In vitro testing is a common first step in assessing combustion generated and engineered nanoparticle related health hazards. Commercially available viability assays are frequently used to compare the toxicity of different particle types and to generate dose response data. Nanoparticles, well known for having large surface areas and chemically active surfaces, may interfere with viability assays, producing a false assessment of toxicity and making it difficult to compare toxicity data. The objective of this study is to measure the extent of particle interference in two common viability assays, the MTT reduction and the lactate dehydrogenase (LDH) release assays. Diesel particles, activated carbon, flame soot, oxidized flame soot, and titanium dioxide particles are assessed for interactions with the MTT and LDH assay under cell-free conditions. Diesel particles, at concentrations as low as 0.05 μg/ml, reduce MTT. Other particle types reduce MTT only at a concentration of 50 μg/ml and higher. The activated carbon, soot, and oxidized soot particles bind LDH to varying extents, reducing the concentration measured in the LDH assay. The interfering effects of the particles explain in part the different toxicities measured in human bronchial epithelial cells (16HBE14o). We conclude that valid particle toxicity assessments can only be assured after first performing controls to verify that the particles under investigation do not interfere with a specific assay at the expected concentrations.

To identify the mechanism through which nanoparticulate zero-valent iron (nZVI; Fe0(s)) damages cells, a series of experiments were conducted in which nZVI in phosphate-buffered saline (PBS) was exposed to oxygen in the presence and absence of human bronchial epithelial cells. When nZVI is added to PBS, a burst of oxidants is produced as Fe0 and ferrous iron (Fe[II]) are converted to ferric iron (Fe[II]). Cytotoxicity and internal reactive oxygen species (ROS) production in cells exposed to nZVI is equivalent to the response observed when cells are exposed to the same concentration of dissolved Fe(II). Experiments conducted in the absence of cells indicate that the oxidant produced during Fe(II) oxidation reacts with methanol and dimethyl sulfoxide, but not with compounds such as tert-butanol and benzoate that react exclusively with hydroxyl radical. The role of reactive oxidants produced during Fe(II) oxidation in cytotoxicity and internal ROS production is further supported by experiments in which cell damage was limited by the addition of ligands that prevented Fe(II) oxidation and by the absence of cell damage when the nanoparticles were oxidized prior to exposure. The behavior of the oxidant produced by nZVI is consistent with an oxidant such as the ferryl ion, rather than hydroxyl radical.

Publication costs assisted by the Petroleum Research Fund The reactions of 0 atoms and OH radicals with acetylene and methylacetyiene were studied in high-intensity room temperature crossed molecular beams. Products were detected using photoionization mass spectrometry. Major and minor observed reactive channels were assigned on the basis of the identity of the products observed and the magnitudes of the product ion signals. The numbers of major observed reactive channels are two for 0 + C2H2, one for OH + C2H2, seven for 0 + C3H4, and four for OH + C3H4. Details on reaction dynamics were obtained using deuterated compounds, and are discussed in detail. Evidence is presented for several types of mechanisms including abstraction, displacement, and addition followed by decomposition.

Electron paramagnetic resonance (EPR) spectroscopy was used to measure the free radicals in the particulate matter (PM) emissions from wood and coal combustion. The intensity of radicals in PM dropped linearly within two months of sample storage and stabilized after that. This factor of storage time was adjusted when comparing radical intensities among different PM samples. An inverse relationship between coal rank and free radical intensities in PM emissions was observed, which was in contrast with the pattern of radical intensities in the source coals. The strong correlation between intensities of free radical and elemental carbon in PM emissions suggests that the radical species may be carbon-centered. The increased g-factors, 2.0029−2.0039, over that of purely carbon-centered radicals may indicate the presence of vicinal oxygen heteroatom. The redox and biology activities of these carbon-centered radicals are worthy of evaluation.

The effect of liquefied natural gas on pollutant emissions was evaluated experimentally with used and new appliances in the laboratory and with appliances installed in residences, targeting information gaps from previous studies. Burner selection targeted available technologies that are projected to comprise the majority of installed appliances over the next decade. Experiments were conducted on 13 cooktop sets, 12 ovens, 5 broiler burners, 5 storage water heaters, 4 forced air furnaces, 1 wall furnace, and 6 tankless water heaters. Air-free concentrations and fuel-based emission factors were determined for carbon monoxide, nitrogen oxides, nitrogen dioxide, and the number of (predominantly ultrafine) particles over complete burns-including transient effects (device warm-up and intermittent firing of burners) following ignition-and during more stable end-of-burn conditions. Formaldehyde was measured over multi-burn cycles. The baseline fuel was Northern California line gas with Wobbe number (a measure of fuel energy delivery rate) of 1320-1340; test fuels had Wobbe numbers of roughly 1390 and 1420, and in some cases 1360. No ignition or operational problems were observed during test fuel use. Baseline emissions varied widely across and within burner groups and with burner operational mode. Statistically significant emissions changes were observed for some pollutants on some burners.

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We developed an inverted, co-flow, methane/air/nitrogen burner that generates a wide range of soot particles sizes and concentrations. By adjusting the flow rates of air, methane, and nitrogen in the fuel, the mean electric mobility diameter and number concentration are varied. Additional dilution downstream of the flame allows us to generate particle concentrations spanning those produced by spark-ignited and diesel engines: particles with mean diameters between 50 and 250 nm and number concentrations from 4.7·10 4 to 10 7 cm -3 . The range of achievable number concentrations, and therefore volume concentrations, can be increased by a factor of 30 by reducing the dilution ratio. These operating conditions make this burner valuable for developing and calibrating diagnostics as well as for other studies involving soot particles.

In vitro exposure to aerosols at the air-liquid interface (ALI) preserves the physical and chemical characteristics of aerosol particles. Although frequently described as being a more physiologic exposure method, ALI exposure has not been directly compared with conventional in vitro exposures where the particles are suspended in medium. We exposed immortalized human bronchial epithelial cells (16HBE14o) to aerosolized diesel exhaust particles at the ALI and to suspensions of collected particles. The response of the cells was determined from measurements of the cell viability and interleukin-8 (IL-8) secretion. The deposited size distribution at the cell surface was measured with transmission electron microscopy to obtain a dose for the ALI exposure. Although exposure by either method caused a slight decrease in cell viability and induced IL-8 secretion, the response to ALI exposure occurred at doses several orders of magnitude lower than exposure to particles in suspension. The most likely sources for the different dose responses are the artifacts introduced during the collection and resuspension of particles for conventional suspension exposures. The number concentration of particles deposited at the ALI is similar to the modeled deposition in the tracheal-bronchial region in a human lung, but the ALI size distribution is skewed toward particles larger than those deposited in the lung.