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It is estimated that greater than 1 million workers are exposed to welding fume (WF) by inhalation daily. The potentially toxic metals found in WF are known to cause multiple adverse pulmonary and systemic effects, including cardiovascular disease, and these metals have also been shown to translocate to the liver. This occupational exposure combined with a high fat (HF) Western diet, which has been shown to cause hyperlipidemia and non-alcoholic fatty liver disease (NAFLD), has the potential to cause significant mixed exposure metabolic changes in the liver. The goal of this study was to use matrix assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS) to analyze the spatial distribution and abundance changes of lipid species in Sprague Dawley rat liver maintained on a HF diet combined with WF inhalation. The results of the MALDI-IMS analysis revealed unique hepatic lipid profiles for each treatment group. The HF diet group had significantly increased abundance of triglycerides and phosphatidylinositol lipids, as well as decreased lysophosphatidic lipids and cardiolipin. Ceramide-1-phosphate was found at higher abundance in the regular (REG) diet WF-exposed group which has been shown to regulate the eicosanoid pathway involved in pro-inflammatory response. The results of this study showed that the combined effects of WF inhalation and a HF diet significantly altered the hepatic lipidome. Additionally, pulmonary exposure to WF alone increased lipid markers of inflammation.

Fluoroacetate (FA) is a tasteless, odorless, water-soluble metabolic poison with severe toxicological effects. Characterized in the mid-1900s, it has been used as a rodenticide but is comparably lethal to all mammals. Many countries have restricted its use, and modern-day accidental human exposures are rare, but recently, concerns have been raised about its application as a chemical weapon with no known antidote. A combined treatment of methylene blue (MB), an antioxidant, and monosodium glutamate (MSG), a precursor of the citric acid cycle substrate alpha-ketoglutarate, has been recommended as an effective countermeasure; however, no peer-reviewed articles documenting the efficacy of this therapy have been published. Using a rodent model, we assessed the effects of MB and MSG on the neurologic, cardiac, and pulmonary systems. Transcriptomic analysis was used to elucidate inflammatory pathway activation and guide bioassays, which revealed the advantages and disadvantages of these candidate countermeasures. Results show that MB and MSG can reduce neurologic signs observed in rats exposed to sodium FA and improve some effects of intoxication. However, while this strategy resolved some signs of intoxication, ultimately it was unable to significantly reduce lethality.

Welding fumes (WF) are a complex mixture of toxic metals and gases, inhalation of which can lead to adverse health effects among welders. The presence of manganese (Mn) in welding electrodes is cause for concern about the potential development of Parkinson's disease (PD)-like neurological disorder. Consequently, from an occupational safety perspective, there is a critical need to prevent adverse exposures to WF. As the fume generation rate and physicochemical characteristics of welding aerosols are influenced by welding process parameters like voltage, current or shielding gas, we sought to determine if changing such parameters can alter the fume profile and consequently its neurotoxic potential. Specifically, we evaluated the influence of voltage on fume composition and neurotoxic outcome. Rats were exposed by whole-body inhalation (40 mg/m(3); 3h/day × 5 d/week × 2 weeks) to fumes generated by gas-metal arc welding using stainless steel electrodes (GMA-SS) at standard/regular voltage (25 V; RVSS) or high voltage (30 V; HVSS). Fumes generated under these conditions exhibited similar particulate morphology, appearing as chain-like aggregates; however, HVSS fumes comprised of a larger fraction of ultrafine particulates that are generally considered to be more toxic than their fine counterparts. Paradoxically, exposure to HVSS fumes did not elicit dopaminergic neurotoxicity, as monitored by the expression of dopaminergic and PD-related markers. We show that the lack of neurotoxicity is due to reduced solubility of Mn in HVSS fumes. Our findings show promise for process control procedures in developing prevention strategies for Mn-related neurotoxicity during welding; however, it warrants additional investigations to determine if such modifications can be suitably adapted at the workplace to avert or reduce adverse neurological risks.

Epidemiologic studies suggest an increased risk of lung cancer with exposure to welding fumes, but controlled animal studies are needed to support this association. Oropharyngeal aspiration of collected “aged” gas metal arc–stainless steel (GMA–SS) welding fume has been shown by our laboratory to promote lung tumor formation in vivo using a two-stage initiation–promotion model. Our objective in this study was to determine whether inhalation of freshly generated GMA-SS welding fume also acts as a lung tumor promoter in lung tumor-susceptible mice. Male A/J mice received intraperitoneal (IP) injections of corn oil or the chemical initiator 3-methylcholanthrene (MCA; 10 μg/g) and 1 week later were exposed by whole-body inhalation to air or GMA-SS welding aerosols for 4 h/d × 4 d/w × 9 w at a target concentration of 40 mg/m(3). Lung nodules were enumerated at 30 weeks post-initiation. GMA-SS fume significantly promoted lung tumor multiplicity in A/J mice initiated with MCA (16.11 ± 1.18) compared to MCA/air-exposed mice (7.93 ± 0.82). Histopathological analysis found that the increased number of lung nodules in the MCA/GMA-SS group were hyperplasias and adenomas, which was consistent with developing lung tumorigenesis. Metal deposition analysis in the lung revealed a lower deposited dose, approximately fivefold compared to our previous aspiration study, still elicited a significant lung tumorigenic response. In conclusion, this study demonstrates that inhaling GMA-SS welding fume promotes lung tumorigenesis in vivo which is consistent with the epidemiologic studies that show welders may be at an increased risk for lung cancer.

Welders are exposed to high concentrations of nanoparticles. Compared to larger particles, nanoparticles have been associated with more toxic effects at the cellular level, including the generation of more reactive oxygen species activity. Current methods for welding-fume aerosol exposures do not differentiate between the nano-fraction and the larger particles. The objectives of this work are to establish a method to estimate the respiratory deposition of the nano-fraction of selected metals in welding fumes and test this method in a laboratory setting. Manganese (Mn), Nickel (Ni), Chromium (Cr), and hexavalent chromium (Cr(VI)) are commonly found in welding fume aerosols and have been linked with severe adverse health outcomes. Inductively coupled plasma mass spectrometry (ICP-MS) and ion chromatography (IC) were evaluated as methods for analyzing the content of Mn, Ni, Cr, and Cr(VI) nanoparticles in welding fumes collected with nanoparticle respiratory deposition (NRD) samplers. NRD samplers collect nanoparticles at deposition efficiencies that closely resemble physiological deposition in the respiratory tract. The limits of detection (LODs) and quantitation (LOQs) for ICP-MS and IC were determined analytically. Mild and stainless steel welding fumes generated with a robotic welder were collected with NRD samplers inside a chamber. LODs (LOQs) for Mn, Ni, Cr, and Cr(VI) were 1.3 μg (4.43 μg), 0.4 μg (1.14 μg), 1.1 μg (3.33 μg), and 0.4 μg (1.42 μg), respectively. Recovery of spiked samples and certified welding fume reference material was greater than 95%. When testing the method, the average percentage of total mass concentrations collected by the NRD samplers was ~30% for Mn, ~50% for Cr, and ~60% for Ni, indicating that a large fraction of the metals may lie in the nanoparticle fraction. This knowledge is critical to the development of toxicological studies aimed at finding links between exposure to welding fume nanoparticles and adverse health effects. Future work will involve the validation of the method in workplace settings. [Supplementary materials are available for this article. Go to the publisher's online edition of Journal of Occupational and Environmental Hygiene for the following free supplemental resource: Digestion, extraction, and analysis procedures for nylon mesh screens.].

Sodium fluoroacetate (1080) is a highly toxic metabolic poison that has the potential because of its lack of defined color, odor, and taste and its high water solubility to be intentionally or unintentionally ingested through food adulteration. Although the mechanism of action for 1080 has been known since the 1950s, no known antidote exists. In an effort to better understand the cardiopulmonary impacts of 1080, we utilized whole-body plethysmography and telemeterized Sprague-Dawley rats which allowed for the real-time measurement of respiratory and cardiac parameters following exposure using a non-invasive assisted-drinking method. Overall, the animals showed marked depression of respiratory parameters over the course of 24 h post-exposure and the development of hemorrhage in the lung tissue. Tidal volume was reduced by 30% in males and 60% in females at 24 h post-exposure, and respiratory frequency was significantly depressed as well. In telemeterized female rats, we observed severe cardiac abnormalities, highlighted by a 50% reduction in heart rate, 75% reduction in systolic blood pressure, and a 3.5-fold lengthening of the QRS interval over the course of 24 h. We also observed a reduction in core body temperature of nearly 15°C. Our study was able to describe the severe and pronounced effects of sodium fluoroacetate poisoning on cardiopulmonary function, the results of which indicate that both tissue specific and systemic deficits contribute to the toxicological progression of 1080 intoxication and will need to be accounted for when developing any potential countermeasure for 1080 poisoning.

Direct-reading instruments have been widely used for characterizing airborne nanoparticles in inhalation toxicology and industrial hygiene studies for exposure/risk assessments. Instruments using electrical mobility sizing followed by optical counting, e.g., scanning or sequential mobility particle spectrometers (SMPS), have been considered as the “gold standard” for characterizing nanoparticles. An SMPS has the advantage of rapid response and has been widely used, but there is little information on its performance in assessing the full spectrum of nanoparticles encountered in the workplace. In this study, an SMPS was evaluated for its effectiveness in producing “monodisperse” aerosol and its adequacy in characterizing overall particle size distribution using three test aerosols, each mimicking a unique class of real-life nanoparticles: singlets of nearly spherical titanium dioxide (TiO2), agglomerates of fiber-like multi-walled carbon nanotube (MWCNT), and aggregates that constitutes welding fume (WF). These aerosols were analyzed by SMPS, cascade impactor, and by counting and sizing of discrete particles by scanning and transmission electron microscopy. The effectiveness of the SMPS to produce classified particles (fixed voltage mode) was assessed by examination of the resulting geometric standard deviation (GSD) from the impactor measurement. Results indicated that SMPS performed reasonably well for TiO2 (GSD = 1.3), but not for MWCNT and WF as evidenced by the large GSD values of 1.8 and 1.5, respectively. For overall characterization, results from SMPS (scanning voltage mode) exhibited particle-dependent discrepancies in the size distribution and total number concentration compared to those from microscopic analysis. Further investigation showed that use of a single-stage impactor at the SMPS inlet could distort the size distribution and underestimate the concentration as shown by the SMPS, whereas the presence of vapor molecules or atom clusters in some test aerosols might cause artifacts by counting “phantom particles.” Overall, the information obtained from this study will help understand the limitations of the SMPS in measuring nanoparticles so that one can adequately interpret the results for risk assessments and exposure prevention in an occupational or ambient environment.

Welding generates complex metal fumes that vary in composition. The objectives of this study were to compare the persistence of deposited metals and the inflammatory potential of stainless and mild steel welding fumes, the two most common fumes used in US industry. Sprague-Dawley rats were exposed to 40 mg/m(3) of stainless or mild steel welding fumes for 3 h/day for 3 days. Controls were exposed to filtered air. Generated fume was collected, and particle size and elemental composition were determined. Bronchoalveolar lavage was done on days 0, 8, 21, and 42 after the last exposure to assess lung injury/inflammation and to recover lung phagocytes. Non-lavaged lung samples were analyzed for total and specific metal content as a measure of metal persistence. Both welding fumes were similar in particle morphology and size. Following was the chemical composition of the fumes-stainless steel: 57% Fe, 20% Cr, 14% Mn, and 9% Ni; mild steel: 83% Fe and 15% Mn. There was no effect of the mild steel fume on lung injury/inflammation at any time point compared to air control. Lung injury and inflammation were significantly elevated at 8 and 21 days after exposure to the stainless steel fume compared to control. Stainless steel fume exposure was associated with greater recovery of welding fume-laden macrophages from the lungs at all time points compared with the mild steel fume. A higher concentration of total metal was observed in the lungs of the stainless steel welding fume at all time points compared with the mild steel fume. The specific metals present in the two fumes were cleared from the lungs at different rates. The potentially more toxic metals (e.g., Mn, Cr) present in the stainless steel fume were cleared from the lungs more quickly than Fe, likely increasing their translocation from the respiratory system to other organs.

Serious questions have been raised by occupational health investigators regarding a possible causal association between neurological effects in welders and the presence of manganese (Mn) in welding fume. Male Sprague-Dawley rats were exposed by inhalation to 40 mg/m(3) of gas metal arc-mild steel (MS) welding fume for 3 h/day for 10 days. Generated fume was collected in the animal chamber during exposure, and particle size, composition, and morphology were characterized. At 1 day after the last exposure, metal deposition in different organ systems and neurological responses in dopaminergic brain regions were assessed in exposed animals. The welding particles were composed primarily of a complex of iron (Fe) and Mn and were arranged as chain-like aggregates with a significant number of particles in the nanometer size range. Mn was observed to translocate from the lungs to the kidney and specific brain regions (olfactory bulb, cortex, and cerebellum) after MS fume inhalation. In terms of neurological responses, short-term MS fume inhalation induced significant elevations in divalent metal ion transporter 1 (Dmt1) expression in striatum and midbrain and significant increases in expression of proinflammatory chemokines (Ccl2, Cxcl2) and cytokines (IL1beta, TNFalpha) in striatum. In addition, mRNA and protein expression of glial fibrillary acidic protein (GFAP) was significantly increased in striatum after MS fume exposure. However, the 10-day MS welding fume inhalation did not cause any changes in dopamine and its metabolites or GABA in dopaminergic brain regions nor did it produce overt neural cell damage as assessed by histopathology. In summary, short-term MS welding fume exposure led to translocation of Mn to specific brain regions and induced subtle changes in cell markers of neuroinflammatory and astrogliosis. The neurofunctional significance of these findings currently is being investigated in longer, more chronic welding fume exposure studies.

Occupational exposure to welding fumes is a known health hazard. To isolate elements in stainless steel welding fumes with high potential for adverse health outcomes, fumes were generated using a robotic gas metal arc system, using four shield gases of varying oxygen content. The objective was to measure Cr(VI) concentrations in a broad spectrum of gas metal arc welding processes, and identify processes of exceptionally high or low Cr(VI) content. The gases used were 95% Ar/5% O(2), 98% Ar/2% O(2), 95% Ar/5%CO(2), and 75% He/25% Ar. The welder was operated in axial spray mode (Ar/O(2), Ar/CO(2)), short-circuit (SC) mode (Ar/CO(2) low voltage and He/Ar), and pulsed axial-spray mode (98% Ar/2% O(2)). Results indicate large differences in Cr(VI) in the fumes, with Ar/O(2) (Pulsed)Ar/O(2)Ar/CO(2)Ar/CO(2) (SC)He/Ar; values were 3000+/-300, 2800+/-85, 2600+/-120, 1400+/-190, and 320+/-290 ppm respectively (means +/- standard errors for 2 runs and 3 replicates per run). Respective rates of Cr(VI) generation were 1.5, 3.2, 4.4, 1.3, and 0.46 microg/min; generation rates were also calculated in terms of microg Cr(VI) per metre of wire used. The generation rates of Cr(VI) increased with increasing O(3) concentrations. Particle size measurements indicated similar distributions, but somewhat higher0.6 microm fractions for the short-circuit mode samples. Fumes were also sampled into 2 selected size ranges, a microspatter fraction (or=0.6 microm) and a fine (0.6 microm) fraction; analysis indicated that Cr(VI) is primarily associated with particles0.6 microm. The conclusion of the study is that Cr(VI) concentrations vary significantly with welding type and shield gas type, and this presents an opportunity to tailor welding practices to lessen Cr(VI) exposures in workplaces by selecting low Cr(VI)-generating processes. Short-circuit processes generated less Cr(VI) than axial-spray methods, and inert gas shielding gave lower Cr(VI) content than shielding with active gases. A short circuit He/Ar shielded process and a pulsed axial spray Ar/O(2) process were both identified as having substantially lower Cr(VI) generation rates per unit of wire used relative to the other processes studied.

Many welders have experienced bronchitis, metal fume fever, lung function changes, and an increase in the incidence of lung infection. Questions remain regarding the possible mechanisms associated with the potential pulmonary effects of welding fume exposure. The objective was to assess the early effects of stainless steel (SS) welding fume inhalation on lung injury, inflammation, and defense responses. Male Sprague-Dawley rats were exposed to gas metal arc-SS welding fume at a concentration of 15 or 40 mg/m{sup 3} x 3 h/day for 1, 3, or 10 days. The control group was exposed to filtered air. To assess lung defense responses, some animals were intratracheally inoculated with 5 x 10{sup 4}Listeria monocytogenes 1 day after the last exposure. Welding particles were collected during exposure, and elemental composition and particle size were determined. At 1, 4, 6, 11, 14, and 30 days after the final exposure, parameters of lung injury (lactate dehydrogenase and albumin) and inflammation (PMN influx) were measured in the bronchoalveolar lavage fluid. In addition, particle-induced effects on pulmonary clearance of bacteria and macrophage function were assessed. SS particles were composed of Fe, Cr, Mn, and Ni. Particle size distribution analysis indicated the mass median aerodynamic diameter of the generatedmore » fume to be 0.255 {mu}m. Parameters of lung injury were significantly elevated at all time points post-exposure compared to controls except for 30 days. Interestingly, no significant difference in lung PMNs was observed between the SS and control groups at 1, 4, and 6 days post-exposure. After 6 days post-exposure, a dramatic increase in lung PMNs was observed in the SS group compared to air controls. Lung bacteria clearance and macrophage function were reduced and immune and inflammatory cytokines were altered in the SS group. In summary, short-term exposure of rats to SS welding fume caused significant lung damage and suppressed lung defense responses to bacterial infection, but had a delayed effect on pulmonary inflammation. Additional chronic inhalation studies are needed to further examine the lung effects associated with SS welding fume exposure.« less

Nine gas metal arc welding (GMAW) processes for stainless steel were assessed for fume generation rates, fume generation rates per g of electrode consumed, and emission rates for hexavalent chromium (Cr(6+)). Elemental manganese, nickel, chromium, iron emissions per unit length of weld, and labor plus consumables costs were similarly measured. Flux-cored arc welding and shielded metal arc (SMAW) processes were also studied. The objective was to identify the best welding processes for reducing workplace exposures, and estimate costs for all processes. Using a conical chamber, fumes were collected, weighed, recovered, and analyzed by inductively coupled atomic emission spectroscopy for metals, and by ion chromatography for Cr(6+). GMAW processes used were Surface Tension Transfer, Regulated Metal Deposition, Cold Metal Transfer, short-circuit, axial spray, and pulsed spray modes. Flux-cored welding used gas shielding; SMAW used E308 rods. Costs were estimated as dollars per m length of a ¼ in (6.3 mm) thick horizontal butt weld; equipment costs were estimated as ratios of new equipment costs to a 250 ampere capacity SMAW welding machine. Results indicate a broad range of fume emission factors for the processes studied. Fume emission rates per g of electrode were lowest for GMAW processes such as pulsed-spray mode (0.2 mg/g), and highest for SMAW (8 mg fume/g electrode). Emission rates of Cr(6+) ranged from 50-7800 µg/min, and Cr(6+) generation rates per g electrode ranged from 1-270 µg/g. Elemental Cr generation rates spanned 13-330 µg/g. Manganese emission rates ranged from 50-300 µg/g. Nickel emission rates ranged from 4-140 µg/g. Labor and consumables costs ranged from $3.15 (GMAW pulsed spray) to $7.40 (SMAW) per meter of finished weld, and were measured or estimated for all 11 processes tested. Equipment costs for some processes may be as much as five times the cost of a typical SMAW welding machine. The results show that all of the GMAW processes in this study can substantially reduce fume, Cr(6+), manganese and costs relative to SMAW, the most commonly used welding process, and several have exceptional capabilities for reducing emissions.

Prolonged, extensive exposure to asphalt fume has been associated with several adverse health effects. Inhaled polycyclic aromatic hydrocarbons (PAHs) from asphalt fume exposure are of concern. The objective of this study was to characterize both qualitative and quantitative differences between fumes generated at 150 degrees C and 180 degrees C using a well-controlled laboratory road paving fume generation system. Fumes were characterized by total volatile and particulate concentration, simulated boiling point profile, and specific PAH content. The mean concentrations of the volatile fractions generated at 180 degrees C and 150 degrees C were 23.3 mg/m3 and 11.2 mg/m3, respectively, demonstrating a statistically significant shift in concentration. The mean concentrations of the particulate fractions generated at 180 degrees C and 150 degrees C were 42.4 mg/m3 and 28.0 mg/m3, respectively. The simulated boiling point profile did not show a significant qualitative difference between the fumes generated at the two temperatures. Naphthalene, acenaphthene, fluorene, phenanthrene, fluoranthene, pyrene, and chrysene were identified and quantified from the fumes.

Asphalt fumes are complex mixtures of various organic compounds, including polycyclic aromatic hydrocarbons (PAHs). PAHs require bioactivation by the cytochrome P-450 monooxygenase system to exert toxic/carcinogenic effects. The present study was carried out to characterize the acute pulmonary inflammatory responses and the alterations of pulmonary xenobiotic pathways in rats exposed to asphalt fumes by inhalation. Rats were exposed at various doses and time periods to air or to asphalt fumes generated at paving temperatures. To assess the acute damage and inflammatory responses, differential cell counts, acellular lactate dehydrogenase (LDH) activity, and protein content of bronchoalveolar lavage fluid were determined. Alveolar macrophage (AM) function was assessed by monitoring generation of chemiluminescence and production of tumor necrosis factor-alpha and interleukin-1. Alteration of pulmonary xenobiotic pathways was determined by monitoring the protein levels and activities of P-450 isozymes (CYP1A1 and CYP2B1), glutathioneS-transferase (GST), and NADPH:quinone oxidoreductase (QR). The results show that acute asphalt fume exposure did not cause neutrophil infiltration, alter LDH activity or protein content, or affect AM function, suggesting that short-term asphalt fume exposure did not induce acute lung damage or inflammation. However, acute asphalt fume exposure significantly increased the activity and protein level of CYP1A1 whereas it markedly reduced the activity and protein level of CYP2B1 in the lung. The induction of CYP1A1 was localized in nonciliated bronchiolar epithelial (Clara) cells, alveolar septa, and endothelial cells by immunofluorescence microscopy. Cytosolic QR activity was significantly elevated after asphalt fume exposure, whereas GST activity was not affected by the exposure. This induction of CYP1A1 and QR with the concomitant down-regulation of CYP2B1 after asphalt fume exposure could alter PAH metabolism and may lead to potential toxic effects in the lung.

To provide quantitative information to choose the best welding processes for minimizing workplace emissions, nine gas metal arc welding (GMAW) processes for mild steel were assessed for fume generation rates, normalized fume generation rates (milligram fume per gram of electrode consumed), and normalized generation rates for elemental manganese, nickel, and iron. Shielded metal arc welding (SMAW) and flux-cored arc-welding (FCAW) processes were also profiled. The fumes were collected quantitatively in an American Welding Society-type fume chamber and weighed, recovered, homogenized, and analyzed by inductively coupled atomic emission spectroscopy for total metals. The processes included GMAW with short circuit, globular transfer, axial spray, pulsed spray, Surface Tension Transfer™, Regulated Metal Deposition™, and Cold Metal Transfer™ (CMT) modes. Flux-cored welding was gas shielded, and SMAW was a single rod type. Results indicate a wide range of fume emission factors for the process variations studied. Fume emission rates per gram of electrode consumed were highest for SMAW (~13 mg fume g(-1) electrode) and lowest for GMAW processes such as pulsed spray (~1.5mg g(-1)) and CMT (~1mg g(-1)). Manganese emission rates per gram of electrode consumed ranged from 0.45 mg g(-1) (SMAW) to 0.08 mg g(-1) (CMT). Nickel emission rates were generally low and ranged from ~0.09 (GMAW short circuit) to 0.004 mg g(-1) (CMT). Iron emission rates ranged from 3.7 (spray-mode GMAW) to 0.49 mg g(-1) (CMT). The processes studied have significantly different costs, and cost factors are presented based on a case study to allow comparisons between processes in specific cost categories. Costs per linear meter of weld were $31.07 (SMAW), $12.37 (GMAW short circuit), and $10.89 (FCAW). Although no single process is the best for minimizing fume emissions and costs while satisfying the weld requirements, there are several processes that can minimize emissions. This study provides information to aid in those choices. Suggestions for overcoming barriers to utilizing new and less hazardous welding processes are also discussed.

We have shown that pulmonary nanoparticle exposure impairs endothelium dependent dilation in systemic arterioles. However, the mechanism(s) through which this effect occurs is/are unclear. The purpose of this study was to identify alterations in the production of reactive species and endogenous nitric oxide (NO) after nanoparticle exposure, and determine the relative contribution of hemoproteins and oxidative enzymes in this process. Sprague-Dawley rats were exposed to fine TiO2 (primary particle diameter approximately 1 microm) and TiO2 nanoparticles (primary particle diameter approximately 21 nm) via aerosol inhalation at depositions of 4-90 microg per rat. As in previous intravital experiments in the spinotrapezius muscle, dose-dependent arteriolar dilations were produced by intraluminal infusions of the calcium ionophore A23187. Nanoparticle exposure robustly attenuated these endothelium-dependent responses. However, this attenuation was not due to altered microvascular smooth muscle NO sensitivity because nanoparticle exposure did not alter arteriolar dilations in response to local sodium nitroprusside iontophoresis. Nanoparticle exposure significantly increased microvascular oxidative stress by approximately 60%, and also elevated nitrosative stress fourfold. These reactive stresses coincided with a decreased NO production in a particle deposition dose-dependent manner. Radical scavenging, or inhibition of either myeloperoxidase or nicotinamide adenine dinucleotide phosphate oxidase (reduced) oxidase partially restored NO production as well as normal microvascular function. These results indicate that in conjunction with microvascular dysfunction, nanoparticle exposure also decreases NO bioavailability through at least two functionally distinct mechanisms that may mutually increase local reactive species.

Nanomaterials are frontier technological products used in different manufactured goods. Because of their unique physicochemical, electrical, mechanical, and thermal properties, single-walled carbon nanotubes (SWCNT) are finding numerous applications in electronics, aerospace devices, computers, and chemical, polymer, and pharmaceutical industries. SWCNT are relatively recently discovered members of the carbon allotropes that are similar in structure to fullerenes and graphite. Previously, we ( 47 ) have reported that pharyngeal aspiration of purified SWCNT by C57BL/6 mice caused dose-dependent granulomatous pneumonia, oxidative stress, acute inflammatory/cytokine responses, fibrosis, and decrease in pulmonary function. To avoid potential artifactual effects due to instillation/agglomeration associated with SWCNT, we conducted inhalation exposures using stable and uniform SWCNT dispersions obtained by a newly developed aerosolization technique ( 2 ). The inhalation of nonpurified SWCNT (iron content of 17.7% by weight) at 5 mg/m3, 5 h/day for 4 days was compared with pharyngeal aspiration of varying doses (5–20 μg per mouse) of the same SWCNT. The chain of pathological events in both exposure routes was realized through synergized interactions of early inflammatory response and oxidative stress culminating in the development of multifocal granulomatous pneumonia and interstitial fibrosis. SWCNT inhalation was more effective than aspiration in causing inflammatory response, oxidative stress, collagen deposition, and fibrosis as well as mutations of K- ras gene locus in the lung of C57BL/6 mice.

Asphalt fumes are complex mixtures of aerosols and vapors containing various organic compounds, including polycyclic aromatic hydrocarbons (PAHs). Previously, we have demonstrated that inhalation exposure of rats to asphalt fumes resulted in dose-dependent induction of CYP1A1 with concomitant down-regulation of CYP2B1 and increased phase II enzyme quinone reductase activity in the rat lung. In the present study, the potential genotoxic effects of asphalt fume exposure due to altered lung microsomal enzymes were studied. Rats were exposed to air or asphalt fume generated under road paving conditions at various concentrations and sacrificed the next day. Alveolar macrophages (AM) were obtained by bronchoalveolar lavage and examined for DNA damage using the comet assay. To evaluate the systemic genotoxic effect of asphalt fume, micronuclei formation in bone marrow polychromatic erythrocytes (PCEs) was monitored. Lung S9 from various exposure groups was isolated from tissue homogenates and characterized for metabolic activity in activating 2-aminoanthracene (2-AA) and benzo[a]pyrene (BaP) mutagenicity using the Ames test with Salmonella typhimurium YG1024 and YG1029. This study showed that the paving asphalt fumes significantly induced DNA damage in AM, as revealed by DNA migration in the comet assay, in a dose-dependent manner, whereas the micronuclei formation in bone marrow PCEs was not detected even at a very high exposure level (1733 mg h/m3). The conversion of 2-AA to mutagens in the Ames test required lung S9-mediated metabolic activation in a dose-dependent manner. In comparison to the controls, lung S9 from rats exposed to asphalt fume at a total exposure level of 479+/-33 mg h/m3 did not significantly enhance 2-AA mutagenicity with either S. typhimurium YG1024 or YG1029. At a higher total asphalt fume exposure level (1150+/-63 mg h/m3), S9 significantly increased the mutagenicity of 2-AA as compared to the control. However, S9 from asphalt fume-exposed rats did not significantly activate the mutagenicity of BaP in the Ames test. These results show that asphalt fume exposure, which significantly altered both phases I and II metabolic enzymes in lung microsomes, is genotoxic to AM and enhances the metabolic activation of certain mutagens through altered S9 content.

As a step to study the health effects of asphalt fume exposure, an analytical method was developed to characterize benzo[a]pyrene and its hydroxy metabolites in the urine of asphalt fume-exposed rats. This method is based on microflow liquid chromatography (LC) coupled to hybrid quadrupole orthogonal acceleration time-of-flight mass spectrometry (Q-TOFMS). Twenty-four female Sprague-Dawley rats were used in the experiment, with 8 as controls and 16 exposed to asphalt fumes in a whole-body inhalation chamber for 10 days (4 h/day). Generated at 150 degrees C, the asphalt fume concentration in the animal exposure chamber ranged 76-117 mg/m(3). In the urine of the asphalt fume-exposed rats, benzo[a]pyrene and its metabolites of 3-hydroxybenzo[a]pyrene, benzo[a]pyrene-7,8-dihydrodiol(+/-), and benzo[a]pyrene-7,8,9,10-tetrahydrotetrol(+/-) were identified, and their concentrations were determined at 2.19 +/- 0.49, 16.17 +/- 0.3, 6.28 +/- 0.36, and 29.35 +/- 0.26 ng/100 mL, respectively. The metabolite concentrations from the controlled group, however, were either under the detection limits or at a relatively very low level (0.19 +/- 0.41 ng/100 mL for benzo[a]pyrene-7,8,9,10-tetrahydrotetrol metabolite). The results clearly indicate that the benzo[a]pyrene and its hydroxy metabolites were significantly elevated (p0.001) in the urine of asphalt fume-exposed rats relative to controls. The study also demonstrated that the combination of microflow LC separation and collision-induced dissociation leading to a characteristic fragmentation pattern by hybrid Q-TOFMS offers a distinct advantage for the identifications and characterizations of the benzo[a]pyrene metabolites.

Asphalt fumes have been reported to produce nasal irritation in road workers. Since inhaled irritants can increase substance P (SP) production in airway neurons, the effects of asphalt fumes on SP production in trigeminal ganglia (TG) sensory neurons innervating the nasal mucosa were investigated. The effects of asphalt fumes on nasal mucosal innervation were examined by measuring SP and calcitonin-gene-related peptide (CGRP) levels in rat TG neurons projecting to the nasal epithelium. Female Sprague-Dawley rats were exposed to asphalt fumes at 16.0 +/- 8.1mg /m3 for 5 consecutive days, 3.5 h/d. Inflammatory cells were measured in nasal cavity lavage fluid. SP and CGRP immunoreactivity (IR) was measured in the cell bodies of trigeminal ganglion sensory neurons projecting to the nasal cavity. A significant increase in neutrophils and macrophages was observed after asphalt fume exposure indicating an inflammatory response in the nasal cavity. The percentage of SP-IR neurons increased significantly in the asphalt-exposed rats, and the proportion of CGRP-IR neurons was also elevated following asphalt exposure. These results indicate that exposure to asphalt fumes produces inflammation and increases the levels of SP and CGRP in TG neurons projecting to the nasal epithelium. The findings are consistent with asphalt-induced activation of sensory C-fibers in the nasal cavity. Enhanced sensory neuropeptide release from nerve terminals in the nasal cavity may produce neurogenic inflammation associated with nasal irritation following exposure to asphalt fumes.

A highly sensitive, selective, and reliable analytical method has been developed and validated for characterization of asphalt fume generated under simulated road paving conditions. A dynamic asphalt fume generation system was modified to provide consistent test atmospheres at simulated asphalt road paving conditions. In the process of fume generation, asphalt was initially preheated in an oven to 170 degrees C, pumped to a large kettle, which maintained the asphalt temperature between 150 and 170 degrees C, and then transferred to the generator. The fume was conducted from the generator to an exposure chamber through a heated transfer line. Characterization of the asphalt fume test atmospheres included the following: (1) determination of the consistency of the asphalt aerosol composition within the generation system; (2) quantification of total organic matter of the asphalt fume by electron impact ionization of isotope dilution gas chromatography/ mass spectrometry); and (3) identification of individual priority polycyclic aromatic hydrocarbons (PAHs) in asphalt fume by selected ion monitoring. With the developed method, asphalt fumes could be characterized into three fractions: (1) filter collection of a large molecular size fraction over a range of mass-to-charge (m/z) ratios of 173-309; (2) XAD-2 trapping of a medium molecular size fraction over a range of m/z ratios of 121-197; and (3) charcoal trapping of a small molecular size fraction that contained mainly the volatile vapor fraction over a range of m/z ratios of 57-141. Total organic matter of the asphalt fume was quantified over the 5 exposure days. Sixteen specific priority PAHs were monitored and identified. These PAHs were determined at trace levels on the filter fraction. A novel approach, which utilizes collision-induced dissociation of fragmentation pathway leading to a characteristic fragmentation pattern by coupling microflow liquid chromatography to atmospheric pressure chemical ionization of quadrupole time-of-flight mass spectrometry, was used to further clarify the trace amount of key components present in simulated road paving asphalt fumes. These results demonstrate that asphalt fume composition could be characterized and specific priority PAHs could be identified by this method. The major advantages of this method are its highly sensitivity, selectivity, and reliability for chemical hazard characterization in a complex mixture. This method is suitable for support toxicity studies using simulated occupational exposure to asphalt fumes.