Exploring a new method for the assessment of metal exposure by analysis of exhaled breath of welders.

PURPOSE: Air monitoring has been the accepted exposure assessment of toxic metals from, e.g., welding, but a method characterizing the actual dose delivered to the lungs would be preferable. Sampling of particles in exhaled breath can be used for the biomonitoring of both endogenous biomarkers and markers of exposure. We have explored a new method for the sampling of metals in exhaled breath from the small airways in a study on welders. METHODS: Our method for particle sampling, Particles in Exhaled Air (PExA®), is based on particle counting and inertial impaction. We applied it on 19 stainless steel welders before and after a workday. In parallel, air monitoring of chromium, manganese and nickel was performed as well as blood sampling after work. RESULTS: Despite substantial exposure to welding fumes, we were unable to show any significant change in the metal content of exhaled particles after, compared with before, exposure. However, the significance might be obscured by a substantial analytical background noise, due to metal background in the sampling media and possible contamination during sampling, as an increase in the median metal contents were indicated. CONCLUSIONS: If efforts to reduce background and contamination are successful, the PExA® method could be an important tool in the investigations of metals in exhaled breath, as the method collects particles from the small airways in contrast to other methods. In this paper, we discuss the discrepancy between our findings and results from studies, using the exhaled breath condensate (EBC) methodology.

Fluorene exposure among PAH-exposed workers is associated with epigenetic markers related to lung cancer.

OBJECTIVES: Exposure to high-molecular-weight polycyclic aromatic hydrocarbons (PAHs) may cause cancer in chimney sweeps and creosote-exposed workers, however, knowledge about exposure to low-molecular-weight PAHs in relation to cancer risk is limited. In this study, we aimed to investigate occupational exposure to the low-molecular-weight PAHs phenanthrene and fluorene in relation to different cancer biomarkers. METHODS: We recruited 151 chimney sweeps, 19 creosote-exposed workers and 152 unexposed workers (controls), all men. We measured monohydroxylated metabolites of phenanthrene and fluorene in urine using liquid chromatography coupled to tandem mass spectrometry. We measured, in peripheral blood, the cancer biomarkers telomere length and mitochondrial DNA copy number using quantitative PCR; and DNA methylation of F2RL3 and AHRR using pyrosequencing. RESULTS: Median PAH metabolite concentrations were higher among chimney sweeps (up to 3 times) and creosote-exposed workers (up to 353 times), compared with controls (p<0.001; adjusted for age and smoking). ∑OH-fluorene (sum of 2-hydroxyfluorene and 3-hydroxyfluorene) showed inverse associations with percentage DNA methylation of F2RL3 and AHRR in chimney sweeps (B (95% CI)=-2.7 (-3.9 to -1.5) for F2RL3_cg03636183, and -7.1 (-9.6 to -4.7) for AHRR_cg05575921: adjusted for age and smoking), but not in creosote-exposed workers. In addition, ∑OH-fluorene showed a 42% mediation effect on the inverse association between being a chimney sweep and DNA methylation of AHRR CpG2. CONCLUSIONS: Chimney sweeps and creosote-exposed workers were occupationally exposed to low-molecular-weight PAHs. Increasing fluorene exposure, among chimney sweeps, was associated with lower DNA methylation of F2RL3 and AHRR, markers for increased lung cancer risk. These findings warrant further investigation of fluorene exposure and toxicity.

Cancer-related proteins in serum are altered in workers occupationally exposed to polycyclic aromatic hydrocarbons: a cross-sectional study.

Exposure to some polycyclic aromatic hydrocarbons (PAH) increases the risk of cancer and is common particularly for workers in occupations such as chimney sweeping. In exposed workers, screening of early cancer-related markers provides important information to identify individuals at risk. Here, we aimed to elucidate the associations between PAH exposure and serum levels of cancer-related proteins in 118 chimney sweeps and 126 occupationally unexposed controls, all non-smoking males from Sweden. Monoydroxylated metabolites of pyrene, phenanthrene, benzo[a]pyrene and benzo[a]anthracene were measured in urine using liquid chromatography coupled to tandem mass spectrometry and 90 cancer-related proteins were measured in serum using a proximity extension assay. Linear regression analysis adjusted for age and body mass index, and false discovery rate (FDR) identified 17 serum proteins that were differentially expressed (16 upregulated and 1 downregulated) in chimney sweeps compared with controls (FDR < 0.05). Concentrations of the peptidase kallikrein 13 (KLK13) showed significant positive associations with urinary concentrations of the PAH metabolites 3-hydroxybenzo[a]pyrene (3-OH-BaP) [B, 95% confidence interval (CI): 0.042, 0.008-0.076] and 3-hydroxybenzo[a]anthracene (3-OH-BaA) (B, 95% CI: 0.068, 0.002-0.134). Moreover, dose-response relationships were observed between KLK13 and 3-OH-BaP (trend test P = 0.027) and 3-OH-BaA (P = 0.035). Pathway and gene ontology analyses showed that cell movement, cell adhesion and cell migration were the predominant molecular functions associated with the top differentially expressed proteins. In conclusion, we found a number of putative cancer-related proteins differentially expressed in workers exposed to PAH. This warrants effective measure to reduce PAH exposure among workers as well as further investigation to confirm these findings.

Filaggrin variations are associated with PAH metabolites in urine and DNA alterations in blood.

Dermal chemical exposure is common in many professions. The filaggrin protein is important for the skin barrier and variations in the filaggrin gene (FLG) may influence the uptake of chemicals via the skin, and consequently, the degree of systemic effects. The aim of this study was to investigate, in chimney sweeps with occupational exposure to polycyclic aromatic hydrocarbons (PAH) from soot, the influence of variation in FLG on internal PAH dose and DNA alterations, including epigenetic, previously linked to cancer and cardiovascular disease. We used TaqMan PCR to genotype 151 chimney sweeps and 152 controls for four FLG null variants (R501X, R2447X, S3247X and 2282del4) which cause impaired skin barrier, and FLG copy number variation (12th repeat, CNV12) which potentially is beneficial for the skin barrier. The internal dose of PAH was represented by urinary PAH metabolites (e.g. 1-hydroxypyrene and 3-hydroxybenzo[a]pyrene) that we measured by LC-MS/MS. We measured epigenetic alterations (methylation of AHRR and F2RL3) in blood by pyrosequencing; and DNA alterations (telomere length and mitochondrial DNA copy number) by real-time PCR. Hypomethylation of AHRR or F2RL3 is a risk factor for lung cancer and shorter telomere length a risk factor for cardiovascular disease. The frequencies of FLG null were 8.6 and 11.8% (p = 0.35), and CNV12 27.8 and 19.7% (p = 0.09) in chimney sweeps and controls, respectively. We found that among chimney sweeps working predominately with soot sweeping (high PAH exposure), CNV12 carriers had lower concentrations of PAH metabolites in urine compared with non-carriers (median 1-hydroxypyrene = 0.37 vs 0.86 µg/g creatinine respectively; p = 0.025 by linear regression models adjusted for age, BMI and smoking) compared to sweeps not carrying CNV12. Further, FLG null was associated with approximately 2.5% higher methylation of F2RL3 (cg03636183, p = 0.019 after adjustment for exposure group, age, BMI and smoking). FLG null was associated with approximately 7% shorter telomere length (p = 0.015, adjusted model). Our results suggest that FLG variations may influence the dose of PAH in highly exposed workers, possibly via dermal uptake. It also suggests that FLG variation may influence the degree of (epi)genotoxicity in the body. FLG variation is common in the working population and should be considered in risk assessment.

DNA methylation of the cancer-related genes F2RL3 and AHRR is associated with occupational exposure to polycyclic aromatic hydrocarbons.

Some polycyclic aromatic hydrocarbons (PAH) are known carcinogens and workplace PAH exposure may increase the risk of cancer. Monitoring early cancer-related changes can indicate whether the exposure is carcinogenic. Here, we enrolled 151 chimney sweeps, 152 controls and 19 creosote-exposed male workers from Sweden. We measured urinary PAH metabolites using LC/MS/MS, the cancer-related markers telomere length (TL) and mitochondrial DNA copy number (mtDNAcn) using qPCR, and DNA methylation of lung cancer-related genes F2RL3 and AHRR using pyrosequencing. The median 1-hydroxypyrene (PAH metabolite) concentrations were highest in creosote-exposed workers (8.0 µg/g creatinine) followed by chimney sweeps (0.34 µg/g creatinine) and controls (0.05 µg/g creatinine). TL and mtDNAcn did not differ between study groups. Chimney sweeps and creosote-exposed workers had significantly lower methylation of AHRR CpG site cg05575921 (88.1 and 84.9%, respectively) than controls (90%). Creosote-exposed workers (73.3%), but not chimney sweeps (76.6%) had lower methylation of F2RL3 cg03636183 than controls (76.7%). Linear regression analyses showed that chimney sweeps had lower AHRR cg05575921 methylation (B = -2.04; P < 0.057, adjusted for smoking and age) and lower average AHRR methylation (B = -2.05; P < 0.035), and non-smoking chimney sweeps had lower average F2RL3 methylation (B = -0.81; P < 0.042, adjusted for age) compared with controls. These cancer-related markers were not associated with urinary concentrations of PAH metabolites. In conclusion, although we found no associations with PAH metabolites in urine (short-term exposure), our results suggest dose-response relationship between PAH exposure and DNA hypomethylation of lung cancer-related loci. These findings indicate that further protective measures should be taken to reduce PAH exposure.

Toluene diisocyanate exposure and autotaxin-lysophosphatidic acid signalling.

Toluene diisocyanate (TDI) is a reactive chemical used in manufacturing plastics. TDI exposure adversely affects workers' health, causing occupational asthma, but individuals differ in susceptibility. We recently suggested a role for signalling mediated by the enzyme autotaxin (ATX) and its product, lysophosphatidic acid (LPA), in TDI toxicity. Here we genotyped 118 TDI-exposed workers for six single-nucleotide polymorphisms (SNPs) in genes encoding proteins implicated in ATX-LPA signalling: purinergic receptor P2X7 (P2RX7), CC motif chemokine ligand 2 (CCL2), interleukin 1ß (IL1B), and caveolin 1 (CAV1). Two P2RX7 SNPs (rs208294 and rs2230911) significantly modified the associations between a biomarker of TDI exposure (urinary 2,4-toluene diamine) and plasma LPA; two IL1B SNPs (rs16944 and rs1143634) did not. CAV1 rs3807989 modified the associations, but the effect was not statistically significant (p = 0.05-0.09). In vitro, TDI-exposed bronchial epithelial cells (16HBE14o-) rapidly released ATX and IL-1ß. P2X7 inhibitors attenuated both responses, but confocal microscopy showed non-overlapping localizations of ATX and IL-1ß, and down-regulation of CAV1 inhibited the ATX response but not the IL-1ß response. This study indicates that P2X7 is pivotal for TDI-induced ATX-LPA signalling, which was modified by genetic variation in P2RX7. Furthermore, our data suggest that the TDI-induced ATX and IL-1ß responses occur independently.

Exposure, respiratory symptoms, lung function and inflammation response of road-paving asphalt workers.

BACKGROUND: Controversy exists as to the health effects of exposure to asphalt and crumb rubber modified (CRM) asphalt, which contains recycled rubber tyres. OBJECTIVE: To assess exposures and effects on airway symptoms, lung function and inflammation biomarkers in conventional and CRM asphalt road pavers. METHODS: 116 conventional asphalt workers, 51 CRM asphalt workers and 100 controls were investigated. A repeated-measures analysis included 31 workers paving with both types of asphalt. Exposure to dust, nitrosamines, benzothiazole and polycyclic aromatic hydrocarbon (PAH) was measured in worksites. Self-reported symptoms, spirometry test and blood sampling were conducted prework and postwork. Symptoms were further collected during off-season for asphalt paving. RESULTS: Dust, PAHs and nitrosamine exposure was highly varied, without difference between conventional and CRM asphalt workers. Benzothiazole was higher in CRM asphalt workers (p<0.001). Higher proportions of asphalt workers than controls reported eye symptoms with onset in the current job. Decreased lung function from preworking to postworking was found in CRM asphalt workers and controls. Preworking interleukin-8 was higher in CRM asphalt workers than in the controls, followed by a decrement after 4 days of working. No differences in any studied effects were found between conventional and CRM asphalt paving. CONCLUSION: CRM asphalt workers are exposed to higher benzothiazole. Further studies are needed to identify the source of nitrosamines in conventional asphalt. Mild decrease in lung function in CRM asphalt workers and work-related eye symptoms in both asphalt workers were observed. However, our study did not find strong evidence for severe respiratory symptoms and inflammation response among asphalt workers.

Occupational exposure to particles and mitochondrial DNA - relevance for blood pressure.

BACKGROUND: Particle exposure is a risk factor for cardiovascular diseases. Mitochondrial DNA (mtDNA) is a primary target for oxidative stress generated by particle exposure. We aimed to elucidate the effects of occupational exposure to particle-containing welding fumes on different biomarkers of mtDNA function, and in turn, explore if they modify the association between particle exposure and cardiovascular response, measured as blood pressure. METHODS: We investigated 101 welders and 127 controls (all non-smoking males) from southern Sweden. Personal sampling of the welders' exposure to respirable dust was performed during work hours (average sampling time: 6.8 h; range: 2.4-8.6 h) and blood pressure was measured once for each subject. We measured relative mtDNA copy number by quantitative PCR and methylation of the mitochondrial regulatory region D-loop and the tRNA encoding gene MT-TF by bisulfite-pyrosequencing. We calculated the relative number of unmethylated D-loop and MT-TF as markers of mtDNA function to explore the modification of mtDNA on the association between particle exposure and blood pressure. General linear models were used for statistical analyses. RESULTS: Welders had higher mtDNA copy number (ß = 0.11, p = 0.003) and lower DNA methylation of D-loop (ß = -1.4, p = 0.002) and MT-TF (ß = -1.5, p = 0.004) than controls. Higher mtDNA copy number was weakly associated with higher personal respirable dust exposure among welders with exposure level above 0.7 mg/m3 (ß = 0.037, p = 0.054). MtDNA function modified the effect of welding fumes on blood pressure: welders with low mtDNA function had higher blood pressure than controls, while no such difference was found in the group with high mtDNA function. CONCLUSION: Increased mtDNA copy number and decreased D-loop and MT-TF methylation were associated with particle-containing welding fumes exposure, indicating exposure-related oxidative stress. The modification of mtDNA function on exposure-associated increase in blood pressure may represent a mitochondria-environment interaction.

Chimney sweeps in Sweden: a questionnaire-based assessment of long-term changes in work conditions, and current eye and airway symptoms.

OBJECTIVES: To explore chimney sweeping work tasks, chimney sweeps' use of protective equipment, and type of fuel used by clients, over time. Further, to assess work-relatedness of current eye and airway symptoms. METHODS: In a cross-sectional study in 2011, male Swedish chimney sweeps (n = 483; age 21-69 years) answered a questionnaire about their occupational history and eye and airway symptoms. RESULTS: Between 1960 and 2010, black-soot-sweeping in private homes was the major task, although it decreased during the time period, for chimney sweeps. Between 1975 and 2010, the use of petroleum oil decreased, whereas the use of pellets and wood increased. Also, the use of gloves and masks increased significantly. Black-soot-sweeping in industry was associated with work-related eye symptoms (prevalence odds ratio POR = 3.76, 95% CI: 1.72-8.24, for every 10% increment of working time, adjusted for age and tobacco smoking). Chimney sweeps also had slightly higher prevalence of cough with increasing black-soot-sweeping (POR = 1.06, 95% CI: 0.99-1.13 for every 10% increment, further adjusted for the use of mask), and the association was more pronounced, although nonsignificant, for black-soot-sweeping in industry (adjusted POR = 1.26, 95% CI: 0.98-1.61). CONCLUSIONS: Chimney sweeping tasks and use of protective equipment as well as type of fuel used by the clients changed significantly over the last 35 years, which may have changed chimney sweeps' exposure to soot. Still, chimney sweeps in Sweden have black-soot-sweeping-related eye and airway symptoms.

Carbon Nanotube Emissions from Arc Discharge Production: Classification of Particle Types with Electron Microscopy and Comparison with Direct Reading Techniques.

INTRODUCTION: An increased production and use of carbon nanotubes (CNTs) is occurring worldwide. In parallel, a growing concern is emerging on the adverse effects the unintentional inhalation of CNTs can have on humans. There is currently a debate regarding which exposure metrics and measurement strategies are the most relevant to investigate workplace exposures to CNTs. This study investigated workplace CNT emissions using a combination of time-integrated filter sampling for scanning electron microscopy (SEM) and direct reading aerosol instruments (DRIs). MATERIAL AND METHODS: Field measurements were performed during small-scale manufacturing of multiwalled carbon nanotubes using the arc discharge technique. Measurements with highly time- and size-resolved DRI techniques were carried out both in the emission and background (far-field) zones. Novel classifications and counting criteria were set up for the SEM method. Three classes of CNT-containing particles were defined: type 1: particles with aspect ratio length:width >3:1 (fibrous particles); type 2: particles without fibre characteristics but with high CNT content; and type 3: particles with visible embedded CNTs. RESULTS: Offline sampling using SEM showed emissions of CNT-containing particles in 5 out of 11 work tasks. The particles were classified into the three classes, of which type 1, fibrous CNT particles contributed 37%. The concentration of all CNT-containing particles and the occurrence of the particle classes varied strongly between work tasks. Based on the emission measurements, it was assessed that more than 85% of the exposure originated from open handling of CNT powder during the Sieving, mechanical work-up, and packaging work task. The DRI measurements provided complementary information, which combined with SEM provided information on: (i) the background adjusted emission concentration from each work task in different particle size ranges, (ii) identification of the key procedures in each work task that lead to emission peaks, (iii) identification of emission events that affect the background, thereby leading to far-field exposure risks for workers other than the operator of the work task, and (iv) the fraction of particles emitted from each source that contains CNTs. CONCLUSIONS: There is an urgent need for a standardized/harmonized method for electron microscopy (EM) analysis of CNTs. The SEM method developed in this study can form the basis for such a harmonized protocol for the counting of CNTs. The size-resolved DRI techniques are commonly not specific enough to selective analysis of CNT-containing particles and thus cannot yet replace offline time-integrated filter sampling followed by SEM. A combination of EM and DRI techniques offers the most complete characterization of workplace emissions of CNTs today.

Toluene diisocyanate: Induction of the autotaxin-lysophosphatidic acid axis and its association with airways symptoms.

Diisocyanates are industrial chemicals which have a wide range of applications in developed and developing countries. They are notorious lung toxicants and respiratory sensitizers. However, the mechanisms behind their adverse effects are not adequately characterized. Autotaxin (ATX) is an enzyme producing lysophosphatidic acid (LPA), and the ATX-LPA axis has been implicated in lung related inflammatory conditions and diseases, including allergic asthma, but not to toxicity of environmental low-molecular-weight chemicals. We investigated effects of toluene diisocyanate (TDI) on ATX induction in human lung epithelial cell models, and we correlated LPA-levels in plasma to biomarkers of TDI exposure in urine collected from workers exposed to <5ppb (parts per billion). Information on workers' symptoms was collected through interviews. One nanomolar TDI robustly induced ATX release within 10min in vitro. A P2X7- and P2X4-dependent microvesicle formation was implicated in a rapid ATX release and a subsequent protein synthesis. Co-localization between purinergic receptors and ATX was documented by immunofluorescence and confocal microscopy. The release was modulated by monocyte chemoattractant protein-1 (MCP-1) and by extracellular ATP. In workers, we found a dose-response relationship between TDI exposure biomarkers in urine and LPA levels in plasma. Among symptomatic workers reporting "sneezing", the LPA levels were higher than among non-symptomatic workers. This is the first report indicating induction of the ATX-LPA axis by an environmental low-molecular-weight chemical, and our data suggest a role for the ATX-LPA axis in TDI toxicity.

Exposure to welding fumes is associated with hypomethylation of the F2RL3 gene: a cardiovascular disease marker.

BACKGROUND: Welders are at risk for cardiovascular disease. Recent studies linked tobacco smoke exposure to hypomethylation of the F2RL3 (coagulation factor II (thrombin) receptor-like 3) gene, a marker for cardiovascular disease prognosis and mortality. However, whether welding fumes cause hypomethylation of F2RL3 remains unknown. METHODS: We investigated 101 welders (median span of working as a welder: 7 years) and 127 unexposed controls (non-welders with no obvious exposure to respirable dust at work), age range 23-60 years, all currently non-smoking, in Sweden. The participants were interviewed about their work history, lifestyle factors and diseases. Personal sampling of respirable dust was performed for the welders. DNA methylation of F2RL3 in blood was assessed by pyrosequencing of four CpG sites, CpG_2 (corresponds to cg03636183) to CpG_5, in F2RL3. Multivariable linear regression analysis was used to assess the association between exposure to welding fumes and F2RL3 methylation. RESULTS: Welders had 2.6% lower methylation of CpG_5 than controls (p<0.001). Higher concentrations of measured respirable dust among the welders were associated with hypomethylation of CpG_2, CpG_4 and CpG_5 (ß=-0.49 to -1.4, p<0.012); p<0.029 adjusted for age, previous smoking, passive smoking, education, current residence and respirator use. Increasing the number of years working as a welder was associated with hypomethylation of CpG_4 (linear regression analysis, ß=-0.11, p=0.039, adjusted for previous smoking). Previous tobacco smokers had 1.5-4.7% (p<0.014) lower methylation of 3 of the 4 CpG sites in F2RL3 (CpG_2, CpG_4 and CpG_5) compared to never-smokers. A non-significant lower risk of cardiovascular disease with more methylation was observed for all CpG sites. CONCLUSIONS: Welding fumes exposure and previous smoking were associated with F2RL3 hypomethylation. This finding links low-to-moderate exposure to welding fumes to adverse effects on the cardiovascular system, and suggests a potential mechanistic pathway for this link, via epigenetic effects on F2RL3 expression.

Maternal exposure to air pollution and type 1 diabetes--Accounting for genetic factors.

BACKGROUND: Genetic and non-genetic factors probably act together to initiate and accelerate development of type 1 diabetes [T1D]. One suggested risk factor contributing to development of T1D is air pollution. OBJECTIVE: The aim of the study was to investigate whether maternal exposure during pregnancy to air pollution, measured as nitrogen oxides [NOx] and ozone, in a low-dose exposure area was associated with the child developing T1D. METHOD: In Scania (Skåne), the most southern county in Sweden, 84,039 infants were born during the period 1999-2005. By the end of April 2013, 324 of those children had been diagnosed with T1D. For each of those T1D children three control children were randomly selected and matched for HLA genotype and birth year. Individually modelled exposure data at residence during pregnancy were assessed for nitrogen oxides [NOx], traffic density and ozone. RESULTS: Ozone as well as NOx exposures were associated with T1D. When the highest exposure group was compared to the lowest group an odds ratios of 1.62 (95% confidence interval [CI] 0.99-2.65) was observed for ozone in the second trimester and 1.58 (95% CI 1.06-2.35) for NOx in the third trimester. CONCLUSION: This study indicates that living in an area with elevated levels of air pollution during pregnancy may be a risk factor for offspring T1D.

Comparison and Evaluation of Multiple Users' Usage of the Exposure and Risk Tool: Stoffenmanager 5.1.

Stoffenmanager is an exposure and risk assessment tool that has a control banding part, with risk bands as outcome and a quantitative exposure assessment part, with the 90th percentile of the predicted exposure as a default outcome. The main aim of the study was to investigate whether multiple users of Stoffenmanager came to the same result when modelling the same scenarios. Other aims were to investigate the differences between outcomes of the control banding part with the measured risk quota and to evaluate the conservatism of the tool by testing whether the 90th percentiles are above the measured median exposures. We investigated airborne exposures at companies in four different types of industry: wood, printing, metal foundry, and spray painting. Three scenarios were modelled and measured, when possible, at each company. When modelled, 13 users visited each company on the same occasion creating individual assessments. Consensus assessments were also modelled for each scenario by six occupational hygienists. The multiple users' outcomes were often spread over two risk bands in the control banding part, and the differences in the quantitative exposure outcomes for the highest and lowest assessments per scenario varied between a factor 2 and 100. Four parameters were difficult for the users to assess and had a large impact on the outcome: type of task, breathing zone, personal protection, and control measures. Only two scenarios had a higher measured risk quota than predicted by the control banding part, also two scenarios had slightly higher measured median exposure value than modelled consensus in the quantitative exposure assessment part. Hence, the variability between users was large but the model performed well.

Detection of Multi-walled Carbon Nanotubes and Carbon Nanodiscs on Workplace Surfaces at a Small-Scale Producer.

BACKGROUND: The industrial use of novel-manufactured nanomaterials such as carbon nanotubes and carbon nanodiscs is increasing globally. Occupational exposure can occur during production, downstream use, and disposal. The health effects of many nanomaterials are not yet fully characterized and to handle nano-objects, their aggregates and agglomerates >100nm (NOAA), a high degree of control measures and personal protective equipment are required. The emission of airborne NOAA during production and handling can contaminate workplace surfaces with dust, which can be resuspended resulting in secondary inhalation exposures and dermal exposures. This study surveys the presence of carbon-based nanomaterials, such as multi-walled carbon nanotubes (MWCNTs) and carbon nanodiscs, as surface contamination at a small-scale producer using a novel tape sampling method. METHODS: Eighteen different surfaces at a small-scale producer were sampled with an adhesive tape sampling method. The surfaces selected were associated with the production and handling of MWCNT powder in the near-field zone. Surfaces in the far-field zone were also sampled. In addition, tape stripping of the skin was performed on one worker. The tape samples were analysed with scanning electron microscopy to detect the carbon-based NOAA. Air sampling with a personal impactor was also performed on a worker who was producing MWCNTs the same day as the tape samples were collected. RESULTS: MWCNTs were detected in 50% of the collected tape samples and carbon nanodiscs in 17%. MWCNTs and carbon nanodiscs were identified in all parts of the workplace, thus, increasing the risk for secondary inhalation and dermal exposure of the workers. Both airborne MWCNTs and carbon nanodiscs were detected in the personal impactor samples. The tape-strip samples from the worker showed no presence of carbon-containing nanoparticles. CONCLUSIONS: Tape sampling is a functional method for detecting surface contamination of carbon-based NOAA and for exposure control during production at potentially any workplace that produces or handles such manufactured nanomaterials. With the tape method, it is possible to monitor if a potential for secondary inhalation exposure or dermal exposure exists through resuspension of dust deposited on workplace surfaces. By means of air sampling, we could confirm that carbon nanodiscs were resuspended into the air at the workplace even though they were not handled during that particular work shift. MWCNTs were detected in the air samples, but can have been derived from either resuspension or from the work tasks with MWCNTs that were performed during the air sampling. Tape sampling is a complementary method to air sampling and together these two methods provide a better view of the hygienic situation in workplaces where NOAA can be emitted into work environments.

The ordinary work environment increases symptoms from eyes and airways in mild steel welders.

PURPOSE: We aimed to follow diary-registered symptoms from eyes and airways in mild steel welders and relate them to different exposure measures. Furthermore, we would clarify the influence of possible effect modifiers. METHODS: Non-smoking welders with (N = 74) and without (N = 32) work-related symptoms the last month were enroled. Symptoms and work tasks each day for three two-week periods during 1 year were obtained. Respirable dust (RD) was measured 1 day each period for each worker. The personal daily exposure was assessed as: (1) days at work, (2) welding time and (3) estimates of RD from welding and grinding, calculated from diary entries and measurements. RESULTS: Only 9.2 % of the particle measurements exceed the Swedish occupational exposure limit (OEL; 5 mg/m(3)). Days at work increased the risk of symptoms studied: eyes: 1.79 (1.46-2.19), nasal: 2.16 (1.81-2.58), dry cough: 1.50 (1.23-1.82) and wheezing and/or dyspnoea: 1.27 (1.03-1.56; odds ratio, 95 % confidence interval). No clear dose-response relationships were found for the other exposure estimates. Eye symptoms increased by number of years welding. Nasal symptoms and dry cough increased having forced expiratory volume in first second below median at baseline. Wheezing and/or dyspnoea increased in winter, by number of years welding, having a negative standard skin-prick test and having a vital capacity above median at baseline. CONCLUSION: The current Swedish OEL may not protect welders against eye and airway symptoms. The results add to the evidence that welders should be offered regular medical surveillance from early in the career.

Exposure and emission measurements during production, purification, and functionalization of arc-discharge-produced multi-walled carbon nanotubes.

BACKGROUND: The production and use of carbon nanotubes (CNTs) is rapidly growing. With increased production, there is potential that the number of occupational exposed workers will rapidly increase. Toxicological studies on rats have shown effects in the lungs, e.g., inflammation, granuloma formation, and fibrosis after repeated inhalation exposure to some forms of multi-walled CNTs (MWCNTs). Still, when it comes to health effects, it is unknown which dose metric is most relevant. Limited exposure data for CNTs exist today and no legally enforced occupational exposure limits are yet established. The aim of this work was to quantify the occupational exposures and emissions during arc discharge production, purification, and functionalization of MWCNTs. The CNT material handled typically had a mean length <5 µm. Since most of the collected airborne CNTs did not fulfil the World Health Organization fibre dimensions (79% of the counted CNT-containing particles) and since no microscopy-based method for counting of CNTs exists, we decided to count all particle that contained CNTs. To investigate correlations between the used exposure metrics, Pearson correlation coefficient was used. METHODS: Exposure measurements were performed at a small-scale producer of MWCNTs and respirable fractions of dust concentrations, elemental carbon (EC) concentrations, and number concentrations of CNT-containing particles were measured in the workers' breathing zones with filter-based methods during work. Additionally, emission measurements near the source were carried out during different work tasks. Respirable dust was gravimetrically determined; EC was analysed with thermal-optical analysis and the number of CNT-containing particles was analysed with scanning electron microscopy. RESULTS: For the personal exposure measurements, respirable dust ranged between <73 and 93 µg m(-3), EC ranged between <0.08 and 7.4 µg C m(-3), and number concentration of CNT-containing particles ranged between 0.04 and 2.0 cm(-3). For the emission measurements, respirable dust ranged between <2800 and 6800 µg m(-3), EC ranged between 0.05 and 550 µg C m(-3), and number concentration of CNT-containing particles ranged between <0.20 and 11cm(-3). CONCLUSIONS: The highest exposure to CNTs occurred during production of CNTs. The highest emitted number concentration of CNT-containing particles occurred in the sieving, mechanical work-up, pouring, weighing, and packaging of CNT powder during the production stage. To be able to quantify exposures and emissions of CNTs, a selective and sensitive method is needed. Limitations with measuring EC and respirable dust are that these exposure metrics do not measure CNTs specifically. Only filter-based methods with electron microscopy analysis are, to date, selective and sensitive enough. This study showed that counting of CNT-containing particles is the method that fulfils those criteria and is therefore the method recommended for future quantification of CNT exposures. However, CNTs could be highly toxic not only because of their length but also because they could contain, for example transition metals and polycyclic aromatic hydrocarbons, or have surface defects. Lack of standardized counting criteria for CNTs to be applied at the electron microscopy analysis is a limiting factor, which makes it difficult to compare exposure data from different studies.

Biomarkers of exposure in Monday morning urine samples as a long-term measure of exposure to aromatic diisocyanates.

PURPOSE: Exposure to diisocyanates is a known occupational hazard. One method for monitoring occupational exposure is by analyzing biomarkers in hydrolyzed urine and plasma. The half-life of the biomarkers in plasma is about 3 weeks, and the urinary elimination is divided into one fast (hours) and one slow phases (weeks). Polymorphism in glutathione S-transferase enzymes (GST) is earlier shown to modify the metabolism. The aim of the study was to assess whether biomarkers of exposure in urine collected after two non-exposed days correlate with levels in plasma and whether they can be used as a measure for long-term exposure to aromatic diisocyanates and further whether polymorphisms in GST influenced the correlations. METHODS: Biomarkers of exposure was analyzed in urine and blood samples collected from 24 workers, exposed to at least one of toluene-, methylenediphenyl- or naphthalene diisocyanate, on a Monday morning after at least two unexposed days. Moreover, genotype was determined for 19 of the workers. RESULTS: The corresponding specific gravity-adjusted biomarkers in urine and plasma levels for the different diisocyanates correlated well (r between 0.689 and 0.988). When taking all samples together, the correlation coefficient was 0.926. Polymorphism in the GSTM1 genotype seemed to modify the association. CONCLUSION: Urine collected after two unexposed days can possibly be used as long-term biomarker of exposure for aromatic diisocyanates.

Exposure to respirable dust and manganese and prevalence of airways symptoms, among Swedish mild steel welders in the manufacturing industry.

PURPOSE: Welding fume consists of metal fumes, e.g., manganese (Mn) and gases, e.g., ozone. Particles in the respirable dust (RD) size range dominate. Exposure to welding fume could cause short- and long-term respiratory effects. The prevalence of work-related symptoms among mild steel welders was studied, and the occupational exposure to welding fumes was quantified by repeated measurements of RD, respirable Mn, and ozone. Also the variance components were studied. METHOD: A questionnaire concerning airway symptoms and occupational history was answered by 79% of a cohort of 484 welders. A group of welders (N = 108) were selected and surveyed by personal exposure measurements of RD and ozone three times during 1 year. RESULTS: The welders had a high frequency of work-related symptoms, e.g., stuffy nose (33%), ocular symptoms (28%), and dry cough (24%). The geometric mean exposure to RD and respirable Mn was 1.3 mg/m(3) (min-max 0.1-38.3 mg/m(3)) and 0.08 mg/m(3) (min-max <0.01-2.13 mg/m(3)), respectively. More than 50% of the Mn concentrations exceeded the Swedish occupational exposure limit (OEL). Mainly, low concentrations of ozone were measured, but 2% of the samples exceeded the OEL. Of the total variance for RD, 30 and 33% can be attributed to within-worker variability and between-company variability, respectively. CONCLUSIONS: Welders had a high prevalence of work-related symptom from the airways and eyes. The welders' exposure to Mn was unacceptably high. To reduce the exposure further, control measures in the welding workshops are needed. Correct use of general mechanical ventilation and local exhaust ventilation can, for example, efficiently reduce the exposure.

Elemental carbon - An efficient method to measure occupational exposure from materials in the graphene family.

Graphene is a 2D-material with many useful properties such as flexibility, elasticity, and conductivity among others. Graphene could therefore become a material used in many occupational fields in the future, which can give rise to occupational exposure. Today, exposure is unknown, due to the lack of efficient measuring techniques for occupational exposure to graphene. Readily available screening techniques for air sampling and -analysis are either nonspecific or nonquantitative. Quantifying materials from the broad graphene family by an easy-to-use method is important for the large-scale industrial application of graphene, especially when for the safety of working environment. Graphene consists primarily of elemental carbon, and the present study evaluates the organic carbon/elemental carbon (OC/EC)-technique for exposure assessment. The purpose of this work is to evaluate the OC/EC analysis technique as an efficient and easy-to-use method for quantification of occupational exposure to graphene. Methods that can identify graphene would be preferable for screening, but they are time consuming and semi-quantitative and therefore not suited for quantitative work environment assessments. The OC/EC-technique is a thermal optical analysis (TOA), that quantitively determines the amount of and distinguishes between two different types of carbon, organic and elemental. The technique is standardised, well-established and among other things used for diesel exposure measurements (ref standard). OC/EC could therefore be a feasible measuring technique to quantitively determine occupational exposure to graphene. The present evaluation of the technique provides an analytical method that works quantitatively for graphene, graphene oxide and reduced graphene oxide. Interestingly, the TOA technique makes it possible to distinguish between the three graphene forms used in this study. The technique was tested in an industrial setting and the outcome suggests that the technique is an efficient monitoring technique to be used in combination with characterisation techniques like for example Raman spectroscopy, scanning electron microscopy and atomic force microscopy.