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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.

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.

The antioxidant activity of natural compounds consists in their ability to modulate gene and protein expression, thus inducing an integrated cell protective response and repair processes against oxidative stress. New screening tools and methodologies are crucial for the actual requirement of new products with antioxidant activity to boost endogenous oxidative stress responsive pathways, Reactive Oxygen Species (ROS) metabolism and immune system activity, preserving human health and wellness. In this study, we performed and tested an integrated oxidative stress analysis, using DPPH assay and PNT2 cells injured with DPPH. We firstly investigated the mechanism of action of the oxidising agent (DPPH) on PNT2 cells, studying the variation in cell viability, oxidative stress genes, inflammatory mediator and ROS levels. The results reveal that DPPH activated ROS production and release of Prostaglandin E2 in PNT2 at low and intermediate doses, while cells switched from survival to cell death signals at high doses of the oxidising agent. This new in vitro oxidative stress model was validated by using Trolox, &beta, carotene and total extract of the green microalga Testraselmis suecica. Only the T. suecica extract can completely counteract DPPH-induced injury, since its chemical complexity demonstrated a multilevel protecting and neutralising effect against oxidative stress in PNT2.

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.

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.

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.

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 (0.01–50 mg/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 (50 mg/L for 5-flourouracil; 30 mg/L for etoposide and 20 mg/L for capecitabine and imatinib). The results indicate that anticancer drugs can affect embryogenesis mechanisms.

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.