Background: Recent data support the hypothesis that genetic damage occurring early in life during childhood can play an important role in the development of chronic diseases in adulthood, including cancer. Objectives: The objective of this paper, part of the MAPEC_LIFE project, is to describe the frequency of micronuclei and meta-nuclear alterations in exfoliated buccal cells of 6-8year-old Italian children recruited in five Italian towns (i.e., Brescia, Torino, Pisa, Perugia and Lecce) with different air pollution levels. Methods: About 200 children per town were recruited from primary schools. Biological samples were collected twice from the same children, in two different seasons (winter 2014-15 and late spring 2015). Cytogenetic damage was evaluated by the buccal micronucleus cytome assay. Results: Overall,n = 1046 children represent the final cohort of the MAPEC_LIFE study. On the whole, the results showed a higher mean MN frequency in winter (0.42 +/- 0.54%o) than late-spring (0.22 +/- 0.34%o). MN frequency observed among the five Italian towns showed a trend that follows broadly the levels of air pollution in Italy: the highest MN frequency was observed in Brescia during both seasons, the lowest in Lecce (winter) and Perugia (late-spring). Conclusions: To the best of our knowledge, the number of recruited children included in the analysis (n = 1046) is the highest compared to previous studies evaluating the frequency of MN in exfoliated buccal cells so far. MN frequency was associated with winter season and living in towns at various levels of air pollution, suggesting an important role of this exposure in determining early cytogenetic effects.

Introduzione. Attualmente la figura del farmacista di comunità sta subendo profonde trasformazioni. Il farmacista, come operatore sanitario del Servizio Sanitario Nazionale, sta ampliando le sue competenze nell’ambito della sanità pubblica, attraverso interventi di prevenzione delle malattie e di promozione della salute. La ricerca ha avuto come obiettivo quello di valutare le conoscenze, gli atteggiamenti e i comportamenti di un campione selezionato di farmacisti titolari di farmacie private e pubbliche della provincia di Perugia e Terni, nei confronti degli interventi di educazione sanitaria e di promozione della salute effettuati. Metodi. Studio cross-sectional condotto mediante l’utilizzo di due strumenti di rilevazione: un questionario anonimo autocompilato contenente 29 domande; una scheda di valutazione ambientale composta da 21 item, compilata da studenti di Scienze farmaceutiche. Risultati. Hanno aderito al progetto il 70% dei farmacisti umbri, che nel 68,9% dei casi ritiene gli interventi di educazione sanitaria molto rilevanti; il 57% è favorevole alla gratuità di tali interventi, con un monte ore settimanale, dedicato a tale attività, di 14,5. Conclusioni. L’indagine mostra una buona sensibilità dei farmacisti nei confronti delle tematiche di educazione sanitaria. Infatti, il farmacista può e deve avere un ruolo essenziale nel coinvolgere responsabilmente tutti i cittadini nella promozione di nuovi comportamenti nei confronti della salute.

Disinfection with performic acid (PFA) represents an emerging technology in wastewater treatment. Many recent studies indicate its effectiveness and suitability as a disinfectant for different applications; several have demonstrated its reliability as an alternative to chlorine for disinfecting secondary effluents from urban wastewater treatment plants (WWTPs). Some disinfection technologies, in relation to their oxidative power, lead to the formation of disinfection by-products (DBPs), some of which are of concern for their toxic and carcinogenic potential. The aim of this study was to investigate potential genotoxic, cytotoxic, and mutagenic effects of this disinfection agent on treated secondary effluent coming from a municipal WWTP. A strategy with multiple short-term tests and different target cells (bacterial, plant, and mammalian) was adopted to explore a relatively wide range of potential genotoxic events. The Ames test (point mutation in Salmonella), the micronucleus (chromosomal damage) and Comet tests (primary DNA damage) on human hepatic cells (HepG2) were conducted to detect mutagenicity and chromosomal DNA alterations. DNA fragmentation and mitochondrial potential assays were conducted to evaluate apoptosis in the same kinds of cells. Mutagenic and clastogenic effect potentials were evaluated by examining micronucleus formation in Allium cepa root cells. In all the in vitro tests, carried out on both disinfected and non-disinfected effluents, negative results were always obtained for mutagenic and genotoxic effects. In the Allium cepa tests, however, some non-concentrated wastewater samples after PFA treatment induced a slight increase in micronucleus frequencies in root cells, but not in a dose-related manner. In conclusion, PFA applied for disinfection to a secondary effluent from a municipal wastewater treatment plant did not contribute to the release of genotoxic or mutagenic compounds. Further studies are required to establish to which extent these findings can be generalized to support PFA for other disinfection applications.

INTRODUCTION Nowadays, the community pharmacist's role is undergoing profound transformations. As a healthcare provider of the National Health Service, pharmacists are expanding their expertise in Public Health through disease prevention and health promotion programme. In relation to health education and health promotion interventions, this research was aimed to evaluate the knowledge, attitudes and behaviours of a selected sample of private and public pharmacists, working in the Province of Perugia and Terni, Umbria region. METHODS Cross-sectional study conducted using two detection tools: a 29-items self-administered, anonymous questionnaire and a 21-items environmental evaluation sheet, compiled by students of Pharmaceutical Sciences. RESULTS 70% of umbrian pharmacists participated in the project, 68.9% of them consider health education interventions "very important", 57% support the gratuity of these interventions with a 14.5 hours/week dedicated to these activities. CONCLUSIONS The survey shows a good pharmacist sensitivity to the issues of health education. Actually, the pharmacist can and should play an essential role in responsibly involving all citizens in promoting new health behaviours in collaboration with physicians.

Many studies have shown the presence of numerous organic genotoxins and carcinogens in drinking water. These toxic substances derive not only from pollution, but also from the disinfection treatments, particularly when water is obtained from surface sources and then chlorinated. Most of the chlorinated compounds in drinking water are nonvolatile and are difficult to characterize. Thus, it has been proposed to study such complex mixtures using short-term genotoxicity tests predictive of carcinogenic activity. Mutagenicity of water before and after disinfection has mainly been studied by the Salmonella/microsome (Ames test); in vitro genotoxicity tests have also been performed in yeasts and mammalian cells; in situ monitoring of genotoxins has also been performed using complete organisms such as aquatic animals or plants (in vivo). The combination of bioassay data together with results of chemical analyses would give us a more firm basis for the assessment of human health risks related to the consumption of drinking water. Tests with different genetic end-points complement each other with regard to sensitivity toward environmental genotoxins and are useful in detecting low genotoxicity levels which are expected in drinking water samples.

OBJECTIVES:Recently published works showed that occupational exposure to antineoplastic drugs (ANPD) is still frequent in hospital settings, despite significant safety policy improvements. The aim of this study was to assess the current level of occupational exposure to ANPD and any potentially associated cytogenetic damages in hospital nurses routinely handling ANPD.METHODS:Occupationally ANPD-exposed (n = 71) and ANPD-unexposed (n = 77; control) nurses were recruited on a voluntary basis from five hospitals in Northern and Central Italy. Evaluation of surface contamination and dermal exposure to ANPD was assessed by determining cyclophosphamide (CP) on selected surfaces (wipes) and on exposed nurses' clothes (pads). The concentration of unmetabolized CP—as a biomarker of internal dose—was measured in end-shift urine samples. Biomonitoring of genotoxic effects (i.e., biological effect monitoring) was conducted by analyzing micronuclei (MN) and chromosome aberrations (CA) in peripheral blood lymphocytes. Genetic polymorphisms for enzymes involved in metabolic detoxification (i.e., glutathione S-transferases) were analyzed as well.RESULTS:We observed a significant increase in MN frequency (5.30 ± 2.99 and 3.29 ± 1.97; mean values ± standard deviation; p < 0.0001) in exposed nurses versus controls, as well as in CA detection (3.30 ± 2.05 and 1.84 ± 1.67; p < 0.0001), exposed subjects versus controls. Our results provide evidence that, despite safety controlled conditions, ANPD handling still represents a considerable genotoxic risk for occupationally exposed personnel.CONCLUSIONS:Because both MN and CA have been described as being predictive of group-increased cancer risk, our findings point to a need for improving specific safety procedures in handling and administering ANPD.

BACKGROUNDElectric arc welding is known to involve considerable exposure to extremely-low-frequency magnetic fields (ELF-MF; 50 Hz). The aim of the present study was to evaluate individual exposure to ELF-MF during arc welding and to assess the eventually associated genotoxic hazard by evaluating primary DNA damage.METHODSThe study group comprised 21 electric arc welders (exposed) and 21 non-exposed control subjects (healthy blood donors). Occupational exposure to ELF-MF was measured using personal dosimeters worn during one complete work-shift (7 am to 5 pm). The extent of primary DNA damage was measured in peripheral blood leukocytes with the standard procedure of the alkaline comet assay.RESULTSTail length showed to have similar values in welders and controls. Whereas, the data showed a significant decrease for tail intensity (p = 0.01) and tail moment (p = 0.02) counts in exposed subjects compared to controls.CONCLUSIONSThe different results of our present study and published investigations from other research groups reporting positive results in the comet assay might be a result of different chromium and/or nickel (or other metals) exposure levels, which lead to DNA-protein cross-links at lower concentrations and DNA single-strand breakages at higher concentrations. Since these results are derived from a small-scale pilot study, a larger scale study should be undertaken.

The present molecular epidemiology study was carried out to evaluate the genotoxic effects of occupational exposure to antineoplastic drugs (ANP). The study was conducted in 52 hospital workers involved in the preparation, handling or administration of ANP in a hospital in Perugia (central Italy) and in 52 non-exposed control subjects matched for age, gender and smoking habits to the exposed subjects. Both comet assay and the micronucleus test were used to evaluate genome damage in peripheral blood lymphocytes in study subjects. The extent of primary DNA damage, as evaluated by the comet assay, was significantly increased in exposed personnel with respect to matched controls. On the other hand, no significant differences in micronuclei frequency was observed between the two groups. Multivariate linear regression analysis showed an association between years of occupational exposure over 10 years and higher extent of primary DNA damage in the exposed group. The results of this study confirm that handling ANP without appropriate precautions carries a genotoxic risk for exposed healthcare workers. These results address the need for regular biological effect monitoring of staff occupationally-exposed to ANP.

ObjectivesPeople who handle antineoplastic drugs, many of which classified as human carcinogens by International Agency for Research on Cancer, are exposed to low doses in comparison with patients; however, the long duration of exposure could lead to health effects. The aim of this work was to evaluate DNA damage in white blood cells from 63 nurses who handle antineoplastic drugs in five Italian hospitals and 74 control participants, using different versions of the Comet assay.MethodsPrimary DNA damage was assessed by using the alkaline version of the assay on leucocytes, whereas to detect DNA oxidative damage and cryptic lesions specifically, the Comet/ENDO III assay and the Comet/araC assay were performed on leucocytes and lymphocytes, respectively.ResultsIn the present study, no significant DNA damage was correlated with the work shift. The exposed population did not differ significantly from the reference group with respect to DNA primary and oxidative damage in leucocytes. Strikingly, in isolated lymphocytes treated with araC, lower data dispersion as well as a significantly lower mean value for the percentage of DNA in the comet tail was observed in exposed participants as compared with the control group (p<0.05), suggesting a potential chronic exposure to crosslinking antineoplastic drugs.ConclusionsAlthough stringent rules were adopted at national and international levels to prevent occupational exposure to antineoplastic drugs, data reported in this study support the idea that a more efficient survey on long-lasting exposures at very low concentrations is needed.

In tunnel construction, workers exposed to dust from blasting, gases, diesel exhausts, and oil mist have shown higher risk for pulmonary diseases. A clear mechanism to explain how these pollutants determine diseases is lacking, and alveolar epithelium's capacity to ingest inhaled fine particles is not well characterized. The objective of this study was to assess the genotoxic effect exerted by fine particles collected in seven tunnels using the cytokinesis-block micronuclei test in an in vitro model on type II lung epithelium A549 cells. For each tunnel, five fractions with different aerodynamic diameters of particulate matter were collected with a multistage cascade sampler. The human epithelial cell line A549 was exposed to 0.2 m(3)/mL equivalent of particulate for 24 h before testing. The cytotoxic effects of particulate matter on A549 cells were also evaluated in two different viability tests. In order to evaluate the cells' ability to take up fine particles, imaging with transmission electron microscopy of cells after exposure to particulate matter was performed. Particle endocytosis after 24 h exposure was observed as intracellular aggregates of membrane-bound particles. This morphologic evidence did not correspond to an increase in genotoxicity detected by the micronucleus test.

Surface waters are increasingly utilized for drinking water because groundwater sources are often polluted. Several monitoring studies have detected the presence of mutagenicity in drinking water, especially from surface sources due to the reaction of natural organic matter with disinfectant. The study aimed to investigate the genotoxic potential of the products of reaction between humic substances, which are naturally present in surface water, and three disinfectants: chlorine dioxide, sodium hypochlorite and peracetic acid. Commercial humic acids dissolved in distilled water at different total organic carbon (TOC) concentrations were studied in order to simulate natural conditions of both ground water (TOC=2.5 mg/L) and surface water (TOC=7.5 mg/L). These solutions were treated with the biocides at a 1:1 molar ratio of C:disinfectant and tested for genotoxicity using the anaphase chromosomal aberration and micronucleus tests in Allium cepa, and the Vicia faba and Tradescantia micronucleus tests. The tests were carried out after different times and with different modes of exposure, and at 1:1 and 1:10 dilutions of disinfected and undisinfected humic acid solutions. A genotoxic effect was found for sodium hypochlorite in all plant tests, at both TOCs considered, while chlorine dioxide gave positive results only with the A.cepa tests. Some positive effects were also detected for PAA (A.cepa and Tradescantia). No relevant differences were found in samples with different TOC values. The significant increase in all genotoxicity end-points induced by all tested disinfectants indicates that a genotoxic potential is exerted even in the presence of organic substances at similar concentrations to those frequently present in drinking water.

Biological Effect Monitoring in Peripheral Blood Lymphocytes from Subjects Occupationally Exposed to Antineoplastic Drugs: Assessment of Micronuclei Frequency: Milena VILLARINI, et al . Department of Medical‐Surgical Specialties and Public Health (Section of Public Health), University of Perugia, Italy— Objectives Antineoplastic drugs (ANPDs) are widely used in the treatment of cancer and some nonneoplastic diseases. However, most if not all of these chemical agents are generally nonselective and, along with tumor cells, normal cells may undergo cytotoxic/genotoxic damage. Italian pharmacists and nurses occupationally exposed to ANPDs during their normal work routines were monitored to evaluate biological effects ( i.e ., cytogenetic damage) eventually associated with exposure. The subjects were also monitored for primary, oxidative and excision repaired DNA damage as evaluated by comet assay (published data). In the present paper, we present the results obtained with the cytokinesis‐block micronucleus (CBMN) test. Methods The CBMN test in peripheral blood lymphocytes was applied because of its ability to detect both clastogenic and aneugenic effects, and because it has recently been reported that micronuclei (MNs) are predictive of cancer risk in human populations. In this study, the evaluation of MN frequency was carried out using the CBMN test in the absence or in the presence of the DNA repair inhibitor Ara‐C (cytosine arabinoside). Results No significant difference was observed for MN frequency comparing nurses handling ANPDs (exposed subjects) and controls; no correlations were found between job seniority, age, smoking habits and MN rates. Conclusions Concerning the aim of this study to evaluate the genotoxic risk arising from occupational exposure to ANPDs, statistically significant differences in MN rates in the subjects under study could not be determined.

Disinfection of surface drinking water, particularly water chlorination, produces by-products with potential genotoxic and/or carcinogenic activity. A study carried out at a pilot plant for drinking water disinfection of lake water revealed mutagenic activity of three different disinfectants (sodium hypochlorite, chlorine dioxide and peracetic acid) in different seasons using in situ mutagenicity assays, both in animal (micronucleus test) and in plant organisms (anaphase chromosomal aberration and micronucleus tests). The effects of the disinfectants appeared to be modulated by the season of exposure. In this study, we tried to understand if (and to what extent) the temperature parameter could actually play an independent role in the registered seasonal variation of mutagenic effects, neglecting the variation of other parameters, e.g. physical conditions and chemical composition of the lake water. Therefore plants (Allium cepa for chromosomal aberration test and Vicia faba for micronucleus test) were exposed to the same disinfected lake-water samples at different temperatures (10℃, 20℃ and 30℃), according the ones registered during the in situ experiment. Long-term expo- sure at the temperatures of 20℃ (both Vicia faba and Allium cepa) and 30℃ (Vicia faba only) to disinfected waters induced clear mutagenic effects. These results show that temperature is an important variable which should be taken into account when in situ exposure of plants is planned for mutagenicity testing. Also, different plant systems clearly show specific temperature ranges suitable for their growth, thereby indicating the need for an accurate selection of the test organism for a specific experimental plan.

Electric arc welding is known to involve considerable exposure to extremely low-frequency magnetic fields (ELF-MF). A cytogenetic monitoring study was carried out in a group of welders to investigate the genotoxic risk of occupational exposure to ELF-MF. This study assessed individual occupational exposure to ELF-MF using a personal magnetic-field dosimeter, and the cytogenetic effects were examined by comparing micronuclei (MN) and sister chromatid exchange (SCE) frequencies in the lymphocytes of the exposed workers with those of non-exposed control subjects (blood donors) matched for age and smoking habit. Cytogenetic analyses were carried out on 21 workers enrolled from two different welding companies in Central Italy and compared to 21 controls. Some differences between the groups were observed on analysis of SCE and MN, whereas replication indices in the exposed were found not to differ from the controls. In particular, the exposed group showed a significantly higher frequency of MN (group mean±SEM: 6.10±0.39) compared to the control group (4.45±0.30). Moreover, the increase in MN is associated with a proportional increase in ELF-MF exposure levels with a dose-response relationship. A significant decrease in SCE frequency was observed in exposed subjects (3.73±0.21) compared to controls (4.89±0.12). The hypothesis of a correlation between genotoxic assays and ELF-MF exposure value was partially supported, especially as regards MN assay. Since these results are derived from a small-scale pilot study, a larger scale study should be undertaken.

Background: Some industrial hygiene studies have assessed occupational exposure to antineoplastic drugs; other epidemiological investigations have detected various toxicological effects in exposure groups labeled with the job title. In no research has the same population been studied both environmentally and epidemiologically. The protocol of the epidemiological study presented here uses an integrated environmental and biological monitoring approach. The aim is to assess in hospital nurses preparing and/or administering therapy to cancer patients the current level of occupational exposure to antineoplastic drugs, DNA and chromosome damage as cancer predictive effects, and the association between the two.Methods/Design: About 80 healthy non-smoking female nurses, who job it is to prepare or handle antineoplastic drugs, and a reference group of about 80 healthy non-smoking female nurses not occupationally exposed to chemicals will be examined simultaneously in a cross-sectional study. All the workers will be recruited from five hospitals in northern and central Italy after their informed consent has been obtained. Evaluation of surface contamination and dermal exposure to antineoplastic drugs will be assessed by determining cyclophosphamide on selected surfaces (wipes) and on the exposed nurses' clothes (pads). The concentration of unmetabolized cyclophosphamide as a biomarker of internal dose will be measured in end-shift urine samples from exposed nurses.Biomarkers of effect and susceptibility will be assessed in exposed and unexposed nurses: urinary concentration of 8-hydroxy-2-deoxyguanosine; DNA damage detected using the single-cell microgel electrophoresis (comet) assay in peripheral white blood cells; micronuclei and chromosome aberrations in peripheral blood lymphocytes. Genetic polymorphisms for enzymes involved in metabolic detoxification (i.e. glutathione S-transferases) will also be analysed.Using standardized questionnaires, occupational exposure will be determined in exposed nurses only, whereas potential confounders (medicine consumption, lifestyle habits, diet and other non-occupational exposures) will be assessed in both groups of hospital workers.Statistical analysis will be performed to ascertain the association between occupational exposure to antineoplastic drugs and biomarkers of DNA and chromosome damage, after taking into account the effects of individual genetic susceptibility, and the presence of confounding exposures.Discussion: The findings of the study will be useful in updating prevention procedures for handling antineoplastic drugs.

A sensitive and rapid method to evaluate toxic and genotoxic properties of drinking water supplied from Lake Trasimeno (Umbria, Central Italy) was worked out analysing bile in Cyprinus carpio exposed for 20 d to lake water treated with 3 different disinfectants, sodium hypochlorite (NaClO), chlorine dioxide (ClO(2)) and peracetic acid (PAA). Fish were sacrificed at 0, 10 and 20 d in order to investigate the time course of these endpoints. An aliquot of bile samples was fractionated by adsorption on C(18) silica cartridges and the genotoxic potential of whole bile and of bile fractions was evaluated by the single-cell microgel-electrophoresis (comet) assay on human colonic adenocarcinoma cells (Caco-2). Bile (both whole and fractionated) from specimens exposed to the three disinfectants always showed a genotoxic activity as compared to the control group. The results of this study provide evidence that all three disinfectants cause an increase in bile genotoxicity of chronically exposed fish.