Kevin R. Smith

In vitro effects of coal fly ashes: hydroxyl radical generation, iron release, and DNA damage and toxicity in rat lung epithelial cells

Oxygen radical generation due to surface radicals, inflammation, and iron release has been suggested as the mechanism of adverse effects of quartz, such as emphysema, fibrosis, and carcinogenic effects. Therefore, we measured iron release, acellular generation of hydroxyl radicals, and oxidative DNA damage and cytotoxicity in rat lung epithelial (RLE) cells by different coal fly ashes (CFA) that contain both quartz and iron. Seven samples of CFA with different particle size and quartz content (up to 14.1%) were tested along with silica (alpha-quartz), ground coal, and coal mine dust (respirable) as positive control particles, and fine TiO(2) (anatase) as a negative control. Five test samples were pulverized fuel ashes (PFA), two samples were coal gasification (SCG) ashes (quartz content <0.1%), and one sample was a ground coal. No marked differences between SCG and PFA fly ashes were observed, and toxicity did not correlate with physicochemical characteristics or effect parameters. Stable surface radicals were only detected in the reference particles silica and coal mine dust, but not in CFA. On the other hand, hydroxyl radical generation by all fly ashes was observed in the presence of hydrogen peroxide, which was positively correlated with iron mobilization and inhibited by deferoxamine, but not correlated with iron or quartz content. Also a relationship between acellular hydroxyl radical generation and oxidative DNA damage in RLE cells by CFA was observed. Differences in hydroxyl radical generation and oxidative damage by the CFA were not related to iron and quartz content, but the respirable ashes (MAT023, 38, and 41) showed a very extensive level of hydroxyl radical generation in comparison to nonrespirable fly ashes and respirable references. This radical generation was clearly related to the iron mobilization from these particles. In conclusion, the mechanisms by which CFA and the positive references (silica, coal mine dust) affect rat lung epithelial cells seem to be different, and the data suggest that quartz in CFA does not act the same as quartz in silica or coal mine dust. On the other hand, the results indicate an important role for size and iron release in generation and subsequent effects of reactive oxygen species caused by CFA.

BIOAVAILABILITY OF IRON FROM COAL FLY ASH: Mechanisms of Mobilization and of Biological Effects

Particulate air pollution contains iron that may be involved in the pathological effects after inhalation. This article reviews work demonstrating that ambient particulate samples (Standard Reference Material [SRM] 1648 and SRM 1649, from the National Institute of Science and Technology) contain iron that can be mobilized from the particle in vitro and inside human lung epithelial (A549) cells. The mobilized iron can then catalyze the formation of reactive oxygen species (ROS). Work is also reviewed on the generation and size fractionation of coal fly ash (CFA) from three commercially important coal types, as well as size fractionation of three types of noncombustion particles. The availability of iron from these particles to A549 cells was measured by citrate mobilization in vitro and induction of the iron storage protein ferritin in particle-treated cells. The amount of bioavailable iron decreased with increasing particle size. The ability of particles to induce synthesis of the proinflammatory cytokine interleukin-8 (IL-8) was also determined. As with the bioavailability of iron, there was an inverse correlation with size. Further work showed that iron in CFA is responsible for IL-8 induction. Mössbauer spectroscopy of a CFA sample before and after desferrioxamine B treatment to remove bioavailable iron showed that the bioavailable iron was associated with the glassy aluminosilicate fraction of the particle. In conclusion, this work shows that bioavailable iron is responsible for ROS production by SRMs and IL-8 induction by CFA in A549 cells. The source of this bioavailable iron in CFA is glassy aluminosilicates, which are found at higher levels in smaller sizes of CFA.

Oxidative injury in the lungs of neonatal rats following short-term exposure to ultrafine iron and soot particles

Greater risk of adverse effects from particulate matter (PM) has been noted in susceptible subpopulations, such as children. However, the physicochemical components responsible for these biological effects are not understood. As critical constituents of PM, transition metals were postulated to be involved in a number of pathological processes of the respiratory system through free radical-medicated damage. The purpose of this study was to examine whether oxidative injury in the lungs of neonatal rats could be induced by repeated short-term exposure to iron (Fe) and soot particles. Sprague Dawley rats 10 d of age were exposed by inhalation to two different concentrations of ultrafine iron particles (30 or 100 µg/m3) in combination with soot particles adjusted to maintain a total particle concentration of 250 µg/m3. Exposure at 10 d and again at 23 d of age was for 6 h/d for 3 d. Oxidative stress was observed at both Fe concentrations in the form of significant elevations in glutathione disulfide (GSSG) and GSSG/glutathione (GSH) ratio and a reduction in ferric/reducing antioxidant power in bronchoalveolar lavage. A significant decrease in cell viability associated with significant increases in lactate dehydrogenase (LDH) activity, interleukin-1-beta (IL-1β), and ferritin expression was noted following exposure to particles containing the highest Fe concentration. Iron from these particles was shown to be bioavailable in an in vitro assay using the physiologically relevant chelator, citrate. Data indicate that combined Fe and soot particle exposure induces oxidative injury, cytotoxicity and pro-inflammatory responses in the lungs of neonatal rats.

Coal Fly Ash and Mineral Dust for Toxicology and Particle Characterization Studies: Equipment and Methods for PM2.5- and PM1-Enriched Samples

Laboratory methods to produce particle samples from known, reproducible sources with sufficient mass to perform both detailed characterization and replicated in vitro toxicological assays are described. These samples are being used to study the ability of inhalable particles to produce abnormal concentrations of intracellular iron, resulting in the production of reactive oxygen species in cultured airway epithelial cells. Bulk samples of size fractionated particles from laboratory-generated coal fly ash and from simulated fugitive mining tailings and road dust were collected as surrogates for important sources of iron-bearing particles in the ambient air. An Andersen cascade impactor was used to produce particle samples enriched in three size ranges: > 10 mu m, 10-2.5 mu m, and < 2.5 mu m aerodynamic diameter. A multijet preseparator and rectangular slot virtual impactor were used to produce a fraction enriched in particles below 1 mu m. Data on the particle production conditions, production rates, and particle sample quality are provided to illustrate the feasibility of the experimental approach. The amount of iron mobilized from particles by a physiologically-relevant chelator does not correlate with the total iron. This supports the hypothesis that particle characteristics and iron speciation are important for the production of abnormal iron concentrations in cultured type A549 human airway epithelial cells. Comparison of results obtained with these surrogate particles to previous work with urban particulate standard reference materials (SRM 1648 and SRM 1649) suggests particle sources and size fractions that should be emphasized for detailed characterization of particle morphology and mineralogy.

Gestational exposure of mice to secondhand cigarette smoke causes bronchopulmonary dysplasia blocked by the nicotinic receptor antagonist mecamylamine.

Background: Cigarette smoke (CS) exposure during gestation may increase the risk of bronchopulmonary dysplasia (BPD)—a developmental lung condition primarily seen in neonates that is characterized by hypoalveolarization, decreased angiogenesis, and diminished surfactant protein production and may increase the risk of chronic obstructive pulmonary disease. Objective: We investigated whether gestational exposure to secondhand CS (SS) induced BPD and sought to ascertain the role of nicotinic acetylcholine receptors (nAChRs) in this response. Methods: We exposed BALB/c and C57BL/6 mice to filtered air (control) or SS throughout the gestation period or postnatally up to 10 weeks. Lungs were examined at 7 days, 10 weeks, and 8 months after birth. Results: Gestational but not postnatal exposure to SS caused a typical BPD-like condition: suppressed angiogenesis [decreased vascular endothelial growth factor (VEGF), VEGF receptor, and CD34/CD31 (hematopoietic progenitor cell marker/endothelial cell marker)], irreversible hypoalveolarization, and significantly decreased levels of Clara cells, Clara cell secretory protein, and surfactant proteins B and C, without affecting airway ciliated cells. Importantly, concomitant exposure to SS and the nAChR antagonist mecamylamine during gestation blocked the development of BPD. Conclusions: Gestational exposure to SS irreversibly disrupts lung development leading to a BPD-like condition with hypoalveolarization, decreased angiogenesis, and diminished lung secretory function. Nicotinic receptors are critical in the induction of gestational SS–induced BPD, and the use of nAChR antagonists during pregnancy may block CS-induced BPD.

Wood Smoke Enhances Cigarette Smoke–Induced Inflammation by Inducing the Aryl Hydrocarbon Receptor Repressor in Airway Epithelial Cells

Our previous studies showed that cigarette smokers who are exposed to wood smoke (WS) are at an increased risk for chronic bronchitis and reduced lung function. The present study was undertaken to determine the mechanisms for WS-induced adverse effects. We studied the effect of WS exposure using four cohorts of mice. C57Bl/6 mice were exposed for 4 or 12 weeks to filtered air, to 10 mg/m(3) WS for 2 h/d, to 250 mg/m(3) cigarette smoke (CS) for 6 h/d, or to CS followed by WS (CW). Inflammation was absent in the filtered air and WS groups, but enhanced by twofold in the bronchoalveolar lavage of the CW compared with CS group as measured by neutrophil numbers and levels of the neutrophil chemoattractant, keratinocyte-derived chemokine. The levels of the anti-inflammatory lipoxin, lipoxin A4, were reduced by threefold along with cyclo-oxygenase (COX)-2 and microsomal prostaglandin E synthase (mPGES)-1 in airway epithelial cells and PGE2 levels in the bronchoalveolar lavage of CW compared with CS mice. We replicated, in primary human airway epithelial cells, the changes observed in mice. Immunoprecipitations showed that WS blocked the interaction of aryl hydrocarbon receptor (AHR) with AHR nuclear transporter to reduce expression of COX-2 and mPGES-1 by increasing expression of AHR repressor (AHRR). Collectively, these studies show that exposure to low concentrations of WS enhanced CS-induced inflammation by inducing AHRR expression to suppress AHR, COX-2, and mPGES-1 expression, and levels of PGE2 and lipoxin A4. Therefore, AHRR is a potential therapeutic target for WS-associated exacerbations of CS-induced inflammation.

Farming, marketing, and changes in the authority of elders among pastoral Rendille and Ariaal.

Pastoral cultures of East Africa include certain institutions that meet requirements for labor organization and control over resources. Institutions that have been adapted to multi-species pastoralism in arid or semi-arid environments include: a patriarchical and gerontocratic control over resources that permits polygyny; an age-system that both dictates work roles and legitimizes control over resources; and marriage practices that permit elder males to control the labor of unmarried junior males. All of these institutions and ideals are interrelated, reinforcing each other and perpetuating the social and ideological order of society. When the economy changes, however, institutions may also change or perhaps become less useful for organizing society. Such is the case for Rendille and Ariaal of northern Kenya who now farm and are more involved in the market economy than their pastoral kin. The result is greater economic opportunity for young men of the warrior age grade, while ways for elders to maintain their authority continue to exist.