Cristina Pignata

Analysis of environmental endocrine disrupting chemicals using the E-screen method and stir bar sorptive extraction in wastewater treatment plant effluents

Endocrine disrupting chemicals (EDCs) have become a major issue in the field of environmental science due to their ability to interfere with the endocrine system. Recent studies show that surface water is contaminated with EDCs, many released from wastewater treatment plants (WWTP). This pilot study used biological (E-screen assay) and chemical (stir bar sorptive extraction-GC-MS) analyses to quantify estrogenic activity in effluent water samples from a municipal WWTP and in water samples of the recipient river, upstream and downstream of the plant. The E-screen assay was performed on samples after solid phase extraction (SPE) to determine total estrogenic activity; the presence of estrogenic substances can be evaluated by measuring the 17-beta-estradiol equivalency quantity (EEQ). Untreated samples were also assayed with an acute toxicity test (Vibrio fischeri) to study the correlation between toxicity and estrogenic disruption activity. Mean EEQs were 4.7 ng/L (+/-2.7 ng/L) upstream and 4.4 ng/L (+/-3.7 ng/L) downstream of the plant, and 11.1 ng/L (+/-11.7 ng/L) in the effluent. In general the WWTP effluent had little impact on estrogenicity nor on the concentration of EDCs in the river water. The samples upstream and downstream of the plant were non-toxic or weakly toxic (0<TU<0.9) while the effluent was weakly toxic or toxic (0.4<TU<7.6). Toxicity and estrogenic activity were not correlated. At most sites, industrial mimics, such as the alkylphenols and phthalates, were present in higher concentrations than natural hormones. Although the concentrations of the detected xenoestrogens were generally higher than those of the steroids, they accounted for only a small fraction of the EEQ because of their low estrogenic potency. The EEQs resulting from the E-screen assay and those calculated from the results of chemical analyses using estradiol equivalency factors were comparable for all samples and closely correlated.

Malondialdehyde-Deoxyguanosine Adduct Formation in Workers of Pathology Wards. The Role of Air Formaldehyde Exposure

Formaldehyde is an ubiquitous pollutant to which humans are exposed. Pathologists can experience high formaldehyde exposure levels. Formaldehyde-among other properties-induce oxidative stress and free radicals, which react with DNA and lipids, leading to oxidative damage and lipid peroxidation, respectively. We measured the levels of air-formaldehyde exposure in a group of Italian pathologists and controls. We analyzed the effect of formaldehyde exposure on leukocyte malondialdehyde-deoxyguanosine adducts (M(1)-dG), a biomarker of oxidative stress and lipid peroxidation. We studied the relationship between air-formaldehyde and M(1)-dG adducts. Air-formaldehyde levels were measured by personal air samplers. M(1)-dG adducts were analyzed by a (32)P-postlabeling assay. Reduction room pathologists were significantly exposed to air-formaldehyde with respect to controls and to the pathologists working in other laboratory areas (p < 0.001). A significant difference for M(1)-dG adducts between exposed pathologists and controls was found (p = 0.045). The effect becomes stronger when the evaluation of air-formaldehyde exposure was based on personal samplers (p = 0.018). Increased M(1)dG adduct levels were only found in individuals exposed to air-formaldehyde concentrations higher than 66 microg/m(3). When the exposed workers and controls were subgrouped according to smoking, M(1)-dG tended to increase in all of the subjects, but a significant association between M(1)-dG and air-formaldehyde was only found in nonsmokers (p = 0.009). Air-formaldehyde played a role positive but not significant (r = 0.355, p = 0.075, Pearson correlation) in the formation of M(1)-dG, only in nonsmokers. Working in the reduction rooms and exposure to air-formaldehyde concentrations higher than 66 microg/m(3) are associated with increased levels of M(1)-dG adducts.

The endocrine disrupting activity of surface waters and of wastewater treatment plant effluents in relation to chlorination

The present study investigated the biological quantification of estrogenic activity in the effluent of a wastewater treatment plant (WWTP) and in the recipient river in north-western Italy. Samples of the WWTP effluent and those of river water upstream and downstream the WWTP were taken from September 2006 to May 2007. The effluent was evaluated in the presence and absence of chlorination. The E-screen assay, with human estrogens receptor-positive MCF-7 BUS breast cancer cells, was performed on samples after solid-phase extraction to determine the total estrogenic activity by measuring the 17beta-estradiol equivalent quantity (EEQ). In order to study the correlation between estrogenicity and toxicity, untreated samples were also assayed with the acute toxicity test Microtox. Furthermore, to determine the efficiency of the chlorination process, all the samples were analysed for disinfection by-products (trihalomethanes) and Escherichia coli. The mean EEQs were 5.0 ng/L (+/-6.1 ng/L) upstream of the plant, 6.7 ng/L (+/-7.4 ng/L) downstream from the plant and 23.3 ng/L (+/-20.4 ng/L) in the WWTP effluent. The difference between upstream and downstream of the treatment plant was not significant. Chlorinated water samples had lower estrogenic activity and E. coli concentrations, but had greater toxicity and higher trihalomethane concentrations. Estrogenic activity was not correlated with toxicity. These results suggest that the WWTP effluent had little impact on the estrogenic activity of the recipient river.

Mutagenic properties of PM2.5 urban pollution in the Northern Italy: the nitro-compounds contribution.

PM2.5 is the breathable fraction of the particulate matter and some adverse health effects, such as respiratory functionality, cardiological diseases and cancer, can be in some measure attributable to this risk factor exposure. Some of the most carcinogen compounds transported by PM2.5 are nitro-compounds. In this study, a strengthened in vitro bioassay--able to predict the mutagenic/carcinogenic activity of the environmental mixtures--was conducted on PM2.5 organic extracts to define the nitro-compounds burden. PM2.5 air pollution was daily monitored, during 2006, in three cities located in the Northern part of Italy (Torino, Pavia and Verona) and the mutagenic properties of the PM2.5 organic extracts were assessed with the Ames test. The bacterial used in this study were three Salmonella typhimurium strains: TA98, nitroreductase-less mutant TA98NR and YG1021 carrying a nitroreductase-producing plasmid. The annual PM2.5 mean level measured in Torino was 46.5 (+/-31.6) microg/m(3), in Pavia 34.8 (+/-25.1) microg/m(3), and in Verona 37.3 (+/-27.8) microg/m(3), while the mutagenicity expressed as TA98 net reverants/m(3) was 28.0 (+/-22.1), 28.3 (+/-24.9), and 34.2 (+/-30.9) respectively. Monthly pool bioassays, conducted with the three different strains, showed a greater mutagenic response of the YG1021 in each city. The relationship among the mutagenic answers for YG1021:TA98:TA98NR was about 6:3:1 (p<0.001). Over nitroreductase activity enhanced the response of 2.2, 2.0 and 1.7 times for Torino, Pavia, and Verona (ANOVA Torino p<0.05) respectively. Without nitroreductase activity the genotoxicity was limited. These biological findings are able to describe a relevant role played by the nitro compounds in the mutagenic properties of the urban PM2.5 in the Padana plain; moreover the bacterial nitroreductase plays a predominant role in DNA interaction primarily for Torino PM2.5 extracts.

Cotinine and N-(2-hydroxyethyl)valine as markers of passive exposure to tobacco smoke in children

Large segments of populations, including children, are exposed to environmental tobacco smoke (ETS), a risk factor for lung cancer and heart, circulatory and respiratory diseases. Recently, ETS was classified as a class A carcinogen by USEPA, as carcinogenic to humans by IARC (group 1) and by the National Toxicology Program of the US National Institutes of Health. Cotinine, a product of the metabolism of nicotine, is measurable in urine and, correlates strictly and directly to ETS exposure, therefore representing a well-known internal dose marker. Another marker of active tobacco smoking is the N-(2-hydroxyethyl) valine (HOEtVal) which results from the reaction between ethylene oxide (EtO) and the N-terminal valine of hemoglobin. The aim of this study was the evaluations of ETS markers, namely urinary cotinine and HOEtVal measured in blood in 100 children with ages ranging between 3 and 13 years. Experimental findings show that cotinine, as a specific internal dose marker, and HOEtVal, as a nonspecific biological effective dose marker, both depend on the passive exposure to ETS as well as on the active habit of smoking.

A review on microbiological decontamination of fresh produce with nonthermal plasma

Food safety is a critical public health issue for consumers and the food industry because microbiological contamination of food causes considerable social and economic burdens on health care. Most foodborne illness comes from animal production, but as of the mid‐1990s in the United States and more recently in the European Union, the contribution of fresh produce to foodborne outbreaks has rapidly increased. Recent studies have suggested that sterilization with nonthermal plasma could be a viable alternative to the traditional methods for the decontamination of heat‐sensitive materials or food because this technique proves capable of eliminating micro‐organisms on surfaces without altering the substrate. In the last 10 years, researchers have used nonthermal plasma in a variety of food inoculated with many bacterial species. All of these experiments were conducted exclusively in a laboratory and, to our knowledge, this technique has not been used in an industrial setting. Thus, the purpose of this review is to understand whether this technology could be used at the industrial level. The latest researches using nonthermal plasma on fresh produce were analysed. These evaluations have focused on the log reduction of micro‐organisms and the treatment time.

Formaldehyde and tobacco smoke as alkylating agents: The formation of N-methylenvaline in pathologists and in plastic laminate workers

OBJECTIVE Aim of this study was to investigate the relationships between the concentration of formaldehyde in air and the alkylation of hemoglobin to form a terminal N-methylenvaline residue in three occupationally exposed groups: a) technicians of pathology wards, b) workers of the plastic laminates industry, and c) a control group. All subjects recruited in this study were also tested on their smoking habits. METHODS Formaldehyde adsorbed on passive air samplers was quantified by HPLC with UV detection (360 nm), cotinine was quantified by GC-MS. Terminal hemoglobin N-methylenvaline was determined by treating globine under reducing conditions with pentafluorophenyl isothiocyanate to yield a derivative, subsequently detected by GC-MS. One-way analysis of variance was performed to compare among the three groups the biomarkers considered in this study. RESULTS For air-FA and N-methylenvaline a difference between the three groups was detected (p < 0.0001) and a significant higher concentration in the two professionally exposed groups was proved. Mean values for FA (μg/m(3)): group a) 188.6, group b) 210.1, and group c) 41.4; mean values for N-methylenvaline (nmol/g of globin): group a) 377.9, group b) 342.8, and group c) 144.8. Conversely, the comparison between the two professionally exposed groups, a) vs b), does not show any significant difference highlighting similar exposition to FA and, consequently, similar biological response. Tobacco smoke proves to have a minor impact on the formation of N-methylenvaline molecular adduct. CONCLUSIONS A positive correlation was demonstrated between professional exposition to air-formaldehyde and hemoglobin alkylation to form N-methylenvaline molecular adduct in two occupationally exposed groups of subjects considered in the present study. In comparison with occupational exposition, tobacco smoke proved to have a minor impact on the formation of N-methylenvaline molecular adduct.

Application of semipermeable membrane device (SPMD) to assess air genotoxicity in an occupational environment.

Semipermeable membrane device (SPMD) is a passive sampler that sequesters lipophilic contaminants, mimicking the bioconcentration in the fatty tissue of organisms. This study was designed to assess the use of SPMD and biological tests (Comet assay and Ames test) for air monitoring. For this purpose an occupational environment with expected polycyclic aromatic hydrocarbons (PAHs) contamination (coke plant) was selected for a case study. The SPMDs were deployed in five occupational contaminated sites and in a control site. The SPMD dialysates were chemically analysed and examined for in vitro DNA-damaging activity in human cells (Jurkat) by Comet assay and for mutagenicity with the Ames test (TA98 strain, w/o S9). Total suspended particulates were also collected and analysed (GC-MS). No biological effect of SPMD extract was revealed in the control site. On the other hand, air samples collected with SPMDs within the coke plant showed variable degrees of genotoxic and mutagenic activity. The highest effects were associated with the highest PAH level recovered in the SPMDs extracts and in particulate samples. Results obtained support the sensitivity of biological tests associated to SPMD sampling for evaluating the health risk of potentially contaminated work environments highlighting the usefulness of SPMDs for environmental air quality monitoring.

Tobacco smoke and formation of N-(2-hydroxyethyl)valine in human hemoglobin

Abstract Human exposure to ethylene oxide (EtO) occurs mainly through inhalation of polluted air in occupational workplaces and/or via tobacco smoke. A significant biochemical reaction of EtO converts the terminal valine of hemoglobin into N-(2-hydroxyethyl)valine (HOEtVal). In the present study, the extent of HOEtVal formation in 360 healthy adults who were not occupationally exposed to EtO was measured with a gas chromatograph/mass spectrometer in the electron-capture negative chemical ionization mode. This parameter was correlated with smoking habits and urinary cotinine concentration, exhibiting a positive relationship between HOEtVal and the number of cigarettes smoked (r 2 = .4416). Urinary cotinine measurements also correlated with HOEtVal and the number of cigarettes smoked. This positive correlation between urinary cotinine and HOEtVal (r 2 = .3893) provides a new perspective on the early stages of carcinogenic processes.

Artificial Turf Football Fields: Environmental and Mutagenicity Assessment

The public has recently raised concerns regarding potential human health and environmental risks associated with tire crumb constituents in the artificial turf of football fields. The aim of the present study was to develop an environmental analysis drawing a comparison between artificial turf football fields and urban areas relative to concentrations of particles (PM10 and PM2.5) and related polycyclic aromatic hydrocarbons (PAHs), aromatic hydrocarbons (BTXs), and mutagenicity of organic extracts from PM10 and PM2.5. No significant differences were found between PM10 concentrations at an urban site and on a turf football field, both during warm and in cold seasons, either with or without on-field activity. PM2.5 concentrations were significantly greater at the urban site in the cold season as was the ratio of PM2.5 to PM10. BTXs were significantly greater at urban sites than on turf football fields on both on warm and cold days. The ratio of toluene to benzene (T/B ratio) was always comparable with that of normal urban conditions. The concentration of PAHs on the monitored football fields was comparable with urban levels during the two different sampling periods, and the contribution of PAHs released from the granular material was negligible. PM10 organic extract mutagenicity for artificial turf football fields was greater, whereas PM2.5 organic extract mutagenicity was lower, compared with the urban site studied. However, both organic extract mutagenicity values were comparable with the organic extract mutagenicity reported in the literature for urban sites. On the basis of environmental monitoring, artificial turf football fields present no more exposure risks than the rest of the city.