Drinking water quality: an in vitro approach for the assessment of cytotoxic and genotoxic load in water sampled along distribution system.

An in vitro approach was performed to assess the quality of drinking water collected at two treatment/distribution networks located near the source (Plant #1) and the mouth of River Po (Plant #2). The water was sampled at different points of each distribution network, before (raw water) and after the chlorine dioxide disinfection, and in two points of the pipeline system to evaluate the influence of the distribution system on the amount and quality of the disinfection by-product. Cytotoxicity and genotoxicity of water extracts were evaluated in human peripheral lymphocytes and Hep-G2 cells by the use of the micronucleus (MN) test and Comet assay.

Different effects of PCB101, PCB118, PCB138 and PCB153 alone or mixed in MCF-7 breast cancer cells.

Polychlorinated biphenyls (PCBs) are ubiquitous, persistent environmental contaminants that can be a potential health hazard. In the present study we analyzed the potential estrogenic effect in MCF-7 cells of four biologically relevant PCB congeners, alone or in mixtures, present in dairy products, vegetable oil and fish: PCB101, PCB118, PCB138 and PCB153. The mixture of four PCB was tested at seven different concentrations.

In vitro cytotoxicity and genotoxicity of chlorinated drinking waters sampled along the distribution system of two municipal networks.

When chlorine is used as a disinfectant for drinking water it may react with organic materials present in or released by the water pipes and thus form by-products that may represent a genotoxic hazard. The aim of this study was to assess the potential genotoxicity and cytotoxicity of extracts of chlorinated drinking water supplied by local aquifers of two Italian towns, Plants 1 and 2, located in the sub-Alpine area and on the Po plain, respectively. The raw water fell within the legal limits with regards to its chemical and physical properties.

Toxicity evaluation of surface water treated with different disinfectants in HepG2 cells.

It is well known that water disinfection through chlorination causes the formation of a mixture of disinfection by-products (DBPs), many of which are genotoxic and carcinogenic. To demonstrate the formation of such compounds, a pilot water plant supplied with water from Lake Trasimeno was set up at the waterworks of Castiglione del Lago (PG, Italy). The disinfectants, continuously added to pre-filtered lake water flowing into three different basins, were sodium hypochlorite, chlorine dioxide and peracetic acid, an alternative disinfectant used until now for disinfecting waste waters, but not yet studied for a possible use in drinking water treatment.

Estrogenic effect of procymidone through activation of MAPK in MCF-7 breast carcinoma cell line.

Procymidone modifies sexual differentiation in vitro and induces estrogenic activity in primary cultured rainbow trout hepatocytes, as shown by an increase in the contents of vitellogenin and heat shock proteins. Since this dicarboximide fungicide is found in human tissues, it was considered of interest to investigate its ability to induce endocrine damage in the MCF-7 human cell line. The mechanism of this estrogenic action was also evaluated.

Study of potential toxic effects on rainbow trout hepatocytes of surface water treated with chlorine or alternative disinfectants.

This study evaluates the effects of disinfection on the formation of toxic compounds in lake water treated with sodium hypochlorite, chlorine dioxide and peracetic acid (a disinfectant not previously used in drinking water processes). Chlorine reacts with the natural organic matter or contaminants in surface waters and produces a complex mixture of disinfection by-products (DBPs), some of which have been shown to be carcinogenic, mutagenic and/or teratogenic in animal studies. To define the potential toxicity on aquatic animals, disinfected drinking waters obtained from a pilot plant fed with water coming from Lake Trasimeno (Perugia) were collected, adsorbed by using silica C18 cartridges, and then eluted in sequence with ethylacetate, dichloromethane and methanol.

Molecular mechanism of the aryl hydrocarbon receptor activation by the fungicide iprodione in rainbow trout (Oncorhynchus mykiss) hepatocytes.

The dicarboximide fungicide iprodione (Ip) causes oxidative damage as a result of the production of free oxygen radicals, and induces cytochrome P4501A3 (CYP1A3) in cultured rainbow trout hepatocytes. The aim of this study was to characterise some of the molecular mechanisms by means of which Ip activates the aryl hydrocarbon receptor (AhR) and subsequently induces the CYP1A3 gene in rainbow trout (Oncorhynchus mykiss). The study was performed using primary hepatocytes and transfected HepG2 cells with a reporter construct, in which luciferase gene expression is under the transcriptional control of a multimerised xenobiotic response elements (4XREs), or a 2.

Estrogenic activity of procymidone in rainbow trout (Oncorhynchus mykiss) hepatocytes: a possible mechanism of action.

It is known that procymidone modifies sexual differentiation in vivo and in vitro, and that it induces vitellogenin (Vtg) synthesis in primary cultured rainbow trout hepatocytes. The aim of this study was to evaluate the mechanism underlying this latter in vitro estrogenic action. The cells were treated for 24 h with procymidone 150 microM (with 17beta-estradiol [E2] 20 microM as a positive control) combined with an estrogen receptor (ER) antagonist (tamoxifen 20 microM or ICI 182,780 1 microM) or, given the drug toxic action on the production of reactive oxygen species (ROS), a free radical scavenger (alpha-tocopherol 30 microM).

[Evaluation of the effects following low doses of inorganic mercury from environmental and occupational exposures].

Aims: This paper reviews the studies, both in vivo and in vitro, carried out for the project on low-dose effects of inorganic mercury, financed by the Italian Ministry of Universities and Scientific and Technological Research.

Results, Comments And Proposal: The results offer both innovative aspects and potential practical applications. Particular attention is drawn to the reliability of biomarkers of exposure [mercury in urine (HgU) and blood (HgB), possibility of speciation] as well as to the availability of guidance values for risk assessment (reference value, action level, biological threshold value).

Early oxidative damage in primary cultured trout hepatocytes: a time course study.

The aim of this study was to evaluate the influence of the two-step hepatocyte isolation procedure on primary cultured trout (Oncorhynchus mykiss) hepatocytes over time. We characterised the possible changes of a variety of some cellular parameters within the first 24-48 h after seeding. We followed the time dependent changes of these parameters during subsequent culture times in order to see if the cells maintained a differentiated status.

Estrogenic activity of procymidone in primary cultured rainbow trout hepatocytes (Oncorhynchus mykiss).

It has been shown that procymidone, a dicarboximide fungicide, alters sexual differentiation in vivo and in vitro. The aim of this study was to evaluate the estrogenic activity of this fungicide using the synthesis of vitellogenin (Vtg) in rainbow trout hepatocyte as a biological marker. The cells were treated for 24 h with procymidone 150 microM, using 17beta-estradiol 20 microM as a positive control.

Response of rainbow trout (Oncorhynchus mykiss) D-11 cell line to 3-methylcholanthrene (3MC) exposure.

The rainbow trout cytochrome P4501A gene subfamily consists of two members, CYP1A1 and CYP1A3, which are induced by polycyclic aromatic hydrocarbons (PAHs). In this study, we investigated the induction of cytochrome P4501A3 in the rainbow trout (Onchorhynchus mykiss) D-11 cell line after 3-methylcholanthrene (3MC) exposure by generating chimeric constructs in which a 2.3 kb fragment or portion of the 5'-flanking region of the trout cytochrome CYP1A3 gene was fused to the firefly luciferase (Luc) gene.

Effect of iprodione, a dicarboximide fungicide, on primary cultured rainbow trout (Oncorhynchus mykiss) hepatocytes.

As is known from literature, iprodione, a dicarboximide fungicide, has a highly specific action, with a capacity to cause oxidative damage through production of free oxygen radicals (ROS), but it does not appear to be species selective. Since this substance is able to diffuse in water, evaluation of its capacity to induce oxidative damage in an aquatic organism such as the rainbow trout (Oncorhynchus mykiss) was considered of particular interest. A study was, therefore, undertaken to investigate the effect of iprodione on free radicals (ROS) and malondialdehyde (MDA) production, reduced glutathione (GSH) content and catalase activity (CAT), in primary cultured trout hepatocytes, following treatment with 0.

Stimulation of in vitro rat hepatocyte proliferation by conditioned medium obtained from an immortalized macrophage cell line.

The hepatomitogenic effect of conditioned medium (CDM), obtained from the N-11 mouse macrophage cell line was analysed in rat hepatocyte primary cultures. CDM concentrations from 0.01% to 100% were used and the stimulating action in terms of mitotic index (MI) was evaluated.

Hyperbaric oxygen increases plasma exudation in rat trachea: involvement of nitric oxide.

This study investigates the microvascular permeability changes in tracheal tissue of rats exposed to hyperbaric oxygen (HBO). Rats, following exposure to HBO or ambient air (control animals) for 1.5, 3 and 6 h, were prepared for recording of nitric oxide exhaled (FENO) in air using a chemiluminescence analyser.

Adaptation to oxidative stress: effects of vinclozolin and iprodione on the HepG2 cell line.

It is well known that the dicarboximide fungicides, vinclozolin and iprodione, induce lipid peroxidation by means of oxygen activation in fungi, but their action on mammalian cells is not yet clear. We therefore investigated the effect of 1- and 24-h treatments with vinclozolin at concentrations of 25, 50, 100 microg/ml and iprodione at concentration of 62.5, 125, 250 microg/ml on malonaldehyde and free radical production and on reduced glutathione levels in the human HepG2 hepatoma cell line.

Carbendazim and n-butylisocyanate: metabolites responsible for benomyl double action on cytochrome P450 in HepG2 cells.

Changes in the cytochrome P450 monooxygenase system were investigated in HepG2 cells treated for 24 h with 1.25, 2.5, 5, 10 and 20 microg/ml of carbendazim (MBC) and n-butylisocyanate (BIC), the principal benomyl metabolites.

Influence of acetylcysteine on aggravation of ischemic damage in ex vivo hearts of rats exposed to hyperbaric oxygen.

Rats were exposed to hyperbaric oxygen (HBO = 100% oxygen; 2.5 atmospheres absolute pressure) for 6 h. Isovolumic left heart preparations from these animals were subjected to global low flow-ischemia (perfusion rate from 12 ml/min to 2 ml/min for 40 min) and reperfusion.

Hyperbaric oxygen worsens myocardial low flow ischemia-reperfusion injury in isolated rat heart.

In these experiments rats were exposed to hyperbaric oxygen (100% oxygen; 2.5 atmospheres absolute pressure) for 1, 3 or 6 h. At the end of these periods the hearts were removed and subjected to low flow ischemia (perfusion rate from 12 ml/min to 2 ml/min for 40 min) and reperfusion.

Benomyl affects the microtubule cytoskeleton and the glutathione level of mammalian primary cultured hepatocytes.

Rat primary hepatocyte cultures have been used to study the effect of Benomyl alone or in combination with Pirimiphos-methyl. The results presented demonstrate that Benomyl alone is responsible for the microtubular disorganization in both a time- and dose-dependent manner, that the effect is reversible after the agent is removed, and that Benomyl is a potent glutathione-depleting agent. Pirimiphos-methyl, alone or combined with Benomyl had no effect on microtubule organization, but reinforced the decrease in glutathione.

Interleukin-1 production after treatment with non-ionic surfactants in a murine keratinocytes cell line.

Detergents are well known irritating agents in human as well as in animal models. Using a murine keratinocyte cell line (HEL30) changes in the interleukin-1alpha profile were characterized in response to three non-ionic detergents, all widely used in the cosmetics industry. The compounds used in this study were the most active (dodoxynol-9, Delta-9), moderate (polyglyceryl-4-lauryl ether, PEL) and mild (PEG-20-glyceryl ricinoleate + ricinoleamide DEA, PEG) in inducing cytotoxicity, measured as lactate dehydrogenase leakage and de novo protein synthesis, on the same cell line after 2 hr of treatment.

Increase of micronucleus frequency in cultured rat hepatocytes treated in vitro with benomyl and pirimiphos-methyl separately and in mixture.

The pesticides benomyl, a benzimidazole fungicide, and pirimiphos-methyl, an organophosphorus insecticide, were tested separately and in combination at a ratio of 6:1, a mixture frequently found in foodstuffs by residual analysis, to determine their possible genotoxic action. The effect was measured by the micronucleus test carried out on cultured rat hepatocytes stimulated to proliferate by epidermal growth factor (EGF). Adult rat hepatocytes were exposed in vitro for 48 h to the substances at increasing non-cytotoxic doses, chosen on the basis of cytotoxicity tests such as LDH and Neutral red assays.

Genotoxicity assay of five pesticides and their mixtures in Saccharomyces cerevisiae D7.

Four organophosphorus pesticides (azinphos-methyl, diazinone, dimethoate, and pirimiphos-methyl), and one carbamate (benomyl) were tested for cytotoxicity, reverse mutation and gene conversion in Saccharomyces cerevisiae D7, with and without the S9 metabolic system. Furthermore, two mixtures of the above compounds, namely benomyl + pirimiphos-methyl (6/1 ratio) and dimethoate + diazinone + azinphos-methyl (10/4/6 ratio) were tested in the same experimental model. Azinphos-methyl, benomyl, and pirimiphos-methyl alone did not induce any genotoxic effect, whereas azinphos-methyl and diazinone were active in inducing reversion and gene conversion.

Intraocular and plasma kinetics of tenoxicam in rabbits.

Non-steroidal anti-inflammatory drugs (NSAID) represent potentially useful agents in the treatment of a number of ocular pathologies, but their intraocular penetration and distribution have not yet been reported. With the aim of clarifying this point, we evaluated the concentrations of the well known NSAID, tenoxicam, in the aqueous and vitreous humors of rabbits treated i.m.