Acyclovir eco-geno-toxicity in freshwater organisms.

This study explores the eco-geno-toxic impact of Acyclovir (ACV), a widely used antiviral drug, on various freshwater organisms, given its increasing detection in surface waters. The research focused on non-target organisms, including the green alga Raphidocelis subcapitata, the rotifer Brachionus calyciflorus, the cladoceran crustacean Ceriodaphnia dubia, and the benthic ostracod Heterocypris incongruens, exposed to ACV to assess both acute and chronic toxicity. The results indicate that while acute toxicity occurs at environmentally not-relevant concentrations, a significant chronic toxicity for C. dubia (EC50 = 0.03 µg/L, NOEC = 0.02·10-2 µg/L), highlighted substantial environmental concern. Furthermore, DNA strand breaks and reactive oxygen species detected in C. dubia indicate significant increase at concentrations exceeding 200 µg/L. Regarding environmental risk, the authors identified chronic exposures to acyclovir causing inhibitory effects on reproduction in B. calyciflorus at hundreds of µg/L and hundredths of µg/L for C. dubia as environmentally relevant environmental concentrations. The study concludes by quantifying the toxic and genotoxic risks of ACV showing a chronic risk quotient higher than the critical value of 1and a genotoxic risk quotient reaching this threshold, highlighting the urgent need for a broader risk assessment of ACV for its significant implications for aquatic ecosystems.

Characterization of Complexes between Imidacloprid and ß-Cyclodextrin: Evaluation of the Toxic Activity in Algae and Rotifers.

The development of new formulations can be driven by the knowledge of host-guest complexes using cyclodextrins which have the ability to include guest molecules within their hydrophobic cavities, improving the physicochemical properties of the guest. To rationally explore new pesticide formulations, the effects of cyclodextrins on the properties of such guest molecules need to be explored. Imidacloprid is a neonicotinoid systemic insecticide used worldwide. In this study, the inclusion complexes of Imidacloprid (IMI) with ß-cyclodextrin (ß-CD) were prepared in the solid state by co-precipitation and the physical mixing method, with a stoichiometry of 1:1 and 1:2 molar ratios. The obtained products, Imidacloprid:ß-cyclodextrin inclusion complex (IMI:ß-CD), were characterized in the solid state by Fourier transform-infrared (FT-IR) spectroscopy and X-ray powder diffractometry (XRD). In solution, the 1:1 stoichiometry for the inclusion complexes was established by the Job plot method, and the binding constant of IMI:ß-CD was determined by UV-vis titration. The toxicity was determined in producers and primary consumers of the freshwater trophic chain, the green alga Raphidocelis subcapitata and the rotifer Brachionus calyciflorus, respectively. The results indicated that Imidacloprid forms inclusion complexes with CDs showing improved physicochemical properties compared to free Imidacloprid. The formation of the inclusion complex reduced the chronic toxicity in rotifers when IMI concentrations were close to those of environmental concern (tenths/hundredths of micromoles/L). Therefore, CD inclusion complexes could provide important advantages to be considered for the future industrial production of new formulations.

Pistacia lentiscus L. fruits showed promising antimutagenic and antigenotoxic activity using both in-vitro and in-vivo test systems.

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 5 µg/ml; sodium azide at 5 and 10 µg/ml; 3-methylcholanthrene (3-MC) at 25 and 50 µg/ml; cyclophosphamide (CP) at 50 and 100 µg/ml; 4-nitroquinoline 1-oxide (4-NQO) at 0.05 µg/ml and 2-amino-anthracene (AA) at 0.2 µg/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.

Low doses of widely consumed cannabinoids (cannabidiol and cannabidivarin) cause DNA damage and chromosomal aberrations in human-derived cells.

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.

Ecotoxic effects of loratadine and its metabolic and light-induced derivatives.

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 µg/L, EC50: 28.14 µg/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.

A New Approach for Improving the Antibacterial and Tumor Cytotoxic Activities of Pipemidic Acid by Including It in Trimethyl-ß-cyclodextrin.

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)-ß-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.

Capsaicin in Hot Chili Peppers: In Vitro Evaluation of Its Antiradical, Antiproliferative and Apoptotic Activities.

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 72 h. Capsaicin induced apoptosis especially in OE19 cell line.

Benzalkonium Chloride and Anticancer Drugs in Binary Mixtures: Reproductive Toxicity and Genotoxicity in the Freshwater Crustacean Ceriodaphnia dubia.

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.

Teratogenic effects of five anticancer drugs on Xenopus laevis embryos.

In recent years, the environmental presence of pharmaceuticals - including anticancer drugs - is an emerging issue. Because of the lack of appropriate critical studies about anticancer drug effects in frogs, the aim of the present study was to investigate lethal and teratogenic effects of five anticancer drugs widely used in large quantities, i.e. 5-flourouracil, capecitabine, cisplatin, etoposide, and imatinib, in the embryos of the South African clawed frog, Xenopus laevis, using FETAX - Frog Embryo Teratogenesis Assay in Xenopus. None of the studied anticancer drugs induced statistically significant mortality within the concentrations tested (0.01-50mg/L, depending on the studied compound), and no growth inhibition of embryos after a 96-h exposure was observed. Except for cisplatin, the other pharmaceuticals induced an increase of developmental malformations such as abdominal edema, axial flexure, head, eyes, gut and heart malformations with statistically significant effects observed at the highest concentrations tested (50mg/L for 5-flourouracil; 30mg/L for etoposide and 20mg/L for capecitabine and imatinib). The results indicate that anticancer drugs can affect embryogenesis mechanisms.

Alpha- and Beta-Cyclodextrin Inclusion Complexes with 5-Fluorouracil: Characterization and Cytotoxic Activity Evaluation.

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.

Mutagenicity, genotoxicity, and estrogenic activity of river porewaters.

We investigated mutagenicity, genotoxicity, and estrogenic activity in the porewaters of two river basins in southern Italy that had different features. Three samples from each site were collected in different seasons from 7 sites for a total of 21 samples. Mutagenicity was measured with the Ames test with and without metabolic activation (S9) using Salmonella typhimurium TA98 and TA100 strains. Genotoxicity was measured with two tests: one involved a chromophore that detected DNA damage in Escherichia coli PQ37 (SOS chromotest), and the other measured micronuclei formation in the root cells of Vicia faba. Estrogenic activity was measured with a yeast-based estrogen receptor assay and an MCF-7 cell-based, estrogen-sensitive proliferation assay. We also applied chemical analyses to detect alkylphenols, pesticides, natural and synthetic hormones, and heavy metals. The porewaters of both river sediments showed mutagenic/genotoxic activity on V. faba test and Ames test, the latter both with and without S9 liver fraction. The SOS chromotest without metabolic activation was not sufficiently sensitive to detect genotoxicity of the porewaters, but the SOS DNA repair system in E. coli PQ37 was activated in the presence of S9 mix. Good correlations were found between mutagenicity/genotoxicity and the concentration of cadmium and between estrogenic activity and the presence of copper. This study assessed the chemical concentrations of some bioavailable pollutants in porewater and detected the overall effects of multiple pollutants that contributed to mutagenicity, genotoxicity, and estrogenic activity of these two basin porewaters, thus increasing our understanding of the environmental consequences of polluted aquatic ecosystems.

ß-Cyclodextrin inclusion complex to improve physicochemical properties of pipemidic acid: characterization and bioactivity evaluation.

The aptitude of cyclodextrins (CDs) to form host-guest complexes has prompted an increase in the development of new drug formulations. In this study, the inclusion complexes of pipemidic acid (HPPA), a therapeutic agent for urinary tract infections, with native ß-CD were prepared in solid state by kneading method and confirmed by FT-IR and 1H NMR. The inclusion complex formation was also characterized in aqueous solution at different pH via UV-Vis titration and phase solubility studies obtaining the stability constant. The 1:1 stoichiometry was established by a Job plot and the inclusion mechanism was clarified using docking experiments. Finally, the antibacterial activity of HPPA and its inclusion complex was tested on P. aeruginosa, E. coli and S. aureus to determine the respective EC50s and EC90s. The results showed that the antibacterial activity of HPPA:ß-CD against E. coli and S. aureus is higher than that of HPPA. Furthermore, HPPA and HPPA:ß-CD, tested on human hepatoblastoma HepG2 and MCF-7 cell lines by MTT assay, exhibited, for the first time, antitumor activities, and the complex revealed a higher activity than that of HPPA. The use of ß-CD allows an increase in the aqueous solubility of the drug, its bioavailability and then its bioactivity.

Imidacloprid: Comparative toxicity, DNA damage, ROS production and risk assessment for aquatic non-target organisms.

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 7 µg/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.

Diclofenac eco-geno-toxicity in freshwater algae, rotifers and crustaceans.

This study assessed the eco-genotoxic impact of diclofenac (DCF) in sentinel species of the freshwater ecosystem. DCF residues are found in freshwater from few ng/L to tens of µg/L due to the inability of conventional wastewater treatment plants to ensure removal efficiency of the drug. An ample body of literature reports on the acute toxicity of DCF in non-target organisms without addressing potential chronic long-term effects on organisms at actual, environmental concentrations. Herein, assessment for acute and chronic toxicity was performed on organisms in vivo exposed to DCF, specifically on the green alga Raphidocelis subcapitata, the rotifer Brachionus calyciflorus and the crustacean Ceriodaphnia dubia. Furthermore, potential DNA damage and expression of antioxidant genes (MnSOD, Cu/ZnSOD and CAT) were evaluated in crustacean neonates. The toxicological risk of DCF was assessed as well as its. GENOTOXIC RISK: The acute toxicity was observed at concentrations far from those of environmental concern. Rotifers and crustaceans were much more chronically sensitive than the algae to DCF, observing besides, the median effect concentrations at tens of µg/L. In crustaceans, DNA damage was noted at units of µg/L, revealing concentrations of environmental concern. The dysregulated activity of SOD and CAT also showed the ability of DCF to provoke oxidative stress. On assessment of environmental risk, the chronic Risk Quotient (RQ) was above the threshold value of 1. Nevertheless, the genotoxic RQ was significantly greater than the chronic RQ, thus, the need of regulatory bodies to acknowledge the genotoxic impact as an environmental risk factor. To our knowledge, this study is the first investigation to perform environmental genotoxic risk assessment of DCF.

Measuring DNA modifications with the comet assay: a compendium of protocols.

The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA-DNA crosslinks, UV-induced cyclobutane pyrimidine dimers and some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species and types of DNA damage, thereby demonstrating its versatility.

Physicochemical characterization and cytotoxic activity evaluation of hydroxymethylferrocene:ß-cyclodextrin inclusion complex.

An inclusion complex of hydroxymethylferrocene (FeMeOH) with ß-cyclodextrin (ß-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 ß-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 ß-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.

Toxic impact of polystyrene microplastic particles in freshwater organisms.

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.

Polystyrene microplastic particles in combination with pesticides and antiviral drugs: Toxicity and genotoxicity in Ceriodaphnia dubia.

Freshwater ecosystems are recognized as non-negligible sources of plastic contamination for the marine environment that is the final acceptor of 53 thousand tons of plastic per year. In this context, microplastic particles are well known to directly pose a great threat to freshwater organisms, they also indirectly affect the aquatic ecosystem by adsorbing and acting as a vector for the transport of other pollutants ("Trojan horse effect"). Polystyrene is one of the most widely produced plastics on a global scale, and it is among the most abundant microplastic particles found in freshwaters. Nevertheless, to date few studies have focused on the eco-genotoxic effects on freshwater organisms caused by polystyrene microplastic particles (PS-MPs) in combination with other pollutants such as pharmaceuticals and pesticides. The aim of this study is to investigate chronic and sub-chronic effects of the microplastic polystyrene beads (PS-MP, 1.0 µm) both as individual xenobiotic and in combination (binary/ternary mixtures) with the acicloguanosine antiviral drug acyclovir (AC), and the neonicotinoid broad-spectrum insecticide imidacloprid (IMD) in one of the most sensitive non-target organisms of the freshwater food chain: the cladoceran crustacean Ceriodaphnia dubia. Considering that the individually selected xenobiotics have different modes of action and/or different biological sites, the Bliss independence was used as reference model for this research. Basically, when C. dubia neonates were exposed for 24 h to the mixtures during Comet assay, mostly an antagonistic genotoxic effect was observed. When neonates were exposed to the mixtures for 7 days, mostly an additive chronic toxic effect occurred at concentrations very close or even overlapping to the environmental ones ranging from units to tens of ng/L for PS-MPs, from tenths/hundredths to units of µg/L for AC and from units to hundreds of µg/L for IMD, revealing great environmental concern.

Cytotoxic evaluation and chemical investigation of tomatoes from plants (Solanum lycopersicum L.) grown in uncontaminated and experimentally contaminated soils.

The aim of this study was to evaluate the cytotoxic activity and the chemical composition of the tomato extracts coming from, Pomodoro Giallo and San Marzano Cirio 3, and then to evaluate the potential changes when plants were grown in soils contaminated by cadmium, chromium and lead. Extracts were investigated by UHPLC-HRMS and UV-Vis. Cell viability (CellTiter-Glo Luminescent assay), enzyme aldehyde dehydrogenase activity (ALDEFLOUR Assay), cell cycle progression (Accuri C6 Flow Cytometer), apoptosis and necrosis (Annexin V-FITC assay) were evaluated on two gastric cancer (AGS and NCI-N87) and two colorectal cancer (HT-29 and HCT 116) cell lines. Different content of polyphenol and carotenoid constituents was observed. Extracts from uncontaminated soil induced cytotoxic activity towards all selected cancer cells, while extracts coming from contaminated soils showed the aberrant phenotype increased in colorectal cancer cells. Chloroform extracts exerted the highest cytotoxic activity. AGS and HT-29 were the most sensitive to cell cycle arrest and to apoptosis. No necrotic effect was observed in HCT 116. The contrasting effects on cancer cells were observed based on tomato variety, the extract polarity, heavy metal identity, and tested cell line. The investigation of potential adverse health effects due to Cd in the fruits should be explored.

Comparative assessment of antimicrobial, antiradical and cytotoxic activities of cannabidiol and its propyl analogue cannabidivarin.

Cannabidiol and cannabidivarin are phytocannabinoids produced by Cannabis indica and Cannabis sativa. Cannabidiol has been studied more extensively than its propyl analogue cannabidivarin. Therefore, we performed a battery of in vitro biological assays to compare the cytotoxic, antiradical and antibacterial activities of both cannabinoids. Potential mitochondrial metabolism alterations, DNA synthesis inhibition, and plasma membrane damage were studied by MTT assay, BrdU-ELISA and LDH assay of cancer and normal human cells exposed to cannabinoids. ABTS and DPPH assays were performed to observe the effects of the cannabinoids on free radicals. Microbial susceptibility tests were performed to study the activity of the cannabinoids in two bacterial species implicated in human infections, Escherichia coli and Staphylococcus aureus. The results showed that the cannabinoids induced medium levels of cytotoxicity in cancer and normal cells at concentrations ranging from 15.80 to 48.63 and from 31.89 to 151.70 µM, respectively, after 72 h of exposure. Cannabinoids did not exhibit a strong antioxidant capacity in scavenging ABTS or DPPH radicals. No evident differences were observed between the two cannabinoids in antimicrobial activity, except with respect to S. aureus, which showed greater susceptibility to cannabidiol than to cannabidivarin after 72 h of exposure.