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Macro and trace elements are important regulators of biological processes, including those ones linked to reproduction. Among them, Ca, Cu, Fe, K, Mg, Mn, Na, Se, and Zn ensure normal spermatic functions. Hence, the aim of this study was to evaluate the concentrations of 26 macro and trace elements (Al, As, Ba, Be, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Li, Mg, Mn, Na, Ni, Pb, Rb, Sb, Se, Sn, Sr, U, V, and Zn) in blood serum and also in semen of healthy young men, homogeneous for age, anthropometric characteristics, and lifestyle, living in three highly polluted areas in Italy. Furthermore, a comparison among three geographical areas was performed to highlight any difference in the investigated parameters and, overall, to speculate any correlations between chemical elements and semen quality. Statistically significant differences (p < 0.05) among the three areas were found for each investigated element, in both semen and serum samples, where inter-area differences were more evident in semen than in blood serum, suggesting human semen as an early environmental marker. Considering the homogeneity of three cohorts, these differences could be due more to environmental conditions in the recruiting areas, suggesting that variations in those involved in reproductive-associated pathways can have an impact on male fertility. Nevertheless, more research is needed to evaluate threshold values for sperm dysfunction and male infertility. Actually, the role of different dietary intake and environmental exposure underlying the observed differences in the recruiting areas is under further investigation for the same cohort.

Air pollution has well-known harmful effects on human beings, causing both acute and chronic diseases. Some data suggest that genetic damage occurring early in life may influence the risk of having cancer and other chronic diseases in adulthood. Therefore, there is a growing interest in studying the genotoxic activity of air pollution, and especially particulate matter. The aim of this study was to analyze airborne particulate matter (PM10) collected in an industrialized town of Northern Italy. PM10 was sampled in six areas of the town, divided in three distinct dimensional classes (

The aim of this paper was to investigate the relationship between micronuclei and DNA damage in children’s buccal mucosa cells and the genotoxicity and mutagenicity of the different sized fractions of particulate matter as well as the concentration of PAHs (polycyclic aromatic hydrocarbons) and metals in particulate matter. Air particulate matter was collected by high volume samplers located near the schools attended by the children on the same days of biological samplings. The mutagenic activity was assessed in different cells in in vitro tests (Ames test on bacteria and comet test on leukocytes). Our study showed weak positive correlations between (a) the mutagenicity of the PM0.5 fraction and PAHs and (b) the micronuclei test of children’s buccal cells and PAHs detected in PM0.5 and PM0.5–3 fractions. A positive correlation was also found between in vitro comet test on leukocytes and PAHs in the PM3–10 fraction. No correlation was observed for metal concentrations in each PM fraction.

The control of the fungal contamination on crops is considered a priority by the sanitary authorities of an increasing number of countries, and this is also due to the fact that the geographic areas interested in mycotoxin outbreaks are widening. Among the different pre- and post-harvest strategies that may be applied to prevent fungal and/or aflatoxin contamination, fungicides still play a prominent role; however, despite of countless efforts, to date the problem of food and feed contamination remains unsolved, since the essential factors that affect aflatoxins production are various and hardly to handle as a whole. In this scenario, the exploitation of bioactive natural sources to obtain new agents presenting novel mechanisms of action may represent a successful strategy to minimize, at the same time, aflatoxin contamination and the use of toxic pesticides. The Aflatox® Project was aimed at the development of new-generation inhibitors of aflatoxigenic Aspergillus spp. proliferation and toxin production, through the modification of naturally occurring molecules: a panel of 177 compounds, belonging to the thiosemicarbazones class, have been synthesized and screened for their antifungal and anti-aflatoxigenic potential. The most effective compounds, selected as the best candidates as aflatoxin containment agents, were also evaluated in terms of cytotoxicity, genotoxicity and epi-genotoxicity to exclude potential harmful effect on the human health, the plants on which fungi grow and the whole ecosystem.

The control of the fungal contamination on crops is considered a priority by the sanitary authorities of an increasing number of countries, and this is also due to the fact that the geographic areas interested in mycotoxin outbreaks are widening. Among the different pre- and post-harvest strategies that may be applied to prevent fungal and/or aflatoxin contamination, fungicides still play a prominent role, however, despite of countless efforts, to date the problem of food and feed contamination remains unsolved, since the essential factors that affect aflatoxins production are various and hardly to handle as a whole. In this scenario, the exploitation of bioactive natural sources to obtain new agents presenting novel mechanisms of action may represent a successful strategy to minimize, at the same time, aflatoxin contamination and the use of toxic pesticides. The Aflatox® Project was aimed at the development of new-generation inhibitors of aflatoxigenic Aspergillus spp. proliferation and toxin production, through the modification of naturally occurring molecules: a panel of 177 compounds, belonging to the thiosemicarbazones class, have been synthesized and screened for their antifungal and anti-aflatoxigenic potential. The most effective compounds, selected as the best candidates as aflatoxin containment agents, were also evaluated in terms of cytotoxicity, genotoxicity and epi-genotoxicity to exclude potential harmful effect on the human health, the plants on which fungi grow and the whole ecosystem.

Background: Human semen quality is affected by lifestyle and environmental factors. Objective: To evaluate the short-term effects of a diet and physical activity intervention on semen quality of healthy young men living in highly polluted areas of Italy. Design, setting, and participants: A randomized controlled trial was conducted. Healthy young men were assigned to an intervention or a control group. Intervention: A 4-mo Mediterranean diet and moderate physical activity program. Outcome measurements and statistical analysis: The primary outcomes were sperm concentration, motility and morphology, concentration of round cells, and semen total antioxidant capacity. Secondary outcomes were adherence to Mediterranean diet and physical activity. All outcomes were measured twice, at the enrollment (t0) and at the end of the intervention (t4). Results and limitations: A total of 263 individuals attended all visits, and underwent examinations and laboratory analyses: 137 in the intervention group and 126 in the control group. The adherence to Mediterranean diet and physical activity level increased more in the intervention group than in the control group from t0 to t4. Sperm concentration, total and progressive motility, and proportion of normal morphology cells increased in the intervention group but decreased in the control group, with statistically significant differences between the two groups at t4. The total antioxidant capacity increased in the intervention group but decreased in the control group, from t0 to t4. Conclusions: Study results showed that an intervention based on Mediterranean diet and regular physical activity can determine an improvement of semen quality in healthy young men. Patient summary: Our study aimed to evaluate the effect of a lifestyle intervention on semen quality of healthy young men. We assigned the 263 enrolled individuals to an intervention or a control group. The intervention group followed a 4-mo Mediterranean diet and moderate physical activity program, at the end of which the participants showed an improvement of semen quality parameters. Our 4-mo lifestyle intervention with a randomized study design showed that an intervention based on Mediterranean diet and regular physical activity can determine an increase of sperm concentration, total and progressive motility, and proportion of spermatozoa with normal morphology, that is, an increase in total antioxidant capacity in healthy young men. These findings underline the effect of lifestyle factors, particularly dietary habits and physical activity, on sperm quality, and show that moderate lifestyle changes recommended for health promotion and chronic disease prevention can also be helpful for preventing male infertility.

Background Human semen quality is affected by metabolic, lifestyle and environmental factors. We planned a randomized controlled trial to evaluate the effects of lifestyle changes on semen quality in healthy young men living in Italian highly polluted areas. We present here some preliminary baseline data on the association between lifestyle and semen quality. Methods 150 non-smokers, non-alcohol drinkers and non-obese young healthy men, aged 18-22 years, have been assigned to an intervention or control group according to a 1:1 randomization plan. The intervention group follows a 4-month nutritional pathway based on a Mediterranean dietary pattern and receives suggestions on practice of physical activity. All subjects undergo urologic examination, measurement of weight, height and abdominal circumference, an interview on lifestyle variables, and provide blood and semen samples in fasting conditions, at the enrollment and at the end of the intervention and follow-up (after 8 months). Adherence to Mediterranean diet is assessed using the “PREvención con DIeta MEDiterránea” (PREDIMED) tool, and physical activity using the “International Physical Activity Questionnaire” (IPAQ). Automated sperm analysis for all semen parameters is performed on the SQA-V GOLD. Results The data of 101 subjects living in a highly industrialized area in North Italy (mean±SD: age: 20.3±1.2 years, BMI: 22.3±2.2) were analyzed. The sperm cell concentration was higher in subjects with IPAQ score > median than those with lower values (68.3±48.6 vs 63.7±47.0 106/mL). Total percent motility was higher in subjects with abdominal circumference < median (50.7±13.6 vs 41.2±18.8, p = 0.004), in subjects with IPAQ score > median (48.2±16.8 vs 44.2±16.8), and in subjects with PREDIMED score > 7 (47.6±18.8 vs 45.0±15.0). Conclusions These preliminary data suggest that semen quality is associated with abdominal fat and physical activity in healthy young men. Key messages The semen quality is a sensitive marker of the health status and is predictive of disease occurrence. Our study suggests that semen quality is associated with lifestyle aspects in healthy young men.

Aflatoxins are secondary fungal metabolites that can contaminate feed and food. They are a cause of growing concern worldwide, because they are potent carcinogenic agents. Thiosemicarbazones are molecules that possess interesting antiaflatoxigenic properties, but in order to use them as crop-protective agents, their cytotoxic and genotoxic profiles must first be assessed. In this paper, a group of thiosemicarbazones and a copper complex are reported as compounds able to antagonize aflatoxin biosynthesis, fungal growth, and sclerotia biogenesis in Aspergillus flavus. The two most interesting thiosemicarbazones found were noncytotoxic on several cell lines (CRL1790, Hs27, HFL1, and U937), and therefore, they were submitted to additional analysis of mutagenicity and genotoxicity on bacteria, plants, and human cells. No mutagenic activity was observed in bacteria, whereas genotoxic activity was revealed by the Alkaline Comet Assay on U937 cells and by the test of chromosomal aberrations in Allium cepa.

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‰) than late-spring (0.22 ± 0.34‰). 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.

Urban air contains many mutagenic pollutants. This research aimed to investigate the presence of mutagens in the air by short-term mutagenicity tests using bacteria, human cells and plants. Inflorescences of Tradescantia were exposed to air in situ for 6h, once a month from January to May, to monitor volatile compounds and micronuclei frequency was computed. On the same days PM10 was collected continuously for 24h. Half of each filter was extracted with organic solvents and studied by means of the Ames test, using Salmonella typhimurium TA98 and TA100 strains, and the comet assay on human leukocytes. A quarter of each filter was extracted with distilled water in which Tradescantia was exposed. PM10 concentration was particularly high in the winter season (> 50 μg/m(3)). In situ exposure of inflorescences to urban air induced a significant increase in micronuclei frequency at all the sites considered, but only in January (p < 0.01). Aqueous extracts collected in January and February induced genotoxic effects in Tradescantia exposed in the laboratory (p < 0.01). Ames test showed that organic extracts of winter urban air were able to induce genetic mutations in S. typhimurium TA98 strain (± S9), but not in TA100 strain, with a revertants/plate number nine times higher than the negative control. Comet assay showed that winter extracts were more toxic and genotoxic than spring extracts. All the mutagenicity tests performed confirmed that urban air in North Italy in winter contains both volatile and non-volatile genotoxic substances able to induce genetic damage in bacteria, human cells and plants.

The issue of food contamination by aflatoxins presently constitutes a social emergency, since they represent a severe risk for human and animal health. On the other hand, the use of pesticides has to be contained, since this generates long term residues in food and in the environment. Here we present the synthesis of a series of chelating ligands based on the thiosemicarbazone scaffold, to be evaluated for their antifungal and antiaflatoxigenic effects. Starting from molecules of natural origin of known antifungal properties, we introduced the thio- group and then the corresponding copper complexes were synthesised. Some molecules highlighted aflatoxin inhibition in the range 67–92% at 100 μM. The most active compounds were evaluated for their cytotoxic effects on human cells. While all the copper complexes showed high cytotoxicity in the micromolar range, one of the ligand has no effect on cell proliferation. This hit was chosen for further analysis of mutagenicity and genotoxicity on bacteria, plants and human cells. Analysis of the data underlined the importance of the safety profile evaluation for hit compounds to be developed as crop-protective agents and at the same time that the thiosemicarbazone scaffold represents a good starting point for the development of aflatoxigenic inhibitors.

In 2015 a IARC Working Group upgraded the classification of PCBs to Group 1 “Carcinogenic to humans”, also on the basis of evidence from epidemiological studies showing an excess risk for melanoma. Increased risks for non-Hodgkin lymphoma (NHL) and breast cancer were also reported though the evidence was limited. However, some recent reviews of studies on PCB exposure and risk of cancer provided discrepant findings. Therefore, we re-evaluated the association between exposure to PCBs and risk of melanoma and NHL by a systematic review and meta-analysis. We retrieved 11 independent cohort studies on occupationally exposed workers. About half of them showed increased standardized mortality or incidence ratios (SMRs or SIRs) for melanoma and none for NHL. The pooled SMRs were 1.32 (95% CI: 1.05–1.64) for melanoma and 0.94 (0.73–1.23) for NHL. Among population-based cohort and case-control studies with individual measures of PCB exposure, one only study was carried out on PCB exposure and melanoma, showing an odds ratio (OR) of 6.0 (2.0–18.2) for the highest compared to lowest quartile of PCB distribution. 13 cohort and case-control studies evaluated the association between NHL and PCB concentration in blood or subcutaneous fat, with summary OR = 1.5 (1.1–1.7) for the highest vs lowest quantile of PCB distribution. However, two cohort studies on people intoxicated by rice oil containing PCBs found no excess of deaths for skin cancer and inconsistent results for NHL. In conclusion, these findings do not provide a strong evidence that PCB exposure can increase the risk of melanoma and NHL in humans.

Polychlorinated biphenyls (PCBs) are persistent organic pollutants produced until the 1980s, which they are still present worldwide. They have been associated with metabolic and endocrine diseases and hypertension in humans, but definite evidence is lacking. A chemical factory producing PCBs caused a heavy pollution in an urban area in Northern Italy. We aimed to evaluate present PCBs serum levels according to demographic and lifestyle variables and their associations with endocrine and metabolic diseases and hypertension in the resident general population. A random sample of 816 adults aged 20–79 years (mean ± SD: 49.1 ± 16.5 years) was enrolled in a cross-sectional population-based study. The participants provided a fasting blood sample for laboratory analysis and were face-to-face interviewed about the presence of chronic diseases. The serum level of total PCBs was computed as the sum of 33 PCB congeners. The median serum level of lipid-adjusted total PCBs was 435.2 and 95th centile was 2154.9 ng/g lipid. Medium and high chlorinated PCBs with immunotoxic and endocrine disrupting activity contributed most to total PCB serum levels, particularly PCBs 138, 153, 170, 180 and 194. The serum levels of total PCBs and of PCB functional groups were positively associated with age and negatively with female gender, education, smoking habit and BMI, and not associated with serum levels of thyroid hormones and TSH and glycaemia and with presence of endocrine diseases, diabetes and hypertension by multivariable analysis. Subgroup analyses according to gender, age and BMI provided similar results. In conclusion, this study shows a long-term persistence of past exposure to PCBs and does not support the hypothesis of an association between PCB exposure and prevalence of endocrine and metabolic diseases and hypertension.

Background Polychlorinated biphenyls (PCBs) have been found to be associated with diabetes in some, but not all, studies performed so far. The aim of this study was to assess the association between PCB serum levels and glycaemia and diabetes in people living in Brescia, a highly industrialised PCB-polluted town in Northern Italy. Design and Methods 527 subjects were enrolled in a cross-sectional population-based study: they were interviewed face-to-face in 2003 and also provided a blood sample under fasting conditions. The concentration of 24 PCB congeners was determined using gas-chromatography (GC/MS). Subsequently, all subjects were included in a follow-up (cohort) study. According to the Local Health Authority health-care database, subjects were considered to be diabetic if they had diabetes at interview time (prevalent cases) or during a 7-year follow-up (incident cases). Results A total of 53 subjects (10.0%) were diabetics: 28 had diabetes at enrolment and other 25 developed the disease subsequently. Diabetes frequency increased according to the serum concentrations of total PCBs and single PCB congeners, but no association was found when estimates were adjusted for education, body mass index, age and gender by logistic regression analysis. Accordingly, glycaemia increased with PCB serum levels, but no association was observed when multiple regression analysis, including confounding factors, was performed. Conclusions This study does not support the hypothesis that PCB environmental exposure is strictly associated with diabetes or glycaemia.

Brescia (North Italy) is located in a highly industrialized area with diffuse environmental pollution by polychlorinated biphenyls (PCBs) and with a high incidence of hepatocellular carcinoma (HCC). We aimed to investigate the concentration of PCBs congeners in serum, liver and adipose tissue of patients with HCC living in this area. We enrolled prospectively 101 HCC incident cases. Lipid-adjusted PCBs concentration of single congeners and total PCBs concentration, computed as the sum of 24 PCBs congeners, were measured in the serum of all subjects and in the liver and abdominal fat of 59 HCC cases undergoing surgery. A high correlation was found between PCBs serum and liver (Spearman r = 0.79), serum and fat ( r = 0.91), and liver and fat ( r = 0.75) concentrations. Serum PCBs concentration in HCC patients did not vary according to demographical, clinical or epidemiological variables, apart from age, as PCBs concentration increased linearly with the subject’s age. This study confirms that serum lipid-adjusted PCBs concentration is a valid measure of PCBs body storage and is highly correlated with PCBs concentration detected in other biological samples.

Aflatoxins represent a serious problem for a food economy based on cereal cultivations used to fodder animal and for human nutrition. The aims of our work are two-fold: first, to perform an evaluation of the activity of newly synthesized thiosemicarbazone compounds as antifungal and anti-mycotoxin agents and, second, to conduct studies on the toxic and genotoxic hazard potentials with a battery of tests with different endpoints. In this paper we report an initial study on two molecules: S-4-isopropenylcyclohexen-1-carbaldehydethiosemicarbazone and its metal complex, bis(S-4-isopropenylcyclohexen-1-carbaldehydethiosemicarbazonato)nickel (II). The outcome of the assays on fungi growth and aflatoxin production inhibition show that both molecules possess good antifungal activities, without inducing mutagenic effects on bacteria. From the assays to ascertain that the compounds have no adverse effects on human cells, we have found that they are cytotoxic and, in the case of the nickel compound, they also present genotoxic effects.

The prevalence of obesity among Italian children has reached such alarming levels as to require detailed studies of the causes of the phenomenon. A cross-sectional study was carried out in order to assess the weight status of 1164 Italian children aged 6–8 years (the Monitoring Air Pollution Effects on Children for Supporting Public Health Policy (MAPEC_LIFE) cohort) and to identify any associations between selected socio-economic and environmental factors and overweight/obesity. The data were obtained by means of a questionnaire given to parents, and any associations were examined by binomial logistic regression analyses. Overweight was found to be positively associated with male gender, parents of non-Italian origin, and parents who smoke, and negatively associated with the parents’ level of education and employment. In addition, the frequency of overweight varied in relation to the geographical area of residence, with a greater prevalence of overweight children in the cities of central-southern Italy. This study highlights the need to implement appropriate obesity prevention programs in Italy, which should include educational measures concerning lifestyle for parents from the earliest stages of their child’s life.

The aim of the MAPEC_LIFE (Monitoring Air Pollution Effects on Children for Supporting Public Health Policy) study is to evaluate the associations between the concentrations of air pollutants and early biological effects in children living in five Italian towns (Brescia, Torino, Lecce, Perugia and Pisa) characterised by varying levels of air pollution. This paper presents the results of micronucleus cytome assays performed on the oral mucosa cells of subjects living in Lecce (Puglia, Italy) and their relationship to factors associated with indoor/outdoor exposure and lifestyles. The study was conducted on 6-8-year-old schoolchildren living in Lecce. The micronucleus cytome assay was performed on exfoliated buccal cells collected from the oral mucosa of children using a soft-bristled toothbrush. Micronuclei were evaluated only in normal differentiated cells. Overall, 43.0% of the samples tested were positive, with an average frequency of 0.28 MN/1000 differentiated cells. Data analysis shows positive associations between the frequency of MN in the children’s buccal mucosa cells and obesity, heavy traffic and smoking mothers, while outdoor sports seem to have the opposite effect. These data will be integrated with data from the other cities involved in the MAPEC_LIFE study and could be used

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.

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.

Background Vallecamonica-Sebino is a community in Northern Italy (99,776 inhabitants) with one of the highest mortality rates for primary liver cancer and cirrhosis in Italy, and voluntary screening for HCV and HBV is widespread. The aim of this study was to estimate the prevalence of chronic liver diseases and their aetiology in the area. Methods We used the following sources of data, linked at an individual level: (1) hospital discharge data; (2) local Viral Hepatitis Services; (3) tests for anti-HCV antibodies and HBsAg from local laboratories; (4) Local Health Authority registry of chronic liver disease patients; (5) drug prescriptions for HBV and HCV treatment; (6) archives of Alcohol Units. Results 3.5% of the residents had chronic liver disease, mainly chronic hepatitis (61.6%), followed by cirrhosis (14.0%) and alcoholic liver disease (11.2%). HCV was the main cause of chronic liver disease in females (46.3%) and males (29.8%), followed by alcohol abuse in males (22.9%) and HBV (10.9% males and 9.2% females). Prevalence of anti-HCV positivity was 3.2%, and increased with age to 8.8% in subjects aged 65 years and over. Conclusion This study shows that an epidemiologic pattern of the prevalence of chronic liver diseases and their aetiology can be obtained using routinely collected data.

Background Polychlorinated biphenyls (PCBs) may interact with several thyroid functions. Severe environmental pollution by PCBs has been observed in Brescia, Italy, due to the presence of a factory that produced these compounds until 1984. Objectives We aimed to investigate the correlation between serum levels of total PCBs and PCB 153 and thyroid hormone serum levels in adults in a cross-sectional population-based study. Methods A total of 527 subjects were enrolled; they were face-to-face interviewed about their history of thyroid disease and provided a blood sample under fasting conditions. The serum levels of free and total triiodothyronine (FT3 and TT3), thyroxine (FT4 and TT4) and thyroid-stimulating hormone (TSH), anti-thyroperoxidase (anti-TPO) and anti-thyroglobulin (anti-TGA), and lipid-adjusted total PCBs and PCB 153 were determined. Results There was no statistically significant difference in total PCB and PCB 153 concentrations in subjects with and without thyroid disease. Inverse, weak, statistically significant correlations were found between total PCBs and FT3 (Spearman's r= −0.09) and between total PCBs and PCB 153 and TSH (Spearman's r =−0.16 and −0.12, respectively). However, multiple regression analysis with FT3 and TSH serum levels as dependent variables and total PCBs and PCB 153 serum levels as predictors, including age, gender and BMI, showed a positive, rather than negative, association of PCBs and PCB 153 with FT3 and no association with TSH. Conclusions This study does not support the hypothesis that relatively high PCB environmental exposure can determine substantial alterations in thyroid function among adult people.

In the last few years chlorine dioxide has been increasingly used for disinfecting drinking water in many countries. Although it does not react with humic substances, chlorine dioxide added to water is reduced primarily to chlorite and chlorate ions, compounds that are under investigation for their potential adverse effects on human health. The aim of this research was to study the genotoxicity of chlorite and chlorate and their mixtures. The end-points included two plant tests (chromosomal aberration test in Allium cepa and micronucleus assay in Tradescantia, carried out at different times of exposure) and two genotoxicity tests in human HepG2 cells (comet assay and cytokinesis-blocked micronucleus test). Preliminary toxicity tests were carried out for both plant and HepG2 assays. The results showed that chlorite and chlorate are able to induce chromosomal damage to plant systems, particularly chromosomal aberrations in A. cepa root tip cells, even at concentrations lower than the limit established by Italian normative law and WHO guidelines. In HepG2 cells increased DNA damage was only observed for chlorate at the lowest concentration. No increase in micronuclei frequency was detected in any of the samples tested in human HepG2 cells.

The Allium cepa assay is an efficient test for chemical screening and in situ monitoring for genotoxicity of environmental contaminants. The test has been used widely to study genotoxicity of many pesticides revealing that these compounds can induce chromosomal aberrations in root meristems of A. cepa. Pesticide residues can be present in fruit and vegetables and represent a risk for human health. The mutagenic and carcinogenic action of herbicides, insecticides and fungicides on experimental animals is well known. Several studies have shown that chronic exposure to low levels of pesticides can cause birth defects and that prenatal exposure is associated with carcinogenicity. This study evaluated the potential application of plant genotoxicity tests for monitoring mutagens in edible vegetables. The presence of pesticides and genotoxic compounds extracted from 21 treated vegetables and eight types of grapes sampled from several markets in Campania, a region in Southern Italy, was monitored concurrently. The extracts were analysed for pesticides by gas chromatography and high-performance liquid chromatography, and for genotoxicity using two plant tests: the micronucleus test and the chromosomal aberration test in A. cepa roots. Thirty-three pesticides were detected, some of which are not approved. Genotoxicity was found in some of the vegetables and grapes tested. Allium cepa tests proved to be sensitive in monitoring genotoxicity in food extracts. The micronucleus test in interphase cells gave a much higher mutagenicity than the chromosomal aberration test in anaphase-telophase cells.

Background: Adrenergic activation has a central role in the development of HF. The function of the β1- and the α2C-adrenergic receptors is influenced by gene polymorphisms: the β1Arg389 variant is associated with increased β1-receptor sensitivity and the α2C-receptor Del322–325 variant is associated with decreased α2C receptor function and increased norepinephrine release. We hypothesised that these polymorphisms could influence the prevalence of heart failure. Methods: The role of the β1- and α2C-adrenergic receptor gene polymorphisms as risk factors for heart failure (HF) was assessed in an Italian white Caucasian population using a case-control study design. Genomic DNA was analysed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RLFP). Results: We compared 260 Caucasian patients with HF and 230 normal subjects. The β1Arg389 allele was frequent both in the patients with HF (69%) and in the normal subjects (73%). The α2CDel322–325 variant was rare in both groups (9% and 8%, respectively). Patients homozygotes for either the β1Arg389 or the α2CDel322–325 alleles had no increased risk of HF (odds ratio [OR], 0.8; 95%CI: 0.5—1.2 and OR, 0.8; 95% CI: 0.4—1.8, respectively). Patients homozygotes for both the β1Arg389 and the α2CDel322–325 alleles had no increased risk of HF as well (OR: 0.6; 95% CI: 0.2—2.1). Conclusions: β1-ARs and α2C-ARs polymorphisms are not associated with an increased risk of HF in an Italian white Caucasian population.

Background We hypothesised that the polymorphisms of the genes encoding for b1and the b2-adrenoceptors may have a role in the pathogenesis of heart failure (HF). We therefore compared the polymorphisms of the b1-adrenoceptor gene (Arg389Gly), the b2-adrenoceptor gene (Arg16Gly, Gln27Glu) and their combinations in patients with HF and normal subjects living in the same area. Methods and results A total of 256 cases with HF (left ventricular ejection fraction 640%) and 230 normal subjects were enrolled. The b1- and b2-adrenoceptor gene polymorphisms were assessed by PCR, followed by restriction enzyme digestion. No differences were observed in the distribution of any of the three genotypes studied in patients with HF and normal subjects. An analysis of the genotype combinations showed a non-significant increase in the risk of HF associated with the Arg389Gly16Gln27 (odds ratio=1.4; 95%CI 0.5‐3.6) and Arg389Gly16 Glu27 (odds ratio=1.2; 95%CI, 0.5‐2.8) homozygous allele combinations. Conclusion None of the three most common polymorphisms of b-adrenoreceptors are associated with an increased risk of HF. � c 2004 Published by Elsevier Ltd on behalf of The European Society of Cardiology.

The Tradescantia/micronuclei test (TRAD/MCN) is a well-validated test for monitoring environmental genotoxicants. These pollutants induce at the early meiotic stage of pollen mother cells chromosome fragments which become micronuclei at the tetrad stage. The standard test protocol requires some hours of exposure of the inflorescences and a recovery time of about 24 hours to reach the early tetrad stage. Since the recovery period represents a critical step of the TRAD/MCN, experiments were performed to establish its length in plants of clone #4430 of the hybrid T. hirsutiflora x T. subacaulis which is widely used in environmental monitoring. The aim of the present research was to ascertain the exact duration of recovery time in order to improve the sensitivity of the TRAD/MCN test. First, studies were performed to select the flowers at the beginning of the meiosis, and then anthers were sampled and studied for a period of 48-86 hours. The complete meiosis in the plants examined required about 80 hours. Second, exposure to genotoxic substances followed by different recovery times was carried out to demonstrate that effectiveness of the TRAD/MCN test is closely related to the duration of the recovery time. The test was carried out by exposing inflorescences to known mutagens (sodium azide and maleic hydrazide) for six hours followed by different recovery times (24-72 hours). The results showed that the frequency of micronuclei in the pollen mother cells increased with the length of the recovery time.

Air pollution has been recognized as a human carcinogen. Children living in urban areas are a high-risk group, because genetic damage occurring early in life is considered able to increase the risk of carcinogenesis in adulthood. This study aimed to investigate micronuclei (MN) frequency, as a biomarker of DNA damage, in exfoliated buccal cells of pre-school children living in a town with high levels of air pollution. A sample of healthy 3-6-year-old children living in Brescia, Northern Italy, was investigated. A sample of the children's buccal mucosa cells was collected during the winter months in 2012 and 2013. DNA damage was investigated using the MN test. Children's exposure to urban air pollution was evaluated by means of a questionnaire filled in by their parents that included items on various possible sources of indoor and outdoor pollution, and the concentration of fine particulate matter (PM10, PM2.5) and NO2 in the 1-3 weeks preceding biological sample collection. 181 children (mean age ± SD: 4.3 ± 0.9 years) were investigated. The mean ± SD MN frequency was 0.29 ± 0.13%. A weak, though statistically significant, association of MN with concentration of air pollutants (PM10, PM2.5 and NO2) was found, whereas no association was apparent between MN frequency and the indoor and outdoor exposure variables investigated via the questionnaire. This study showed a high MN frequency in children living in a town with heavy air pollution in winter, higher than usually found among children living in areas with low or medium-high levels of air pollution.

Internet and e-commerce have profoundly changed society, the economy, and the world of health care. The web offers opportunities to improve health, but it may also represent a big health hazard since it is a basically unregulated market with very low consumer protection. In this paper we analyze marketing and pricing strategies of online pharmacies (OPs). Our analysis shows that OPs use strategies that would be more suitable for a commodity market than for drugs. These strategies differentiate according to variety (brand or generic), quality, quantity, and target group. OPs are well aware that the vacuum in the legislation allows them to reach a target of consumers that pharmacies cannot normally reach, such as those who would like to use the drug without consulting a physician (or, even worse, against the physician's advice). In this case, they usually charge a higher price, reassure the users by minimizing on the side effects, and induce them to bulk purchase through sensible price discounts. This analysis suggests that the selling of drugs via the Internet can turn into a “public health risk”, as has been pointed out by the US Food and Drug Administration.

The potential migration of genotoxic compounds into mineral water stored in polyethylene terephthalate (PET) bottles was evaluated by an integrated chemical/biological approach using short-term toxicity/genotoxicity tests and chemical analysis. Six commercial brands of still and carbonated mineral water bottled in PET and in glass were stored at 40 °C for 10 days in a stove according to the standard EEC total migration test (82/711/EEC), or at room temperature in the dark. After treatment, the samples were analysed using gas-chromatography/mass spectrometry (GC/MS) to detect volatile and non-volatile compounds, the Microtox ® test to evaluate potential toxicity of the samples, and three mutagenicity tests – Tradescantia and Allium cepa micronucleus tests and the Comet assay on human leukocytes – to detect their genotoxic activity. GC/MS analysis did not detect phthalates or acetaldehyde in the water samples. The Microtox ® test found no toxic effects. Mutagenicity tests detected genotoxic properties of some samples in both PET and glass bottles. Statistical analyses showed a positive association between mineral content and mutagenicity (micronuclei in A. cepa and DNA damage in human leukocytes). No clear effect of treatment and PET bottle was found. These results suggest the absence of toxic compounds migrating from PET regardless of time and conditions of storage. In conclusion, bottle material and stove treatment were not associated with the genotoxic properties of the water; the genotoxic effects detected in bottled water may be related to the characteristics of the water (minerals and CO 2 content).

Background/aims The aims were to estimate among patients with hepatitis C virus (HCV) infection the prevalence of alcohol and coffee intake and smoking habit, the reliability of these self-reported data and the possible change of patients’ habit after their first contact with a Viral Hepatitis Service. Methods 229 patients were initially interviewed personally at the Viral Hepatitis Service and after 6 months they were re-interviewed by phone in regard to their alcohol, coffee drinking and smoking habits. Results Alcohol drinkers were 55.5% of males and 35.3% of females. Most subjects drank coffee daily, both men (90.0%) and women (84.9%). The proportion of current smokers was higher in males (43.6%) than females (26.9%). We found a fair to good reliability of self-reported data regarding patients’ habits, alcohol and coffee intake, and number of cigarettes smoked daily. We observed a statistically significant decrease in alcohol and coffee intake and cigarettes smoked between baseline and follow-up interviews. Conclusion We found a fairly high proportion of HCV-infected patients who regularly drink alcohol and coffee beverages and smoke cigarettes, especially among males. The reliability of self-reported data on these habits seems satisfactory. More decisive action to modify these habits, especially alcohol intake, is required in these patients.

An integrated chemical analytical and biological approach was used to detect the presence of genotoxins in the drinking water of four Italian cities which obtain their water supply from different sources (superficial or source waters). A battery of rapid and sensible in-vitro and in-vivo tests were used to detect genotoxic compounds, and chemical analytical methods to detect disinfection by-products. The aim was to provide information useful for routine monitoring of drinking water and recommendations for improving the management of disinfection and distribution establishments.