Combustion conditions influence toxicity of flame-generated soot to ocular (ARPE-19) cells.

Soot is a prevalent aerosol found both indoors and outdoors that has several sources, such as natural (e.g., wildfires), civilian (e.g., cooking), or military (e.g., burn pit operation). Additionally, within the sources, factors that influence the physicochemical properties of the soot include combustion temperature, oxygen availability, and fuel type. Being able to reproduce soot in the laboratory and systematically assess its toxicity is important in the pursuit of elucidating pathologies associated with its exposure. Of the organs of interest, we targeted the eye given the scant attention received. Yet, air pollution constituents such as soot have been linked to diseases such as age-related macular degeneration and proliferative vitreoretinopathy. We developed a bench-scale system to synthesize different types of soot, that is, soot with a systematically varied physical attributes or chemical composition. We used common analytical techniques to probe such properties, and used statistical analyses to correlate them with toxicity in vitro using ARPE-19 cells. Within the range of flame conditions studied, we find that soot toxicity increases with increasing oxygen concentration in fuel-rich premixed flames, and weakly increases with decreasing flame temperature. Additionally, soot particles produced in premixed flames are generally smaller in size, exhibit a lesser fractal structure, and are considerably more toxic to ARPE-19 cells than soot particles produced in non-premixed flames.

Protection levels of N95-level respirator substitutes proposed during the COVID-19 pandemic: safety concerns and quantitative evaluation procedures.

OBJECTIVE: The COVID-19 pandemic has precipitated widespread shortages of filtering facepiece respirators (FFRs) and the creation and sharing of proposed substitutes (novel designs, repurposed materials) with limited testing against regulatory standards. We aimed to categorically test the efficacy and fit of potential N95 respirator substitutes using protocols that can be replicated in university laboratories. SETTING: Academic medical centre with occupational health-supervised fit testing along with laboratory studies. PARTICIPANTS: Seven adult volunteers who passed quantitative fit testing for small-sized (n=2) and regular-sized (n=5) commercial N95 respirators. METHODS: Five open-source potential N95 respirator substitutes were evaluated and compared with commercial National Institute for Occupational Safety and Health (NIOSH)-approved N95 respirators as controls. Fit testing using the 7-minute standardised Occupational Safety and Health Administration fit test was performed. In addition, protocols that can be performed in university laboratories for materials testing (filtration efficiency, air resistance and fluid resistance) were developed to evaluate alternate filtration materials. RESULTS: Among five open-source, improvised substitutes evaluated in this study, only one (which included a commercial elastomeric mask and commercial HEPA filter) passed a standard quantitative fit test. The four alternative materials evaluated for filtration efficiency (67%-89%) failed to meet the 95% threshold at a face velocity (7.6 cm/s) equivalent to that of a NIOSH particle filtration test for the control N95 FFR. In addition, for all but one material, the small surface area of two 3D-printed substitutes resulted in air resistance that was above the maximum in the NIOSH standard. CONCLUSIONS: Testing protocols such as those described here are essential to evaluate proposed improvised respiratory protection substitutes, and our testing platform could be replicated by teams with similar cross-disciplinary research capacity. Healthcare professionals should be cautious of claims associated with improvised respirators when suggested as FFR substitutes.

Size-Dependent Filtration Efficiency of Alternative Facemask Filter Materials.

The use of facemasks is proven to mitigate the spread of the coronavirus and other biological agents that cause disease. Various forms of facemasks, made using different materials, are being used extensively, and it is important to determine their performance characteristics. The size-dependent filtration efficiency and breathing resistance of household sterilization wrap fabrics, and isolation media (American Society for Testing and Materials (ASTM)- and non-ASTM-rated), were measured in filter-holder- and mannequin-in-chamber-based systems, focusing on particles sizes between 20 nm and 2 µm. Double-layer MERV-14 (Minimum Efficiency Reporting Values with rating 14) showed the highest filtration efficiency (94.9-73.3%) amongst household filter media, whereas ASTM-rated isolation masks showed the highest filtration efficiencies (95.6-88.7) amongst all the masks considered. Filtration efficiency of 3D-printed masks with replaceable filter media was found to depend on the degree of sealing around the media holder, which depended on the material's compressibility. Filtration efficiencies of triple-layer combinations (95.8-85.3%) follow a profile similar to single layers but with improved filtration efficiencies.

Susceptibility to inhaled flame-generated ultrafine soot in neonatal and adult rat lungs.

Over a quarter of the U.S. population is exposed to harmful levels of airborne particulate matter (PM) pollution, which has been linked to development and exacerbation of respiratory diseases leading to morbidity and mortality, especially in susceptible populations. Young children are especially susceptible to PM and can experience altered anatomic, physiologic, and biological responses. Current studies of ambient PM are confounded by the complex mixture of soot, metals, allergens, and organics present in the complex mixture as well as seasonal and temporal variance. We have developed a laboratory-based PM devoid of metals and allergens that can be replicated to study health effects of specific PM components in animal models. We exposed 7-day-old postnatal and adult rats to a single 6-h exposure of fuel-rich ultrafine premixed flame particles (PFPs) or filtered air. These particles are high in polycyclic aromatic hydrocarbons content. Pulmonary cytotoxicity, gene, and protein expression were evaluated at 2 and 24 h postexposure. Neonates were more susceptible to PFP, exhibiting increased lactate dehydrogenase activity in bronchoalveolar lavage fluid and ethidium homodimer-1 cellular staining in the lung in situ as an index of cytotoxicity. Basal gene expression between neonates and adults differed for a significant number of antioxidant, oxidative stress, and proliferation genes and was further altered by PFP exposure. PFP diminishes proliferation marker PCNA gene and protein expression in neonates but not adults. We conclude that neonates have an impaired ability to respond to environmental exposures that increases lung cytotoxicity and results in enhanced susceptibility to PFP, which may lead to abnormal airway growth.

Age specific responses to acute inhalation of diffusion flame soot particles: cellular injury and the airway antioxidant response.

Current studies of particulate matter (PM) are confounded by the fact that PM is a complex mixture of primary (crustal material, soot, metals) and secondary (nitrates, sulfates, and organics formed in the atmosphere) compounds with considerable variance in composition by sources and location. We have developed a laboratory-based PM that is replicable, does not contain dust or metals and that can be used to study specific health effects of PM composition in animal models. We exposed both neonatal (7 days of age) and adult rats to a single 6-h exposure of laboratory generated fine diffusion flame particles (DFP; 170 µg/m(3)), or filtered air. Pulmonary gene and protein expression as well as indicators of cytotoxicity were evaluated 24 h after exposure. Although DFP exposure did not alter airway epithelial cell composition in either neonates or adults, increased lactate dehydrogenase activity was found in the bronchoalveolar lavage fluid of neonates indicating an age-specific increase in susceptibility. In adults, 16 genes were differentially expressed as a result of DFP exposure whereas only 6 genes were altered in the airways of neonates. Glutamate cysteine ligase protein was increased in abundance in both DFP exposed neonates and adults indicating an initiation of antioxidant responses involving the synthesis of glutathione. DFP significantly decreased catalase gene expression in adult airways, although catalase protein expression was increased by DFP in both neonates and adults. We conclude that key airway antioxidant enzymes undergo changes in expression in response to a moderate PM exposure that does not cause frank epithelial injury and that neonates have a different response pattern than adults.

GAS-PHASE FLAME SYNTHESIS AND PROPERTIES OF MAGNETIC IRON OXIDE NANOPARTICLES WITH REDUCED OXIDATION STATE.

Iron oxide nanoparticles of reduced oxidation state, mainly in the form of magnetite, have been synthesized utilizing a new continuous, gas-phase, nonpremixed flame method using hydrocarbon fuels. This method takes advantage of the characteristics of the inverse flame, which is produced by injection of oxidizer into a surrounding flow of fuel. Unlike traditional flame methods, this configuration allows for the iron particle formation to be maintained in a more reducing environment. The effects of flame temperature, oxygen-enrichment and fuel dilution (i.e. the stoichiometric mixture fraction), and fuel composition on particle size, Fe oxidation state, and magnetic properties are evaluated and discussed. The crystallite size, Fe(II) fraction, and saturation magnetization were all found to increase with flame temperature. Flames of methane and ethylene were used, and the use of ethylene resulted in particles containing metallic Fe(0), in addition to magnetite, while no Fe(0) was present in samples synthesized using methane.

Generation of oxidant response to copper and iron nanoparticles and salts: Stimulation by ascorbate.

The present work describes a two-stage approach to analyzing combustion-generated samples for their potential to produce oxidant stress. This approach is illustrated with the two commonly encountered transition metals, copper and iron. First, their abilities to generate hydroxyl radical were measured in a cell-free, phosphate-buffered saline solution containing ascorbate and/or citrate. Second, their abilities to induce heme oxygenase-1 in cultured human epidermal keratinocytes were assessed in cell culture. Combustion-generated copper oxide nanoparticles were active in both assays and were found to be soluble in culture medium. Depletion of glutathione in the cells or loading the cells with ascorbate greatly increased heme oxygenase-1 induction in the presence of copper. By contrast, iron oxide nanoparticles were active in the phosphate-buffered saline but not in cell culture, and they aggregated in culture medium. Soluble salts of copper and iron exhibited the same contrast in activities as the respective combustion-generated particles. The results suggest that the capability of combustion-generated environmental samples to produce oxidant stress can be screened effectively in a two step process, first in phosphate-buffered saline with ascorbate and subsequently in epithelial cell culture for those exhibiting activity initially. The results also point to an unanticipated interaction in cells of oxidant stress-generating metals with an antioxidant (ascorbate) that is usually missing in culture medium formulations. Thus, ascorbate supplementation of cultured human cells is likely to improve their ability to model the in vivo effects of particulate matter containing copper and other redox-active metals.

Environmental aging of polycyclic aromatic hydrocarbons on soot and its effect on source identification.

Soot-associated PAHs were exposed to simulated sunlight to investigate disappearance rates under environmental aging conditions and to examine the robustness of diagnostic ratios for PAH source apportionment. Naphthalene, acenaphthylene, acenaphthene, and fluorene showed an obvious two-phase disappearance in all experiments while phenanthrene and anthracene exhibited this behavior for all but the highest soot loading. The first phase loss is 5-40 times faster than the second phase loss and occurred within 3h for naphthalene, acenaphthylene, acenaphthene, and fluorene and within 10h for phenanthrene and anthracene. Two-phase disappearance was not observed for any of the higher molecular weight PAHs with 4-6 rings. Each PAH has a unique loss rate via photodegradation and volatilization and these rates of some PAHs were affected by soot loadings; phenanthrene and anthracene showed similar rates in the first phase and increased loss rates in the second phase as soot loading increased. In the absence of light, the loss of PAHs was related to both temperature and molecular characteristics. Due to differences in disappearance rates of individual PAHs under illumination over extended times, prolonged exposure to sunlight could change the interpretation of some diagnostic ratios used previously for PAH source identification. This result indicates that more consistent and accurate methods that take into consideration the longevity of particulate PAHs are needed for reliable source apportionment.

Redox Dynamics of Mixed Metal (Mn, Cr, and Fe) Ultrafine Particles.

The impact of particle composition on metal oxidation state, and on changes in oxidation state with simulated atmospheric aging, are investigated experimentally in flame-generated nanoparticles containing Mn, Cr, and Fe. The results demonstrate that the initial fraction of Cr(VI) within the particles decreases with increasing total metal concentration in the flame. In contrast, the initial Mn oxidation state was only partly controlled by metal loading, suggesting the importance of other factors. Two reaction pathways, one reductive and one oxidative, were found to be operating simultaneously during simulated atmospheric aging. The oxidative pathway depended upon the presence of simulated sunlight and O(3), whereas the reductive pathway occurred in the presence of simulated sunlight alone. The reductive pathway appears to be rapid but transient, allowing the oxidative pathway to dominate with longer aging times, i.e. greater than ∼8 hours. The presence of Mn within the particles enhanced the importance of the oxidative pathway, leading to more net Cr oxidation during aging implying that Mn can mediate oxidation by removal of electrons from other particulate metals.