Transcriptional activation of inflammasome components by Libby amphibole and the role of iron.

The induction of the NALP3 inflammasome complex is shown to be necessary for the development of fibrosis after asbestos exposure. Libby amphibole (LA) induces lung inflammation and fibrosis, while complexation of iron (Fe) on fibers inhibits inflammation. In this study we examined the ability of LA to induce the inflammasome cascade and the role of Fe in modulating inflammasome activity. Spontaneously hypertensive rats were exposed intratracheally to either saline (300 µl), deferoxamine (Def) (1 mg), FeCl(3) (21 µg), LA (0.5 mg), Fe-loaded LA (Fe + LA), or LA + Def. Activities of oxidative stress-sensitive enzymes, expression of inflammasome-specific genes, and cytokine proteins in bronchoalveolar lavage fluid were analyzed. Lung enzymes at 4 h and 24 h post-exposure were unchanged. LA increased lung expression of genes including interleukin-1ß (IL-1ß), cathepsin-B, ASC, NALP3, interleukin (IL)-6 and NFκB. LA+Fe significantly reduced IL-1ß and NFκB with a trend of reduction in ASC, NALP3, cathepsin-B and IL-6 expression. Def treatment did not reverse the inhibitory effect of Fe on IL-1ß and ASC but reversed IL-6 expression. CCL-7, CCL-12, CXCL-3 and COX-2 were induced by LA while LA+Fe tended to reduce these responses. Phosphorylation of ERK but not MEK was increased at 4 h after LA but not LA+Fe exposure. In conclusion, components of the NALP3 inflammasome are transcriptionally activated acutely during LA-induced inflammation. The key inflammatory regulators IL-1ß and NFκB were inhibited in the presence of surface-complexed Fe possibly through decreased ERK signaling upstream of the NALP3 inflammasome. The inflammasome activation by LA may contribute to fibrosis, and Fe may reduce this response and alter compensatory mechanisms in individuals exposed to LA.

Toxicity and recovery in the pregnant mouse after gestational exposure to the cyanobacterial toxin, cylindrospermopsin.

Cylindrospermopsin (CYN) is a tricyclic alkaloid toxin produced by fresh water cyanobacterial species worldwide. CYN has been responsible for both livestock and human poisoning after oral exposure. This study investigated the toxicity of CYN to pregnant mice exposed during different segments of gestation. The course of recovery and individual responses to the toxin were evaluated. Adverse effects of CYN were monitored up to 7 weeks post-dosing by clinical examination, histopathology, biochemistry and gene expression. Exposure on gestational days (GD) 8-12 induced significantly more lethality than GD13-17 exposure. Periorbital, gastrointestinal and distal tail hemorrhages were seen in both groups. Serum markers indicative of hepatic injury (alanine amino transferase, aspartate amino transferase and sorbitol dehydrogenase) were increased in both groups; markers of renal dysfunction (blood urea nitrogen and creatinine) were elevated in the GD8-12 animals. Histopathology was observed in the liver (centrilobular necrosis) and kidney (interstitial inflammation) in groups exhibiting abnormal serum markers. The expression profiles of genes involved in ribosomal biogenesis, xenobiotic and lipid metabolism, inflammatory response and oxidative stress were altered 24 h after the final dose. One week after dosing, gross, histological and serum parameters had returned to normal, although increased liver/body weight ratio and one instance of gastrointestinal bleeding was found in the GD13-17 group. Gene expression changes persisted up to 2 weeks post-dosing and returned to normal by 4 weeks. Responses of individual animals to CYN exposure indicated highly significant inter-animal variability within the treated groups.

Irrigation and fertilization effects on seed number, size, germination and seedling growth: implications for desert shrub establishment.

Plants with limited resources adjust partitioning among growth, survival, and reproduction. We tested the effects of water and nutrient amendments on seed production, size, and quality in Sarcobatus vermiculatus (greasewood) to assess the magnitude and importance of changes in reproductive partitioning. In addition, we assessed interactions among the environment of seed-producing plants (adult plant scale), seed size, and seedling microenvironment (seedling scale) on successful seedling establishment. Interactions of these factors determine the scale of resource heterogeneity that affects seedling establishment in deserts. Both total number of seeds produced per plant and seed quality (weight and germination) increased significantly in the enriched treatment in a 3-year field experiment. Seedling length 3 days after germination and seed N concentration, other measures of seed quality, were higher for seed from both irrigated and enriched plants than for seed from control plants. Field S. vermiculatus seed production and quality can be substantially increased with irrigation and nutrient enrichment at the adult plant scale and this allows management of seed availability for restoration. However, based on a greenhouse study, seedling environment, not the environment of the seed-producing plant or seed size, was the most important factor affecting seedling germination, survival, and growth. Thus it appears that production of more seed may be more important than improved seed quality, because higher quality seed did not compensate for a low-resource seedling environment. For both natural establishment and restoration this suggests that heterogeneity at the scale of seedling microsites, perhaps combined with fertilization of adult shrubs (or multi-plant patches), would produce the greatest benefit for establishing seedlings in the field.

Importance of internal hydraulic redistribution for prolonging the lifespan of roots in dry soil.

Redistribution of water within plants could mitigate drought stress of roots in zones of low soil moisture. Plant internal redistribution of water from regions of high soil moisture to roots in dry soil occurs during periods of low evaporative demand. Using minirhizotrons, we observed similar lifespans of roots in wet and dry soil for the grapevine 'Merlot' (Vitis vinifera) on the rootstock 101-14 Millardet de Gramanet (Vitis riparia x Vitis rupestris) in a Napa County, California vineyard. We hypothesized that hydraulic redistribution would prevent an appreciable reduction in root water potential and would contribute to prolonged root survivorship in dry soil zones. In a greenhouse study that tested this hypothesis, grapevine root systems were divided using split pots and were grown for 6 months. With thermocouple psychrometers, we measured water potentials of roots of the same plant in both wet and dry soil under three treatments: control (C), 24 h light + supplemental water (LW) and 24 h light only (L). Similar to the field results, roots in the dry side of split pots had similar survivorship as roots in the wet side of the split pots (P = 0.136) in the C treatment. In contrast, reduced root survivorship was directly associated with plants in which hydraulic redistribution was experimentally reduced by 24 h light. Dry-side roots of plants in the LW treatment lived half as long as the roots in the wet soil despite being provided with supplemental water (P < 0.0004). Additionally, pre-dawn water potentials of roots in dry soil under 24 h of illumination (L and LW) exhibited values nearly twice as negative as those of C plants (P = 0.034). Estimates of root membrane integrity using electrolyte leakage were consistent with patterns of root survivorship. Plants in which nocturnal hydraulic redistribution was reduced exhibited more than twice the amount of electrolyte leakage in dry roots compared to those in wet soil of the same plant. Our study demonstrates that besides a number of ecological advantages to protecting tissues against desiccation, internal hydraulic redistribution of water is a mechanism consistent with extended root survivorship in dry soils.

Influence of temporal heterogeneity in nitrogen supply on competitive interactions in a desert shrub community.

Soil nutrients in arid systems are supplied to plants in brief pulses following precipitation inputs. While these resource dynamics have been well documented, little is known about how this temporal heterogeneity influences competitive interactions. We examined the impacts of the temporal pattern of N supply on competitive intensity and ability in an N-limited desert shrub community. At our field site, the three codominant shrubs, Atriplex confertifolia, A. parryi, and Sarcobatus vermiculatus, differ in seasonal growth patterns, with A. confertifolia and S. vermiculatus achieving higher growth rates earlier in the growing season than A. parryi. We predicted that these timing differences in maximum growth rate may interact with temporal variation in N supply to alter competitive abilities over time. Seedlings of the two Atriplex species were planted either individually in field plots or as target plants surrounded by neighbor seedlings. After one year of establishment, the same amount of (15)N was applied to plots either as early spring pulses, mid spring pulses or continuously through the second growing season. Competitive effects were observed under continuous and pulsed N supply. Averaged across all target-neighbor treatments, competitive intensity was approximately 1.8-fold greater when N was pulsed compared to when N was supplied continuously, but overall, the outcome of competitive interactions was not influenced by N pulse timing. While the timing of resource supply did not differentially influence the competitive abilities of coexisting species in this system, the temporal pattern of resource supply did alter the intensity of competitive interactions among species. While additional studies in other systems are needed to evaluate the generality of these results, this study suggests that competitive intensity may not necessarily be a direct function of productivity or resource availability as traditionally assumed. Instead, the intensity of competitive interactions in resource-poor systems may depend upon the temporal pattern of resource supply.

High apoplastic solute concentrations in leaves alter water relations of the halophytic shrub, Sarcobatus vermiculatus.

Predawn plant water potential (Psi(w)) is used to estimate soil moisture available to plants because plants are expected to equilibrate with the root-zone Psi(w). Although this equilibrium assumption provides the basis for interpreting many physiological and ecological parameters, much work suggests predawn plant Psi(w) is often more negative than root-zone soil Psi(w). For many halophytes even when soils are well-watered and night-time shoot and root water loss eliminated, predawn disequilibrium (PDD) between leaf and soil Psi(w) can exceed 0.5 MPa. A model halophyte, Sarcobatus vermiculatus, was used to test the predictions that low predawn solute potential (Psi(s)) in the leaf apoplast is a major mechanism driving PDD and that low Psi(s) is due to high Na+ and K+ concentrations in the leaf apoplast. Measurements of leaf cell turgor (Psi(p)) and solute potential (Psi(s)) of plants grown under a range of soil salinities demonstrated that predawn symplast Psi(w) was 1.7 to 2.1 MPa more negative than predawn xylem Psi(w), indicating a significant negative apoplastic Psi(s). Measurements on isolated apoplastic fluid indicated that Na+ concentrations in the leaf apoplast ranged from 80 to 230 mM, depending on salinity, while apoplastic K+ remained around 50 mM. The water relations measurements suggest that without a low apoplastic Psi(s), predawn Psi(p) may reach pressures that could cause cell damage. It is proposed that low predawn apoplastic Psi(s) may be an efficient way to regulate Psi(p) in plants that accumulate high concentrations of osmotica or when plants are subject to fluctuating patterns of soil water availability.

Plant N capture from pulses: effects of pulse size, growth rate, and other soil resources.

In arid ecosystems, the ability to rapidly capture nitrogen (N) from brief pulses is expected to influence plant growth, survival, and competitive ability. Theory and data suggest that N capture from pulses should depend on plant growth rate and availability of other limiting resources. Theory also predicts trade-offs in plant stress tolerance and ability to capture N from different size pulses. We injected K15NO3, to simulate small and large N pulses at three different times during the growing season into soil around the co-dominant Great Basin species Sarcobatus vermiculatus, Chrysothamnus nauseosus ssp. consimilis, and Distichlis spicata. Soils were amended with water and P in a partial factorial design. As predicted, all study species showed a comparable decline in N capture from large pulses through the season as growth rates slowed. Surprisingly, however, water and P availability differentially influenced the ability of these species to capture N from pulses. Distichlis N capture increased up to tenfold with water addition while Chrysothamnus N capture increased up to threefold with P addition. Sarcobatus N capture was not affected by water or P availability. Opposite to our prediction, Sarcobatus, the most stress tolerant species, captured less N from small pulses but more N from large pulses relative to the other species. These observations suggest that variation in N pulse timing and size can interact with variable soil water and P supply to determine how N is partitioned among co-existing Great Basin species.

Nitrogen addition increases fecundity in the desert shrub Sarcobatus vermiculatus.

Nutrients, in addition to water, limit desert primary productivity, but nutrient limitations to fecundity and seed quality in desert ecosystems have received little attention. Reduced seed production and quality may affect recruitment, population, and community processes. At the Mono Basin, CA, USA where the alkaline, sandy soil has very low availability of N, P, and most other nutrients, seed production, recruitment, and dominance of the desert shrub Sarcobatus vermiculatus decrease over a dune successional sequence. Concurrently, Sarcobatus leaf N, P, and Ca/Mg ratio decline from early to later successional dunes. At two later successional dune sites, we fertilized adult Sarcobatus shrubs for 2 years and determined which nutrient(s) limited growth, seed production, and seed quality. We also tested whether nutrient addition at these older sites made these fitness-related variables equivalent to a younger, high-fecundity site. Nitrogen addition, alone, increased Sarcobatus leaf N, growth, and seed production per shoot module. Any treatment including P, Ca, Mg, or micronutrients but not N had an insignificant effect on growth and fecundity. Nitrogen addition also increased filled seed weight, a predictor of potential seedling survival, at one of the sites. Nitrogen-limited seed production and seed mass may reduce Sarcobatus fitness and contribute to the observed successional changes in plant community composition in this alkaline desert ecosystem.

Phosphorus in periphyton mats provides the best metric for detecting low-level P enrichment in an oligotrophic wetland.

Growing concern over the ecological consequence of phosphorus (P) enrichment in freshwater wetlands has elicited considerable debate over the concentration of water column P associated with eutrophication. In the oligotrophic Everglades, the displacement of native communities by enriched ones is widespread and has occurred at sites experiencing only minimal elevations in P input. To help define regulatory criteria for P inputs to the Everglades, we constructed an experiment that mimics P input to the natural system by continuously delivering P at concentrations elevated 5, 15 and 30 microgl(-1) above ambient to 100-m long flow-through channels. We compared patterns of P accumulation in the water, periphyton, detritus and soils among the channel treatments and also along a 16 km transect from an enriched canal that inflows to the interior of the same marsh. Water column TP and SRP were unrelated to input TP concentration in both the experiment and the marsh transect. However, concentrations of TP in periphyton mats were significantly elevated at all levels of experimental enrichment and as far as 2 km downstream from water inputs into the marsh. Elevated periphyton TP was associated with significant loss of periphyton biomass. In oligotrophic wetlands, traditional measures of water column SRP and TP will substantially underestimate P loading because biotically incorporated P is displaced from the water column to benthic surfaces. Using periphyton TP as a metric of P enrichment is uncomplicated and analogous to pelagic TP assessments in lakes where most P is sequestered in phytoplankton.

Extensive summer water pulses do not necessarily lead to canopy growth of Great Basin and northern Mojave Desert shrubs.

Plant species and functionally related species groups from arid and semi-arid habitats vary in their capacity to take up summer precipitation, acquire nitrogen quickly after summer precipitation, and subsequently respond with ecophysiological changes (e.g. water and nitrogen relations, gas exchange). For species that respond ecophysiologically, the use of summer precipitation is generally assumed to affect long-term plant growth and thus alter competitive interactions that structure plant communities and determine potential responses to climate change. We assessed ecophysiological and growth responses to large short-term irrigation pulses over one to three growing seasons for several widespread Great Basin and northern Mojave Desert shrub species: Chrysothamnus nauseosus, Sarcobatus vermiculatus, Atriplex confertifolia, and A. parryi. We compared control and watered plants in nine case studies that encompassed adults of all four species, juveniles for three of the species, and two sites for two of the species. In every comparison, plants used summer water pulses to improve plant water status or increase rates of functioning as indicated by other ecophysiological characters. Species and life history stage responses of ecophysiological parameters (leaf N, delta15N, delta13C, gas exchange, sap flow) were consistent with several previous short-term studies. However, use of summer water pulses did not affect canopy growth in eight out of nine comparisons, despite the range of species, growth stages, and site conditions. Summer water pulses affected canopy growth only for C. nauseosus adults. The general lack of growth effects for these species might be due to close proximity of groundwater at these sites, co-limitation by nutrients, or inability to respond due to phenological canalization. An understanding of the connections between short-term ecophysiological responses and growth, for different habitats and species, is critical for determining the significance of summer precipitation for desert community dynamics.

Genetics of drought adaptation in Arabidopsis thaliana: I. Pleiotropy contributes to genetic correlations among ecological traits.

We examined patterns of genetic variance and covariance in two traits (i) carbon stable isotope ratio delta13C (dehydration avoidance) and (ii) time to flowering (drought escape), both of which are putative adaptations to local water availability. Greenhouse screening of 39 genotypes of Arabidopsis thaliana native to habitats spanning a wide range of climatic conditions, revealed a highly significant positive genetic correlation between delta13C and flowering time. Studies in a range of C3 annuals have also reported large positive correlations, suggesting the presence of a genetically based trade-off between mechanisms of dehydration avoidance (delta13C) and drought escape (early flowering). We examined the contribution of pleiotropy by using a combination of mutant and near-isogenic lines to test for positive mutational covariance between delta13C and flowering time. Ecophysiological mutants generally showed variation in delta13C but not flowering time. However, flowering time mutants generally demonstrated pleiotropic effects consistent with natural variation. Mutations that caused later flowering also typically resulted in less negative delta13C and thus probably higher water use efficiency. We found strong evidence for pleiotropy using near-isogenic lines of Frigida and Flowering locus C, cloned loci known to be responsible for natural variation in flowering time. These data suggest the correlated evolution of delta13C and flowering time is explained in part by the fixation of pleiotropic alleles that alter both delta13C and time to flowering.

Night-time conductance in C3 and C4 species: do plants lose water at night?

Significant night-time stomatal conductance and transpiration were found for 11 out of 17 species with a range of life histories (herbaceous annual, perennial grass, shrub, tree), photosynthetic pathways (C(3), C(4)), and habitats in the western United States. Across species and habitats, higher night-time conductance and transpiration were associated with higher daytime values. The prevalence, mechanisms and ecological implications of substantial night-time water loss deserve further investigation.

Axial ligand modulation of the electronic structures of binuclear copper sites: analysis of paramagnetic 1H NMR spectra of Met160Gln Cu(A).

Cu(A) is an electron-transfer copper center present in heme-copper oxidases and N2O reductases. The center is a binuclear unit, with two cysteine ligands bridging the metal ions and two terminal histidine residues. A Met residue and a peptide carbonyl group are located on opposite sides of the Cu2S2 plane; these weaker ligands are fully conserved in all known Cu(A) sites. The Met160Gln mutant of the soluble subunit II of Thermus thermophilus ba3 oxidase has been studied by NMR spectroscopy. In its oxidized form, the binuclear copper is a fully delocalized mixed-valence pair, as are all natural Cu(A) centers. The faster nuclear relaxation in this mutant suggests that a low-lying excited state has shifted to higher energies compared to that of the wild-type protein. The introduction of the Gln residue alters the coordination mode of His114 but does not affect His157, thereby confirming the proposal that the axial ligand-to-copper distances influence the copper-His interactions (Robinson, H.; Ang, M. C.; Gao, Y. G.; Hay, M. T.; Lu, Y.; Wang, A. H. Biochemistry 1999, 38, 5677). Changes in the hyperfine coupling constants of the Cys beta-CH2 groups are attributed to minor geometrical changes that affect the Cu-S-C(beta)-H(beta) dihedral angles. These changes, in addition, shift the thermally accessible excited states, thus influencing the spectral position of the Cys beta-CH2 resonances. The Cu-Cys bonds are not substantially altered by the Cu-Gln160 interaction, in contrast to the situation found in the evolutionarily related blue copper proteins. It is possible that regulatory subunits in the mitochondrial oxidases fix the relative positions of thermally accessible Cu(A) excited states by tuning axial ligand interactions.

Diffuse alveolar damage after exposure to an oil fly ash.

Epidemiological investigation has established an association between exposure to particulate matter (PM) and both human mortality and diverse indices of human morbidity. However, attributing adverse health effects of specific individuals to PM exposure in these studies is not possible. Consequently, their clinical presentation remains ill-defined. We describe a 42-yr-old male with both respiratory damage, abnormal blood end points, and cardiac effects following an exposure to an emission source air pollution particle aerosolized during the cleaning of his domestic oil-burning stove. Early symptoms of shortness of breath and wheezing progressed over 2 wk to hypoxic respiratory failure necessitating mechanical ventilation. Blood indices were abnormal. Thoracoscopic biopsy demonstrated particle-laden macrophages and diffuse alveolar damage. Symptomatic and objective improvement rapidly followed initiation of corticosteroids. He developed typical anginal symptoms within 2 wk of discharge; however, coronary angiography did not identify any significant narrowing of the epicardial coronary arteries. This patient presents with the aggregate of potential injuries described by epidemiological methods to be associated with air pollution particle exposure.

Local adaptation across a climatic gradient despite small effective population size in the rare sapphire rockcress.

When assigning conservation priorities in endangered species, two common management strategies seek to protect remnant populations that (i) are the most genetically divergent or (ii) possess the highest diversity at neutral genetic markers. These two approaches assume that variation in molecular markers reflects variation in ecologically important traits and ignore the possibility of local adaptation among populations that show little divergence or variation at marker loci. Using common garden experiments, we demonstrate that populations of the rare endemic plant Arabis fecunda are physiologically adapted to the local microclimate. Local adaptation occurs despite (i) the absence of divergence at almost all marker loci and (ii) very small effective population sizes, as evidenced by extremely low levels of allozyme and DNA sequence polymorphism. Our results provide empirical evidence that setting conservation priorities based exclusively on molecular marker diversity may lead to the loss of locally adapted populations.

Pulsed EPR/ENDOR characterization of perturbations of the Cu(A) center ground state by axial methionine ligand mutations.

The effect of axial ligand mutation on the Cu(A) site in the recombinant water soluble fragment of subunit II of Thermus thermophilus cytochrome c oxidase ba(3) has been investigated. The weak methionine ligand was replaced by glutamate and glutamine which are stronger ligands. Two constructs, M160T0 and M160T9, that differ in the length of the peptide were prepared. M160T0 is the original soluble fragment construct of cytochrome ba(3) that encodes 135 amino acids of subunit II, omitting the transmembrane helix that anchors the domain in the membrane. In M160T9 nine C-terminal amino acids are missing, including one histidine. The latter has been used to reduce the amount of a secondary T2 copper which is most probably coordinated to a surface histidine in M160T0. The changes in the spin density in the Cu(A) site, as manifested by the hyperfine couplings of the weakly and strongly coupled nitrogens, and of the cysteine beta-protons, were followed using a combination of advanced EPR techniques. X-band ( approximately 9 GHz) electron-spin-echo envelope modulation (ESEEM) and two-dimensional (2D) hyperfine sublevel correlation (HYSCORE) spectroscopy were employed to measure the weakly coupled (14)N nuclei, and X- and W-band (95 GHz) pulsed electron-nuclear double resonance (ENDOR) spectroscopy for probing the strongly coupled (14)N nuclei and the beta-protons. The high field measurements were extremely useful as they allowed us to resolve the T2 and Cu(A) signals in the g( perpendicular) region and gave (1)H ENDOR spectra free of overlapping (14)N signals. The effects of the M160Q and M160E mutations were: (i) increase in A( parallel)((63,65)Cu), (ii) larger hyperfine coupling of the weakly coupled backbone nitrogen of C153, (iii) reduction in the isotropic hyperfine interaction, a(iso), of some of the beta-protons making them more similar, (iv) the a(iso) value of one of the remote nitrogens of the histidine residues is decreased, thus distinguishing the two histidines, and finally, (v) the symmetry of the g-tensor remained axial. These effects were associated with an increase in the Cu-Cu distance and subtle changes in the geometry of the Cu(2)S(2) core which are consistent with the electronic structural model of Gamelin et al. (Gamelin, D. R.; Randall, D. W.; Hay, M. T.; Houser, R. P.; Mulder, T. C.; Canters, G. W.; de Vries, S.; Tolman, W. B.; Lu, Y.; Solomon, E. I. J. Am. Chem. Soc. 1998, 120, 5246-5263).

Intra-specific variation in xylem cavitation in interior live oak (Quercus wislizenii A. DC.).

Xylem cavitation induced by water stress reduces plant hydraulic conductance and can indicate the habitat a species evolved in and its phylogenetic background. Species differ widely in cavitation resistance, but less is known about intra-specific variation. Cavitation resistance was assessed for field-collected adult and sapling size classes from three populations of interior live oak (Quercus wislizenii A. DC.) in California, USA. Root and stem cavitation resistance of two-year old seedlings from a greenhouse experiment was also measured. Cavitation resistance curves were determined by injecting air into the vascular system to induce cavitation and measuring the subsequent decline in hydraulic conductance. Based on the air-seeding hypothesis, the absolute value of the air pressures should be equivalent to the tensions that cause cavitation under dehydrating conditions. Conductance declined exponentially with applied pressure for both roots and stems. Comparisons between populations did not reveal significant differences despite good statistical power. The 50% loss in conductance point occurred between 1.0-1.6 MPa; conductance declined more slowly thereafter. Conductance was 21-30% of maximum at 4.0 MPa and 7-14% at 8.0 MPa. Saplings exhibited a nearly identical pattern compared with adults except at 4.0 MPa, where saplings exhibited slightly less cavitation (7%). Greenhouse seedling stems were more resistant compared with both field-collected adults and with seedling roots. The 50% loss in conductance point occurred at 0.83 and 2.6 MPa for seedling roots and stems, respectively. Seedling stems maintained conductance of 20.9% at 8.0 MPa while most roots were fully cavitated between 5.0-8.0 MPa.

Effect of ozone on diesel exhaust particle toxicity in rat lung.

Ambient particulate matter (PM) concentrations have been associated with mortality and morbidity. Diesel exhaust particles (DEP) are present in ambient urban air PM. Coexisting with DEP (and PM) is ozone (O(3)), which has the potential to react with some components of DEP. Some reports have shown increased lung injury in rats coexposed to PM and O(3), but it is unclear whether this increased injury was due to direct interaction between the pollutants or via other mechanisms. To examine whether O(3) can directly react with and affect PM bioactivity, we exposed DEP to O(3) in a cell-free in vitro system and then examined the bioactivity of the resultant DEP in a rat model of lung injury. Standard Reference Material 2975 (diesel exhaust PM) was initially exposed to 0.1 ppm O(3) for 48 h and then instilled intratracheally in Sprague-Dawley rats. Rat lung inflammation and injury was examined 24 h after instillation by lung lavage. The DEP exposed to 0.1 ppm O(3) was more potent in increasing neutrophilia, lavage total protein, and LDH activity compared to unexposed DEP. The increased DEP activity induced by the O(3) exposure was not attributable to alteration by air that was also present during the O(3) exposure. Exposure of DEP to a higher O(3) concentration (1.0 ppm) led to a decreased bioactivity of the particles. In contrast, carbon black particles, low in organic content relative to DEP, did not exhibit an increase in any of the bioactivities examined after exposure to 0.1 ppm O(3). DEP incorporated O(3) (labeled with (18)O) in a linear fashion. These data suggest that ambient concentrations of O(3) can increase the biological potency of DEP. The ozonized DEP may play a role in the induction of lung responses by ambient PM.

Iron disequilibrium in the rat lung after instilled blood.

STUDY OBJECTIVES: The extravasation of erythrocytes into the human lung occurs in a myriad of pulmonary disorders. Metal that is initially included in hemoglobin has been postulated to precipitate a disequilibrium in iron metabolism, to present an oxidative stress, and to contribute to tissue injury in several lung diseases. The objective of this study is to test the hypothesis that the tracheal instillation of blood in an animal model would have significant effects on iron equilibrium and would be associated with an injury to the lower respiratory tract. DESIGN: Rats were intratracheally instilled with either 1.0 mL saline solution (n = 36) or 1.0 mL blood (n = 36). Biochemical end points and histochemistry were obtained at times between 20 min and 14 days after the exposure to saline solution or blood. RESULTS: Total and nonheme iron concentrations in tracheal lavage fluid increased after the instillation of the blood. The percentage of neutrophils in the lavage fluid was elevated 1 day after the instillation of blood and remained at that level for at least 4 days following exposure, while protein concentrations were significantly increased at 1 day and 2 days only. Erythrocytes in the lung tissue were stained for hemoglobin immediately after exposure, but by 4 days after exposure, there was none. Ferritin was elevated between 1 day and 4 days after exposure, but by 7 days after exposure, the expression of this storage protein had returned to baseline values. CONCLUSIONS: We conclude that intratracheal instillation of whole blood in the rat can induce a neutrophilic lung injury that is associated with a disruption of normal iron metabolism. This disruption of the iron equilibrium is made evident by quantifying iron and staining for hemoglobin and ferritin. All indexes of biological effect had corrected by 7 days after exposure.