Prenatal and early life exposure to air pollution induced hippocampal vascular leakage and impaired neurogenesis in association with behavioral deficits.

Exposure to traffic-related air pollution (TRAP) is associated with a range of neurodevelopmental disorders in human populations. In rodent models, prenatal TRAP exposure increased depressive behaviors and increased brain microglial activity. To identify cellular mechanisms, we examined adult neurogenesis and the blood-brain barrier (BBB) in relation to cognition and motivated behaviors in rats that were exposed to a nano-sized TRAP subfraction from gestation into adulthood. At age 5 months, exposed male rats had 70% fewer newly generated neurons in the dentate gyrus (DG) of the hippocampus. Microglia were activated in DG and CA1 subfields (35% more Iba1). The BBB was altered, with a 75% decrease of the tight junction protein ZO-1 in the CA1 layer, and twofold more iron deposits, a marker of microhemorrhages. The exposed rats had impaired contextual memory (novel object in context), reduced food-seeking behavior, and increased depressive behaviors (forced swim). Deficits of de novo neurogenesis were inversely correlated with depressive behavior, whereas increased microbleeds were inversely correlated with deficits in contextual memory. These findings give the first evidence that prenatal and early life exposure to TRAP impairs adult hippocampal neurogenesis and increases microbleeds in association with behavioral deficits.

Particulate air pollutants, APOE alleles and their contributions to cognitive impairment in older women and to amyloidogenesis in experimental models.

Exposure to particulate matter (PM) in the ambient air and its interactions with APOE alleles may contribute to the acceleration of brain aging and the pathogenesis of Alzheimer's disease (AD). Neurodegenerative effects of particulate air pollutants were examined in a US-wide cohort of older women from the Women's Health Initiative Memory Study (WHIMS) and in experimental mouse models. Residing in places with fine PM exceeding EPA standards increased the risks for global cognitive decline and all-cause dementia respectively by 81 and 92%, with stronger adverse effects in APOE ɛ4/4 carriers. Female EFAD transgenic mice (5xFAD+/-/human APOE ɛ3 or ɛ4+/+) with 225 h exposure to urban nanosized PM (nPM) over 15 weeks showed increased cerebral ß-amyloid by thioflavin S for fibrillary amyloid and by immunocytochemistry for Aß deposits, both exacerbated by APOE ɛ4. Moreover, nPM exposure increased Aß oligomers, caused selective atrophy of hippocampal CA1 neurites, and decreased the glutamate GluR1 subunit. Wildtype C57BL/6 female mice also showed nPM-induced CA1 atrophy and GluR1 decrease. In vitro nPM exposure of neuroblastoma cells (N2a-APP/swe) increased the pro-amyloidogenic processing of the amyloid precursor protein (APP). We suggest that airborne PM exposure promotes pathological brain aging in older women, with potentially a greater impact in ɛ4 carriers. The underlying mechanisms may involve increased cerebral Aß production and selective changes in hippocampal CA1 neurons and glutamate receptor subunits.

Efficient determination of vehicle emission factors by fuel use category using on-road measurements: downward trends on Los Angeles freight corridor I-710.

To evaluate the success of vehicle emissions regulations, trends in both fleet-wide average emissions as well as high-emitter emissions are needed, but it is challenging to capture the full spread of vehicle emission factors (EFs) with chassis dynamometer or tunnel studies, and remote sensing studies cannot evaluate particulate compounds. We developed an alternative method that links real-time on-road pollutant measurements from a mobile platform with real-time traffic data, and allows efficient calculation of both the average and the spread of EFs for light-duty gasoline-powered vehicles (LDG) and heavy-duty diesel-powered vehicles (HDD). This is the first study in California to report EFs under a full range of real-world driving conditions on multiple freeways. Fleet average LDG EFs were in agreement with most recent studies and an order of magnitude lower than observed HDD EFs. HDD EFs reflected the relatively rapid decreases in diesel emissions that have recently occurred in Los Angeles/California, and on I-710, a primary route used for goods movement and a focus of additional truck fleet turnover incentives, HDD EFs were often lower than on other freeways. When freeway emission rates (ER) were quantified as the product of EF and vehicle miles traveled (VMT) per time per mile of freeway, despite a twoto three-fold difference in HDD fractions between freeways, ERs were found to be generally similar in magnitude. Higher LDG VMT on low HDD fraction freeways largely offset the difference. Therefore, the conventional assumption that free ways with the highest HDD fractions are significantly worse sources of total emissions in Los Angeles may no longer be true.

A comparative assessment of PM2.5 exposures in light-rail, subway, freeway, and surface street environments in Los Angeles and estimated lung cancer risk.

According to the U.S. Census Bureau, 570000+ commuters in Los Angeles travel for over 60 minutes to work. Studies have shown that a substantial portion of particulate matter (PM) exposure can occur during this commute. This study represents the integration of the results from five commute environments in Los Angeles. Personal PM exposures are discussed for the: (1) METRO gold line, a ground-level light-rail route, (2) METRO red line, a subway line, (3) the 110, a high volume freeway with low heavy-duty vehicle (HDV) fraction, (4) the 710, a major corridor for HDVs from the Port of Los Angeles, and (5) Wilshire/Sunset Boulevards, major surface streets. Chemical analysis including total and water-soluble metals and trace elements, elemental and organic carbon (EC/OC), and polycyclic aromatic hydrocarbons (PAHs) was performed. The focus of this study is to compare the composition and estimated lung cancer risk of PM2.5 (dp < 2.5 µm) for the five differential commute environments. Metals associated with stainless steel, notably Fe, Cr, and Mn, were elevated for the red line (subway), most likely from abrasion processes between the rail and brakes; elements associated with tire and brake wear and oil additives (Ca, Ti, Sn, Sb, and Pb) were elevated on roadways. Elemental concentrations on the gold line (light-rail) were the lowest. For water-solubility, metals observed on the red line (subway) were the least soluble. PAHs are primarily derived from vehicular emissions. Overall, the 710 exhibited high levels of PAHs (3.0 ng m−3), most likely due to its high volume of HDVs, while the red and gold lines exhibited low PAH concentrations (0.6 and 0.8 ng m−3 for red and gold lines, respectively). Lastly, lung cancer risk due to inhalation of PAHs was calculated based on a commuter lifetime (45 years for 2 hours per workday). Results showed that lung cancer risk for the 710 is 3.8 and 4.5 times higher than the light-rail (gold line) and subway (red line), respectively. With low levels of both metal and PAH pollutants, our results indicate that commuting on the light-rail (gold line) may have potential health benefits when compared to driving on freeways and busy roadways.

Organic compound characterization and source apportionment of indoor and outdoor quasi-ultrafine particulate matter in retirement homes of the Los Angeles Basin.

UNLABELLED: Quasi-ultrafine (quasi-UF) particulate matter (PM(0.25)) and its components were measured in indoor and outdoor environments at four retirement communities in Los Angeles Basin, California, as part of the Cardiovascular Health and Air Pollution Study (CHAPS). The present paper focuses on the characterization of the sources, organic constituents and indoor and outdoor relationships of quasi-UF PM. The average indoor/outdoor ratios of most of the measured polycyclic aromatic hydrocarbons (PAHs), hopanes, and steranes were close to or slightly lower than 1, and the corresponding indoor-outdoor correlation coefficients (R) were always positive and, for the most part, moderately strong (median R was 0.60 for PAHs and 0.74 for hopanes and steranes). This may reflect the possible impact of outdoor sources on indoor PAHs, hopanes, and steranes. Conversely, indoor n-alkanes and n-alkanoic acids were likely to be influenced by indoor sources. A chemical mass balance model was applied to both indoor and outdoor speciated chemical measurements of quasi-UF PM. Among all apportioned sources of both indoor and outdoor particles, vehicular emissions was the one contributing the most to the PM(0.25) mass concentration measured at all sites (24-47% on average). PRACTICAL IMPLICATIONS: Although people (particularly the elderly retirees of our study) generally spend most of their time indoors, a major portion of the PM(0.25) particles they are exposed to comes from outdoor mobile sources. This is important because, an earlier investigation, also conducted within the Cardiovascular Health and Air Pollution Study (CHAPS), showed that indoor-infiltrated particles from mobile sources are more strongly correlated with adverse health effects observed in the elderly subjects living in the studied retirement communities compared with other particles found indoors (Delfino et al., 2008).

Inhaled ultrafine particulate matter affects CNS inflammatory processes and may act via MAP kinase signaling pathways.

In addition to evidence that inhalation of ambient particulate matter (PM) can increase cardiopulmonary morbidity and mortality, the brain may also constitute a site adversely effected by the environmental presence of airborne particulate matter. We have examined the association between exposure to PM and adverse CNS effects in apolipoprotein E knockout (ApoE-/-) mice exposed to two levels of concentrated ultrafine particulate matter in central Los Angeles. Mice were euthanized 24h after the last exposure and brain, liver, heart, lung and spleen tissues were collected and frozen for subsequent bioassays. There was clear evidence of aberrant immune activation in the brains of exposed animals as judged by a dose-related increase in nuclear translocation of two key transcription factors, NF-kappaB and AP-1. These factors are involved in the promotion of inflammation. Increased levels of glial fibrillary acidic protein (GFAP) were also found consequent to particulate inhalation suggesting that glial activation was taking place. In order to determine the mechanism by which these events occurred, levels of several MAP kinases involved in activation of these transcription factors were assayed by Western blotting. There were no significant changes in the proportion of active (phosphorylated) forms of ERK-1, IkB and p38. However, the fraction of JNK in the active form was significantly increased in animals receiving the lower concentration of concentrated ambient particles (CAPs). This suggests that the signaling pathway by which these transcription factors are activated involves the activation of JNK.

Detection of changes in alveolar macrophage iron status induced by select PM2.5-associated components using iron-response protein binding activity.

The extent of adverse health effects, including induction/exacerbation of infectious lung disease, arising from entrainment of equivalent amounts (or exposure to a fixed increment) of fine particulate matter (PM2.5) can vary from region to region or city to city in a region. To begin to explain how differing effects on host resistance might arise after exposure to PM2.5 from various sites, we hypothesized that select metals (e.g., V, Al, and Mn) in each PM2.5 caused changes in alveolar macrophage (AM) Fe status that, ultimately, would lead to altered antibacterial function. To test this, iron-response protein (IRP) binding activity in a rat AM cell line was assessed after exposure to Fe alone and in conjunction with V, Mn, and/or Al at ratios of V:Fe, Al:Fe, or Mn:Fe encountered in PM2.5 samples from New York City, Los Angeles, and Seattle. Results indicated that V and Al each significantly altered IRP activity, though effects were not consistently ratio-(i.e., dose-) dependent; Mn had little impact on activity. We conclude that the reductions in Fe status detected here via the IRP assay arose, in part, from effects on transferrin-mediated Fe3+ delivery to the AM. Ongoing studies using this assay are allowing us to better determine: (1) whether mass (and/or molar) relationships between Fe and V, Al, and/or Mn in any PM2.5 sample consistently govern the extent of change in AM Fe status; (2) how much any specified PM2.5 constituent (metal or nonmetal) contributes to the overall disruption of Fe status found induced by an intact parent sample; and (3) whether induced changes in binding activity are relatable to other changes expected to occur in the AM, that is, in IRP-dependent mRNA/levels of ferritin/transferrin receptor and Fe-dependent functions. These studies demonstrate that pollutant-induced effects on lung cell Fe status can be assessed in a reproducible manner using an assay that can be readily performed by investigators who might otherwise have no access to other very costly analytical equipment, such as graphite atomic absorption or x-ray fluorescence spectro(photo)meters.

Particulate matter in polluted air may increase biomarkers of inflammation in mouse brain.

The etiology of neurodegenerative disorders is at present unknown. However, many of these disorders are associated with an increase in oxidative and inflammatory events. Although a small percentage of these disorders are familial cases linked to specific genetic defects, most are idiopathic. Thus, environmental factors are thought to play an important role in the onset and progression of such disorders. We have demonstrated that exposure (4 h, 5 days per week for 2 weeks) to concentrated airborne particulate matter increases inflammatory indices in brain of ovalbumin-sensitized BALB/c mice. Animals were divided into three exposure groups: filtered air (control), ultrafine particles, or fine and ultrafine particles. The levels of proinflammatory cytokines interleukin-1 alpha (IL-1alpha) and tumor necrosis factor alpha (TNF-alpha) were increased in brain tissue of mice exposed to particulate matter compared to that of control animals. Levels of the immune-related transcription factor NF-kappaB were also found to be substantially elevated in the brain of exposed groups compared with the control group. These data indicate that components of inhaled particulate matter may trigger a proinflammatory response in nervous tissue that could contribute to the pathophysiology of neurodegenerative diseases.

Ambient fine and coarse particle suppression of alveolar macrophage functions.

Alveolar macrophages (AM) are part of the innate immunological defense system and are among the first cells to respond to the effects of inhaled particles. Study of macrophage responses to particles is, therefore, relevant to understanding the mechanisms by which inhaled particles can adversely affect health. Size-fractionated ambient particles were collected at traffic-dominated sites in The Netherlands using a mobile high volume slit impactor system. AM were obtained by bronchoalveolar lavage from adult as well as aged rats and were incubated with for 4 h with collected particles at concentrations of 25-1000 pg per cell. Free radical generation by AM was measured with and without stimulation of AM with phorbol myristate acetate (PMA). There were dose-dependent decreases in macrophage production of superoxide radicals as measured by the chemiluminescent method. Coarse particles were more toxic than were fine particles. Suppression of free radical production did not seem to be related to the presence of bioavailable iron or to endotoxin associated with the particles. There were no statistically significant differences related to age or strain of the rats tested. We conclude that in vitro tests using AM is a useful and rapid method for delineating differences in toxicity between environmental samples of size fractionated ambient particles.

Development and evaluation of a continuous coarse (PM10-PM2.5) particle monitor.

In this paper, we describe the development and laboratory and field evaluation of a continuous coarse (2.5-10 microm) particle mass (PM) monitor that can provide reliable measurements of the coarse mass (CM) concentrations in time intervals as short as 5-10 min. The operating principle of the monitor is based on enriching CM concentrations by a factor of approximately 25 by means of a 2.5-microm cut point round nozzle virtual impactor while maintaining fine mass (FM)--that is, the mass of PM2.5 at ambient concentrations. The aerosol mixture is subsequently drawn through a standard tapered element oscillating microbalance (TEOM), the response of which is dominated by the contributions of the CM, due to concentration enrichment. Findings from the field study ascertain that a TEOM coupled with a PM10 inlet followed by a 2.5-microm cut point round nozzle virtual impactor can be used successfully for continuous CM concentration measurements. The average concentration-enriched CM concentrations measured by the TEOM were 26-27 times higher than those measured by the time-integrated PM10 samplers [the micro-orifice uniform deposit impactor (MOUDI) and the Partisol] and were highly correlated. CM concentrations measured by the concentration-enriched TEOM were independent of the ambient FM-to-CM concentration ratio, due to the decrease in ambient coarse particle mass median diameter with an increasing FM-to-CM concentration ratio. Finally, our results illustrate one of the main problems associated with the use of real impactors to sample particles at relative humidity (RH) values less than 40%. While PM10 concentrations obtained by means of the MOUDI and Partisol were in excellent agreement, CM concentrations measured by the MOUDI were low by 20%, and FM concentrations were high by a factor of 5, together suggesting particle bounce at low RH.

Factors affecting the stability of the performance of ambient fine-particle concentrators.

This article describes a systematic evaluation of factors affecting the stability of the performance of Harvard ambient fine-particle concentrators, an essential requirement for controlled animal and human exposure studies that utilize these technologies. Phenomenological problems during the operation of the concentrator, including pressure drop increase and decrease in concentration enrichment, were statistically correlated with ambient air parameters such as temperature, relative humidity, PM2.5 mass concentration, and mass median diameter. The normalized hourly pressure drop across the concentrator was strongly associated (R2 = .81) with the product of ambient PM2.5 mass concentration and the difference between the vapor pressure downstream of the impactor nozzle and the saturation vapor pressure at the adiabatic expansion temperature (i.e., the temperature of the aerosol immediately downstream of the virtual impactors). From multiple regression analysis, the average enrichment factor was predicted reasonably well (R2 = .67) by aerosol mass median diameter and the normalized hourly pressure drop. Based on these results, we can anticipate in any given day whether an exposure study can be conducted without a considerable increase in the concentrator pressure drop, which might lead to an abrupt or premature termination of the exposure. As particle mass concentration and ambient dewpoint are the two main parameters responsible for raising the pressure drop across the concentrator, efforts should be made to either desiccate the ambient aerosol at days of high dewpoints, or to dilute the ambient PM at days of high concentrations, prior to drawing the aerosol through the virtual impactors. The latter approach is recommended on days of severe ambient pollution conditions because it is simpler and also makes it possible to maintain the appropriate concentration level delivered to the exposure chamber.

Urban air particulate inhalation alters pulmonary function and induces pulmonary inflammation in a rodent model of chronic bronchitis.

Epidemiological studies have reported increased morbidity in human populations following inhalation of elevated levels of urban particulate matter. These responses are especially prevalent in populations with chronic obstructive pulmonary diseases, including chronic bronchitis. Toxicological studies have reported altered pulmonary function and increased pulmonary inflammation following particulate inhalation in the laboratory setting. However, most of these studies have utilized artificial particles that may not accurately mimic outdoor air pollutant conditions. Few studies have utilized actual urban air particle samples in inhalation studies. In the present study, the effects of inhaled concentrated urban air particulates on pulmonary function and pulmonary inflammation are addressed. Normal rats and rats with chronic bronchitis induced by approximately 200 ppm SO(2) for 6 wk were subsequently subjected to filtered air or concentrated air particles (CAPs). Twelve rats per group in 4 groups (48 rats total) were exposed for 5 h/day for 3 consecutive days. The CAPs aerosol levels were 206, 733, and 607 microg/m(3) (MMAD = 0.18 microm, sigma(g) = 2.9) on days 1, 2, and 3, respectively. Following the final day of exposure, pulmonary function parameters, including peak expiratory flow (PEF), tidal volume (TV), respiratory frequency (RF), and minute volume (MV), were measured and compared to preexposure baseline levels. Twenty-four hours following the final day of exposure, bronchoalveolar lavage was performed for total cell counts, differential cell counts, and total lavage protein levels. Pulmonary responses to CAPs in chronic bronchitic animals indicated a significant increase in tidal volume as well as peak expiratory flow. In CAPs-exposed animals without underlying bronchitis, significantly increased tidal volume was observed. Significant pulmonary inflammation was observed in the CAPs-exposed animals, particularly those with chronic bronchitis. Significant increases in neutrophils, lymphocytes, and total lavage protein were observed. These results suggest two distinct mechanistic responses to inhaled particles: a stress-type pulmonary function response marked by increases in flow and volume, that is, deeper breathing; and acute pulmonary inflammation marked by cellular influx, particularly neutrophils. From these data it is concluded that inhaled urban air particles alter pulmonary breathing parameters and increase pulmonary inflammation.

A pilot study to characterize fine particles in the environment of an automotive machining facility.

The main goal of this study was to characterize fine particles (e.g., smaller than about 3 microns) in an automotive machining environment. The Toledo Machining Plant of Chrysler Corporation was selected for this purpose. The effect of local mechanical processes as aerosol sources was a major part of this investigation. To determine the size-dependent mass concentration of particles in the plant, the Micro-Orifice Uniform Deposit Impactor (MOUDI Model 100, MSP Corp., Minneapolis, Minnesota) was used. The MOUDI was placed at central locations in departments with sources inside the plant, so that the obtained information on the size distribution realistically represents the aerosol to which plant workers are exposed. Sampling was conducted over a 4-day period, and during three periods per day, each matching the work shifts. A special effort was made to place the MOUDI at a central location of a department with relatively homogeneous particle sources. The selected sampling sites included welding, grinding, steel machining, and heat treating processes. The average 24-hour mass concentrations of particles smaller than 3.2 microns in aerodynamic diameter were 167.8, 103.9, 201.7, and 112.7 micrograms/m3 for welding, grinding, mild steel, and heat treating processes, respectively. Finally, the mass median diameters of welding, heat treatment, machining, and grinding operations were approximately 0.5, 0.5, 0.6, and 0.8 micron, respectively.

Alveolar macrophage interaction with air pollution particulates.

We applied flow cytometric analysis to characterize the in vitro response of alveolar macrophages (AM) to air pollution particulates. Normal hamster AM were incubated with varying concentrations of residual oil fly ash (ROFA) or concentrated ambient air particulates (CAP). We found a dose-dependent increase in AM-associated right angle light scatter (RAS) after uptake of ROFA (e.g., mean channel number 149.4 +/- 6.5, 102.5 +/- 4.1, 75.8 +/- 3.5, and 61.0 +/- 4.6 at 200, 100, 50, and 25 mg/ml, respectively) or CAP. A role for scavenger-type receptors (SR) in AM uptake of components of ROFA and CAP was identified by marked inhibition of RAS increases in AM pretreated with the specific SR inhibitor polyinosinic acid. We combined measurement of particle uptake (RAS) with flow cytometric analysis of intracellular oxidation of dichlorofluorescin. Both ROFA and CAP caused a dose-related intracellular oxidant stress within AM, comparable to that seen with phorbol myristate acetate (PMA) (e.g., fold increase over control, 6.6 +/- 0.4, 3.6 +/- 0.4, 4.6 +/- 0.5, 200 mg/ml ROFA, 100 mg/ml ROFA, and 10(-7) M PMA, respectively). We conclude that flow cytometry of RAS increases provides a useful relative measurement of AM uptake of complex particulates within ROFA and CAP. Both ROFA and CAP cause substantial intracellular oxidant stress within AM, which may contribute to subsequent cell activation and production of proinflammatory mediators.

Evaluation of the TEOM method for measurement of ambient particulate mass in urban areas.

Increased interest in the health effects of ambient particulate mass (PM) has focused attention on the evaluation of existing mass measurement methodologies and the definition of PM in ambient air. The Rupprecht and Patashnick Tapered Element Oscillating MicroBalance (TEOM) method for PM is compared with time-integrated gravimetric (manual) PM methods in large urban areas during different seasons. Comparisons are conducted for both PM10 and PM2.5 concentrations. In urban areas, a substantial fraction of ambient PM can be semi-volatile material. A larger fraction of this component of PM10 may be lost from the TEOM-heated filter than the Federal Reference Method (FRM). The observed relationship between TEOM and FRM methods varied widely among sites and seasons. In East Coast urban areas during the summer, the methods were highly correlated with good agreement. In the winter, correlation was somewhat lower, with TEOM PM concentrations generally lower than the FRM. Rubidoux, CA, and two Mexican sites (Tlalnepantla and Merced) had the highest levels of PM10 and the largest difference between TEOM and manual methods. PM2.5 data from collocation of 24-hour manual samples with the TEOM are also presented. As most of the semi-volatile PM is in the fine fraction, differences between these methods are larger for PM2.5 than for PM10.

A technique to expose animals to concentrated fine ambient aerosols.

This paper presents the development and evaluation of an ambient particle concentrator for conducting animal inhalation exposure studies. The system utilizes the principle of virtual impactors to concentrate ambient particles in the size range 0.1-2.5 microns (aerodynamic diameter; dp) by drawing them through a series of three virtual impactors. Each impactor contains the majority of ambient fine mass (dp < 2.5 microns aerodynamic diameter) in a bleed flow (minor flow) that is 20% of the total flow entering the virtual impactor. The virtual impactors have been characterized using indoor air samples as test aerosols. Fine mass and sulfate concentrations at the outlet of the concentrating system were compared to the ambient fine mass and sulfate levels, which were determined using Harvard-Marple impactors. In each of the stages, particle concentration was increased by a factor of approximately 3. Thus, an overall concentration factor of about 25-30 was achieved. The main goal of this study was to demonstrate the feasibility of conducting animal exposures using the newly developed ambient fine particle concentrator.