but only for the freshly drawn milk that increase was significantly different. The co-incubation of lactobacilli with the yeast determined a stronger antioxidant activity especially for the freshly drawn milk. The stability of the tested strains during storage in standard conditions was determined in UHT fermented milk to enable a possible use in probiotic food production. The results showed that the yeast incubated with lactobacilli determined an improvement of the viability of bacteria. © 2012 The Authors. International Journal of Food Science and Technology © 2012 Institute of Food Science and Technology.

An inclusion complex of hydroxymethylferrocene (FeMeOH) with β-cyclodextrin (β-CD) was prepared in the solid state by different techniques such as physical mixture, coprecipitation, kneading and freeze-drying. The formation of the inclusion complex was confirmed by X-ray Powder Diffractometry and Fourier Transform-Infrared spectroscopy. In aqueous solution, the 1:1 stoichiometry was established by a Job plot. The inclusion complex formation was also investigated by NMR and the stability constant (Kb) of the complex was determined to be 478 M−1, which is in agreement with that obtained with UV-Vis tritation (Kb = 541.3 M−1). The phase solubility study showed a diagram classified as BS type and that the solubility of FeMeOH was slightly increased in the presence of β-CD. Furthermore, utilizing phase solubility diagram data, the Kb was estimated to be equal to 528.0 M−1. The cytotoxic activity of FeMeOH and its complexation product with β-CD was determined using the MTT-assay on MDA-MB-231 cell line, showing that the inclusion complex has a higher capability of inhibiting cell growth compared to that of pure FeMeOH.

Pipemidic acid (HPPA) is a quinolone antibacterial agent used mostly to treat gram-negative infections of the urinary tract, but its therapeutic use is limited because of its low solubility. Thus, to improve drug solubility, natural cyclodextrins (CDs) are used for their ability of including guest molecules within their cavities. The aim of this work was to evaluate the antibacterial activity and the preliminary anticancer activity of HPPA included into Heptakis (2,3,6-tri-O-methyl)-β-cyclodextrin (TRIMEB) as a possible approach for a new innovative formulation. The inclusion complex of HPPA with TRIMEB was prepared in solid state by the kneading method and confirmed by FT-IR and powered X-ray diffraction. The association in aqueous solutions of pipemidic acid with TRIMEB was investigated by UV-Vis spectroscopy. Job’s plots have been drawn by UV-visible spectroscopy to confirm the 1:1 stoichiometry of the host–guest assembly. The antibacterial activity of HPPA, TRIMEB and of their complex was tested on Escherichia coli, Pseudomonas aeruginosa, and Staphilococcus aureus. The complex was able to increase 47.36% of the median antibacterial activity of the free HPPA against E. coli (IC50 = 249 µM vs. 473 µM). Furthermore, these samples were tested on HepG-2 and MCF-7. After 72 h, the median tumoral cytotoxicity exerted by the complex was increased by 78.08% and 94.27% for HepG-2 and MCF-7 respectively, showing a stronger bioactivity of the complex than the single HAPPA.

In this study, a comparison of two carbamic pesticides, chlorpropham and phenisopham, was carried out in terms of both photodegradability and ecotoxicity. The photochemical behaviour of the two pesticides was investigated under environmental-like conditions (aqueous media, UVB or solar irradiation). The photochemical kinetic parameters were calculated by irradiating 5 × 10-5 M solutions (H2O-CH3CN, 9:1 v/v) using UVB lamps. For chlorpropham and phenisopham similar half-life times (39.0 and 55.0 min) were determined. Irradiation by sunlight leads to longer degradation half-life times (about 3 months), while it is possible to observe the formation of the same photoproducts. The well-known dechlorination reaction to a hydroxyphenylcarbamate was observed for chlorpropham. Phenisopham undergoes photo-Fries reaction to give rearranged products (hydroxybenzamides) and fragmentation products (hydroxyphenylcarbamate and N-ethylaniline). Acute and chronic toxicity tests of pesticides and their photoproducts were performed on organisms from two levels of the freshwater aquatic chain, the anostraca crustacean Thamnocephalus platyurus, the rotifer Brachionus calyciflorus and the alga Pseudokirchneriella subcapitata. The acute results showed that chlorpropham had median lethal concentrations for the crustacean T. platyurus and the rotifer B. calyciflorus of 10.16 and 35.19 mg L-1, respectively, and phenisopham did not show any acute toxicity as the derivatives up to 10 mg L-1. The only exception was N-ethylaniline which exhibited an acute LC50 value of 0.46 mg L-1. Phenisopham was the most toxic in the long term exposure while its five derivatives showed lower chronic potential for rotifers and algae. The same trend was observed for chlorpropham except for rotifers. This journal is © the Partner Organisations 2014.

The combined genotoxic effects of four anticancer drugs (5-fluorouracil [5-FU], cisplatin [CDDP], etoposide [ET] and imatinib mesylate [IM]) were studied testing their binary mixtures on two crustaceans of the freshwater aquatic chain. The in vivo comet assay on cells from Daphnia magna and Ceriodaphnia dubia was used in single and combined exposure experiments. Assessment was based on two distinct effect sizes determined from dose-response experiments. Doses for single and combined exposures expected to result in these effect sizes were computed based on Bliss independence as reference model. Statistical comparison by analysis of variance of single and combined toxicities allowed accepting or rejecting the independency hypothesis. The results in D. magna showed independent action for all mixtures except for IM+5-FU that showed an antagonistic interaction. In C. dubia, most mixtures had antagonist interactions except IM+5-FU and IM+CDDP that showed Bliss independence. Despite the antagonistic interactions our results demonstrated that combinations of anticancer drugs could be of environmental concern because effects occur at very low concentrations that are in the range of concentrations encountered in aquatic systems. The combined genotoxic effects of four anticancer drugs (5-fluorouracil [5-FU], cisplatin [CDDP], etoposide [ET], and imatinib mesylate [IM]) were studied testing their binary mixtures in two crustaceans that are part of the freshwater food chain, namely Daphnia magna and Ceriodaphnia dubia. Genotoxicity was assessed using the in vivo comet assay. Assessment was based on two distinct effect sizes determined from dose-response experiments. Doses for single and combined exposures expected to result in these effect sizes were computed based on Bliss independence as reference model. Statistical comparison by analysis of variance of single and combined toxicities allowed accepting or rejecting the independency hypothesis. The results obtained for D. magna showed independent action for all mixtures except for IM+5-FU that showed an antagonistic interaction. In C. dubia, most mixtures had antagonist interactions except IM+5-FU and IM+CDDP that showed Bliss independence. Despite the antagonistic interactions, our results demonstrated that combinations of anticancer drugs could be of environmental concern because effects occur at very low concentrations that are in the range of concentrations encountered in aquatic systems.

Caffeine is ubiquitous in surface and ground waters and it has been proposed as a marker of the anthropogenic pressure on the environment. Sewage treatment plants based on active sludges seem to be not very efficient in its complete removal from effluents while addnl. disinfection treatments by chlorination are able to do it. In a simulation of the chlorination step herein we report that caffeine is transformed in six byproducts: 8-???chlorocaffeine, 1,???3-???dimethyl-???5-???azabarbituric acid, N,???N'-???dimethylparabanic acid, N,???N'-???dimethyloxalamide, N-???methylurea and N,???N'-???dimethylurea. The ecotoxicity of caffeine and identified compds. was evaluated on the rotifer Brachionus calyciflorus and the alga Pseudokirchneriella subcapitata to assess acute and chronic toxicity, while SOS Chromotest and Ames Test were used to detect the genotoxic potential of the investigated compds. Moreover, we assessed the possible antigenotoxic effect of the selected compds. using SOS Chromotest after co-???incubation with the std. genotoxin, 4-???nitroquinoline 1-???oxide. Chronic exposure to these compds. caused inhibition of growth population on the rotifer while the algae seemed to be unaffected. Results indicated that caffeine (1)???, N,???N'-???dimethyloxamide (4) and N,???N'-???dimethylparabanic acid (5) reduced ??-???galactosidase activity in comparison with pos. control, both at 1 and 5 mg???/L of 4-???NQNO with a good dose-???response.

The toxicity and genotoxicity of the cationic surfactant benzalkonium chloride (BAC) were studied using Daphnia magna and Ceriodaphnia dubia as model systems. Acute and chronic toxicity testing were performed according to the international standard guidelines and the genotoxicity was detected through the comet assay on cells from whole organisms in vivo exposed. Acute effects occurred at concentrations in the order of tens of μg/L in D. magna and hundreds of μg/L in C. dubia. Chronic effects were found at one order of magnitude less than short-term effects maintaining the same difference in sensitivity between D. magna and C. dubia. BAC induced relevant DNA damage, in both cladocerans; the lowest adverse effect levels were 0.4 and 4 ng/L for D. magna and C. dubia, respectively. As these effective concentrations are far lower than BAC occurrence in surface waters (units of μg/L) a concerning environmental risk cannot be excluded. The findings of this study showed that D. magna and C. dubia, could be used as model organisms to detect acute and chronic toxicity as well as genotoxicity at the whole organism level. The toxicity and genotoxicity of the cationic surfactant benzalkonium chloride (BAC) were studied using Daphnia magna and Ceriodaphnia dubia as model systems. Acute and chronic toxicity testing were performed according to the international standard guidelines and the genotoxicity was detected through the comet assay on cells from whole organisms in vivo exposed. Acute effects occurred at concentrations in the order of tens of mu g/L in D. magna and hundreds of mu g/L in C. dubia. Chronic effects were found at one order of magnitude less than short-term effects maintaining the same difference in sensitivity between D. magna and C. dubia. BAC induced relevant DNA damage, in both cladocerans; the lowest adverse effect levels were 0.4 and 4 ng/L for D. magna and C. dubia, respectively. As these effective concentrations are far lower than BAC occurrence in surface waters (units of mu g/L) a concerning environmental risk cannot be excluded. The findings of this study showed that D. magna and C. dubia, could be used as model organisms to detect acute and chronic toxicity as well as genotoxicity at the whole organism level. (C) 2015 Elsevier Ltd. All rights reserved.

Chlorination experiments on two drugs (sildenafil and tadalafil) were performed mimicking the conditions of a typical wastewater treatment process. The main transformation products were isolated by chromatographic techniques (Thin Layer Chromatography (TLC), Column Chromatography (CC), High Performance Liquid Chromatography (HPLC)) and fully characterized employing Nuclear Magnetic Resonance (NMR) and Mass Spectrometry (MS) analyses. The environmental effects of the parent compounds and transformation products were evaluated using an overall toxicity approach that considered aquatic acute and chronic toxicity on Brachionus calyciflorus and Ceriodaphnia dubia as well as mutagenesis and genotoxicity on bacterial strains. The results revealed that both parent drugs did not show high acute and chronic toxicity for the organisms utilized in the bioassays while, chronic exposure to chlorine derivatives caused inhibition of growth population on rotifers and crustaceans. A mutagenic potential was found for all the compounds investigated. © 2013 Elsevier B.V.

Olive mill wastewaters (OMW) can be a severe problem when disposed of as untreated because of their high organic load, elevated concentration of polyphenols, and moderately low biodegradability. In the present study, the acute toxicity of 15 compounds with low molecular weight (

The occurrence of pharmaceuticals in the environment is of great concern and only few data are available about the adverse effects of such molecules and their derivatives on non-target aquatic organisms. This study was designed to assess the toxic potential of Naproxen, a nonsteroidal anti-inflammatory, Naproxen Na, its freely water soluble sodium salt and their photoproducts in the aquatic environment. Bioassays were performed on algae, rotifers and microcrustaceans to assess acute and chronic toxicity. Furthermore, possible genotoxic effects of photoderivatives were investigated using SOS chromotest and Ames fluctuation test. The results showed that photoproducts were more toxic than the parent compounds both for acute and chronic values, while genotoxic and mutagenic effects were not found. These findings suggested the opportunity to consider derivatives in ecotoxicology assessment of drugs. © 2005 Elsevier B.V. All rights reserved.

The ecotoxicity of the following six antibiotics on aquatic organisms was investigated: Erythromycin, Oxytetracyclin, Sulfamethoxazole, Ofloxacin, Lincomycin and Clarithromycin. Bioassays were performed on bacteria, algae, rotifers, microcrustaceans and fish to assess acute and chronic toxicity, while SOS Chromotest and Ames test were used to detect the genotoxic potential of the investigated drugs. For risk assessment, the environmental impact was calculated by MEC/PNEC ratio using the available data from the literature regarding their occurrence in the aquatic environment and the toxicity data obtained from the bioassays performed. The ecotoxicological results showed that acute toxicity was in the order of mg/L while, for the chronic data the antibiotics were bioactive at concentrations in the order of μg/L, mainly for the algae. Drugs investigated were one or two order of magnitude less active against rotifers and crustaceans. Ofloxacin was the only genotoxic compound and Sulfamethoxazole, Ofloxacin and Lincomycin were mutagenic. As for environmental risk, the macrolides were found to be the most harmful for the aquatic environment. © 2004 Elsevier B.V. All rights reserved.

Olive-oil-mill wastewater (OMW) has significant polluting properties due to its high levels of chemical oxygen demand (COD), biochemical oxygen demand (BOD), and phenols. In the present study, different commercial bacterial formulations were used in the biological treatment of OMW. COD and toxicity testing using primary consumers of the aquatic food chain (the rotifer Brachionus calyciflorus and the crustacean Daphnia magna) were employed to evaluate abatement of the organic load and reduction of the toxic potential. In addition, the four most active formulations were tested mixed pair-wise on the basis of their unique characteristics in order to evaluate the improvement of treatment. The effect of treatment was assessed by measuring COD removal, reduction of total phenols, and decreased toxicity. The results obtained with the mixed formulations showed that the maximum removal of the organic load was about 85%, whereas phenols were reduced by about 67%. The toxicity for rotifers decreased by 43% and for crustaceans by about 83%. © Springer-Verlag 2004.

The aim of the present study was to investigate the in vitro estrogenic and the cytotoxic activity of six cytostatics (5-fluorouracil, capecitabine, cisplatin, doxorubicin, etoposide, and imatinib) belonging to the five classes of Anatomical Therapeutic Classification (ATC) detected in wastewater systems. The estrogenic activity was assessed by YES-assay on Saccharomyces cerevisiae-RMY326 and E-screen on MCF-7 cells. The cytotoxic activity was assessed by MTT Cell Proliferation Assay on the MCF-7 and the MDA-MB-231 cells.The results of estrogenic activity, detected by E-screen and expressed as EC50, showed a high potential of imatinib (10-7μM) followed by cisplatin and 5-fluorouracil. Capecitabine was poorly estrogenic while etoposide and doxorubicin EC50 values were not possible to determine. Cytotoxicity was found at concentrations far from those detected in effluents. The potential endocrine activity of the most active drugs could be associated with human and wildlife risk when considering their occurrence in the environment. © 2014.

Anticancer drugs, interfering with DNA in every living organism, may pose a threat to aquatic environment, even more when they occur as complex mixtures. We investigated the combined long term toxic potential of four anti-neoplastic drugs (5-fluorouracil [5-FU], cisplatin [CDDP], etoposide [ET] and imatinib mesylate [IM]) testing their binary mixtures on two primary consumers of the freshwater aquatic chain with close phylogenetic relationship: Daphnia magna and Ceriodaphnia dubia . The combined toxicities were assessed using two distinct effect sizes that should be observed if Bliss independence holds. Direct statistical comparison by analysis of variance of single and combined toxicities under the assumption of Bliss independence allowed to accept or reject the independency hypothesis. Independency was confirmed for all mixtures both in D. magna and in C. dubia , except for IM+ ET and IM + CDDP in D. magna and for ET + CDDP and ET + 5-FU in C. dubia which at the highest concentrations showed an antagonistic interaction. A synergic tendency was found testing IM + CDDP on C. dubia at the lowest concentration selected. Thus, the chronic ecotoxicological data evaluated in this study show not only a potential environmental risk of anticancer drugs, especially considering their potential synergistic effects, but also the necessity to integrate statistical models with experimental data to establish the real environmental impact of such compounds.

The aim of the present work is the study of the migration from active LDPE polymer films of ?-tocopherol adsorbed onto purely siliceous and amino-functionalized SBA-15 mesoporous silica. In particular, the effect of the pore size and the chemical functionality of the internal walls of the mesophase was thoroughly investigated. Co-condensation of tetraethylorthosilicate and aminopropyltriethoxysilane was exploited to bind amino groups on the pore surface of SBA-15. The synthesized mesoporous powders were characterized by means of X-ray diffraction and N2 adsorption/desorption at 77 K. Powders loaded with tocopherol were characterized by infrared spectroscopy and thermogravimetric analysis. Results show that the maximum of the pore size distribution reduces from 90 Å for purely siliceous SBA-15 to 73 Å for amino-functionalized SBA-15. Infrared analysis shows that tocopherol interacts with the amino groups of functionalized SBA-15. Migration tests were performed at 25 °C, using 96% v/v ethanol as fatty food simulant, on both polymer films containing about 1% wt/wt of free tocopherol and about 3% wt/wt of tocopherol loaded onto purely siliceous and amino-functionalized SBA-15. The tocopherol diffusivity of films containing functionalized mesoporous silica decreased of about 60% with respect to films containing free tocopherol. This is due to the decrease in the pore size and to the increase in diffusion resistance caused by the functionalization of the internal pore walls with the amino groups. Moreover, the oxygen radical absorbing capacity assay of the produced active polymer films proved the antioxidant effectiveness of tocopherol released from samples after manufacturing process.

Nicotine, the main alkaloid of tobacco, is a non-prescription drug to which all members of a tobacco-smoking society are exposed either through direct smoke inhalation or through second-hand passive 'smoking'. Nicotine is also commercially available in some pharmaceutical products and is used worldwide as a botanical insecticide in agriculture. Nicotine dynamics in indoor and outdoor environments as well as the human excretions and the manufacturing process are responsible for its entry in the environment through municipal and industrial wastewater discharges. The presence of nicotine in surface and ground waters points out that it survives a conventional treatment process and persists in potable-water supplies. Complete removal of nicotine is instead reported when additional chlorination steps are used. In this paper a simulation of STP chlorination of nicotine and a genotoxic evaluation of its main degradation products are reported. Under laboratory conditions removal of nicotine seems not to be due to mineralization but to transformation in oxidized and chlorinated products. The by-products have been isolated after fractionation by diverse chromatographic procedures and their structures determined using mass spectrometry and H-1 and C-13 NMR spectroscopy. Preliminary genotoxic SOS Chromotests with Escherichia coil PQ37 evidence no toxicity of the products. (C) 2012 Elsevier B.V. All rights reserved.

Toxicity and endocrine interference of influent and effluent waters from domestic and industrial wastewater treatment plants were determined. In addition, chemical analyses were performed to detect the presence of 17β-estradiol, 17α-ethinyl estradiol, nonylphenol, 4-octylphenol, and p-t-octylphenol as well as lead, copper, and cadmium in these matrices. The results showed that despite low acute toxic potential, most of the samples tested showed both genotoxicity and endocrine interference. Furthermore, to establish whether the observed effects were caused by the alkylphenols and the heavy metals detected, toxic, genotoxic, and endocrine interference tests also were performed on pure chemicals. The acute toxicity was measured on the crustacean Daphnia magna. The estrogenic activity was determined by using the yeast estrogen screen with Saccharomyces cerevisiae RMY326, whereas the SOS Chromotest and Ames test detected the genotoxicity on Escherichia coli PQ37 and Salmonella typhimurium TA98 and TA100, respectively. The results showed that the toxicity found in the matrices did not match the values found for pure chemicals, but a clear correlation was found between alkylphenols and genotoxicity. Both heavy metals and alkylphenols took part in the endocrine interference activity. © 2007 SETAC.

In the last few years many concerns have been raised regarding the environmental safety of alkylphenol polyethoxylate surfactants (APnEOs).They are widely used in detergents, paints, herbicides and many other formulated products. It has been estimated that 60% of APnEOs end up in the aquatic environment; they are biodegradable and transformed into alkylphenols, such as nonylphenol and octylphenol that are hydrophobic and tend to accumulate. In the present study, acute and chronic aquatic toxicity and the estrogenic activity of the following eight alkylphenols were assessed: 4-nonylphenol, 4-octylphenol, 4-nonylphenol-10-ethoxylate, 4-tert-octylphenol, POE (1 to 2)-nonylphenol, POE (6)-nonylphenol, POE (3)-tert-octylphenol and POE (9 to 10)-tert-octylphenol. The toxic potential was measured on the crustaceans Daphnia magna and Ceriodaphnia dubia, while the estrogenic activity was determined by using the YES-test with the strain Saccharomyces cerevisiae RMY326. The results showed that the exposure of crustaceans to the eight xenoestrogens investigated caused both acute and chronic effects. The EC50 values found for C. dubia at 48 h were compared to D. magna at 24 h and, gave a first indication about the toxic activity of the compounds investigated, that is better expressed in the long-term. In fact, chronic data showed a strong increase in toxicity with EC50 values one or two orders of magnitude lower than the acute values. The results of the YES-test showed that nonylphenol, octylphenol and 4-tert-octylphenol were the most estrogenic and the bioassay was able to detect their estrogenicity at very low concentrations (ng-μg/l). © 2005 Elsevier Ltd. All rights reserved.