Are 6-8 year old Italian children moving away from the Mediterranean diet?

BACKGROUND: The Mediterranean diet (MD) is considered one of the healthiest dietary models, as it decreases the risk of chronic diseases and may modulate the organism's early response to environmental pollution. In recent decades, Mediterranean countries have been replacing their traditional diet with other less healthy eating habits, especially among children and teenagers. OBJECTIVE: The aim of this study was to evaluate the MD and the level of adherence to it in 6-8 year old Italian children, in relation to residence, lifestyle, and social and family contexts. METHODS: A questionnaire was administered to the children's parents in two seasons in 5 Italian towns. The diet section contained 116 questions investigating the frequency of consumption of different types of food. The Italian Mediterranean Index (IMI) was calculated according to the intake of 6 typical Mediterranean and 4 non-Mediterranean foods. On the basis of IMI score, MD adherence was classified as low (≤ 3 IMI score), medium (4-5) and high (≥ 6). Total energy load and diet composition in micro- and macronutrients were calculated from consumption frequency. RESULTS: Diet analysis was computed on 1164 subjects with two complete questionnaires. Body mass index, calculated for each subject, showed that 28.9% of the children were overweight, the figure varying slightly with area of residence. Our findings showed that 59.0% of the children had a low score for MD adherence. CONCLUSIONS: The results of this study showed that most Italian children did not follow the MD and socio-economic characteristics appeared not to be associated with type of diet.

In memory of Professor Giuseppina Scassellati Sforzolini.

This special issue of the journal "Annali di Igiene: Medicina Preventiva e di Comunità" is dedicated to the memory of Professor Giuseppina Scassellati Sforzolini (Mogadishu 1929 ­ Perugia 2014), Emeritus Professor of Public Health at the University of Perugia

Feasibility and reliability of a questionnaire for evaluation of the exposure to indoor and outdoor air pollutants, diet and physical activity in 6-8-year-old children.

INTRODUCTION: The MAPEC-Life project aims to study the biological effects of early exposure to air pollutants on the oral mucosa cells of school-age children in five Italian cities. A questionnaire was created to evaluate the association between outdoor and indoor airborne pollutants, lifestyle, diet and biomarker effects. The feasibility and reliability of the questionnaire were evaluated. METHODS: A questionnaire was drawn up to be filled in by the parents of 6-8-year-old children. It consisted of 148 questions on the children's health, physical activity, environmental exposures and the frequency of food consumption at the main meals. First we conducted a questionnaire feasibility study involving 53 volunteer parents. We then performed a reliability study by administering the questionnaire to a further 156 parents and again one month later (test/retest method). The correlations between answers at the first and second administration of the questionnaire were evaluated using the Kappa statistic and Spearman's coefficient. RESULTS: After verifying the feasibility of the questionnaire, we conducted a reliability analysis on 132 completed questionnaires. The percentage of agreement between the first and the second responses given was over 70%, all K values being greater than 0.6. The analysis of calories and macronutrients also showed good agreement. CONCLUSIONS: The questionnaire drawn up for the study proved to be sufficiently reliable for gathering information about the factors of interest in our study of the relationship between air pollution and early biological effects in children.

Micronucleus assays with Tradescantia pollen tetrads: an update.

Micronucleus (MN) assays with early pollen tetrad cells of Tradescantia (Trad-MN assays) are at present the most widely used bioassays with plants for the detection of genotoxins in the environment. So far, ∼ 160 chemicals have been tested and ∼ 100 articles that concern complex environmental mixtures were published. This article summarises the results of Trad-MN studies, which have been carried out during the last 15 years with individual compounds and investigations concerning the pollution of environmental compartments (soil, water and air). The evaluation shows that the effects of certain genotoxins such as heavy metals, radionuclides, pesticides and air pollutants can be easily detected with this test. Comparisons with results obtained in MN studies with mitotic (root tip) cells indicate that meiotic tetrad cells are in general more sensitive. Important issues for future research concern the evaluation of the suitability of wildlife Tradescantia species that are sometimes used instead of specific clones (such as #4430 for which standardised protocols have been developed) as well as the assessment of the predictive value of Trad-MN results in regard to the prediction of cancer hazards in humans and adverse effects at the ecosystem level. The fact that the genotoxic effects of certain compound such as metals, which can be detected with plant bioassays, in particular with the Trad-MN assay but not in other commonly used bioassays (e.g. in bacterial tests) makes them an essential element in the batteries for environmental monitoring.

[Assessment of the genotoxic effects of gaseous emissions and percolates from three municipal landfills using plant bioassays]. / Valutazione dei rischi genotossici derivanti dalla produzione di biogas e percolati in tre discariche controllate mediante test su vegetali.

The biomonitoring of genotoxic effects in environmental complex mixtures using higher plants is very useful for hazard evaluation. In this study we evaluated the potential application of plant genotoxicity tests in monitoring mutagens in landfill environment. The clastogenic effects of gaseous emissions (biogas)from three municipal landfills were evaluated by in situ monitoring using the Tradescantia micronucleus assay. The cytotoxicity and genotoxicity of leachates were studied using the Allium cepa test. We found no significant differences in micronuclei frequency in pollen cells of Tradescantia. Leachate samples showed elevated toxicity that inhibited root tip development in Allium cepa. Genotoxicity of the leachates was evaluated in diluted samples only. We found a significant increase in chromosomal metaphase aberrations only in one of the samples analyzed. In conclusion, biogas was not shown to be a real hazard, whereas leachates were found to display elevated toxicity. It would be advisable to treat leachates before releasing them into the environment as they can cause ecological damages. Since plant bioassays are very useful for the in situ monitoring of environmental genotoxins they are important for the prevention of environmental pollution resulting from the disposal of solid waste.

Evaluation of chlorite and chlorate genotoxicity using plant bioassays and in vitro DNA damage tests.

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.

Genotoxicity of the disinfection by-products resulting from peracetic acid- or hypochlorite-disinfected sewage wastewater.

Wastewater disinfection is routinely carried out to prevent the spread of human pathogens present in wastewater effluents. To this aim, chemical and physical treatments are applied to the effluents before their emission in water bodies. In this study, the influence of two widely used disinfectants, peracetic acid (PAA) and sodium hypochlorite (NaClO), on the formation of mutagenic by-products was investigated. Wastewater samples were collected before and after disinfection, in winter and in summer, at a pilot plant installed in a municipal wastewater-treatment plant. Samples were adsorbed using silica C18 cartridges and the concentrates were tested for mutagenicity in the Salmonella typhimurium reversion test with strains TA98 and TA100. Non-concentrated water samples were tested with two plant genotoxicity assays (the Allium cepa root anaphase aberration test and the Tradescantia/micronucleus test). Mutagenicity assays in bacteria and in Tradescantia showed borderline mutagenicity in some of the wastewater samples, independent of the disinfection procedure applied. Negative results were obtained in the A. cepa anaphase aberration test. These results indicate that, in the conditions applied, wastewater disinfection with PAA and NaClO does not lead to the formation of significant amounts of genotoxic by-products.

Monitoring of volatile and non-volatile urban air genotoxins using bacteria, human cells and plants.

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.

Monitoring nitrite, N-nitrosodiethanolamine, and mutagenicity in cutting fluids used in the metal industry.

We carried out an integrated environmental/biological monitoring program to evaluate cancer hazards among metal industry workers exposed to cutting fluids. Several cutting fluids were sampled according to response to a semiquantitative nitrite rapid test in metal factories in central Italy. The nitrite-positive samples were analyzed for nitrite and nitrosodiethanolamine (NDELA) content and mutagenic activity. The nitrite-negative samples were analyzed only for mutagenicity. Of the total samples, 20.6% were nitrite positive, and all contained NDELA. However, nitrite content was not quantitatively predictive of the NDELA content, which varied enormously among samples (0.3-1900 mg/kg). Nitrite-negative samples were always nonmutagenic. Mutagenicity was found in half the NDELA-containing samples but was not related to nitrite or NDELA content. Nitrite screening of cutting fluids in the field is an interesting method for identifying samples that potentially contain NDELA and other unknown mutagens and, when performed with short-term mutagenicity tests, nitrite screening seems to be a valid tool by which industrial managers and health officers could minimize the health hazards associated with occupational exposure to cutting fluids.

Biological monitoring of workers exposed to N-nitrosodiethanolamine in the metal industry.

Biological monitoring of occupational hazards was performed in workers using cutting fluids containing N-nitrosodiethanolamine (NDELA). The study involved a group of 25 male subjects from some metal factories in central Italy who used cutting fluids with an NDELA content of > or = 5 mg/l (high-exposure group) and a group of 37 males exposed to cutting fluids with an NDELA content < 5 mg/l (low-exposure group). For comparison, we recruited a control group consisting of 37 subjects living in the same area. For all subjects, internal dose (urinary excretion of NDELA, mutagens, and thioethers), early biological effects (sister chromatid exchanges in blood peripheral lymphocytes), and urinary excretion of D-glucaric acid (DGA) as an endpoint product in the glucuronidation pathway were assessed. The results showed that only the workers using cutting fluids with NDELA concentrations of > or = 5 mg/l excreted trace amounts of NDELA in their urine. Urine excretion of mutagens was similar in the two exposure groups and in the controls. High-exposure subjects had a higher mean value of urinary thioethers than low-exposure and control subjects, but no differences were found in urinary DGA or lymphocyte sister chromatid exchange among the three groups. Smoking status increased the mean values of all the biomarkers, and coffee drinking was associated with urinary DGA excretion.

Biological monitoring of genotoxic hazard in workers of the rubber industry.

Biological monitoring of genotoxic hazard in the rubber industry was performed in 19 male workers and 20 age-matched controls in a local health unit in northern Italy. Peripheral blood lymphocytes were analyzed for the presence of DNA damage (single-cell microgel-electrophoresis, or comet assay) and for cytogenetic parameters (sister chromatid exchanges and micronuclei frequency, and proliferative rate index). The following bioassays were performed in urine samples: a) mutagenicity test and concentration of thioethers as markers of exposure, and b) excretion of D-glucaric acid and 6-beta-hydroxycortisol (related to 17-hydroxycorticosteroid excretion) as indicators of the inductive status of the microsomal enzyme system (phase-I). The exposed subjects showed statistically higher mean values of 17-hydroxycorticosteroids and micronuclei and lower values of 6-beta-hydroxycortisol than controls, when taking cigarette smoking into account. The comet assay showed higher values for migration distance in exposed subjects than controls, although the differences were not significant at a p-value of 0.05. These findings suggest that industrial exposure in the rubber processing industry may cause genetic damage and may modify the activity level of some enzymes; these results should be considered with caution due to the small number of subjects enrolled.

Urine mutagenicity and biochemical parameters as markers of exposure to petroleum pitch using a rat model.

A petroleum pitch sample collected in a carbon electrode factory was studied using a series of in vivo assays for genotoxicity and enzymatic induction capability. Rats were treated with the petroleum derivative in three doses: 100, 50, and 10 mg/kg body weight. The treatment produced a rapid excretion of mutagenic substances in the urines of the first 24 hr only in rats treated with high doses (100 and 50 mg/kg). No faecal mutagenic activity was observed. Analyses of urinary thioethers showed that urinary metabolites derived from the compounds present in the pitch-sample at the lowest dose-administered (10 mg/kg) were eliminated primarily as cysteine conjugates. The pitch sample was found to be a good inducer of pulmonary and hepatic aryl hydrocarbon hydroxylase, especially after a 50 mg/kg dose. Urinary D-glucaric acid content was always statistically increased in treated animals compared with controls, confirming the enzymatic induction activity. Hepatic glutathione-S-transferase activity increased following treatment with 50 and 10 mg/kg doses.

Genotoxicity of surface water treated with different disinfectants using in situ plant tests.

Disinfection of surface drinking water, in particular water chlorination, results in many by-products with potential genotoxic and/or carcinogenic activity. In the present study, we evaluated the genotoxicity of surface water after treatment with different disinfectants by means of in situ plant genotoxicity assays (micronucleus and chromosomal aberration tests) which can detect both clastogenic and aneugenic effects. The study was carried out at a pilot plant using lake water after sedimentation and filtration. This water supplied four stainless steel basins: three basins were disinfected with sodium hypochlorite, chlorine dioxide, and peracetic acid and the fourth basin containing untreated lake water was used as a control. Plants were exposed in situ in the basins. The study was carried out using water collected in different seasons over a period of about 1 year in order to assess the treatments in different physical and chemical lake water conditions. The micronucleus test in root cells of Vicia faba (Vicia faba/MCN test) revealed genotoxicity in many samples of disinfected water. The micronucleus test in Tradescantia pollen cells and the chromosome aberration test in root cells of Allium cepa showed genotoxic effects only in some disinfected samples, but also revealed genotoxicity in raw water. The results of the study indicated that the Vicia faba/MCN test was the most sensitive plant assay for disinfected water and that peracetic acid disinfection produced similar or lower genotoxicity than sodium hypochlorite or chlorine dioxide treatment.

Mutagenicity and clastogenicity of gas stove emissions in bacterial and plant tests.

The aim of this research was to study the gaseous and particulate emissions of genotoxic substances during cooking with two types of methane stoves (a new one and an old one). The particulates were sampled both with a cascade impactor air sampler and an impinger with ice trap and analyzed by two bacterial mutagenicity tests (Ames and Kado tests) and by HPLC for polycyclic aromatic hydrocarbons (PAH). Gaseous emissions were studied in situ using the Ames test, a clastogenicity plant test (Tradescantia-micronucleus test), and in an automated system for chemical analyses. Clear indirect mutagenicity was found only with the Kado test (TA98-S9) in extracts of particulates emitted from the old methane stove and collected with the impinger. Similar mutagenicity (TA98+S9) was also found for the finest fraction of particulates (<0.5 um) collected from both stoves. Gaseous emissions of both stoves caused clastogenicity in the in situ experiments with the Tradescantia-micronucleus test. The physico-chemical analyses of the emissions showed also the presence of very fine particulates and trace amounts of PAH. The exposure of these genotoxins could be particularly important for occupationally exposed individuals in homes and businesses and for susceptible subjects living indoors for long periods (infants, children, the sick, and the elderly).

Factors associated with non-participation of women in a breast cancer screening programme in a town in northern Italy.

STUDY OBJECTIVE: The aim was to investigate the reasons for the high percentage of women refusing to attend a breast cancer screening programme in the Health District of Brescia, Italy. DESIGN: This was a survey of a sample of non-attenders to the programme, who were interviewed using a structured questionnaire. SETTING: Non-attenders all lived in a central area of the town near the screening centre. PARTICIPANTS: Of the 612 non-attenders eligible for interview, 183 could not be interviewed: one had died, 86 were away from home at two different visits, 32 were no longer resident at the known address, eight had serious health problems, 17 had undergone mastectomy, and 39 refused the interview. Overall, a total of 429 of the 612 eligible women were interviewed (70.1%). MEASUREMENTS AND MAIN RESULTS: Attenders and non-attenders were compared with respect to demographic and socioeconomic factors, use of preventive medicine, and prevalence of risk factors for breast cancer. The response was higher among less educated women, married and widowed women, and those born in the province than among more educated, single or divorced, and immigrant women. Most of the women interviewed gave practical reasons for non-participation, but lack of interest/distrust and fear/worry/anxiety also seemed important. The number of non-attenders who had had a Papanicolaou test within the previous three years was three times higher than those who had had mammography, suggesting that non-attenders were more interested in types of preventive medicine other than screening for breast cancer by mammography. Attenders and non-attenders appeared similar as regards distribution of conventional breast cancer risk factors. CONCLUSIONS: Greater effort in the information campaign might increase the participation rate in screening for breast cancer, although to a lesser extent than expected: if non-attenders potentially recruitable in our screening were added to attenders, overall compliance of the programme was about 75%, lower than that observed in some programmes in northern Europe.

Patterns and covariates of alcohol drinking among high school students in 10 towns in Italy: a cross-sectional study.

A self-report questionnaire on alcohol and tobacco use was administered anonymously to 5221 high-school students in grade 9 (aged 14-15) and to 4154 students in grade 13 (aged 18-19) in 10 Italian towns. Alcohol consumption was greater among males than females and among 13th graders with respect to 9th graders. Students who drank alcoholic beverages every day were, in males and females, respectively, 22.5% and 9.0% among 9th grade and 31.3% and 10.0% among 13th grade students; those who drank 7 or more alcohol units a week were 26.8% and 11.1% among 9th grade and 42.7% and 13.9% among 13th grade students; finally, those who felt drunk at least once in the last 12 months were 19.8% and 13.9% among 9th grade and 41.4% and 19.0% among 13th grade students. Both alcohol consumption and frequency of drunkenness were found to be associated with both experimental and regular cigarette smoking and with psychoactive drug consumption, but not with socio-economic, cultural and demographic factors. Two main distinct drinking patterns were found among Italian adolescents: on the one hand, the daily drinking of moderate amount of wine with meals, together with parents, which was more common among males than females, as in other 'Latin' countries; and on the other hand, the occasional intake of alcoholic beverages in excess, especially beer and spirits, out of the family but together with peers.

Urine mutagenicity and biochemical effects of the drinking water mutagen, 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-furanone (MX), following repeated oral administration to mice and rats.

Mutagenicity analysis of urine from rats treated by oral gavage with MX at a dose of 64 mg/kg for 14 days revealed that only 0.3% of the administered compound was excreted in a genotoxically active form. At lower doses, mutagenicity was not detectable. No evidence of micronucleus induction in peripheral blood erythrocytes was observed in mice treated similarly. These findings indicate that MX is extensively detoxified in vivo and is unlikely to cause genetic damage in systemic tissues except at relatively high doses where detoxification pathways become saturated. In a separate experiment, significant depressions were observed in D-glucaric acid and thioether excretion and in levels of several liver enzymes involved in xenobiotic metabolism. The mechanism for these metabolic alterations and their relevance to the in vivo metabolism of the compound require further investigation.

In vitro genotoxic effects of the insecticide deltamethrin in human peripheral blood leukocytes: DNA damage ('comet' assay) in relation to the induction of sister-chromatid exchanges and micronuclei.

Deltamethrin, a synthetic dibromo-pyrethroid insecticide, is extensively used in agriculture, forestry and in household products because of its high activity against a broad spectrum of insect pests (both adults and larvae), its low animal toxicity and its lack of persistence in the environment. Data on the genotoxicity and carcinogenicity of deltamethrin are rather controversial, depending on the genetic system or the assay used. The aim of this study was to further evaluate the potential genotoxic activity of deltamethrin. The in vitro genotoxicity of deltamethrin has been evaluated by assessing the ability of the insecticide to damage DNA (as evaluated using the single-cell microgel-electrophoresis or 'comet' assay) or induce sister-chromatid exchanges (SCE) and micronuclei (MN) in human peripheral blood leukocytes. All treatments were conducted with and without the presence of an external bioactivation source (+/- S9mix). The results indicate that deltamethrin, in the presence of metabolic activation (+ S9mix), is able to induce DNA damage (double- and single-strand breaks, alkali-labile sites and open excision repair sites) as revealed by the increasing tail moment values observed with increasing doses. The frequency of SCE and MN were not statistically increased in deltamethrin-treated cells as compared to controls, both with and without S9mix. However, lower deltamethrin doses were tested, as compared to 'comet' assay, because of cytotoxicity.

Monitoring of mutagens in urban air samples.

This research was designed to examine the presence of mutagenic/carcinogenic compounds in urban airborne particulates sampled with the inhalable PM-10 high volume sampler in two different streets of Brescia, a heavily industrialized town in northern Italy, using the Tradescantia/micronucleus test and a bacterial mutagenicity test (Kado test, a more sensitive version of the Ames test). In addition, the Tradescantia/micronucleus test was used for in situ monitoring of gaseous pollutants in other urban areas of Brescia and in two car tunnels, one with heavy car traffic in Perugia, a town in central Italy, and one in Brescia with moderate traffic. The Tradescantia-micronucleus test carried out on extracts of airborne particulates gave positive results only for the sample collected in the traffic-congested street where also higher bacterial mutagenicity was found. The in situ monitoring of the urban areas with the Tradescantia/micronucleus test always gave negative results. Monitoring carried out in the two car tunnels showed a significant increase in micronuclei frequency only in flowers exposed in the smaller and more polluted tunnel.

Monitoring airborne genotoxicants in the rubber industry using genotoxicity tests and chemical analyses.

This research was designed to examine the presence of mutagenic/carcinogenic compounds in airborne pollutants in the rubber industry using an integrated chemical/biological approach. Inhalable airborne particulate matter (PM-10: <10 microm) was collected in four rubber factories using a high-volume sampler equipped with a cascade impactor for particle fractionation. The organic extracts of two different fractions (0.5-10 microm and <0.5 microm) were examined for mutagenicity with the Ames test and for in vitro DNA-damaging activity in human leukocytes by single-cell microgel electrophoresis (Comet assay). The extracts were also studied by gas chromatography/mass spectrometry (GC/MS) for polycyclic aromatic hydrocarbon (PAH) content. Nitrosamines in ambient air were sampled on cartridges and analysed by GC with a thermal energy analyser (TEA) detector. Airborne volatile genotoxins were monitored in situ using a clastogenicity plant test (Tradescantia/micronuclei test). The results showed that airborne particulates were mainly very fine (<0.5 microm) and that trace amounts of genotoxic nitrosamines (N-nitrosodimethylamine: 0.10-0.98 microg/m(3); N-nitrosomorpholine: 0.77-2.40 microg/m(3)) and PAH (total PAH: 0.34-11.35 microg/m(3)) were present in air samples. Some extracts, particularly those obtained from the finest fractions, were mutagenic with the Ames test and genotoxic with the Comet assay. In situ monitoring of volatile mutagens using the Tradescantia/micronuclei test gave positive results in two working environments. The results showed the applicability of this integrated chemical-biological approach for detecting volatile and non-volatile genotoxins and for monitoring genotoxic hazards in the rubber industry.