Elemental status on early mediaeval skeletal remains: Diet reconstruction

S196

Abstracts / Toxicology Letters 205S (2011) S180–S300

Method: Samples were taken every 15 days from August 2009 to August 2010 from a wastewater treatment plant. Trace elements were measured by Absorption Atomic Spectrometry (AAS) after sample incineration and acid digestion. Results: Mean values in mg·kg−1 dry weight and standard deviation were: Zn 460.03 ± 115.20; Cu 75.81 ± 13.17; Ni 13.93 ± 2.12; Cr 33.22 ± 3.31; Pb 46.71 ± 5.40 and Cd 2.34 ± 0.91. Monitoring shows that sewage sludges collected during the autumn and winter seasons present higher metal concentrations than those from the spring and summer seasons. All metal levels in the studied sewage sludges are below the values allowed by European and Spanish legislation. Sewage sludges from this wastewater treatment plant present ideal characteristics to be used as a fertilizer for agriculture, forests, green areas and reclaimed lands, and would not produce environmental health problems on soils. In addition, this study suggests a possibility of commercial benefit for the studied water treatment plant. doi:10.1016/j.toxlet.2011.05.675

P2052 The modifying effect of NOS3 variants on the association between blood lead and hypertension F. Scinicariello 1,∗ , A. Yesuprya 2 , B.A. Fowler 1 , M. Chang 2 , N. Dowling 2 1

Division of Toxicology, CDC/ATSDR, Atlanta, USA, 2 CDC, Atlanta, USA

Nitric oxide (NO) has an important role in the regulation of blood pressure, and there have been reports of significant negative correlations of NO activity and blood lead level (BLL). Previous studies have found that carriers of the NOS3 rs2070744C allele who have been exposed to lead have significantly lower NO activity. This decreased NO activity may increase arterial stiffness and, consequently, the risk of hypertension. We aim to assess whether variants (rs2070744 and rs1799983) in the NOS3 gene modify the association between blood lead levels and risk of hypertension. Multivariable regressions models were used to examine if the NOS3 variants modify the association between BLLs and risk of hypertension in the three major U.S. racial/ethnic groups in the NHANES III DNA bank, 1991–1994 (n = 6016). In non-Hispanic blacks, within the highest quartile of lead (3.8–52.9 ␮g/dL), the prevalence of hypertension was significantly higher among carriers of the NOS3 rs2070744C allele (POR = 2.02, p = 0.02) compared to NOS3 rs2070744 TT homozygote individuals. An inverse correlation was found between BLL and hypertension and systolic blood pressure (SBP) in non-Hispanic whites (NHW) carrying the T allele of NOS3 rs1799983, compared to those homozygous for the G allele. The SBP difference was significantly greater only at lower levels of BLL (≤1.00 ␮g/dL). At the lowest lead quartile (0.7–1.4 ␮g/dL), NHW carrying the T allele had a significantly higher prevalence of hypertension compared to individuals homozygous for the G allele (p = 0.004). Our data suggest that NOS3 variants may modify the association between BLL and hypertension. doi:10.1016/j.toxlet.2011.05.676

P2053 Evaluation of DNA damage by comet assay in metal exposed Turkish population T. Söylemezo˘glu 1,∗ , Z. Kayaalti 2 , E. Söylemez 3 , A. Bacaksız 3 , E. Tutkun 4 1 Forensic Chemistry and Forensic Toxicology, Forensic Sciences, Ankara, Turkey, 2 Forensic Genetic, Forensic Sciences, Ankara, Turkey, 3 Forensic Sciences, Ankara, Turkey, 4 Ankara Occupational Diseases, Ankara, Turkey

Metals are known to be genotoxic and potential mediators of DNA damage such as DNA strand breaks, DNA cross-links with proteins and radical products. Individuals who work the metal industries exposed to metals throughout their working lifetime. DNA single-strand breaks can be determined by the Comet assay which is a rapid and sensitive method. The aim of this study was to assess the occupational risk of workers exposed to metals. For this purpose, 122 whole blood samples from 61 workers and 61 control volunteers were studied by comet assay and evaluated according to four image-analysis parameters including tail intensity (TI), tail DNA (DNAt ), tail moment (TM) and olive tail moment (OTM). According to our results, mean of TI, DNAt , TM and OTM in exposed group had higher than in control group and statistically significant association was found between the exposed workers and control group (p < 0.01). Smoker-exposed group had significantly higher DNA damage than smoker-control groups (p < 0.01). Mean of TI, DNAt , TM and OTM in non-smoker exposed group had higher than in non-smoker control group and all of them were significant (p < 0.01). When the smoker exposed group and non-smoker exposed group compared, only OTM was statistically significant (p < 0.05). In control group, all of the studied comet parameters were statistically significant between the smoker and non-smoker (p < 0.01). Consequently, our results showed that the comet assay is a suitable biomarker in the determination of DNA damage and a markedly increased DNA damage was observed in the exposed group according to the controls. doi:10.1016/j.toxlet.2011.05.677

P2054 Elemental status on early mediaeval skeletal remains: Diet reconstruction D. Sutlovic 1,∗ , A. Stipisic 2 , M. Versic 3 , Z. Knezovic 2 , J. Marusic 2 1

Forensic Medicine, University Hospital and School of Medicine Split, Split, Croatia, 2 Public Health Institute of Split-Dalmatian County, Split, Croatia, 3 Forensic Medicine, School of Medicine Split, Split, Croatia The total absorbed metals dose could be objectively determined by checking the element status in biological samples. Content of heavy metals in a diet could correlate with heavy metals content in human bones; therefore, determining heavy metals concentrations, and their relationship in human bone, could be used to reconstruct the basic diet. One of the goals of this study was to testing metal concentration levels of the bone material excavated from ancient burials for better understanding of medieval living habits. The aim of our study was to determine metal content of 100 mediaeval individuals excavated from Ostrovica and Naklice burial sites (Southern Croatia) and 30 recent human bones. After microwave digestion (CEM, USA Model Mars 5-2004) element content of manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), calcium (Ca) and stron-

Abstracts / Toxicology Letters 205S (2011) S180–S300

tium (Sr) were measured with an Atomic Absorption Spectrometer. Mercury concentration was determined by direct consecutive measured by mercury analyzer. When comparing results from ancient to the modern bone heavy metal concentrations, it is obvious that Cu, Ca and Sr greatly exceeded the values while concentration of Pb, Cd and Hg was lower. Concentration of Sr and Ca was a few times higher than in modern bone samples. In correlation with elemental contents of various main food components known to have been used during the Early Middle Ages, we could reconstruct a menu. With exception of milk, all main dietary components, such as roots and tubers, meat, cereals, legumes and leafy vegetables are rich in Zn, especially the vegetables. doi:10.1016/j.toxlet.2011.05.678

P2055 Increase in blood heavy metals in early pregnancy induces hypertension in pregnant women M. Vigeh 1,∗ , K. Yokoyama 2 , A. Atsuko Shinohara 2 , M. Afshinrokh 3 1

Department of Hazard Assessment, National Institute of Occupational Safety and Health, Kawasaki, Japan, 2 Department of Epidemiology and Environmental Health, Juntendo University Faculty of Medicine, Tokyo, Japan, 3 Deputy For Health, University of Tehran, Tehran, Iran Purpose: Many studies have focused on the health effects of low levels of lead and other toxic metals after the exposure amount decreased in the past decades. To clarify the effects of trace metals on the pregnancy outcomes we have conducted a longitudinal study on apparently healthy pregnant women since 2007. Methods: Pregnant women, who referred to the three research hospitals for prenatal care were asked to participate in the survey. As pregnancy induced-hypertension develops several weeks before its clinical manifestation, we collected blood samples as early as possible (8–12 weeks of gestation) to examine the causal relationship between increased in metal concentrations and development of pregnancy hypertension. Three blood samples (one for each trimester) of 364 singleton pregnant women, aged 16–35 years, were collected and then analyzed using inductively coupled plasma-mass spectrometry. Results: Average whole blood level of lead and antimony were 38.1 and 5.2 ␮g/L, respectively. Blood antimony levels were significantly correlated with systolic blood pressure either at first trimester or at the time of delivery. This finding is inconsistent with our pervious study (2003–2004) and suggests that heavy metal (antimony) could induce pregnancy hypertension. doi:10.1016/j.toxlet.2011.05.679

S197

Methods in toxicology P2056 A novel dynamic gas mixing system to assess the inflammatory response of BTXE exposure on human lung cells at low concentrations S. Achard 1,∗ , M. Delaval 1 , J. Dugay 2 , V. Pichon 2 , T. Krebs 3 , I. Momas 1 , N. Seta 1 1

Laboratoire De Santé Publique Et Environnement Ea4064, Université Paris Descartes, Paris, France, 2 Laboratoire Sciences Analytiques, Bioanalytiques Et Miniaturisation, Umr Pesca 7195, Ecole Supérieure de Physique et Chimie Industrielles, Paris, France, 3 Vitrocell Systems, Waldkirch, Germany Purpose: Toluene is frequently found in domestic environments at low levels. To study the potential of inflammatory response of epithelial lung cells to toluene exposure the generation of stable atmospheres with reproducible results is the first essential step. Our aim was to validate the use of the Vitrocell® Spiking System (VSS), a novel gas mixing system which was developed for our purpose. The system was used to expose human lung cells to toluene atmospheres at environmental levels (20–200 ␮g/m3 ) in a controlled manner. Method: The VSS allows the generation of the gaseous atmosphere dynamically and automatically by means of a digital syringe pump. The pollutant in liquid form is injected in a heated air mixing block with high accuracy. Programmable temperature control allows the vaporization of the tested compound according to its boiling point. A secondary dilution with synthetic clean air allows obtaining low concentrations. The obtained gas mixture was captured on Tenax® adsorbent resins at the exit of the VSS. After thermal desorption, the sample was introduced by means of a GERSTEL MultiPurpose Sampler into a gas chromatograph combined with mass spectrometry detection. The atmosphere generated by the VSS was then used to expose A549 cells in the Vitrocell® air/liquid interface exposure module as described previously. Results: Our preliminary results indicate that generation of controlled toluene atmosphere at environmental levels using the VSS is feasible, simple and adapted to air/liquid cellular exposure. Further testing pollutant generation for reproducibility and reliable dose–response establishment is in progress. doi:10.1016/j.toxlet.2011.05.680

P2057 Determination of morphine, 6-acetylmorphine, amphetamines, tramadol, and metabolites in hair by LC–MS–MS M.G. Ahmed Forensic Lab, Medicolegal Authority, Ministry of Justic, Egypt, Cairo, Egypt Determination of morphine, 6-acetylmorphine, amphetamines, tramadol, and metabolites in hair by LC–MS–MS Mohammed Gaber Ahmed Medicolegal Authority, Ministry of Justice, Egypt Hair testing was developed to understand the history of a drug-abuser. It could also be applied for public health purpose. Because the lower detection limit, developing a hair testing method using liquid chromatography/tandem mass spectrometry (LC–MS–MS) is better than gas chromatography/mass spectrometry. We have suc-