Half of Italian children suffered major DNA damage from breathing toxic particulates, a 2014-2015 study revealed. In the same period, Italy’s overall death rate jumped 11.3% and spraying/geoengineering deployment massively increased.

Scientists tested 1200 six to eight year olds for DNA damage – which can cause cancer and be passed down to future generations. Out of the 220 children tested in Turin, 53% showed mutated DNA caused by breathing toxic particulates.

The image above is a NASA satellite image of heavy spraying over Italy, December 22nd, 2016.

This follows news that Italy’s overall death rate leapt 11.3% between 2014 & 2015.

This is an automatic translation of the study.

effects of air pollution on children – MAPEC_LIFE Project – fotocomunicato and press kit
12/20/2016

Turin – 20 December 2016 – The project MAPEC_LIFE “Monitoring Air Pollution Effects on Children for supporting public health policy”, approved in 2013 by the European Commission under the LIFE + 2012 Program, Environment Policy and Governance, is the first large multicenter study on early biological effects of airborne pollutants on buccal cells of children of 6-8 years, residents in 5 Italian cities (Turin, Brescia, Lecce, Perugia and Pisa), in relation to the concentration of certain pollutants and socio-demographic characteristics and the children’s lifestyles.

The University of Turin – Section of Environmental Health, Department of Public Health and Pediatric – is partner in the project that started January 1, 2014 and ending on 31 December 2016. The project coordinator is the University Brescia. The other partners are: the University of Perugia, Pisa University, the University of Salento, the City of Brescia and the Service Centre Multisettoriale and Technology of Brescia.

“The MAPEC_LIFE is a project of Public Health – explains Prof. Giorgio Gilli responsible for the project dissemination activities for the University of Turin – whose main purpose is the protection of public health, through the collection of scientific data that might be supportive to the policy makers in the promotion of new and better environmental policies. In the course of the project we found interest and support from the various components of civil society: local authority institutions and leaderships school, from teachers to families and children, an indication of a now widespread awareness and shared on issues of environment, health and prevention. ”
Methods

The buccal mucosa cells of all recruited children have been collected by a slight extension of the cheek brushing and analyzed in the laboratory for the presence of micronuclei, as an indicator of damage to cellular DNA. This effect is a biomarker of early biological damage not directly related to an individual’s risk, but indicative of a population exposure to risk factors.

To assess the real exposure of children to air pollution, during the days of the biological sample, it proceeded to the sampling of atmospheric particulate matter (PM0,5), by means of high volume air samplers placed in schoolyards frequented by children recruited. The PM0,5 thus collected was analyzed to assess the concentration of polycyclic aromatic hydrocarbons (PAHs) and their nitro-compounds (nitroIPA), the toxicity of human lung cells in culture (A549 cells) genotoxicity by the micronucleus test and the comet assay on the same cell type (A549 cells) and the mutagenicity by the Ames bacterial cell assay.

For a more complete assessment of air quality to which children were exposed they were collected throughout the sampling period the Regional Agencies for Environment Protection Data relating to the levels of the main airborne pollutants (PM10, PM2.5 , NO, NO2, CO, SO2, O3 and benzene). In addition, parents of children, a questionnaire was administered to collect information on the state of health of the child, outdoor and indoor exposures, socioeconomic characteristics, lifestyle and nutrition.

To study the relationship between exposure to pollutants and biological effects of the biological and environmental samples were collected both in winter and in late spring, periods, in the urban reality of our country, respectively high and low levels of several air pollutants. In addition, to assess the intra-subject biological variability in children recruited in Brescia is a third biological sampling was carried out in winter, after one year from the previous year.

The data thus collected were analyzed by the use of multivariate statistical analysis models to evaluate the associations between DNA damage, levels of pollutants and other factors investigated.

The project includes also the development of a package of teaching aids to help cope with the children and the schools the main themes of the project: air pollution, effects on health and healthy lifestyles.

Results

1. suspected Population

The environmental and biological sampling were completed in January 2016. In the five cities were involved 26 primary schools, for a total of 139 classes; in Turin participated in the 7 project schools (24 classes) owned by 2 institutes Inclusive (N. Tommaseo and Ilaria Alpi) and a Learning Circle (P. Gobetti).

In total, 1149 children (220 in Turin) were included in the study and examined in both seasons: winter (November 2014-March 2015) and the spring (April-June 2015). The children involved are half male and on average have a high socio-economic level. Of them, one child in eight is exposed to secondhand smoke at home, one in three is overweight or obese, and a child of 2 has a power supply that does not follow the principles of the Mediterranean diet. Overall, the data of the questionnaires completed by parents show that the weight status of children is comparable to that of other studies on Italian children of the same age and confirm a higher proportion of overweight children in the regions of Central and Southern Italy.

2. Measurements of air pollutants

The data collected by the Regional Agencies for Environmental Protection showed urban pollution levels higher in winter than in the spring and in the northern Italian cities (Turin and Brescia) than those in the center-south (Pisa, Perugia and Lecce) . In particular, the average PM10 levels observed in the winter sampling period (November 2014-March 2015) are: 50 mg / m3 in Turin, 45 g / m3 in Brescia, 29 mg / m3a Pisa and Perugia, 27 micrograms / m3 Lecce. The levels of PM10 is significantly lower in the spring season (April-June 2015): 24 mg / m3 in Turin, 26 g / m3 in Brescia, 22 g / m3 in Pisa, 17 g / m3 in Perugia and 20 ug / m3 Lecce.

The same seasonal pattern and geographical features all suspects pollutants, except ozone, instead, is more concentrated in the spring season and has similar concentrations in different cities.

Even PAH concentrations and nitroIPA detected in atmospheric particulate matter PM0,5 championship at the schools involved in the project follow the same trend: the spring levels are up to 10 times lower than in winter and northern cities have the highest concentrations in both seasons.

3. toxic and genotoxic activity of airborne particulate samples

The results of laboratory tests performed by MAPEC_LIFE project showed that the samples of atmospheric particulate PM0.5 induce toxic effects, genotoxic and carcinogenic, albeit contained, in cell culture. Again the effects of the seasonal flu: the samples collected in winter induce greater effects than those collected in the spring. As for the non-specific toxicity and the promotion of carcinogenicity, the Brescia particulate air pollution is what gave him greater effects. In mutagenicity tests, however, that of Turin proved to be the most active particles, followed by that of Brescia, Pisa, Perugia and finally Lecce. The ability to induce mutations is correlated to the concentration of PAHs and nitro-PAHs in PM0,5.

“In a general context that has a trend in reduction of pollution levels by particulate air pollution – intervenes Prof. Elisabetta Carraro, head of the University of Turin – the study confirmed the seasonality of this pollutant and a gradient North -South, with the highest concentrations observed in winter in the cities of northern Italy. The in vitro evaluation of cells in the laboratory, of the PM0,5 ability to determine DNA damage showed an overall modest effect associated with the season, the city and the concentration of polycyclic aromatic hydrocarbons on PM0,5 “.

4. genotoxic effects in buccal cells of children

In winter 52.7% of children it has been shown to have at least one micronucleus in buccal mucosa cells (mean value: 0.44 MN / 1000 cells). There is however a low correlation between the levels of micronuclei detected in the winter season and those measured in spring in the same child.

Comparing the children of 5 cities, it is noted that those of Brescia have on average a greater number of micronuclei than the other (0.56 MN / 1000 cells). Follow the children of Pisa (0.50 MN / 1000 cells), Perugia, Torino and Lecce (0.41, 0.39 and 0.32 MN / 1000 cells, respectively). In the spring, we observe a biological effect halving in all cities (average value: 0.22 MN / 1000 cells) with an even decrease in the percentage of children with at least micronuclei in buccal cells (35.9%).

Applying advanced models of multivariate statistical analysis, it is seen that the levels of some pollutants (benzene, PM2.5, ozone, SO2 and PAH) are associated with the frequency of micronuclei in cells of children. In particular, the increase in the risk of having micronuclei in buccal cells for the increase of a pollutant unit is: 20.1% for benzene (1 g / m3), 1.1% for PM2.5 (1 ug / m3), 1.3% for ozone (1 mg / m3), 4.2% for SO2 (1 g / m3) and 1.7% for PAH in PM0,5 (1 ng / m3), while in the limits of the estimates of uncertainty due to the high variability of the phenomenon. The relative importance of these pollutants also depends on the variability of concentrations in different cities.

Finally, it is seen that the passive smoke exposure and overweight in children tend to increase the risk of having micronuclei, while the healthy diet tends to decrease.

“The project MAPEC_LIFE – intervenes Prof. Giorgio Gilli – has also provided an opportunity to speak in schools of air pollution, its effects on health and healthy lifestyles, with an educational project that produced fact sheets for teachers and videogames for children. All this material is freely downloadable from the website of the project. The interest and the effectiveness of the activities undertaken demonstrate the importance of these issues since primary school. ”

conclusions

This study highlighted the ability of the ultrafine fraction of particulate matter (PM0,5) to induce toxic, mutagenic and carcinogenic, albeit modest, in the cells treated in the laboratory.

The early biological effect, highlighted in buccal cells of children as the presence of micronuclei, was associated with:

Season – the biological effect measured is significantly higher in winter than in the spring;
City – the children of Brescia showed the greatest effect, followed in order by those of Pisa, Perugia, Torino and Lecce;
concentration of benzene, PM2.5, ozone, SO2 and IPA – the increase in these pollutants was moderately associated with an increase in micronuclei in the children’s cells;
characteristics of children – healthy diet has been shown to attenuate the effect, while in passive smoking exposure and aggravate overweight.

In considering these results, it should be noted that the 2014-2015 winter season was characterized by a relatively low average levels of airborne pollutants, compared to previous years, probably because of mild temperatures and high rainfall. Therefore, it is possible that the biological effect present in other winter seasons is higher than that measured in the present study.

“I MAPEC_LIFE study represents the first large multicenter study on early biological effects of airborne pollutants on buccal cells of children of 6-8 years – explains Prof. Elizabeth Carraro – Therefore, the results of MAPEC_LIFE study are not comparable with those of other research conducted to date in the world on the biological effects of air pollution, because the latter, mostly small, were carried out with different methods and subjects of different ages. ”

“In conclusion – says Prof. Giorgio Gilli – the level of biological markers we studied was moderately associated with concentrations of some airborne pollutants and other factors, and may be an indicator of possible, future adverse health effects. These effects, in the light of current knowledge, are detectable at the population level, but are not predictive of the onset of disease in the individual. ”

Here’s a pdf of the associated PowerPoint presentation.

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