Long-term Ozone Exposure Research Articles

Long-Term Ozone Exposure and Cardiovascular Mortality in a Large-Scale Prospective Study

There are suggestions that ground-level ozone (O3) may be associated with adverse cardiovascular impacts, including death. However, results from long-term epidemiological studies are scarce and con...

Large-scale protein analysis of European beech trees following four vegetation periods of twice ambient ozone exposure.

This study provides useful information on how European beech, an economically and ecologically important tree species, reacts on the molecular level to increased ozone concentrations expected in the near future. The main emphasis in the present study was placed on identifying differentially abundant proteins after long-term ozone exposure under climatically realistic settings, rather than short-term responses or reactions under laboratory conditions. Additionally, using nursery-grown beech trees, we took into account the natural genotypic variation of this species. As such, the results presented here provide information on molecular responses to ozone in an experimental plant system at very close to natural conditions. Furthermore, this proteomic approach was supported by previous studies on the present experiment. Ultimately, the combination of this proteomic approach with several approaches including transcriptomics, analysis of non-structural carbohydrates, and morphological effects contributes to a more global picture of how beech trees react under increased ozone concentrations.

High tolerance of subalpine grassland to long-term ozone exposure is independent of N input and climatic drivers

In a seven-year study, we tested effects of increased N and O3 deposition and climatic conditions on biomass of subalpine grassland. Ozone risk was assessed as exposure (AOT40) and as stomatal flux (POD0,1). We hypothesized that productivity is higher under N- and lower under O3 deposition, with interactions with climatic conditions.Aboveground biomass was best correlated with growing-degree days for May (GDDMay). Nitrogen deposition increased biomass up to 60% in the highest treatment, and 30% in the lowest addition. Also belowground biomass showed a positive N-response. Ozone enrichment had no effect on biomass, and no interaction between O3 and N was observed. Growth response to N deposition was not correlated to GDDMay or precipitation, but indicated a cumulative effect over time.Productivity of subalpine grassland is tolerant to increasing ozone exposure, independent of N input and climatic drivers. N deposition rates at current critical loads, strongly increase the grassland yield.

Ozone and Survival in Four Cohorts with Potentially Predisposing Diseases

Time series studies have reported associations between ozone and daily deaths. Only one cohort study has reported the effect of long-term exposures on deaths, and little is known about effects of chronic ozone exposure on survival in susceptible populations. We investigated whether ozone was associated with survival in four cohorts of persons with specific diseases in 105 United States cities, treating ozone as a time varying exposure. We used Medicare data (1985–2006), and constructed cohorts of persons hospitalized with chronic conditions that might predispose to ozone effects: chronic obstructive pulmonary disease, diabetes, congestive heart failure, and myocardial infarction. Yearly warm-season average ozone was merged to the individual follow-up in each city. We applied Cox proportional hazard model for each cohort within each city, adjusting for individual risk factors, temperature, and city-specific long-term trends. We found significant associations with a hazard ratio for mortality of 1.06 (95% confidence interval [CI], 1.03–1.08) per 5-ppb increase in summer average ozone for persons with congestive heart failure; of 1.09 (95% CI, 1.06–1.12) with myocardial infarction; of 1.07 (95% CI, 1.04–1.09) with chronic obstructive pulmonary disease; and of 1.07 (95% CI, 1.05–1.10) for diabetics.We also found that the effect varied by region, but that this was mostly explained by mean temperature, which is likely a surrogate of air conditioning use, and hence exposure. This is the first study that follows persons with specific chronic conditions, and shows that long-term ozone exposure is associated with increased risk of death in these groups.

Changes in the proteome of juvenile European beech following three years exposure to free-air elevated ozone

Tropospheric ozone, one of the most phytotoxic air pollutants, may specially impose in long-lived forest trees substantial reduction in productivity and biomass. European beech saplings grown in lysimeter around areas were used to monitor proteomic changes upon elevated ozone concentrations following four vegetation periods of exposure. A proteome study based on highly sensitive two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) was performed to identify protein changes in European beech, the most important deciduous tree in Central Europe. Main emphasis was on identifying differentially expressed proteins after long-time period of ozone exposure under natural conditions rather than short-term responses or reactions under controlled conditions. Our results clearly demonstrate a response of European beech saplings to long-term ozone fumigation at the protein level. We indicate changes in the protein abundance of 142 protein spots; among them 59 were increased and 83 decreased following three years of elevated ozone exposure. As the first step, 40 proteins were identified by a homology driven mass spectrometric approach. Some of the identified proteins have been previously described in the context of short-term ozone responses in plants, indicating, at least for certain cellular functions, the congruence of plant reactions following short- and long-term ozone exposure. Under elevated ozone exposure, abundance of proteins related to the Calvin cycle and photosynthetic electron transport chain were decreased whereas the abundance of proteins regarding the carbon metabolism/catabolism were increased.

The Global Burden of Ozone on Respiratory Mortality: No Clear Evidence for Association

Anenberg et al. (2010) estimated the global burden of respiratory mortality attributable to long-term ozone exposure based on a single observational study by Jerrett et al. (2009). Because no other study has clearly demonstrated impacts of chronic ozone exposure on deaths from respiratory-related causes, we believe that reliance on the study by Jerrett et al. to establish causality and global impact is misplaced and that the conclusions of Anenberg et al. are likely unfounded. Jerrett et al. (2009) carried out a follow-up analysis of the American Cancer Society (ACS) cohort. Other ACS studies reported no associations between long-term ozone exposure and cardiopulmonary mortality that are robust to model inclusion of co-pollutants (e.g., Krewski et al. 2000; Pope et al. 2002). In addition, other long-term studies of ozone-related respiratory or cardiopulmonary mortality did not report positive associations (Goodman 2010; Health Effects Institute 2009). Anenberg et al. (2010) suggested that long-term respiratory mortality is plausible because short-term ozone mortality has been documented, but inconsistent evidence for an association between short-term ozone exposure and respiratory mortality indicates that this relationship is not well established. Jerrett et al. (2009) did not provide “clear” evidence of an association between long-term ozone exposure and respiratory mortality, as Anenberg et al. (2010) stated in their article. Jerrett et al. (2009) did not adequately control for potential confounding effects of particulate matter ≤ 2.5 μm in aerodynamic diameter (PM2.5) for several reasons. Jerrett et al. (2009) used only 2 years of data for PM2.5 (1999–2000) but ozone concentrations from 1977–2000. Although ozone and PM2.5 levels decreased considerably from 1977 to 2000, they used higher ozone levels observed in the past but only the more recent PM2.5 levels. Furthermore, their ozone metric focused on daily maximum hourly levels in the warm seasons, whereas they used annual average PM2.5 concentrations. As noted by Jerrett et al. (2009), this approach likely increased the potential to observe an association between ozone and mortality and decreased the ability to observe potential PM2.5 confounding of this association. In addition, confounding by other co-pollutants (e.g., sulfur dioxide), a clear issue in earlier ACS analyses (Krewski et al. 2000), was not examined. Accordingly, Jerrett et al. did not demonstrate an association between ozone and respiratory mortality that is independent of other co-pollutants. Another aspect of the Jerrett et al. (2009) study that is inconsistent with an association between long-term ambient ozone exposure and respiratory mortality is the biologically implausible, inverse associations of ozone with cardiovascular and all-cause mortality. The magnitude of these associations is the same—although opposite in direction—as the risk estimate for respiratory mortality; thus, it is likely that associations of this magnitude are not indicative of a causal relationship. It was inappropriate for Jerrett et al. (2009) to combine data across cities for a U.S. national risk estimate, given the known geographic heterogeneity of ozone-mortality findings (Goodman 2010). In addition, socioeconomic data (a potential confounder) was collected in 1982–1983 for the ACS study but never updated. For these reasons, the U.S. national risk estimate reported by Jerrett et al. (2009) should not be extrapolated globally. The analysis by Anenberg et al. (2010) was based on an uncorroborated study that likely misinterpreted the findings regarding ozone effects. The utility of estimating the global burden of an effect based on a single study, for which no causal association has been established in other studies, is not apparent. Conclusions drawn from such an analysis should be interpreted with caution.

Minor effects of long-term ozone exposure on boreal peatland species Eriophorum vaginatum and Sphagnum papillosum

The effects of long-term ozone fumigation on two common peatland plant species, a sedge Eriophorum vaginatum L. and a moss Sphagnum papillosum Lindb., were studied applying peatland microcosms. The peat cores with intact vegetation were cored from an oligotrophic pine fen and partially embedded into the soil of an open-air experimental field for four growing seasons. The open-air ozone exposure field consists of eight circular plots of which four were fumigated with elevated ozone concentration (doubled ambient) and four were ambient controls. The results showed that E. vaginatum and S. papillosum can tolerate ozone better than expected. Elevated ozone concentration did not affect overall relative length growth of E. vaginatum or S. papillosum. The leaf cross-section area of E. vaginatum leaves was 8% bigger in the ozone treatment compared to that in the ambient control. Ultrastuctural variables did not show any significant treatment effect in E. vaginatum or in S. papillosum. Total chlorophyll (a + b) concentration tended to increase in early growing season under ozone exposure. During the first growing season, elevated ozone concentration decreased methanol-extractable, UV-absorbing compounds in E. vaginatum. The results suggest that E. vaginatum and S. papillosum are ozone tolerant plant species and are likely able to cope with expected increase in tropospheric ozone concentration.

Promotion of cardiovascular disease by exposure to the air pollutant ozone

in the decades following the discovery of the chemical nature of environmental air pollutants and the biochemical reactions they could undergo, it was viewed that the primary tissue target of air pollution would be our lungs. This made sense, as the principal constituents of photochemical air

Long-Term Ozone Exposure and Mortality

Although many studies have linked elevations in tropospheric ozone to adverse health outcomes, the effect of long-term exposure to ozone on air pollution-related mortality remains uncertain. We examined the potential contribution of exposure to ozone to the risk of death from cardiopulmonary causes and specifically to death from respiratory causes. Data from the study cohort of the American Cancer Society Cancer Prevention Study II were correlated with air-pollution data from 96 metropolitan statistical areas in the United States. Data were analyzed from 448,850 subjects, with 118,777 deaths in an 18-year follow-up period. Data on daily maximum ozone concentrations were obtained from April 1 to September 30 for the years 1977 through 2000. Data on concentrations of fine particulate matter (particles that are < or = 2.5 microm in aerodynamic diameter [PM(2.5)]) were obtained for the years 1999 and 2000. Associations between ozone concentrations and the risk of death were evaluated with the use of standard and multilevel Cox regression models. In single-pollutant models, increased concentrations of either PM(2.5) or ozone were significantly associated with an increased risk of death from cardiopulmonary causes. In two-pollutant models, PM(2.5) was associated with the risk of death from cardiovascular causes, whereas ozone was associated with the risk of death from respiratory causes. The estimated relative risk of death from respiratory causes that was associated with an increment in ozone concentration of 10 ppb was 1.040 (95% confidence interval, 1.010 to 1.067). The association of ozone with the risk of death from respiratory causes was insensitive to adjustment for confounders and to the type of statistical model used. In this large study, we were not able to detect an effect of ozone on the risk of death from cardiovascular causes when the concentration of PM(2.5) was taken into account. We did, however, demonstrate a significant increase in the risk of death from respiratory causes in association with an increase in ozone concentration.

Long-term ozone exposure of potato: Free radical content and leaf injury analysed by Q-band ESR spectroscopy and image analysis

This paper presents Q-band electron spin resonance (ESR) studies on free radicals (FR) generated in potato leaves exposed to different O3 levels in open-top chambers (OTC), together with a quantitative study of the relationship between FR signal intensity and area of potato leaf damage. The advantages of Q-band when compared to X-band ESR spectroscopy are analysed, the main advantage being an absence of overlapping between Mn(II) and FR signals, allowing a quantitative study of FR signal intensity. This study also reports on a graphical method developed to quantitatively measure the damaged area on leaves caused by ozone exposure. Results indicate a direct relationship between FR signal intensity (measured as area under the signal) and percentage of O3 damage and clearly demonstrate a close relationship between visible ozone-induced symptoms and permanent FR concentration in potato leaves.

Long-Term Ozone Exposure Attenuates 1-Nitronaphthalene–Induced Cytotoxicity in Nasal Mucosa

1-Nitronaphthalene (1-NN) and ozone are cytotoxic air pollutants commonly found as components of photochemical smog. The mechanism of toxicity for 1-NN involves bioactivation by cytochrome P450s and subsequent adduction to proteins. Previous studies have shown that 1-NN toxicity in the lung is considerably higher in rats after long-term exposure to ozone compared with the corresponding filtered air-exposed control rats. The aim of the present study was to establish whether long-term exposure to ozone alters the susceptibility of nasal mucosa to the bioactivated toxicant, 1-NN. Adult male Sprague-Dawley rats were exposed to filtered air or 0.8 ppm ozone for 8 hours per day for 90 days, followed by a single treatment with 0, 12.5, or 50.0 mg/kg 1-NN by intraperitoneal injection. The results of the histopathologic analyses show that the nasal mucosa of rats is a target of systemic 1-NN, and that long-term ozone exposure markedly lessens the severity of injury, as well as the protein adduct formation by reactive 1-NN metabolites. The antagonistic effects were primarily seen in the nasal transitional epithelium, which corresponds to the main site of histologic changes attributed to ozone exposure (goblet cell metaplasia and hyperplasia). Long-term ozone exposure did not appear to alter susceptibility to 1-NN injury in other nasal regions. This study shows that long-term ozone exposure has a protective effect on the susceptibility of nasal transitional epithelium to subsequent 1-NN, a result that clearly contrasts with the synergistic toxicological effect observed in pulmonary airway epithelium in response to the same exposure regimen.

Shift in birch leaf metabolome and carbon allocation during long-term open-field ozone exposure

Shift in birch leaf metabolome and carbon allocation during long-term open-field ozone exposure

Effects of long-term ozone exposure on citrus: Chlorophyll a fluorescence and gas exchange

Three-years-old trees of Satsuma mandarin (Citrus unshiu [Mak.] Marc.) cv. Okitsu were exposed to O3 fumigation during long term (one year) in open-top chambers. As a result of the treatment, chlorophyll a fluorescence and gas exchange parameters were modified with respect to trees growing in O3-free conditions. Net photosynthetic rate and stomatal conductance decreased and intercellular CO2 concentration increased according to a reduction of the non-cyclic electron flow and a lower capacity to reduce the quinone pool. O3 also reduced the development of non-photochemical quenching preventing the dissipation of excess excitation energy and, therefore, generated several alterations in photosynthetic apparatus. All these effects were obtained in long-term exposure and higher O3 concentration. In O3 ambient conditions, the effects were minor.

Structural changes in the vascular bundles of light-exposed and shaded spruce needles suffering from Mg deficiency and ozone pollution

The correlation between structural changes of the vascular bundles and needle yellowing was examined for needles of damaged spruce (Picea abies (L.) Karst.) growing at a Mg-deficient and ozone polluted mountain site in the Central Black Forest (840m a.s.l.). In the previous year's sun-exposed needles, the following sequence of events was observed: (1) rapid needle yellowing, (2) hypertrophy and anomalous divisions of cambium cells, (3) phloem collapse, and, (4) production of atypical xylem tracheids. Under defined shade (reduction of the photosynthetically active photon flux density of the ambient light by 85-90%), the needles remained green, while the phloem collapsed completely within the first 6 weeks of shading; subsequently, a reversal of the collapse was observed. Under both light conditions, the content of Mg not bound to chlorophyll (Mg(free)) was in the range of 0.1 mg g(-1) needle dry matter, and hardly changed throughout the investigation period. After Mg fertilization, the Mg(free) level of the previous year's needles increased to 0.2 mg g(-1) dry matter, the light-exposed needles remained green, and the vascular bundles developed no anomalies. The data show that the rapid needle yellowing of ozone-exposed Mg-deficient needles did not depend on the collapse of the phloem. Mg deficiency played a key role in the development of anomalous vascular bundles under light, and also appears to explain the transient changes in sieve cell structure under shade. The role of Mg deficiency, rather than ozone pollution, in the damage of the sieve cells was confirmed in a long-term ozone exposure experiment with young clonal spruce growing under defined conditions.

In Vivo Effects of Ozone Exposure on Protein Adduct Formation by 1-Nitronaphthalene in Rat Lung

The incidence of serious photochemical smog events is steadily growing in urban environments around the world. The electrophilic metabolites of 1-nitronaphthalene (1-NN), a common air pollutant in urban areas, have been shown to bind covalently to proteins. 1-NN specifically targets the airway epithelium, and the toxicity is synergized by prior long-term ozone exposure in rat. In this study we investigated the formation of 1-NN protein adducts in the rat airway epithelium in vivo and examined how prior long-term ozone exposure affects adduct formation. Eight adducted proteins, several involved in cellular antioxidant defense, were identified. The extent of adduction of each protein was calculated, and two proteins, peroxiredoxin 6 and biliverdin reductase, were adducted at high specific activities (0.36-0.70 and 1.0 nmol adduct/nmol protein). Furthermore, the N-terminal region of calreticulin, known as vasostatin, was adducted only in ozone-exposed animals. Although vasostatin was adducted at relatively low specific activity (0.01 nmol adduct/nmol protein), the adduction only in ozone-exposed animals makes it a candidate protein for elucidating the synergistic toxicity between ozone and 1-NN. These studies identified in vivo protein targets for reactive 1-NN metabolites that are potentially associated with the mechanism of 1-NN toxicity and the synergistic effects of ozone.

Long-term ozone exposure and ozone uptake of grapevines in open-top chambers

Ozone uptake of pot-grown grapevines, fruit yield and sugar concentrations in juice were studied over the course of 4 years. In three of the study years, plants were exposed to contrasting ozone levels in open-top chambers. Results showed that juice quality was more sensitive to ozone exposure than grape yield. Ozone-induced reductions in grape yield were best described by the level of ozone exposure in the 2 years prior to harvest, whereas sugar content of juice was mainly affected by current and previous year ozone exposure. Regression models were developed to facilitate quantitative assessments of the effects of ozone on yield and quality. The observed effects on grape yield and on carbohydrate accumulation in fruit were slightly better explained when a threshold of 6 nmol m −2 s −1 was applied to cumulative uptake of ozone (CUO6) compared with AOT40 exposure or other CUO thresholds. Curvilinear regression models were developed which explained up to 75% of variance of the datasets. These models indicate a critical level of 1.1 mmol m −2 ozone (CUO6 from June to September) to protect carbohydrate accumulation in fruit with ozone taken up by the plants over consecutive years. For grape yield the CUO6 threshold was 2.2 mmol m −2 ozone. Exceedance of this threshold would be expected to reduce grape yield and sugar translocation to fruit by 10% with substantial degree of uncertainty (95% confidence interval 1–19%). For individual years, the critical level could be increased to 2.3 mmol m −2 for carbohydrate accumulation and 3.5 mmol m −2 CUO6 for grape yield if the CUO6 of 0 in the subsequent year was not exceeded.

Ozone Impacts on Vegetation

During the past 50 years, phytotoxic effects of tropospheric ozone on plants have been observed over large areas of the northern hemisphere. The present review highlights some important aspects of the interaction between tropospheric ozone and plants. These include the transfer of ozone from the air to the site of action inside the leaves, primary reactions at the cell membrane, metabolic disturbances, and resulting effects on plant growth. Based on experimental exposure-response relationships, the ozone risk for vegetation can be assessed; in Europe, these assessments suggest widespread impacts of short-term and long-term ozone exposure on plants under well-irrigated conditions.

Biochemical changes in needles of Pinus radiata D. Don trees in relationship to long-term ozone exposure indices

Ambient concentrations of ozone in Europe are high enough to cause negative effects on vegetation. Therefore, many efforts have been made to determine exposure indices and critical levels for protection of vegetation. In this context, the choice of a suitable attribute to determine the pollutant effect is of paramount importance. Until now, much of the work has been done with attributes such as biomass or growth. In the present work correlation factors have been established between biochemical parameters (peroxidase activity, ascorbate and sulfhydryl contents) of Pinus radiata trees and exposure indices of ozone. Our results show that peroxidase cannot be used as an indicator of effects of long-term exposure to ozone but still remains as an excellent indicator of short-term ozone fluctuations in the field. Ascorbate may act as an intermediate indicator responding to both short fluctuations and long-term exposures to ozone. Finally, sulfhydryl may be used as a long-term indicator in relation to the AOT (average over threshold) exposure index. Our results also point to the fact that Pinus radiata may be affected by ozone at AOT values lower than 10 ppm.h as already observed with other tree species.

Long-Term Residence in Areas of High Ozone: Associations with Respiratory Health in a Nationwide Sample of Nonsmoking Young Adults

Few studies have examined the respiratory effects of multiyear ozone exposures in human populations. We examined associations between current respiratory health status and long-term ozone exposure histories in 520 Yale College (New Haven, CT) students who never smoked. Questionnaires addressed current respiratory symptoms, respiratory disease history, residential history, and other factors. The symptoms of cough, phlegm, wheeze apart from colds, and a composite respiratory symptom index (RSI) were selected as outcome measures. Forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV(1)), forced expiratory flow rate between 25 and 75% of FVC (FEF(25-75)), and forced expiratory flow rate at 75% of FVC (FEF(75)) were obtained by forced expiration into spirometers. Ozone exposure was treated as a dichotomous variable, where subjects were assigned to the high-exposure group if they lived for 4 or more years in a U.S. county with 10-year average summer-season daily 1-hr maximum ozone levels [greater/equal to] 80 ppb. Lung function and respiratory symptoms were analyzed by multiple linear and logistic regression on ozone exposure, controlling for covariates. Lung function was lower in the group with high ozone exposures: differences were statistically significant for FEV(1) [-3.1%; 95% confidence interval (CI), -0.2 to -5.9%] and FEF(25-75) (-8.1%; CI, -2.3 to -13.9%), and nearly so for FEF(75) (-6.7%; CI, 1.4 to -14.8). Gender-specific analyses revealed stronger associations for males than for females. The symptoms of chronic phlegm, wheeze apart from colds, and RSI were increased in the ozone-exposed group, with odds ratios of 1.79 (CI, 0.83-3.82), 1.97 (CI, 1.06-3.66), and 2.00 (CI, 1.15-3.46), respectively. We conclude that living for 4 or more years in regions of the country with high levels of ozone and related copollutants is associated with diminished lung function and more frequent reports of respiratory symptoms.

Long-term residence in areas of high ozone: associations with respiratory health in a nationwide sample of nonsmoking young adults [dsee comments

Few studies have examined the respiratory effects of multiyear ozone exposures in human populations. We examined associations between current respiratory health status and long-term ozone exposure histories in 520 Yale College (New Haven, CT) students who never smoked. Questionnaires addressed current respiratory symptoms, respiratory disease history, residential history, and other factors. The symptoms of cough, phlegm, wheeze apart from colds, and a composite respiratory symptom index (RSI) were selected as outcome measures. Forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV(1)), forced expiratory flow rate between 25 and 75% of FVC (FEF(25-75)), and forced expiratory flow rate at 75% of FVC (FEF(75)) were obtained by forced expiration into spirometers. Ozone exposure was treated as a dichotomous variable, where subjects were assigned to the high-exposure group if they lived for 4 or more years in a U.S. county with 10-year average summer-season daily 1-hr maximum ozone levels [greater/equal to] 80 ppb. Lung function and respiratory symptoms were analyzed by multiple linear and logistic regression on ozone exposure, controlling for covariates. Lung function was lower in the group with high ozone exposures: differences were statistically significant for FEV(1) [-3.1%; 95% confidence interval (CI), -0.2 to -5.9%] and FEF(25-75) (-8.1%; CI, -2.3 to -13.9%), and nearly so for FEF(75) (-6.7%; CI, 1.4 to -14.8). Gender-specific analyses revealed stronger associations for males than for females. The symptoms of chronic phlegm, wheeze apart from colds, and RSI were increased in the ozone-exposed group, with odds ratios of 1.79 (CI, 0.83-3.82), 1.97 (CI, 1.06-3.66), and 2.00 (CI, 1.15-3.46), respectively. We conclude that living for 4 or more years in regions of the country with high levels of ozone and related copollutants is associated with diminished lung function and more frequent reports of respiratory symptoms.