Platinum levels in nasal lavage fluid as a biomarker for traffic-related exposure and inflammation in children.

Platinum (Pt) is a well-known constituent of particles emitted by catalytic converters during car operation. To evaluate Pt as a potential marker for traffic related particle exposure, we investigated Pt content along with metals vanadium (V) and chromium (Cr) in coarse and fine particulate matter (PM), sampled in four areas with different traffic density, as well as in the nasal lavage (NAL) of 67 children (average age: 6 years) living in these areas. The different sites were characterised by significant differences in air pollutants including PM, NO, NO(2), CO and Cr, but differences in V or Pt were absent. No significant differences in neutrophil and epithelial cell counts or concentrations of the neutrophil chemoattractant interleukin-8 (IL-8) were found in the NAL of children living in the different areas. In addition, the concentrations of V, Cr and Pt, which were detectable in 64%, 73% and 93% of the individuals, respectively, did not differ between the different locations. However, in the NAL of the children, a significant correlation between Pt and the number of neutrophils/ml (r=0.40, p<0.001) as well as of epithelial cells/ml (r=0.41, p<0.001) was found. No relation was present between nasal inflammation and nasal Cr levels, whereas a relatively weak association was observed between V and epithelial cells counts (r=0.30, p=0.018). In conclusion, our data suggests a role for nasal lavage Pt as a candidate biomarker for traffic-related PM, which is able to induce inflammation in the upper respiratory tract.

Temporal variation of hydroxyl radical generation and 8-hydroxy-2'-deoxyguanosine formation by coarse and fine particulate matter.

AIMS: To determine the induction of 8-hydroxy-2'-deoxyguanosine (8-OHdG) by fine (<2.5 microm) and coarse (10-2.5 microm) particulate matter (PM) sampled over time at one sampling location, and to relate the observed effects to the hydroxyl radical (*OH) generating activities and transition metal content of these samples, and to meteorological parameters. METHODS: Weekly samples of coarse and fine PM were analysed for H(2)O(2) dependent *OH formation using electron spin resonance (ESR) and formation of 8-OHdG in calf thymus DNA using an immuno-dotblot assay. Immunocytochemistry was used to determine 8-OHdG formation in A549 human epithelial lung cells. To determine temporal effects, samples from six weeks in summer and six weeks in autumn/winter were compared using ESR and the dotblot assay. Concentrations of leachable V, Cr, Fe, Ni, and Cu were determined by inductively coupled plasma mass spectrometry. RESULTS: Both PM fractions elicited *OH generation as well as 8-OHdG formation in calf thymus DNA and in A549 cells. 8-OHdG formation in the naked DNA was significantly related to *OH generation, but not to metal concentrations except for copper. A significantly higher *OH generation was observed for coarse PM, but not fine PM collected during the autumn/winter season; this was not due to differences in sampled mass or metal content. Specific weather conditions under which increased *OH formation in the coarse mode was observed suggest that other, as yet unknown, anthropogenic components might affect the radical generating capacity of PM. CONCLUSIONS: Both coarse and fine PM are able to generate *OH, and induce formation of 8-OHdG. When considered at equal mass, *OH formation shows considerable variability with regard to the fraction of PM, as well as the sampling season. The toxicological implications of this heterogeneity in *OH formation by PM, as can be easily determined by ESR, need further investigation.

Concentrations of lead in blood, hair and saliva of German children living in three different areas of traffic density.

Lead contents in hair, whole blood and saliva were determined for 245 healthy children (121 male, 124 female, age: 8-10 years) from three residential areas of Düsseldorf (North-Rhine-Westphalia, Germany) with different traffic densities. The geometric mean for the lead content in hair was found to be 0.87 microg/g (range: 0.2-9.9 microg/g) for the entire test group. While the levels of lead in hair in the suburban population were significantly lower than in the two city centre populations, no significant difference concerning the lead content in hair could be detected in the latter. The geometric mean for lead concentration in whole blood amounted to 25.0 microg/l (range: 8.0-154 microg/l). There was no significant difference between the sub-groups. The lead concentrations found in saliva were rather low (range: < 1.5-47.0 microg/l). Of the values, 89% were below the detection limit of 1.5 microg/l. Due to reduced levels of lead in fuel, the present study exposes that the amount of lead in the children examined has further decreased compared to preceding surveys. The correlation between the lead content in hair and the road traffic density was not corroborated by the findings with regard to amounts of lead found in blood, indicating that residual lead from fuel does not result in a substantial burden of lead found in the whole body. In contrast to levels of lead found in blood, levels of lead found in hair may be influenced more by environmental conditions. Saliva is not a suitable material for biological monitoring with respect to lead exposure in children.

Tolerance to nickel: oral nickel administration induces a high frequency of anergic T cells with persistent suppressor activity.

We adapted our mouse model of allergic contact hypersensitivity to nickel for the study of tolerance. Sensitization in this model is achieved by the administration of nickel ions with H(2)O(2); nickel ions alone are unable to prime naive T cells, but can restimulate primed ones. A 4-wk course of oral or i.p. administration of 10 mM NiCl(2) to naive mice induced tolerance, preventing the induction of hypersensitivity for at least 20 wk; long term desensitization of nickel-sensitized mice, however, required continuous NiCl(2) administration. When splenic T cells of orally tolerized donors, even after a treatment-free interval of 20 wk, were transferred to naive recipients, as with lymph node cells (LNC), they specifically prevented sensitization of the recipients. The LNC of such donors were anergic, because upon in vivo sensitization with NiCl(2) in H(2)O(2) and in vitro restimulation with NiCl(2), they failed to show the enhanced proliferation and IL-2 production as seen with LNC of mice not tolerized before sensitization. As few as 10(2) bulk T cells, consisting of both CD4(+) and CD8(+) cells, were able to specifically transfer tolerance to nickel. A hypothesis is provided to account for this extraordinarily high frequency of nickel-reactive, suppressive T cells; it takes into account that nickel ions fail to act as classical haptens, but form versatile, unstable metal-protein and metal-peptide complexes. Furthermore, a powerful amplification mechanism, such as infectious tolerance, must operate which allows but a few donor T cells to tolerize the recipient.

Determination of dichloroanilines in human urine by GC-MS, GC-MS-MS, and GC-ECD as markers of low-level pesticide exposure.

Methods for the determination of 3,4-dichloroaniline (3,4-DCA) and 3,5-dichloroaniline (3,5-DCA) as common markers of eight non-persistent pesticides in human urine are presented. 3,5-DCA is a marker for the exposure to the fungicides vinclozolin, procymidone, iprodione, and chlozolinate. Furthermore the herbicides diuron, linuron, neburon, and propanil are covered using their common marker 3,4-DCA. The urine samples were treated by basic hydrolysis to degrade all pesticides, metabolites, and their conjugates containing the intact moieties completely to the corresponding dichloroanilines. After addition of the internal standard 4-chloro-2-methylaniline, simultaneous steam distillation extraction (SDE) followed by liquid-liquid extraction (LLE) was carried out to produce, concentrate and purify the dichloroaniline moieties. Gas chromatography (GC) with mass spectrometric (MS) and tandem mass spectrometric (MS-MS) detection and also detection with an electron-capture detector (ECD) after derivatisation with heptafluorobutyric anhydride (HFBA) were employed for separation, detection, and identification. Limit of detection of the GC-MS-MS and the GC-ECD methods was 0.03 and 0.05 microg/l, respectively. Absolute recoveries obtained from a urine sample spiked with the internal standard, 3,5-, and 3,4-DCA, ranged from 93 to 103% with 9-18% coefficient of variation. The three detection techniques were compared concerning their performance, expenditure and suitability for their application in human biomonitoring studies. The described procedure has been successfully applied for the determination of 3,4- and 3,5-DCA in the urine of nonoccupationally exposed volunteers. The 3,4-DCA levels in these urine samples ranged between 0.13 and 0.34 microg/g creatinine or 0.11 and 0.56 microg/l, while those for 3,5-DCA were between 0.39 and 3.33 microg/g creatinine or 0.17 and 1.17 microg/l.

Internal platinum, palladium, and gold exposure in environmentally and occupationally exposed persons.

In a pilot study the urinary platinum (Pt), palladium (Pd), and gold (Au) excretion was analyzed in 27 dental technicians, 17 road construction workers and 17 school-leavers using sector field inductively coupled plasma mass spectrometry (SF-ICP-MS). Detection limits in urine were 0.24 ng/l for Pt and Au and 0.17 ng/l regarding Pd. A standardized questionnaire was used to assess information about kind and degree of contact to these metals, the physical condition of the volunteers and confounding factors. Significant differences between the three study groups were found. The mean Pt, Pd, and Au excretions of the dental technicians were significantly higher than those of the road construction workers and school-leavers. This indicates that the occupational treatment of dental alloys leads to an internal exposure to these metals which is distinctly higher than that from automobile exhaust exposure. Significant differences between Monday morning (pre-shift) and Thursday afternoon (post-shift) urine samples of the dental technicians were not found. The Pt excretion of road construction workers working near a much traveled highway was comparable with that of school-leavers being less (only environmentally) exposed to automobile exhaust. Regarding Pd and Au the road construction workers showed a tendency to higher levels in urine when compared with the school-leavers, but statistically significant differences were not found. The tendency to higher urinary Pd and Au levels in the road-construction workers may be explained by their slightly greater number of noble metal containing artificial dentures, which may cause an additional exposure. A statistically significant effect of age on the urinary noble metal excretion was not detectable.

Determination of selected microbial volatile organic compounds by diffusive sampling and dual-column capillary GC-FID--a new feasible approach for the detection of an exposure to indoor mould fungi?

A new, analytically valid procedure is described to assess the exposure of human beings to the so-called microbial volatile organic compounds (MVOCs) in air. The method can be used routinely for large sample numbers and is especially valuable as a basis for further research on the correlation between single MVOCs and indoor mould growth. The procedure is based on the fact that fungi produce a variety of volatile organic compounds, such as 3-methylbutan-1-ol, 3-methylbutan-2-ol, fenchone, heptan-2-one, hexan-2-one, octan-3-one, octan-3-ol, pentan-2-ol, alpha-terpineol, and thujopsene, which they emit into the indoor environment. Using diffusive samplers, these MVOCs are adsorbed onto charcoal during a sampling interval of four weeks. The described method is thus superior to existing methods which use short-term active sampling. After desorption with carbon disulfide, the MVOCs were determined by dual-column gas chromatography with flame ionization detection using the large-volume injection technique for sample introduction. The detection limits ranged between 0.15 and 0.53 microgram m-3, within-series precision was found to range between 6.5 and 19.0%, and recovery was between 77 and 118%. The procedure has been successfully applied in the context of a large field study to measure the indoor MVOC exposure in children's rooms of 132 dwellings. The objective of the study was to examine the relation between indoor mould growth, the indoor MVOC exposure and the prevalence of adverse health effects. Information about mould formation has been obtained by a questionnaire and by the determination of colony forming units of mould fungi in mattress dust. With the exception of 3-methylbutan-2-ol, fenchone, nonan-2-one, octan-2-one, and thujopsene, indoor air concentrations of all MVOCs under investigation were significantly higher inside damp and mouldy dwellings. From the primary MVOCs under investigation, 3-methylbutan-1-ol, hexan-2-one, heptan-2-one, and octan-3-ol were found to be most reliable indicators for mould formation. A correlation was also found between selected MVOCs and the occurrence of mould species in mattress dust. Aspergillus sp. correlated with heptan-2-one, hexan-2-one, octan-3-ol, octan-3-one, and alpha-terpineol, while the occurrence of Eurotium sp. was correlated with higher indoor air concentrations of 3-methylbutan-1-ol, 3-methylbutan-2-ol, heptan-2-one, hexan-2-one, octan-3-ol, and thujopsene. Children living in dwellings with elevated MVOC levels had a higher prevalence of asthma, hay fever, wheezing, and irritations of the eyes. These positive associations persisted after controlling for confounding factors such as age, sex, body-mass index, number of siblings, social status, passive smoking, type of heating, and ventilation habits. However, they were not statistically significant. This lack of significance may be a result of the small number of investigated samples.

Long term urinary platinum, palladium, and gold excretion of patients after insertion of noble metal dental alloys.

The aim of this study was to investigate to which extent noble-metal dental alloys contribute to the total platinum (Pt), palladium (Pd), and gold (Au) body burden of the general population. The urinary Pt, Pd, and Au excretion was determined in three non-occupationally exposed volunteers before and up to 3 months after insertion of a highgold dentalalloy. The in-vitro release of Pt, Pd, and Au from four different types of dental alloys into either artificial saliva or 1% lactic acid solution was additionally investigated. The Pt, Pd, and Au concentrations were determined by sector field inductively coupled plasma mass spectrometry (SF-ICP-MS). Before insertion of the high-gold dental alloy, the Pt excretion of the patients ranged between 1.0 and 7.4 ng l-1 (0.6-3.3 ng g-1 creatinine). In the immediate post-insertion phase the Pt excretion rose to 10.5-59.6 ng l-1 (14.5-33.2 ng g-1 creatinine). This is a mean increase by a factor of 12 compared with the average Pt excretion before insertion. Three months after insertion, the Pt excretion was still elevated by a factor of 7. Contrary to Pt, the Au and Pd excretion in urine was not significantly increased after insertion of this type of high-gold dental alloy. Our in-vitro investigations confirm the assumption that Pt, Pd, and Au are released from noble metal containing dental alloys by corrosion. Under the applied conditions, the release was in the lower ng cm-2 range. It can be concluded that the Pt release from dental alloys can predominantly contribute to the Pt exposure of non-occupationally exposed persons. It can exceed the exposure from all other environmental sources including the Pt release from automobile exhaust catalysts.

Determination of physiological levels of volatile organic compounds in blood using static headspace capillary gas chromatography with serial triple detection.

A static capillary gas chromatographic method using three different detectors [photoionization detector (PID), electron capture detector (ECD) and flame ionization detector (FID)] switched in series is presented for the determination of volatile organic compounds (VOCs) in sub microgram l-1 levels. The method was applied for the analysis of selected environmentally and occupationally relevant non-halogenated and chlorinated aromatic hydrocarbons (e.g., benzene, toluene, xylenes, dichlorobenzenes) as well as chlorinated aliphatic hydrocarbons (e.g., trichloroethene, tetrachloroethene) in blood samples. Detailed investigations, in respect to the figures of merit were carried out. For most of the selected VOCs detection limits (calculated as the three-fold standard deviation of low level calibration standards) in the range from 26 (benzene) to 67 ng l-1 (m/p-xylene) were achieved which are comparable with those reported for dynamic headspace techniques in combination with mass spectrometric detection. For the individual VOCs the within-series precision varied from 4 to 19% and the day-to-day precision from 11 to 28%. Regarding PID as well as FID the calibration graphs for all substances were linear up to at least 10 micrograms l-1 while the ECD response was linear up to concentrations of about 0.6 microgram l-1 for the halogenated compounds. Our method is applicable for the quantitative determination of VOCs in blood in the occupationally as well as in the physiologically relevant (normal) concentration range.

Determination of benzene, toluene, ethylbenzene and xylenes in indoor air at environmental levels using diffusive samplers in combination with headspace solid-phase microextraction and high-resolution gas chromatography-flame ionization detection.

An improved analytical method for passive air sampling is presented based on a combination of commercially available diffusive samplers with headspace solid-phase microextraction and high-resolution gas chromatography with flame ionization detection (HRGC-FID). This procedure is targeted for short-term BTEX (benzene, toluene, ethylbenzene and o-, m- and p-xylenes) determinations at environmental concentrations and can be applied for sampling intervals between 30 min and 24 h. The analytes are adsorbed onto the charcoal pad of a passive sampler and then extracted with carbon disulphide-methanol. After removal of the carbon disulphide by xanthation, the BTEXs are enriched on a Carboxen SPME fiber, thermally desorbed and analysed by HRGC-FID. Detection limits for a sampling interval of 2 h are between 0.4 and 2 micrograms/m3, within-series precision ranges between 6.6 and 12.8%, day-to-day precision is between 11.1 and 15.2%. The results obtained with this procedure are validated by comparison with active sampling. Detection limits and a further reduction of the sampling time are limited by blanks of the chemicals and the diffusive samplers. Procedures to eliminate these blanks are described in detail. Applications such as the determination of BTEXs in indoor air inside buildings, inside a train and a car are presented, indicating the usefulness of the described procedure for short-term measurements of environmental BTEX concentrations. An advantage of passive samplers is the storage stability for at least six months, which is essential for its use in large epidemiological studies.

Determination of polychlorinated biphenyls and chlorinated pesticides in human body fluids and tissues.

A fast and reliable method for the determination of polychlorinated biphenyls (PCBs) and chlorinated pesticides in human body fluids and tissues is presented. Sample clean-up and selective enrichment of analytes are carried out by liquid column chromatography without a prior solvent extraction step. Analytes are determined either by high-resolution capillary gas chromatography with mass spectrometric detection, or by dual-column GC followed by electron-capture detection. A procedure is described for the simultaneous determination of environmental levels of the PCB congeners 28, 52, 101, 138, 153 and 180 in human cord serum. The method allows the simultaneous determination of chlorinated pesticides such as aldrin, DDT, DDE, BHC and HCB and is also applicable to other biological matrices, such as bone marrow or tissues.

Internal and external tetrachloroethene exposure of persons living in differently polluted areas of Northrhine-Westphalia (Germany).

An epidemiological study was performed to measure the internal and external tetrachloroethene exposure of persons living in two differently polluted areas of Northrhine-Westphalia (Germany). Tetrachloroethene concentrations were determined in venous blood samples of 5- to 7-year-old children (n = 81) and 55-year-old women (n = 91) living in Essen, an industrial city located in the Ruhr area. 103 children und 131 women of the same age living in Borken, a small town north of the Ruhr area, served as reference group. Outdoor air samples were collected on passive samplers (sampling period: 4 weeks) from 70 measurement points per study area (about 2 km2, mean distance 100 m). In the course of a year these measurements were repeated three times to cover seasonal variations. Parallel to the outdoor measurement periods, indoor air concentrations were determined in the homes of those women from Essen and Borken, who donored a blood sample. Tetrachloroethene levels in blood were generally low with a geometric mean of 0.05 microgram/L in women and 0.021 microgram/L in children. Nevertheless, children and women living in the industrial area were found to have significantly higher tetrachloroethene levels in blood than those of the reference group. In both study areas blood levels of women exceeded those of children by a factor of 2. Participants living in the neighbourhood of a dry-cleaning shop had distinctly elevated blood levels. The same applied to persons who stored dry-cleaned clothes at home. Like the internal exposure, external exposure was also higher in Essen than in Borken. In both areas tetrachloroethene concentrations indoors exceeded those outdoors. Outdoor tetrachloroethence concentrations were significantly increased during the cold season, while the opposite was true for indoor levels. The correlation between indoor and outdoor exposure was found to be significant, while those between blood levels and outdoor exposure became only significant when people living next to a dry-cleaning shop were excluded. No significant relationship was observed between blood and indoor tetrachlorethene levels. It is concluded that the higher tetrachloroethene blood levels of the urban population result from the higher atmospheric concentrations in industrial areas with tetrachloroethene emitting sources like metal and textile industry. The fact that indoor air tetrachloroethene levels exceeded those outdoors can only be explained by the presence of additional indoor sources. Provided that women spend on average more time indoors than children the higher indoor air concentrations may be the reason for the higher blood tetrachloroethene levels found in women. Persons living near a dry-cleaning shop or storing dry-cleaned clothes at home showed a higher internal and external exposure to tetrachloroethene than other persons. In individual cases it can by far exceed the average exposure of the general population, so that health impairments can not be generally excluded.

Long-term mercury excretion in urine after removal of amalgam fillings.

The long-term urinary mercury excretion was determined in 17 28- to 55-year-old persons before and at varying times (up to 14 months) after removal of all (4-24) dental amalgam fillings. Before removal the urinary mercury excretion correlated with the number of amalgam fillings. In the immediate post-removal phase (up to 6 days after removal) a mean increase of 30% was observed. Within 12 months the geometric mean of the mercury excretion was reduced by a factor of 5 from 1.44 micrograms/g (range: 0.57-4.38 micrograms/g) to 0.36 microgram/g (range: 0.13-0.88 microgram/g). After cessation of exposure to dental amalgam the mean half-life was 95 days. These results show that the release of mercury from dental amalgam contributes predominantly to the mercury exposure of non-occupationally exposed persons. The exposure from amalgam fillings thus exceeds the exposure from food, air and beverages. Within 12 months after removal of all amalgam fillings the participants showed substantially lower urinary mercury levels which were comparable to those found in subjects who have never had dental amalgam fillings. A relationship between the urinary mercury excretion and adverse effects was not found. Differences in the frequency of effects between the pre- and the post-removal phase were not observed.

Internal lead and cadmium exposure in 6-year-old children from western and eastern Germany.

Lead and cadmium levels in blood and deciduous teeth (shed incisors only) of 6-year-old German children were determined in 1991 in a large epidemiological study carried out in rural and urban areas of western Germany (Duisburg, Essen, Gelsenkirchen, Dortmund, Borken) and eastern Germany (Leipzig, Halle, Magdeburg, Osterburg, Gardelelegen, Salzwedel). In total, blood lead and cadmium levels of 2311 German children and tooth lead and cadmium levels of 790 German children were analyzed. Blood lead levels were generally low in all study areas with geometric means between 39.3 micrograms/l and 50.8 micrograms/l in the western German and between 42.3 micrograms/l and 68.1 micrograms/l in the eastern German study areas. The mean blood lead level of Turkish children (n = 213) living in the western German study areas was 50.1 micrograms/l and thus 5.6 micrograms/l higher than the overall geometric mean of the western German children. The higher exposure may be explained by a higher oral uptake from food and different living conditions. These children were excluded from multiple regression analysis because they were all living in the western study areas. The mean tooth lead levels ranged between 1.50 and 1.74 micrograms/g in the western and between 1.51 micrograms/g and 2.72 micrograms/g in the eastern study areas. Thus, they show a distribution pattern similar to blood. Blood and tooth lead levels were higher in urban than in rural areas and higher in the eastern German than in the western German study areas. With regard to the blood and tooth cadmium concentrations, no significant differences between the study areas could be found.(ABSTRACT TRUNCATED AT 250 WORDS)

Occupational chronic exposure to organic solvents. XIII. Glycolether exposure during the production of varnishes.

Seventeen persons (2 women and 15 men), who were exposed to glycolethers in a varnish production plant, were examined according to their external and internal solvent exposure. The workers in the production plant (n = 12) were exposed to average concentrations of ethoxyethanol, ethoxyethyl acetate, butoxyethanol, 1-methoxypropanol-2, 2-methoxypropyl-1-acetate and xylene of 2.8; 2.7; 1.1; 7.0; 2.8 and 1.7 ppm. In the air of the store (n = 3) and in the laboratory (n = 2) only minor concentrations of xylene respectively xylene and ethoxyethyl acetate could be measured. Internal exposure was estimated by measuring butoxyethanol (BE) in blood as well as ethoxyacetic acid (EAA) and butoxyacetic acid (BAA) in urine samples. Urine samples were taken pre- and post-shift. As expected, the highest values were found in the varnish production. The average post shift concentrations of BE, EAA and BAA were 121.3 micrograms/l; 167.8 and 10.5 mg/l. The relatively high concentrations of EAA and BAA in pre-shift samples can be explained by the long half-lives of these metabolites. According to our findings most of the glycolethers were taken up through the skin. Comparing our results with those reported in the literature we think that a future tolerable limit value for the concentration of ethoxyacetic acid in urine should be in the order of 100 to 200 mg/l.