The effects of the exposure to neurotoxic elements on Italian schoolchildren behavior.

Neurodevelopmental disorders are constantly increasing on a global scale. Some elements like heavy metals are known to be neurotoxic. In this cross-sectional study we assessed the neurobehavioral effect of the exposure to trace elements including lead, mercury, cadmium, manganese, arsenic and selenium and their interactions among 299 schoolchildren residing in the heavily polluted Taranto area in Italy. Whole blood, urine and hair were collected for metal analyses, while the Child Behavior Checklist and the Social Responsiveness Scale, administered to the main teacher and the mothers were considered to identify behavioral problems in children. Blood lead mainly influenced social problems, aggressive behavior, externalizing and total problems. Urinary arsenic showed an impact on anxiety and depression, somatic problems, attention problems and rule breaking behavior. A significant interaction between lead and arsenic was observed, with a synergistic effect of the two metals increasing the risk of attention problems, aggressive behavior, externalizing problems and total problems. Overall, we were able to test that higher blood lead, urinary arsenic concentrations and their interaction increase the risk of neurobehavioral problems. This is in line with the U.S. Environmental Protection Agency's priority list of hazardous substances where arsenic and lead are ranked as first and second respectively.

Neurocognitive impact of metal exposure and social stressors among schoolchildren in Taranto, Italy.

BACKGROUND: Metal exposure is a public health hazard due to neurocognitive effects starting in early life. Poor socio-economic status, adverse home and family environment can enhance the neurodevelopmental toxicity due to chemical exposure. Disadvantaged socio-economic conditions are generally higher in environmentally impacted areas although the combined effect of these two factors has not been sufficiently studied. METHODS: The effect of co-exposure to neurotoxic metals including arsenic, cadmium, manganese, mercury, lead, selenium, and to socio-economic stressors was assessed in a group of 299 children aged 6-12 years, residing at incremental distance from industrial emissions in Taranto, Italy. Exposure was assessed with biological monitoring and the distance between the home address and the exposure point source. Children's cognitive functions were examined using the Wechsler Intelligence Scale for Children (WISC) and the Cambridge Neuropsychological Test Automated Battery (CANTAB). Linear mixed models were chosen to assess the association between metal exposure, socio-economic status and neurocognitive outcomes. RESULTS: Urinary arsenic, cadmium and hair manganese resulted inversely related to the distance from the industrial emission source (ß - 0.04; 95% CI -0.06, - 0.01; ß - 0.02; 95% CI -0.05, - 0.001; ß - 0.02 95% CI -0.05, - 0.003) while the WISC intellectual quotient and its sub-scores (except processing speed index) showed a positive association with distance. Blood lead and urinary cadmium were negatively associated with the IQ total score and all sub-scores, although not reaching the significance level. Hair manganese and blood lead was positively associated with the CANTAB between errors of spatial working memory (ß 2.2; 95% CI 0.3, 3.9) and the reaction time of stop signal task (ß 0.05; 95% CI 0.02, 0.1) respectively. All the other CANTAB neurocognitive tests did not show to be significantly influenced by metal exposure. The highest socio-economic status showed about five points intellectual quotient more than the lowest level on average (ß 4.8; 95% CI 0.3, 9.6); the interaction term between blood lead and the socio-economic status showed a significant negative impact of lead on working memory at the lowest socio-economic status level (ß - 4.0; 95% CI -6.9, - 1.1). CONCLUSIONS: Metal exposure and the distance from industrial emission was associated with negative cognitive impacts in these children. Lead exposure had neurocognitive effect even at very low levels of blood lead concentration when socio-economic status is low, and this should further address the importance and prioritize preventive and regulatory interventions.

Hexavalent chromium in tattoo inks: Dermal exposure and systemic risk.

BACKGROUND: The presence of hexavalent chromium (Cr[VI]), which is carcinogenic to humans and a dermal sensitizer, in tattoo inks may represent a serious health concern. The level of this impurity is limited to 0.2 mg/kg in tattoo inks by the European Resolution ResAP(2008)1. OBJECTIVES: To analyse 29 tattoo inks, produced in Europe and the United States, of different colours and brands, for Cr(VI) to assess their conformity with ResAP(2008)1 and to characterize dermal and systemic risks. METHODS: Ion chromatography and inductively coupled plasma mass spectrometry were used to determine the levels of Cr(VI) in inks; risk characterization was performed by calculating the systemic exposure dosage (SED) and margin of safety (MoS). RESULTS: Ninety per cent of inks contained Cr(VI) (range: 0.22-4.09 mg/kg), ie, above the maximum allowed level, and no information appeared on the label. More than 1 mg/kg Cr(VI) was detected in 27.6% of inks; these might represent a possible cause of dermal adverse reactions. Exposure to Cr(VI) in inks resulted in negligible SED values and MoS values of >100 (safety threshold), indicating no appreciable systemic risk. CONCLUSIONS: The minimization of Cr(VI) contamination and the use of technology compliant with good manufacturing practices is recommended to increase the safety of tattoo inks.

ICP-MS based methods to characterize nanoparticles of TiO2 and ZnO in sunscreens with focus on regulatory and safety issues.

This study sought to develop analytical methods to characterize titanium dioxide (TiO2) and zinc oxide (ZnO) nanoparticles (NPs), including the particle size distribution and concentration, in cream and spray sunscreens with different sun protection factor (SPF). The Single Particle Inductively Coupled Plasma-Mass Spectrometry (SP ICP-MS) was used as screening and fast method to determine particles size and number. The Asymmetric Flow-Field Flow Fractionation (AF4-FFF) as a pre-separation technique was on-line coupled to the Multi-Angle Light Scattering (MALS) and ICP-MS to determine particle size distributions and size dependent multi-elemental concentration. Both methods were optimized in sunscreens in terms of recovery, repeatability, limit of detection and linear dynamic range. Results showed that sunscreens contained TiO2 particles with an average size of ≤107 nm and also a minor number of ZnO particles sized ≤98 nm. The higher fraction of particles <100 nm was observed in sunscreens with SPF 50+ (ca. 80%); the lower percentage (12-35%) in sunscreens with lower SPF values. Also the higher TiO2 (up to 24% weight) and ZnO (ca. 0.25% weight) concentrations were found in formulations of SPF 50+. Creamy sunscreens could be considered safe containing TiO2 and ZnO NPs less than the maximum allowable concentration of 25% weight as set by the European legislation. On the contrary, spray products required additional considerations with regard to the potential inhalation of NPs. The developed methods can contribute to the actual demand for regulatory control and safety assessment of metallic NPs in consumers' products.

Determination of mercury in hair: Comparison between gold amalgamation-atomic absorption spectrometry and mass spectrometry.

Mercury is a heavy metal that causes serious health problems in exposed subjects. The most toxic form, i.e., methylmercury (MeHg), is mostly excreted through human hair. Numerous analytical methods are available for total Hg analysis in human hair, including cold vapour atomic fluorescence spectrometry (CV-AFS), inductively coupled plasma mass spectrometry (ICP-MS) and thermal decomposition amalgamation atomic absorption spectrometry (TDA-AAS). The aim of the study was to compare the TDA-AAS with the ICP-MS in the Hg quantification in human hair. After the washing procedure to minimize the external contamination, from each hair sample two aliquots were taken; the first was used for direct analysis of Hg by TDA-AAS and the second was digested for Hg determination by the ICP-MS. Results indicated that the two data sets were fully comparable (median; TDA-AAS, 475ngg-1; ICP-MS, 437ngg-1) and were not statistically different (Mann-Whitney test; p=0.44). The two techniques presented results with a good coefficient of correlation (r=0.94) despite different operative ranges and method limits. Both techniques satisfied internal performance requirements and the parameters for method validation resulting sensitive, precise and reliable. Finally, the use of the TDA-AAS can be considered instead of the ICP-MS in hair analysis in order to reduce sample manipulation with minor risk of contamination, less time consuming due to the absence of the digestion step and cheaper analyses.

Associations among exposure to methylmercury, reduced Reelin expression, and gender in the cerebellum of developing mice.

Genetic risk factors acting during pregnancy or early after birth have been proposed to account for the exponential increase of autism diagnoses in the past 20 years. In particular, a potential link with exposure to environmental mercury has been suggested. Male sex constitutes a second risk factor for autism. A third potential genetic risk factor is decreased Reelin expression. Male heterozygous reeler (rl(+/-)) mice show an autism-like phenotype, including Purkinje cells (PCs) loss and behavioral rigidity. We evaluated the complex interactions between 3 risk factors, i.e. genetic status, sex, and exposure to methylmercury (MeHg), in rl(+/-) mice. Mice were exposed to MeHg during the prenatal and early postnatal period, either at a subtoxic dose (2 ppm in Dams' drinking water), or at a toxic dose (6 ppm Dams' drinking water), based on observations in other rodent species and mice strains. We show that: (a) 2 ppm MeHg does not cause PCs loss in the different animal groups, and does not enhance PCs loss in rl(+/-) males; consistent with a lack of overt neurotoxicity, 2 ppm MeHg per se does not cause behavioral alterations (separation-induced ultrasonic calls in newborns, or sociability and social preference in adults); (b) in stark contrast, 6 ppm MeHg causes a dramatic reduction of PCs number in all groups, irrespective of genotype and sex. Cytochrome C release from mitochondria of PCs is enhanced in 6 ppm MeHg-exposed groups, with a concomitant increase of µ-calpain active subunit. At the behavioral level, 6 ppm MeHg exposure strongly increases ultrasonic vocalizations in all animal groups. Notably, 6 ppm MeHg significantly decreases sociability in rl(+/-) male mice, while the 2 ppm group does not show such as decrease. At a subtoxic dose, MeHg does not enhance the autism-like phenotype of male rl(+/-) mice. At the higher MeHg dose, the scenario is more complex, with some "autism-like" features (loss of sociability, preference for sameness) being evidently affected only in rl(+/-) males, while other neuropathological and behavioral parameters being altered in all groups, independently from genotype and sex. Mitochondrial abnormalities appear to play a crucial role in the observed effects.

Priority persistent contaminants in people dwelling in critical areas of Campania Region, Italy (SEBIOREC biomonitoring study).

To investigate if protracted living in degraded environments of the Caserta and Naples provinces (Campania Region, Italy) had an impact on exposure of local people, highly toxic persistent contaminants were measured in blood, blood serum, and human milk of a large number of healthy donors. Sampling was carried out from 2008 to 2009. Blood was collected from over 850 20-64-year old donors; by pooling, 84 blood and 84 serum samples were obtained. Milk was donated by 52 mothers: specimens were pooled into six samples. Polychlorodibenzodioxins (PCDDs), polychlorodibenzofurans (PCDFs), and polychlorobiphenyls (PCBs, dioxin-like (DL) and non-dioxin-like (Σ6PCBs)), arsenic (As), cadmium (Cd), mercury (Hg), and lead (Pb) were measured in serum (organic biomarkers) and blood (metals); these chemicals and polybromobiphenyl ethers (Σ9PBDEs) were analyzed in milk. PCDD+PCDF, DL-PCB, TEQTOT, and Σ6PCB concentration ranges (medians) in serum were 6.26-23.1 (12.4), 3.42-31.7 (11.5), 10.0-52.8 (23.9) pgTEQ97/g fat, and 55.5-647 (219) ng/g fat, respectively, while in milk concentration ranges were 5.99-8.77, 4.02-6.15, 10.0-14.2 pgTEQ97/g fat, and 48.7-74.2 ng/g fat. Likewise, As, Cd, Hg, and Pb findings in blood spanned 2.34-13.4 (5.83), 0.180-0.930 (0.475), 1.09-7.60 (2.60), 10.2-55.9 (28.8) µg/L, respectively; only Pb could be measured in milk (2.78-5.99 µg/L). Σ9PBDE levels in milk samples were 0.965-6.05 ng/g fat. Biomarkers' concentrations were found to be compatible with their current values in European countries and in Italy, and consistent with an exposure primarily determined by consumption of commercial food from the large distribution system. Based on relatively higher biomarker values within the hematic biomonitoring database, the following municipalities were flagged as possibly deserving attention for health-oriented interventions: Brusciano and Caivano (As), Giugliano (Hg), Pianura (PCDDs+PCDFs), and Qualiano-Villaricca (As, Hg). The analysis of samples' qualitative variability indicated that biomarker composition was sensitive at municipality level, a feature that can potentially drive interventions for future local risk assessment and/or management measures.

Human biomonitoring for Cd, Hg and Pb in blood of inhabitants of the Sacco Valley (Italy).

INTRODUCTION: The Sacco Valley (Lazio, Italy) is characterized by high density population and several industrial chemical productions that during the time had led to a substantial amount of by-products. The result was a severe environmental pollution of the area and in particular of the river Sacco. In 1991, the analysis of water and soils samples of three industrial landfills revealed the presence of organochlorine compounds and heavy metals. A research project named "Health of residents living in Sacco Valley area", coordinated by the regional Department of Epidemiology, was undertaken and financed to evaluate the state of health of the population living near those polluted areas. MATERIALS AND METHODS: Cd, Hg and Pb were quantified in 246 blood samples of potentially exposed residents of the Sacco Valley by quadrupole inductively coupled plasma mass spectrometry (Q-ICP-MS). RESULTS: Individuals who agreed to be sampled did not exhibit high levels of the elements. The distance from the river does not seem to be directly connected with the elements levels in blood. The contribution of these contaminants to the total intake due to ingestion of food was difficult to evaluate. The unclear trend of data would require a characterization of the polluted site with environmental sampling of different matrices.

Blood thiamine, zinc, selenium, lead and oxidative stress in a population of male and female alcoholics: clinical evidence and gender differences.

INTRODUCTION: Long term alcohol abuse is associated with deficiencies in essential nutrients and minerals that can cause a variety of medical consequences including accumulation of toxic metals. AIM: The aim of this research is to get evidence-based data to evaluate alcohol damage and to optimize treatment. Thiamine and thiamine diphosphate (T/TDP), zinc (Zn), selenium (Se), lead (Pb) and oxidative stress in terms of reactive oxygen metabolites (ROMs) were examined in blood samples from 58 alcohol dependent patients (17 females and 41 males). RESULTS: T/TDP concentration in alcoholics resulted significantly lower than controls (p < 0.005) for both sexes. Serum Zn and Se did not significantly differ from reference values. Levels of blood Pb in alcoholics resulted significantly higher (p < 0.0001) than Italian reference values and were higher in females than in males. ROMs concentration was significantly higher than healthy population only in female abusers (p = 0.005). CONCLUSION: Alcoholics show a significant increase in blood oxidative stress and Pb and decrease in thiamine. Impairment occurs mainly in female abusers confirming a gender specific vulnerability.

[Epidemiologic studies on the health status of the population living in the Sacco River Valley]. / Indagini epidemiologiche sullo stato di salute della popolazione residente nell'area della Valle del Sacco.

UNLABELLED: OBIETTIVO: to analyze the health status of the population living in an area close to the Colleferro industrial plant. SETTING AND PARTICIPANTS: the area of the Sacco River Valley, Central Italy nearby Rome, has been heavily polluted over the years by industrial wastes deriving from the chemical industrial plant in Colleferro. In 2006, it was discovered that the herds of livestock were contaminated by beta-hexachlorocycloexane (ß-HCH, an industrial waste belonging, as well as lindane, to the group of hexachlorocycloexane isomers). MAIN OUTCOME MEASURES: the analyses of mortality and morbidity were carried out for the 1998-2007 period (calculation of standardized mortality ratios, SMR), and for the period 2003-2007 (calculation of standardized hospitalization ratios, SHR), respectively. The general population in the Lazio Region has been considered as reference. In addition, a biomonitoring study was conducted on a sample of the population living in 4 areas of the Sacco River Valley with different levels of exposure and the following persistent organic pollutants were measured in the blood (α, ß and γ-HCH, HCB p,p'-DDT and p,p'- DDE, 6 NDL-PCB congeners and 12 DL-PCBs, PCDDs and PCDFs), and heavy metals (Cd, Hg, Pb). RESULTS: cancer mortality in men was increased in the area (SMR=1.20), especially for specific cancer sites (stomach, larynx, lungs, pleura, myeloma); in women an excess of mortality from diabetes was detected (SMR=1.44). The analysis of morbidity indicated an excess of hospitalization for various cancers (larynx, myeloma) in men, for respiratory illness and asthma in both genders and for thyroid disease in women. The biomonitoring study found high mean concentration of ß-HCH (mean: 99.05 ng/g fat, SD=121.3), with higher levels in the population living along the river (mean=150 ng/g fat; SD=153.5), likely occurred through water and local food. CONCLUSION: the area of Colleferro has been polluted by multiple sources and the human population has been exposed to industrial chemicals, toxic substances in the workplace, and to the cumulative accumulation of organic pesticides especially through water and food.

[Exploratory biomonitoring study among workers of livestock farms of the Taranto Province]. / Studio esplorativo di biomonitoraggio tra gli allevatori delle masserie della Provincia di Taranto.

OBJECTIVE: to conduct a survey on blood levels of metals and organochlorine compounds (dioxins and PCBs) among workers of livestock farms, and to identify the factors able to explain the magnitude and variability of the measured concentrations. DESIGN: exploratory human biomonitoring study. SETTING AND PARTICIPANTS: a sample of 45 workers from farms (masserie) located in the Province of Taranto was included in the study. The farms, located between 6 and 43 km from the industrial area, are family owned and animals (mainly sheep and goats) are locally reared outdoors. The study subjects have similar eating habits, including food consumption resulting from their production. RESULTS: manganese concentrations and, to a lesser extent, arsenic, cadmium and lead are in the medium-high range of levels observed in the Italian general population. This study does not identify factors able to explain the variability in the blood levels of metals. The amount of dioxins and PCBs, on the other hand, is strongly associated both with subjects age and the distance of farm from the industrial site. CONCLUSION: the results of this survey are suggestive of research hypotheses that need to be confirmed by biomonitoring studies of adequate design and size, in particular the hypothesis that the blood level of dioxins and PCBs decreases with increasing distance from the industrial site. We recommend ad hoc studies to better characterize the exposure of farmers (also associated with agricultural activities), and of the inhabitants of Taranto, to environmental persistent pollutants, also in terms of food matrices locally produced and consumed.

Method validation for determination of cadmium and lead in offal by means of quadrupole inductively coupled plasma mass spectrometry.

Offal includes viscera and internal organs that have been removed from butchered animals, that may be either directly eaten or processed for the production of other foodstuff. Such organs are able to accumulate high concentrations of potentially toxic heavy metals posing a risk for human health when ingested. Because high levels of Cd and Pb may produce damages to humans, Commission Regulation no. 1881/2006 and its amendment established maximum levels for those two elements in edible bovine, porcine, and ovine offal. In the present study, a method based on microwave acid-assisted digestion and quadrupole inductively coupled plasma mass spectrometric quantification of Cd and Pb in offal was validated according to the EU common standards. The main parameters evaluated in the validation process were: the detection and the quantification limits (LoD, LoQ), the recovery, the repeatability, the within-laboratory reproducibility, the linearity range, and the standard measurement uncertainty. The results obtained for LoD and LoQ in microg kg(-1) were, respectively, Cd, 1.8 and 5.4; Pb, 5.1 and 15.5; meanwhile, the mean recovery was about 98% for Cd and 103% for Pb. Repeatability was around 5% for Cd and 4% for Pb. The expanded standard measurement uncertainty, expressed as percentage and with a coverage factor of 2, was estimated as follows: Cd, 4.9%; Pb, 8.7%. For both elements, the main contribution was due to the within-laboratory reproducibility of the measurements.

Calcium, copper, iron, magnesium, silicon and zinc content of hair in Parkinson's disease.

The aetiology of Parkinson's disease (PD) is still unknown, but some hypotheses have focused on the imbalances in body levels of metals as co-factors of risk. To assess whether hair could be a reliable marker of possible changes, calcium (Ca), copper (Cu), iron (Fe), magnesium (Mg), silicon (Si) and zinc (Zn) were determined in hair from 81 patients affected by PD and 17 age-matched controls. Care was taken to eliminate external contamination of the hair by thorough washing. Digestion of the matrix was achieved by an acid-assisted microwave procedure. Quantification of the elements was performed by inductively coupled plasma atomic emission spectrometry. Results indicated significantly lower levels of Fe in the hair of patients (p=0.018) compared with controls. Ca and Mg levels were slightly lower while Zn levels were higher in patients, although these differences were not significant; neither were variations in Cu and Si. Ca and Mg were at least 1.5 times higher in females than in males in both controls and patients. In addition, Ca correlated positively with Mg in both groups and in both sexes (p-value always less than 0.03), and negatively with age in patients (p<0.01). Finally, element levels did not correlate with either the duration or the severity of the disease or with anti-Parkinson treatment.

The network of the Italian laboratories: a proficiency test on the quantification of trace elements in serum.

A proficiency test on the quantification of Al, Cu, Mn, Se and Zn in serum was carried out to verify the performance of about 30 regional laboratories of the network of Italian laboratories. The exercise consisted of four runs in which the laboratories were free in choosing analytical methods to determine trace elements in freeze-dried animal serum. Laboratories performances were evaluated by the study of statistical functions as Coefficients of Variation (CV), Youden plot and Z-score value. As for Al, the results were generally characterized by good accuracy and precision, in spite of the low levels of the element (5-7 microg l(-1)). Copper determination had some problems only at low concentration (about 160 microg l(-1)--first run), in which an elevated number of anomalous data were found. Better data were achieved for Zn, for which anomalous values were mainly stragglers than outliers. Due to the low number of data for Mn (concentrations from 0.6 to 60 microg l(-1)) and Se (concentration from 45 to 106 microg l(-1)), a restricted statistical treatment was applied; for these elements high CV values were found (range from 30 to 80%). The results of this trial confirmed that participation in a proficiency test represents a benefit for all analytical laboratories. In fact, with few exceptions, most of the participants improved their performances in terms of Z-score values.

Development of methods for the quantification of essential and toxic elements in human biomonitoring.

Analytical methods using sector field ICP-MS and ICP-AES were developed for the determination of Al, Ba, Be, Bi, Ca, Cd, Co, Cr, Cu, Fe, Hg, Li, Mg, Mn, Mo, Ni, Pb, Sb, Si, Sn, Sr, Tl, V, W, Zn and Zr in cerebrospinal fluid, urine, serum and blood. Sample treatment procedures merging high sample throughput, simplicity and low contamination risk were set up. Method performances were evaluated in terms of detection limits, accuracy and precision. The limits were below 0.05 ng ml(-1) for all the elements, except for Al (all matrices), Hg (blood), Pb (blood) and Sn (serum and blood). The accuracy varied from 86% to 110% and the precision was always below 6%.

Monitoring of chemical elements and oxidative damage in patients affected by Alzheimer's disease.

The haematic concentration of 26 metals and the oxidative damage in 60 patients (20 males and 40 females) affected by Alzheimer's disease and 44 healthy individuals (33 males and 11 females) were compared. In patients, the following significant (p < or = 0.05) discrepancies were found: i) increment of Ca, Cd, Hg, Mg, Si and Sn, and decrement of Al, Co, Fe and Zn in serum; ii) higher concentrations of Cu, Li, Mn, Sn and Zr and lower of Fe, Hg, Mo in blood; iii) overproduction of oxidant species (SOS) and decrease of the anti-oxidant capacity (SAC) (p < or = 0.001, for both). Variables that, joined, better discriminated between patients and controls resulted to be Si, SOS, SAC, Co, Ca, Al in serum (94% of cases correctly classified) and Cu, Zr, Mo and Fe in blood (90% of cases properly categorized).