Harmonizing the assessment of allostatic load across cycles of the Canadian Health Measures Survey: Variable selection and calculation method.

Background: The availability of measures to operationalize allostatic load - the cumulative toll on the body of responding to stressor demands - in population health surveys may differ across years or surveys, hampering analyses on the entire sampled population. Here, impacts of variable selection and calculation method were evaluated to generate an allostatic load index applicable across all cycles of the Canadian Health Measures Survey (CHMS). Methods: Data from CHMS cycles 1 to 4 were used to compare allostatic load scores when replacing the most prevalent risk factor, waist-to-hip ratio - available in cycles 1 to 4 but not 5 and 6 - with body mass index (BMI), waist circumference, waist circumference within BMI groups (classified as normal, overweight, or obese), or waist-to-height ratio. Indexes were generated using clinical or sex-specific empirically defined risk thresholds and as count-based or continuous scores. Logistic regression models that included age and sex were used to relate each potential index to socioeconomic indicators (educational attainment, household income). Results: Of the variables assessed, waist-to-height ratio and waist circumference were closest to waist-to-hip ratio according to an individual's percentile ranking and in classifying "at risk" using either clinical or empirically defined cut-offs. Allostatic load profiles generated using waist-to-height ratios most closely resembled profiles constructed using waist-to-hip ratios. Sex-dependent associations with educational attainment and household income were maintained across constructs whether indexes were count-based or continuous. Interpretation: Allostatic load profiles and associations with socioeconomic indicators were robust to variable substitution and method of calculation, supporting the use of a harmonized index across survey cycles to assess the cumulative toll on health of stressor exposure.

Modulation of insulin signaling pathway genes by ozone inhalation and the role of glucocorticoids: A multi-tissue analysis.

Air pollution is associated with increased risk of metabolic diseases including type 2 diabetes, of which dysregulation of the insulin-signaling pathway is a feature. While studies suggest pollutant exposure alters insulin signaling in certain tissues, there is a lack of comparison across multiple tissues needed for a holistic assessment of metabolic effects, and underlying mechanisms remain unclear. Air pollution increases plasma levels of glucocorticoids, systemic regulators of metabolic function. The objectives of this study were to 1) determine effects of ozone on insulin-signaling genes in major metabolic tissues, and 2) elucidate the role of glucocorticoids. Male Fischer-344 rats were treated with metyrapone, a glucocorticoid synthesis inhibitor, and exposed to 0.8 ppm ozone or clean air for 4 h, with tissue collected immediately or 24 h post exposure. Ozone inhalation resulted in distinct mRNA profiles in the liver, brown adipose, white adipose and skeletal muscle tissues, including effects on insulin-signaling cascade genes (Pik3r1, Irs1, Irs2) and targets involved in glucose metabolism (Hk2, Pgk1, Slc2a1), cell survival (Bcl2l1), and genes associated with diabetes and obesity (Serpine1, Retn, Lep). Glucocorticoid-dependent regulation was observed in the liver and brown and white adipose tissues, while effects in skeletal muscle were largely unaffected by metyrapone treatment. Gene expression changes were accompanied by altered phosphorylation states of insulin-signaling proteins (BAD, GSK, IR-ß, IRS-1) in the liver. The results show that systemic effects of ozone inhalation include tissue-specific regulation of insulin-signaling pathway genes via both glucocorticoid-dependent and independent mechanisms, providing insight into mechanisms underlying adverse effects of pollutants.

Randomized Cross-Over Study of In-Vehicle Cabin Air Filtration, Air Pollution Exposure, and Acute Changes to Heart Rate Variability, Saliva Cortisol, and Cognitive Function.

To determine how traffic-related air pollution (TRAP) exposures affect commuter health, and whether cabin air filtration (CAF) can mitigate exposures, we conducted a cross-over study of 48 adults exposed to TRAP during two commutes with and without CAF. Measurements included particulate air pollutants (PM2.5, black carbon [BC], ultrafine particles [UFPs]), volatile organic compounds, and nitrogen dioxide. We measured participants' heart rate variability (HRV), saliva cortisol, and cognitive function. On average, CAF reduced concentrations of UFPs by 26,232 (95%CI: 11,734, 40,730) n/cm3, PM2.5 by 6 (95%CI: 5, 8) µg/m3, and BC by 1348 (95%CI: 1042, 1654) ng/m3, or 28, 30, and 32%, respectively. Each IQR increase in PM2.5 was associated with a 28% (95%CI: 2, 60) increase in high-frequency power HRV at the end of the commute and a 22% (95%CI: 7, 39) increase 45 min afterward. IQR increases in UFPs were associated with increased saliva cortisol in women during the commute (18% [95%CI: 0, 40]). IQR increases in UFPs were associated with strong switching costs (19% [95%CI: 2, 39]), indicating a reduced capacity for multitasking, and PM2.5 was associated with increased reaction latency, indicating slower responses (5% [95%CI: 1, 10]). CAF can reduce particulate exposures by almost a third.

Establishing an air-liquid interface exposure system for exposure of lung cells to gases.

OBJECTIVE: Growing interest in non-animal-based models has led to the development of devices to expose cells to airborne substances. Cells/tissues grown at the air-liquid interface (ALI) are more representative of lung cells/tissues in vivo compared to submerged cell cultures. Additionally, airborne exposures should allow for closer modeling of human lung toxicity. However, such exposures present technical challenges, including maintaining optimal cell health, and establishing consistent exposure monitoring and control. We aimed to establish a reliable system and procedures for cell exposures to gases at the ALI. METHODS: We tested and adapted a horizontal-flow ALI-exposure system to verify and optimize temperature, humidity/condensation, and control of atmosphere delivery. We measured temperature and relative humidity (RH) throughout the system, including at the outlet (surrogate measures) and at the well, and evaluated viability of lung epithelial A549 cells under control conditions. Exposure stability, dosimetry, and toxicity were tested using ozone. RESULTS: Temperatures measured directly above wells vs. outflow differed; using above-well temperature enabled determination of near-well RH. Under optimized conditions, the viability of A549 cells exposed to clean air (2 h) in the ALI system was unchanged from incubator-grown cells. In-well ozone levels, determined through reaction with potassium indigotrisulfonate, confirmed dosing. Cells exposed to 200 ppb ozone at the ALI presented reduced viability, while submerged cells did not. CONCLUSION: Our results emphasize the importance of monitoring near-well conditions rather than relying on surrogate measures. Rigorous assessment of ALI exposure conditions led to procedures for reproducible exposure of cells to gases.

Hair cortisol as a viable tool for the assessment of an association between environmental noise exposure and chronic stress.

Entrenched in the well-established link between stress and health, noise exposure as a potential contributor to stress-related health effects receives tremendous attention. Indeed, exposure to noise can act as a stressor as evidenced through increased heart rate, blood pressure, adrenaline, epinephrine, and cortisol. Cortisol is secreted from the adrenal glands in response to stressor-induced activation of the hypothalamic-pituitary-adrenal axis. For assessment of environmental noise and stress, repeated sampling in blood, saliva, or urine is necessary to evaluate the association between environmental noise exposure and protracted changes in cortisol. Controlling for the many variables that influence the secretion of cortisol at discrete sampling intervals is challenging. Studies suggest that systemically produced cortisol integrates and remains in hair as it grows, providing a measure that integrates a cortisol response over a longer period, circumventing several limitations associated with multiple sampling. Robust evidence supports the integration of cortisol into hair, yet recent studies call into question the notion that cortisol is retained with growth. The current paper discusses the strengths and limitations of hair cortisol analysis with an emphasis on its utility as a measure of chronic stress in environmental noise studies.

Residential greenness and indicators of stress and mental well-being in a Canadian national-level survey.

BACKGROUND: Residential proximity to greenness in urban areas has been shown to confer a number of health benefits, including improved mental health. We investigated whether greenness was associated with self-reported stress, distress, and mental health among adult participants of multiple cycles of a national Canadian health survey, and whether these associations varied by sex, age, income, and neighbourhood characteristics. METHODS: Our study population included 397,900 participants of the Canadian Community Health Survey, 18 years of age or older, who lived in census metropolitan areas between 2000 and 2015. We used the Normalized Difference Vegetation Index (NDVI) to characterize participants' exposure to greenness within 250 m, 500 m, and 1 km buffers from a representative location of their postal code. Health outcomes included: self-reported perceptions of life stress, psychological distress, and self-rated mental health. We used multiple regression models, adjusted for relevant individual and neighbourhood-level variables to estimate associations (and 95% confidence intervals) between each outcome and exposure to greenness. FINDINGS: In models with all participants, we observed 6% lower odds of poor self-rated mental health per increase in the interquartile range (i.e., 0.12) of NDVI within a 500 m buffer. Across the three outcomes, we found substantial heterogeneity in effect size across categories of sex, age, and community-level indicators of deprivation and urban form. For example, each incremental increase in greenness exposure was associated with a reduction of 0.61 (95% CI: 0.81 to -0.51) on the K10 psychological distress score among those living in the active core of cities, and with an increase of 0.07 (95% CI: 0.03-0.12) on this score among those living in the most suburban areas. CONCLUSIONS: Our results indicate that the potential benefits of residential greenness on mental health vary across personal and neighbourhood-level characteristics and are sensitive to how the outcome is measured. Additional research is needed to understand which features of greenness are most relevant to different sub-groups of the population to maximize these health benefits.

Air pollution exposure during pregnancy and childhood and brain morphology in preadolescents.

BACKGROUND: Studies investigating the relationship between exposure to air pollution and brain development using magnetic resonance images are emerging. However, most studies have focused only on prenatal exposures, and have included a limited selection of pollutants. Here, we aim to expand the current knowledge by studying pregnancy and childhood exposure to a wide selection of pollutants, and brain morphology in preadolescents. METHODS: We used data from 3133 preadolescents from a birth cohort from Rotterdam, the Netherlands (enrollment: 2002-2006). Concentrations of nitrogen oxides, coarse, fine, and ultrafine particles, and composition of fine particles were estimated for participant's home addresses in pregnancy and childhood, using land use regression models. Structural brain images were obtained at age 9-12 years. We assessed the relationships of air pollution exposure, with brain volumes, and surface-based morphometric data, adjusting for socioeconomic and life-style characteristics, using single as well as multi-pollutant approach. RESULTS: No associations were observed between air pollution exposures and global volumes of total brain, and cortical and subcortical grey matter. However, we found associations between higher pregnancy and childhood air pollution exposures with smaller corpus callosum, smaller hippocampus, larger amygdala, smaller nucleus accumbens, and larger cerebellum (e.g. -69.2mm3 hippocampal volume [95%CI -129.1 to -9.3] per 1ng/m3 increase in pregnancy exposure to polycyclic aromatic hydrocarbons). Higher pregnancy exposure to air pollution was associated with smaller cortical thickness while higher childhood exposure was associated with predominantly larger cortical surface area. CONCLUSION: Higher pregnancy or childhood exposure to several air pollutants was associated with altered volume of several brain structures, as well as with cortical thickness and surface area. Associations showed some similarity to delayed maturation and effects of early-life stress.

Ozone increases plasma kynurenine-tryptophan ratio and impacts hippocampal serotonin receptor and neurotrophic factor expression: Role of stress hormones.

Air pollution is associated with adverse impacts on the brain, including cognitive decline and increased incidence of dementia, depression and anxiety; however, underlying mechanisms remain unclear. We have shown that both ozone and particulate matter activate the hypothalamic-pituitary-adrenal (HPA) axis, increasing plasma glucocorticoids and altering mRNA profiles in multiple tissues including the brain. HPA axis dysregulation has been associated with central nervous system impacts, including key effects in the hippocampus; accordingly, we hypothesized that pollutant-dependent increases in glucocorticoid levels impact biological pathways relevant to brain health. Fischer-344 rats were treated with metyrapone (0 or 50 mg/kg), a glucocorticoid synthesis inhibitor, and exposed to ozone (0 or 0.8 ppm) for 4 h (n = 5/group) to investigate the role of glucocorticoids in ozone-dependent effects on tryptophan metabolism and expression of serotonin receptors and neurotrophic factors. Ozone increased plasma levels of the tryptophan metabolite kynurenine (~2-fold) and decreased tryptophan levels (~1.2 fold). Hippocampal expression of serotonin receptors exhibited differential regulation following exposure, and expression of key neurotrophic factors (brain-derived neurotrophic factor, vascular endothelial growth factor A, insulin-like growth factor-1, tyrosine kinase receptor B, b-cell lymphoma 2) was decreased. Some, but not all effects were abrogated by metyrapone treatment, suggesting both glucocorticoid-dependent and -independent regulation. Exposure to exogenous corticosterone (10 mg/kg) followed by clean air reproduced the ozone effects that were blocked with metyrapone, confirming the specificity of effects to glucocorticoids. These results indicate that ozone can modify pathways relevant to brain health and establish a role for the HPA axis in mediating these effects.

Effect of industrial point-source air pollutants on fractional exhaled nitric oxide in healthy volunteers.

BACKGROUND: Few studies have examined the effects of industrial, fixed-site sources of air pollution on lung inflammation in nearby residents. We investigated the effects of short-term exposure to ambient air near a steel plant on the fractional exhaled concentration of nitric oxide (FeNO), a measure of airway inflammation, in healthy volunteers. METHODS: A cross-over study design was used. Fifty-nine non-smoking participants (mean age 24 years) were randomly assigned to each of two 5-day exposure scenarios: breathing ambient air adjacent to a steel plant or 5 km away at a college campus site. FeNO and on-site air pollutants were measured daily. Mixed effects linear regression models were used for data analysis, adjusting for sex, temperature, humidity and day of week. RESULTS: Compared with the college site, PM 2.5, ultrafine PM, SO2, NO2 and CO levels were significantly greater near the steel plant. FeNO was 15.3% (95% CI, 6.6%, 24.8%) higher near the plant compared to the college site. CONCLUSIONS: Exposure to ambient air near a steel plant was associated with increased airway inflammation as measured by exhaled nitric oxide.

Self-rated stress, distress, mental health, and health as modifiers of the association between long-term exposure to ambient pollutants and mortality.

BACKGROUND: Individual and neighbourhood-scale socioeconomic characteristics modify associations between exposure to air pollution and mortality. The role of stress, which may integrate effects of social and environmental exposures on health, is unknown. We examined whether an individual's perspective on their own well-being, as assessed using self-rated measures of stress and health, modifies the pollutant-mortality relationship. METHODS: The Canadian Community Health Survey (CCHS)-mortality cohort includes respondents from surveys administered between 2001 and 2012 linked to vital statistics and postal codes from 1981 until 2016. Annual fine particulate matter (PM2.5), nitrogen dioxide (NO2), and ozone (O3) exposure estimates were attached to a sample of cohort members aged 30-89 years (n = 398,300 respondents/3,848,400 person-years). We examined whether self-rated stress, distress, mental health, and general health modified associations between long-term exposure to each pollutant (three-year moving average with one-year lag) and non-accidental mortality using Cox survival models, adjusted for individual- (i.e. socioeconomic and behavioural) and neighbourhood-scale covariates. RESULTS: In fully-adjusted models, the relationship between exposure to pollutants and mortality was stronger among those with poor self-rated mental health, including a significant difference for NO2 (hazard ratio (HR) = 1.15, 95% CI 1.06-1.25 per IQR) compared to those with very good/excellent mental health (HR = 1.05, 95% CI 1.01-1.08; Cochran's Q = 4.01; p < 0.05). Poor self-rated health was similarly associated with higher pollutant-associated HRs, but only in unadjusted models. Stress and distress did not modify pollutant-mortality associations. CONCLUSIONS: Poor self-rated mental and general health were associated with increased mortality attributed to exposure to ambient pollutants.

The association between ambient air pollution concentrations and psychological distress.

BACKGROUND: A growing number of epidemiological studies have linked air pollution exposure to psychological conditions. Laboratory studies indicate that air pollutants can activate the neuroendocrine stress axis and modulate stress hormone levels, which could contribute to the development or exacerbation of psychological distress. The present study examined the spatial associations between air pollutants (fine particulate matter [PM2.5], nitrogen dioxide [NO2] and ground-level ozone [O3]) and psychological distress among subjects in the most populous provinces in Canada. DATA AND METHODS: Subjects were sampled from the Canadian Community Health Survey in three regions (Quebec in 2005 [n=25,800], British Columbia and Alberta in 2005 [n=23,000], and Ontario in 2011 [n=36,000]), and were assigned estimates of annual exposure to three ambient air pollutants (PM2.5, NO2 and O3) for the same years. Individual psychological distress was assessed using the Kessler Psychological Distress Scale (K10), based on anxiety and depressive symptoms in the past month. Regression models (both ordinary least squares and simultaneous autoregressive models) were applied to estimate associations between K10 distress scores and each air pollutant, after adjusting for individual (demographic, socioeconomic and behavioural) and neighbourhood covariates.. RESULTS: Psychological distress was positively associated with PM2.5 and NO2 in all three regions, and with O3 in Quebec. However, after further adjusting for individual and neighbourhood covariates, the associations between distress and air pollution remained statistically significant only in Quebec. DISCUSSION: Some evidence for positive associations between psychological distress and ambient air pollution after adjusting for spatial autocorrelation was found.

Corticosterone determination in bronchoalveolar lavage fluid and its relationship to free and total plasma corticosterone.

Endogenous glucocorticoids modulate airway and lung inflammation in various respiratory diseases and after exposure to airborne contaminants. Although bronchoalveolar lavage fluid (BALF) is commonly used to evaluate the inflammatory and immune response in the lungs, limited information is available on determination of endogenous glucocorticoid levels in BALF. Here we describe a simple method to determine corticosterone in BALF, and evaluate the relationship between BALF and plasma corticosterone.

Oxidative and Toxicological Evolution of Engineered Nanoparticles with Atmospherically Relevant Coatings.

The health impacts associated with engineered nanoparticles (ENPs) released into the atmosphere have not been adequately assessed. Such impacts could potentially arise from the toxicity associated with condensable atmospheric secondary organic material (SOM), or changes in the SOM composition induced by ENPs. Here, these possibilities are evaluated by investigating the oxidative and toxicological evolution of TiO2 and SiO2 nanoparticles which have been coated with SOM from the O3 or OH initiated oxidation of α-pinene. It was found that pristine SiO2 particles were significantly more cytotoxic compared to pristine TiO2 particles. TiO2 in the dark or under UV irradiation catalytically reacted with the SOM, increasing its O/C by up to 55% over photochemically inert SiO2 while having negligible effects on the overall cytotoxicity. Conversely, the cytotoxicity associated with SiO2 coated with SOM was markedly suppressed (by a factor of 9, at the highest exposure dose) with both increased SOM coating thickness and increased photochemical aging. These suppressing effects (organic coating and photo-oxidation of organics) were attributed to a physical hindrance of SiO2-cell interactions by the SOM and enhanced SOM viscosity and hydrophilicity with continued photo-oxidation, respectively. These findings highlight the importance of atmospheric processes in altering the cytotoxicity of ENPs.

Stress hormones as potential mediators of air pollutant effects on the brain: Rapid induction of glucocorticoid-responsive genes.

Air pollution is associated with adverse effects on brain health including cognitive decline, dementia, anxiety, depression, and suicide. While toxicological studies have demonstrated the potential for repeated or chronic pollutant exposure to lead to disease states, characterisation of initial biological responses to exposure is needed to better understand underlying mechanisms. The brain is highly sensitive to glucocorticoids (primarily cortisol in humans, corticosterone in rodents), stress hormones that play important roles in cognition and mental health. We tested whether glucocorticoids could be implicated in central nervous system (CNS) effects of pollutant exposure by examining glucocorticoid-dependent signaling across brain regions after exposure to the common pollutant ozone. Male Fischer-344 rats were exposed for 4 h to air or 0.8 ppm ozone ±â€¯metyrapone (50 mg/kg), a drug that blocks corticosterone synthesis (n = 5/group). Key glucocorticoid-responsive genes (serum- and glucocorticoid-inducible kinase, SGK; glucocorticoid-inducible leucine zipper, GILZ), and a gene responsive to both glucocorticoids and oxidative stress (metallothionein (MT)-1), were increased by ozone in all brain regions (olfactory bulb, frontal lobe, cortex, midbrain, hippocampus, cerebellum, brainstem), correlating with plasma corticosterone levels. Metyrapone prevented the increase in SGK and GILZ, and reduced but did not eliminate the effect on MT-1, suggesting glucocorticoid-dependent and -independent regulation. Administering exogenous corticosterone (10 mg/kg) to air-exposed rats reproduced the ozone effects, confirming specificity. The results demonstrate that early pollutant effects include stress hormone-dependent signaling. As both ozone and particulate matter activate the hypothalamic-pituitary-adrenal axis, and elevated glucocorticoids are implicated in brain pathologies, stress hormones could contribute to CNS impacts of air pollutants.

Cumulative toll of exposure to stressors in Canadians: An allostatic load profile.

BACKGROUND: The cumulative toll of exposure to stressors (psychosocial, chemical, physical) can contribute to disease processes. The concept of allostatic load, essentially the cost of maintaining physiological stability in response to environmental demands, may be useful in assessing broad population health impacts of stressors beyond morbidity and mortality. In the present study, allostatic load scores were generated for Canadians and associations with age, sex, education and household income were examined. DATA AND METHODS: Data from cycles 1, 2, and 3 (2007 to 2013) of the Canadian Health Measures Survey (CHMS) were used to generate a composite index of cumulative health burden (allostatic load score) for adults aged 20 to 79 (n=8,678) based on risk thresholds for nine biological measures: diastolic blood pressure, systolic blood pressure, heart rate, high-density lipoprotein (HDL), total cholesterol, glycated hemoglobin (HBA1c), waist-to-hip ratio, C-reactive protein (CRP), and albumin. Logistic regression models that included age (continuous), sex, education and household income were fit to generate model-adjusted predicted allostatic load scores. RESULTS: The most prevalent individual risk factors were elevated waist-to-hip ratio, elevated CRP, total cholesterol, and low HDL. Allostatic load scores increased with age. Males generally exhibited higher scores than females. Lower educational attainment and lower household income were found to be significantly associated with higher allostatic load scores after taking account of the effects of age and sex. DISCUSSION: Age and socioeconomic gradients are associated with differences in allostatic load scores in the Canadian population. This composite measure of multisystem dysfunction, generated from a nationally representative survey that includes measurement of numerous health-relevant behaviours, biomarkers, and chemical levels, can be used in future to quantify sub-clinical impacts on health.

Ozone modifies the metabolic and endocrine response to glucose: Reproduction of effects with the stress hormone corticosterone.

Air pollution is associated with increased incidence of metabolic disease (e.g. metabolic syndrome, obesity, diabetes); however, underlying mechanisms are poorly understood. Air pollutants increase the release of stress hormones (human cortisol, rodent corticosterone), which could contribute to metabolic dysregulation. We assessed acute effects of ozone, and stress axis involvement, on glucose tolerance and on the metabolic (triglyceride), endocrine/energy regulation (insulin, glucagon, GLP-1, leptin, ghrelin, corticosterone), and inflammatory/endothelial (TNF, IL-6, VEGF, PAI-1) response to exogenous glucose. Male Fischer-344 rats were exposed to clean air or 0.8 ppm ozone for 4 h in whole body chambers. Hypothalamic-pituitary-adrenal (HPA) axis involvement in ozone effects was tested through subcutaneous administration of the glucocorticoid synthesis inhibitor metyrapone (50 mg/kg body weight), corticosterone (10 mg/kg body weight), or vehicle (40% propylene glycol) prior to exposure. A glucose tolerance test (2 g/kg body weight glucose) was conducted immediately after exposure, with blood samples collected at 0, 30, 60, 90, and 120 min. Ozone exposure impaired glucose tolerance, an effect accompanied by increased plasma triglycerides but no impairment of insulin release. Ozone diminished glucagon, GLP-1, and ghrelin responses to glucose, but did not significantly impact inflammatory/endothelial analytes. Metyrapone reduced corticosterone but increased glucose and triglycerides, complicating evaluation of the impact of glucocorticoid inhibition. However, administration of corticosterone reproduced the profile of ozone effects, supporting a role for the HPA axis. The results show that ozone-dependent changes in glucose tolerance are accompanied by altered metabolic and endocrine responses to glucose challenge that are reproduced by exogenous stress hormone.

Stress axis variability is associated with differential ozone-induced lung inflammatory signaling and injury biomarker response.

Ozone (O3), a ubiquitous urban air pollutant, causes adverse pulmonary and extrapulmonary effects. A large variability in acute O3-induced effects has been observed; however, the basis for interindividual differences in susceptibility is unclear. We previously demonstrated a role for the hypothalamic-pituitary-adrenal (HPA) stress axis and glucocorticoid response in acute O3 toxicity. Glucocorticoids have important anti-inflammatory actions, and have been shown to regulate lung inflammatory responses. We hypothesised that a hyporesponsive HPA axis would be associated with greater O3-dependent lung inflammatory signaling. Two genetically-related rat strains with known differences in stress axis reactivity, highly-stress responsive Fischer (F344) and less responsive Lewis (LEW), were exposed for 4 h by nose-only inhalation to clean air or 0.8 ppm O3, and euthanized immediately after exposure. As expected, baseline (air-exposed) plasma corticosterone was significantly lower in the hypo-stress responsive LEW. Although O3 exposure increased plasma corticosterone in both strains, corticosterone remained significantly lower in LEW when compared to F334. LEW exhibited greater O3-induced inflammatory cytokine/chemokine signaling compared to F344, consistent with the lower corticosterone levels. Since we observed strain-specific differences in inflammatory signaling, we further investigated injury biomarkers (total protein, albumin and lactate dehydrogenase). Although the hyper-responsive F344 exhibited lower inflammatory signaling in response to O3 compared with LEW, they had greater levels of lung injury biomarkers. Our results indicate that stress axis variability is associated with differential O3-induced lung toxicity. Given the large variability in stress axis reactivity among humans, stress axis regulation could potentially be a determining factor underlying O3 sensitivity.

Cardiovascular and inflammatory mechanisms in healthy humans exposed to air pollution in the vicinity of a steel mill.

BACKGROUND: There is a paucity of mechanistic information that is central to the understanding of the adverse health effects of source emission exposures. To identify source emission-related effects, blood and saliva samples from healthy volunteers who spent five days near a steel plant (Bayview site, with and without a mask that filtered many criteria pollutants) and at a well-removed College site were tested for oxidative stress, inflammation and endothelial dysfunction markers. METHODS: Biomarker analyses were done using multiplexed protein-array, HPLC-Fluorescence, EIA and ELISA methods. Mixed effects models were used to test for associations between exposure, biological markers and physiological outcomes. Heat map with hierarchical clustering and Ingenuity Pathway Analysis (IPA) were used for mechanistic analyses. RESULTS: Mean CO, SO2 and ultrafine particles (UFP) levels on the day of biological sampling were higher at the Bayview site compared to College site. Bayview site exposures "without" mask were associated with increased (p < 0.05) pro-inflammatory cytokines (e.g IL-4, IL-6) and endothelins (ETs) compared to College site. Plasma IL-1ß, IL-2 were increased (p < 0.05) after Bayview site "without" compared to "with" mask exposures. Interquartile range (IQR) increases in CO, UFP and SO2 were associated with increased (p < 0.05) plasma pro-inflammatory cytokines (e.g. IL-6, IL-8) and ET-1(1-21) levels. Plasma/saliva BET-1 levels were positively associated (p < 0.05) with increased systolic BP. C-reactive protein (CRP) was positively associated (p < 0.05) with increased heart rate. Protein network analyses exhibited activation of distinct inflammatory mechanisms after "with" and "without" mask exposures at the Bayview site relative to College site exposures. CONCLUSIONS: These findings suggest that air pollutants in the proximity of steel mill site can influence inflammatory and vascular mechanisms. Use of mask and multiple biomarker data can be valuable in gaining insight into source emission-related health impacts.

Exposure to air pollution near a steel plant is associated with reduced heart rate variability: a randomised crossover study.

BACKGROUND: Epidemiological studies have shown that as ambient air pollution (AP) increases the risk of cardiovascular mortality also increases. The mechanisms of this effect may be linked to alterations in autonomic nervous system function. We wished to examine the effects of industrial AP on heart rate variability (HRV), a measure of subtle changes in heart rate and rhythm representing autonomic input to the heart. METHODS: Sixty healthy adults were randomized to spend five consecutive 8-h days outdoors in one of two locations: (1) adjacent to a steel plant in the Bayview neighbourhood in Sault Ste Marie Ontario or (2) at a College campus, several kilometers from the plant. Following a 9-16 day washout period, participants spent five consecutive days at the other site. Ambient AP levels and ambulatory electrocardiogram recordings were collected daily. HRV analysis was undertaken on a segment of the ambulatory ECG recording during a 15 min rest period, near the end of the 8-h on-site day. Standard HRV parameters from both time and frequency domains were measured. Ambient AP was measured with fixed site monitors at both sites. Statistical analysis was completed using mixed-effects models. RESULTS: Compared to the College site, HRV was statistically significantly reduced at the Bayview site by 13% (95%CI 3.6,19.2) for the standard deviation of normal to normal, 8% (95%CI 0.1, 4.9) for the percent normal to normal intervals differing by more than 50 ms, and 15% (95%CI 74.9, 571.2) for low frequency power. Levels of carbon monoxide, sulphur dioxide, nitrogen dioxide, and fine and ultrafine particulates were slightly, but statistically significantly, elevated at Bayview when compared to College. Interquartile range changes in individual air pollutants were significantly associated with reductions in HRV measured on the same day. The patterns of effect showed a high degree of consistency, with nearly all pollutants significantly inversely associated with at least one measure of HRV. CONCLUSIONS: The significant associations between AP and changes in HRV suggest that ambient AP near a steel plant may impact autonomic nervous system control of the heart.

Responses of A549 human lung epithelial cells to cristobalite and α-quartz exposures assessed by toxicoproteomics and gene expression analysis.

In this study, we used cytotoxicity assays, proteomic and gene expression analyses to examine the difference in response of A549 cells to two silica particles that differ in physical properties, namely cristobalite (CR) and α-quartz (Min-U-Sil 5, MI). Cytotoxicity assays such as lactate dehydrogenase release, 5-bromo-2'-deoxyuridine incorporation and cellular ATP showed that both silica particles could cause cell death, decreased cell proliferation and metabolism in the A549 human lung epithelial cells. While cytotoxicity assays revealed little difference between CR and MI exposures, proteomic and gene expression analyses unveiled both similar and unique molecular changes in A549 cells. For instance, two-dimensional gel electrophoresis data indicated that the expression of proteins in the cell death (e.g., ALDH1A1, HTRA2 and PRDX6) and cell proliferation (e.g., FSCN1, HNRNPAB and PGK1) pathways were significantly different between the two silica particles. Reverse transcription-polymerase chain reaction data provided additional evidence supporting the proteomic findings. Preliminary assessment of the physical differences between CR and MI suggested that the extent of surface interaction between particles and cells could explain some of the observed biological effects. However, the differential dose-response curves for some other genes and proteins suggest that other physical attributes of particulate matter can also contribute to particulate matter-related cellular toxicity. Our results demonstrated that toxicoproteomic and gene expression analyses are sensitive in distinguishing subtle toxicity differences associated with silica particles of varying physical properties compared to traditional cytotoxicity endpoints. Copyright © 2016 Her Majesty the Queen in Right of Canada. Journal of Applied Toxicology published by John Wiley & Sons, Ltd.