The stress-responsive cytotoxic effect of diesel exhaust particles on lymphatic endothelial cells.

Diesel exhaust particles (DEPs) are very small (typically < 0.2 µm) fragments that have become major air pollutants. DEPs are comprised of a carbonaceous core surrounded by organic compounds such as polycyclic aromatic hydrocarbons (PAHs) and nitro-PAHs. Inhaled DEPs reach the deepest sites in the respiratory system where they could induce respiratory/cardiovascular dysfunction. Additionally, a previous study has revealed that a portion of inhaled DEPs often activate immune cells and subsequently induce somatic inflammation. Moreover, DEPs are known to localize in lymph nodes. Therefore, in this study we explored the effect of DEPs on the lymphatic endothelial cells (LECs) that are a constituent of the walls of lymph nodes. DEP exposure induced cell death in a reactive oxygen species (ROS)-dependent manner. Following exposure to DEPs, next-generation sequence (NGS) analysis identified an upregulation of the integrated stress response (ISR) pathway and cell death cascades. Both the soluble and insoluble components of DEPs generated intracellular ROS. Three-dimensional Raman imaging revealed that DEPs are taken up by LECs, which suggests internalized DEP cores produce ROS, as well as soluble DEP components. However, significant cell death pathways such as apoptosis, necroptosis, ferroptosis, pyroptosis, and parthanatos seem unlikely to be involved in DEP-induced cell death in LECs. This study clarifies how DEPs invading the body might affect the lymphatic system through the induction of cell death in LECs.

Interplay of Climate Change and Air Pollution- Projection of the under-5 mortality attributable to ambient particulate matter (PM2.5) in South Asia.

Ambient fine particulate matter (PM2.5) pollution is a leading health risk factor for children under- 5 years, especially in developing countries. South Asia is a PM2.5 hotspot, where climate change, a potential factor affecting PM2.5 pollution, adds a major challenge. However, limited evidence is available on under-5 mortality attributable to PM2.5 under different climate change scenarios. This study aimed to project under-5 mortality attributable to long-term exposure to ambient PM2.5 under seven air pollution and climate change mitigation scenarios in South Asia. We used a concentration-risk function obtained from a previous review to project under-5 mortality attributable to ambient PM2.5. With a theoretical minimum risk exposure level of 2.4 µg/m3, this risk function was linked to gridded annual PM2.5 concentrations from atmospheric modeling to project under-5 mortality from 2010 to 2049 under different climate change mitigation scenarios. The scenarios were developed from the Aim/Endues global model based on end-of-pipe (removing the emission of air pollutants at the source, EoP) and 2 °C target measures. Our results showed that, in 2010-2014, about 306.8 thousand under-5 deaths attributable to PM2.5 occurred in South Asia under the Reference (business as usual) scenario. The number of deaths was projected to increase in 2045-2049 by 36.6% under the same scenario and 7.7% under the scenario where EoP measures would be partially implemented by developing countries (EoPmid), and was projected to decrease under other scenarios, with the most significant decrease (81.2%) under the scenario where EoP measures would be fully enhanced by all countries along with the measures to achieve 2 °C target (EoPmaxCCSBLD) across South Asia. Country-specific projections of under-5 mortality varied by country. The current emission control strategy would not be sufficient to reduce the number of deaths in South Asia. Robust climate change mitigation and air pollution control policy implementation is required.

Differential Pattern of Cell Death and ROS Production in Human Airway Epithelial Cells Exposed to Quinones Combined with Heated-PM2.5 and/or Asian Sand Dust.

The combined toxicological effects of airborne particulate matter (PM), such as PM2.5, and Asian sand dust (ASD), with surrounding chemicals, particularly quinones, on human airway epithelial cells remain underexplored. In this study, we established an in vitro combination exposure model using 1,2-naphthoquinones (NQ) and 9,10-phenanthroquinones (PQ) along with heated PM (h-PM2.5 and h-ASD) to investigate their potential synergistic effects. The impacts of quinones and heated PM on tetrazolium dye (WST-1) reduction, cell death, and cytokine and reactive oxygen species (ROS) production were examined. Results revealed that exposure to 9,10-PQ with h-PM2.5 and/or h-ASD dose-dependently increased WST-1 reduction at 1 µM compared to the corresponding control while markedly decreasing it at 10 µM. Higher early apoptotic, late apoptotic, or necrotic cell numbers were detected in 9,10-PQ + h-PM2.5 exposure than in 9,10-PQ + h-ASD or 9,10-PQ + h-PM2.5 + h-ASD. Additionally, 1,2-NQ + h-PM2.5 exposure also resulted in an increase in cell death compared to 1,2-NQ + h-ASD and 1,2-NQ + h-PM2.5 + h-ASD. Quinones with or without h-PM2.5, h-ASD, or h-PM2.5 + h-ASD significantly increased ROS production, especially with h-PM2.5. Our findings suggest that quinones, at relatively low concentrations, induce cell death synergistically in the presence of h-PM2.5 rather than h-ASD and h-PM2.5 + h-ASD, partially through the induction of apoptosis with increased ROS generation.

Metabolic outcomes and changes in innate immunity induced by diesel exhaust particles airway exposure and high-fat high-sucrose diet.

AIMS: Epidemiological studies have shown that exposure to diesel exhaust particles (DEP) is associated with metabolic diseases. We used mice with nonalcoholic fatty liver disease (NAFLD) caused by a high-fat, high-sucrose diet (HFHSD), which mimics a Western diet, to investigate the mechanism of NAFLD exacerbation via changes in innate immunity in the lungs by airway exposure to DEP. MAIN METHODS: Six-week-old C57BL6/J male mice were fed HFHSD, and DEP was administered endotracheally once a week for eight weeks. The histology, gene expression, innate immunity cells in the lung and liver, and the serum inflammatory cytokine levels, were investigated. KEY FINDINGS: Under the HFHSD, DEP increased blood glucose levels, serum lipid levels, and NAFLD activity scores, and also the expression of genes associated with inflammation in the lungs and liver. DEP caused an increase in ILC1s, ILC2s, ILC3s, and M1 macrophages in the lungs and a marked increase in ILC1s, ILC3s, M1 macrophages, and natural killer cells in the liver, while ILC2 levels were not changed. Furthermore, DEP caused high levels of inflammatory cytokines in the serum. SIGNIFICANCE: Chronic exposure to DEP in HFHSD-fed mice increased inflammatory cells involved in innate immunity in the lungs and raised local inflammatory cytokine levels. This inflammation spread throughout the body, suggesting the association with the progression of NAFLD via increased inflammatory cells involved in innate immunity and inflammatory cytokine levels in the liver. These findings contribute to a better understanding of the role of innate immunity in air pollution-related systemic diseases, especially metabolic diseases.

Association between Long-Term Ambient PM2.5 Exposure and under-5 Mortality: A Scoping Review.

Studies have established a link between exposure to fine particles (PM2.5) and mortality in infants and children. However, few studies have explored the association between post-birth exposure to PM2.5 and under-5 mortality. We conducted a scoping review to identify relevant epidemiological evidence on the association between post-birth ambient PM2.5 exposure and under-5 mortality. We searched PubMed and Web of Science for articles published between 1970 and the end of January 2022 that explicitly linked ambient PM2.5 and under-5 mortality by considering the study area, study design, exposure window, and child age. Information was extracted on the study characteristics, exposure assessment and duration, outcomes, and effect estimates/findings. Ultimately, 13 studies on infant and child mortality were selected. Only four studies measured the effect of post-birth exposure to PM2.5 on under-5 mortality. Only one cohort study mentioned a positive association between post-birth ambient PM2.5 exposure and under-5 mortality. The results of this scoping review highlight the need for extensive research in this field, given that long-term exposure to ambient PM2.5 is a major global health risk and child mortality remains high in some countries.

Oral Exposure to Polystyrene Microplastics of Mice on a Normal or High-Fat Diet and Intestinal and Metabolic Outcomes.

BACKGROUND: Microplastics (MPs) are small particles of plastic (≤5mm in diameter). In recent years, oral exposure to MPs in living organisms has been a cause of concern. Leaky gut syndrome (LGS), associated with a high-fat diet (HFD) in mice, can increase the entry of foreign substances into the body through the intestinal mucosa. OBJECTIVES: We aimed to evaluate the pathophysiology of intestinal outcomes associated with consuming a high-fat diet and simultaneous intake of MPs, focusing on endocrine and metabolic systems. METHODS: C57BL6/J mice were fed a normal diet (ND) or HFD with or without polystyrene MP for 4 wk to investigate differences in glucose tolerance, intestinal permeability, gut microbiota, as well as metabolites in serum, feces, and liver. RESULTS: In comparison with HFD mice, mice fed the HFD with MPs had higher blood glucose, serum lipid concentrations, and nonalcoholic fatty liver disease (NAFLD) activity scores. Permeability and goblet cell count of the small intestine (SI) in HFD-fed mice were higher and lower, respectively, than in ND-fed mice. There was no obvious difference in the number of inflammatory cells in the SI lamina propria between mice fed the ND and mice fed the ND with MP, but there were more inflammatory cells and fewer anti-inflammatory cells in mice fed the HFD with MPs in comparison with mice fed the HFD without MPs. The expression of genes related to inflammation, long-chain fatty acid transporter, and Na+/glucose cotransporter was significantly higher in mice fed the HFD with MPs than in mice fed the HFD without MPs. Furthermore, the genus Desulfovibrio was significantly more abundant in the intestines of mice fed the HFD with MPs in comparison with mice fed the HFD without MPs. Muc2 gene expression was decreased when palmitic acid and microplastics were added to the murine intestinal epithelial cell line MODE-K cells, and Muc2 gene expression was increased when IL-22 was added. DISCUSSION: Our findings suggest that in this study, MP induced metabolic disturbances, such as diabetes and NAFLD, only in mice fed a high-fat diet. These findings suggest that LGS might have been triggered by HFD, causing MPs to be deposited in the intestinal mucosa, resulting in inflammation of the intestinal mucosal intrinsic layer and thereby altering nutrient absorption. These results highlight the need for reducing oral exposure to MPs through remedial environmental measures to improve metabolic disturbance under high-fat diet conditions. https://doi.org/10.1289/EHP11072.

Effects of styrene monomer on a mouse model of atopic dermatitis.

Aim: Styrene monomer (SM) is a basic chemical used as a raw material for polystyrene and unsaturated polyester resins and in the production of synthetic resins, synthetic rubbers, paints, and adhesives. To date, it is unclear whether SM is associated with the aggravation of atopic dermatitis. The aim was to investigate the effects of SM on atopic dermatitis-like skin lesions induced by mite allergen in NC/Nga mice.Methods: Male mice were injected intradermally with mite allergen on their right ears. In the presence of an allergen, SM (3.5 or 350 µg/animal/week) was administered by intraperitoneal injection. We evaluated clinical scores, ear thickening, histologic findings, and the protein expressions of cytokines and chemokines.Results: Macroscopic and microscopic examinations demonstrated that exposure to SM at a dose of 3.5 µg caused an exacerbation of atopic dermatitis-like skin lesions related to mite allergen. These changes were consistent with the level of histamine in the ear tissue as an overall trend. In contrast, 350-µg SM did not show significant enhancement effects.Conclusion: These results indicate that SM exacerbated atopic dermatitis-like skin lesions at hundred-fold lower levels than the level that causes no observed adverse effects as determined by histologic changes in rodent livers. SM could be at least partly responsible for the recent increase in atopic dermatitis.Impact statementStyrene monomer (SM) is classified as an International Agency for Research on Cancer group 2B carcinogen and includes neurotoxicity and respiratory disorders. However, the effects of SM as a chemical substance on existing allergic pathophysiology have not been elucidated yet. This study demonstrated that SM exacerbated murine atopic dermatitis-like skin lesions at hundred-fold lower levels than the level that causes no observed adverse effects as determined by histologic changes in rodent livers, which was concomitant with the local level of histamine. These data hasten a need for comprehensive research to clarify the chemical pollutants' effects of doses much lower than NOAEL on vulnerable pathophysiologies such as allergy/atopy.

Effect modification by temperature on the association between O3 and emergency ambulance dispatches in Japan: A multi-city study.

Numerous epidemiological studies have reported that ozone (O3) and temperature are independently associated with health outcomes, but modification of the effects of O3 on health outcomes by temperature, and vice versa, has not been fully described. This study aimed to investigate effect modification by temperature on the association between O3 and emergency ambulance dispatches (EADs) in Japan. Data on daily air pollutants, ambient temperature, and EADs were obtained from eight Japanese cities from 2007 to 2015. A distributed lag non-linear model combined with Poisson regression was performed with temperature as a confounding factor and effect modifier to estimate the effects of O3 on EADs at low (<25th percentile), moderate (25th-75th percentile), and high (>75th percentile) temperature for each city. The estimates obtained from each city were pooled by random-effects meta-analysis. When temperature was entered as a confounder, the estimated effects of O3 on EADs for all acute, cardiovascular, and respiratory illnesses were largest at lag 0 (current-day lag). Therefore, this lag was used to further estimate the effects of O3 on EADs in each temperature category. The estimated effects of O3 on EADs for all acute, cardiovascular, and respiratory illnesses in all eight Japanese cities increased with increasing temperature. Specifically, a 10 ppb increase in O3 was associated with 0.80 % (95 % CI: 0.25 to 1.35), 0.19 % (95 % CI: -0.85 to 1.25), and 1.14 % (95 % CI: -0.01 to 2.31) increases in the risk of EADs for all acute, cardiovascular, and respiratory illnesses, respectively, when city-specific daily temperature exceeded the 75th percentile. Our findings suggest that the association between O3 and EADs for all acute, cardiovascular, and respiratory illnesses is the highest during high temperature. Finding of this study can be used to develop potential mitigation measures against O3 exposure in high temperature environment to reduce its associated adverse health effects.

Effects of Streamer Discharge on PM2.5 Containing Endotoxins and Polyaromatic Hydrocarbons and Their Biological Responses In Vitro.

Experimental and epidemiological studies have demonstrated that fine particulate matter with a diameter of <2.5 µm (PM2.5) affects both the respiratory and immune systems. However, effective approaches to reduce PM2.5-induced hazardous effects have not been discovered yet. Streamer discharge is a category of plasma discharge in which high-speed electrons collide with oxygen and nitrogen molecules. Although streamer discharge can reportedly eliminate bacteria, molds, chemical substances, and allergens, its ability to decontaminate PM2.5 has not been previously demonstrated. The present study explored whether streamer discharge treatment could reduce PM2.5-induced inflammatory responses by employing an in vitro system. PM2.5 was collected under four conditions (Bangkok (Sep.−Dec.), Bangkok (Dec.−Mar.), Singapore, and Taipei). Airway epithelial cells and antigen-presenting cells exposed to non-treated PM2.5 in several conditions resulted in inflammatory responses. Streamer-discharged PM2.5 (Bangkok (Sep.−Dec.)) decreased the expression of interleukin (IL)-6 and IL-8 compared to non-treated PM2.5. Moreover, composition analysis demonstrated that streamer discharge reduced some compounds, such as endotoxins and polycyclic aromatic hydrocarbons, included in PM2.5 that can elicit inflammatory responses. Streamer discharge treatment can reduce endotoxins, polycyclic aromatic hydrocarbons, and the subsequent inflammatory responses induced by PM2.5 in vitro.

Estimation of hospital visits for respiratory diseases attributable to PM10 from vegetation fire smoke and health impacts of regulatory intervention in Upper Northern Thailand.

The air quality in Upper Northern Thailand (UNT) deteriorates during seasonal vegetation fire events, causing adverse effects especially on respiratory health outcomes. This study aimed to quantitatively estimate respiratory morbidity from vegetation fire smoke exposure, and to assess the impact of a burning ban enforced in 2016 on morbidity burden in UNT. We computed daily population exposure to fire-originated PM10 and estimated its health burden during a 5-year period from 2014 to 2018 using daily fire-originated PM10 concentration and the concentration-response function for short-term exposure to PM10 from vegetation fire smoke and respiratory morbidity. In subgroups classified as children and older adults, the health burden of respiratory morbidity was estimated using specific effect coefficients from previous studies conducted in UNT. Finally, we compared the health burden of respiratory morbidity before and after burning ban enforcement. Approximately 130,000 hospital visits for respiratory diseases were estimated to be attributable to fire-originated PM10 in UNT from 2014 to 2018. This estimation accounted for 1.3% of total hospital visits for respiratory diseases during the 5-year period, and 20% of those during burning events. Age-specific estimates revealed a larger impact of PM10 in the older adult group. The number of hospital visits for respiratory diseases attributable to fire-originated PM10 decreased from 1.8% to 0.5% after the burning ban policy was implemented in the area. Our findings suggest that PM10 released from vegetation fires is a health burden in UNT. The prohibition of the burning using regulatory measure had a positive impact on respiratory morbidity in this area.

Post-staining Raman analysis of histological sections following decolorization.

Hematoxylin and eosin (HE) staining of tissue sections is a powerful tool for observing changes in the tissue structure and is used as the most fundamental and vital technique in histology. However, xenobiotics such as polymers and inorganic or organic materials have low dyeability, making it difficult to observe the distribution of materials across tissues. Raman spectroscopy is an advantageous technique for identifying materials in tissues using spectroscopic fingerprints by laser irradiation without staining. In this study, we developed a combined method for morphological observation and Raman spectral acquisition on the identical tissue slide by employing a decolorization step to remove eosin-induced fluorescence in HE-stained samples. Our method eliminated the fluorescence background and allowed the identical-field pathological observation, enabling simultaneous identification of biological responses and materials in tissues.

Effects of Oral Exposure to Low-Dose Bisphenol S on Allergic Asthma in Mice.

Bisphenol S (BPS) is increasingly being used as an alternative for bisphenol A; however, its health effects remain unclear. We investigated the effects of oral exposure to low-dose BPS on allergic asthma. C3H/HeJ male mice were intratracheally administered with allergen (ovalbumin (OVA), 1 µg/animal) every 2 weeks from 6 to 11 weeks old. BPS was ingested by drinking water at doses equivalent to 0.04, 0.4, and 4 µg/kg/day. We then examined pulmonary inflammation, airway hyperresponsiveness, serum OVA-specific immunoglobulin (Ig) levels, Th2 cytokine/chemokine production, and mediastinal lymph node (MLN) cell activities. Compared with OVA alone, moderate-dose BPS (BPS-M) with OVA significantly enhanced pulmonary inflammation, airway hyperresponsiveness, and OVA-specific IgE and IgG1. Furthermore, interleukin (IL)-5, IL-13, IL-33, and CCL11/Eotaxin protein levels in the lungs increased. Conversely, these allergic responses were reduced in the high-dose BPS+OVA group. In MLN cells, BPS-M with OVA increased the total cell count and activated antigen-presenting cells including conventional dendritic cell subset (cDC2). After OVA restimulation, cell proliferation and Th2 cytokine production (IL-4, IL-5, and IL-13) in the culture supernatant also increased. Therefore, oral exposure to low-dose BPS may exacerbate allergic asthmatic responses by enhancing Th2-polarized responses and activating the MLN cells.

Effect of a vegetation fire event ban on hospital visits for respiratory diseases in Upper Northern Thailand.

BACKGROUND: Upper Northern Thailand (UNT) has been episodically affected by air pollution from vegetation burning, which causes adverse respiratory health effects. However, no study has evaluated the effect of regulatory actions to prohibit vegetation burning on respiratory morbidity. We examined the effect of a burning ban enforced in May 2016 on hospital visits for respiratory diseases in UNT. METHODS: This study used data from eight provinces in UNT. Analyses were conducted for January to April of 2014-2016 (before ban enforcement) and January to April of 2017-2018 (after ban enforcement). Particulate matter of 10 microns in diameter or smaller (PM10) concentrations, numbers of satellite fire hotspots and age-standardized rates of hospital visits for respiratory diseases before and after ban enforcement were compared. The effect of the ban on hospital visits for respiratory diseases was evaluated using an interrupted time-series analysis controlled for season-specific temporal trends, day of week, public holiday, temperature, relative humidity, number of hospitals and offset population, with gastrointestinal diseases as a negative control. A meta-analysis was performed to pool province-specific effect estimates. RESULTS: The daily average PM10 concentration and the number of fire hotspots decreased after ban enforcement in all provinces in UNT, with percent changes ranging from 5.3 to 34.3% and 14.3 to 81.5%, respectively. The adjusted pooled effect estimates of hospital visits for respiratory diseases decreased by 9.1% (95% CI: 5.1, 12.9), whereas a null association was observed for gastrointestinal diseases. CONCLUSION: The burning ban had a positive impact on both air pollution levels and rates of hospital visits for respiratory diseases in UNT.

Dietary Exposure to Flame Retardant Tris (2-Butoxyethyl) Phosphate Altered Neurobehavior and Neuroinflammatory Responses in a Mouse Model of Allergic Asthma.

Tris (2-butoxyethyl) phosphate (TBEP) is an organophosphate flame retardant and used as a plasticizer in various household products such as plastics, floor polish, varnish, textiles, furniture, and electronic equipment. However, little is known about the effects of TBEP on the brain and behavior. We aimed to examine the effects of dietary exposure of TBEP on memory functions, their-related genes, and inflammatory molecular markers in the brain of allergic asthmatic mouse models. C3H/HeJSlc male mice were given diet containing TBEP (0.02 (TBEP-L), 0.2 (TBEP-M), or 2 (TBEP-H) µg/kg/day) and ovalbumin (OVA) intratracheally every other week from 5 to 11 weeks old. A novel object recognition test was conducted in each mouse at 11 weeks old. The hippocampi were collected to detect neurological, glia, and immunological molecular markers using the real-time RT-PCR method and immunohistochemical analyses. Mast cells and microglia were examined by toluidine blue staining and ionized calcium-binding adapter molecule (Iba)-1 immunoreactivity, respectively. Impaired discrimination ability was observed in TBEP-H-exposed mice with or without allergen. The mRNA expression levels of N-methyl-D aspartate receptor subunits Nr1 and Nr2b, inflammatory molecular markers tumor necrosis factor-α oxidative stress marker heme oxygenase 1, microglia marker Iba1, and astrocyte marker glial fibrillary acidic protein were significantly increased in TBEP-H-exposed mice with or without allergen. Microglia and mast cells activation were remarkable in TBEP-H-exposed allergic asthmatic mice. Our results indicate that chronic exposure to TBEP with or without allergen impaired object recognition ability accompanied with alteration of molecular expression of neuronal and glial markers and inflammatory markers in the hippocampus of mice. Neuron-glia-mast cells interaction may play a role in TBEP-induced neurobehavioral toxicity.

Long-term air pollution exposure and decreased kidney function: A longitudinal cohort study in Bangkok Metropolitan Region, Thailand from 2002 to 2012.

BACKGROUND: Kidney dysfunction is considered a cardiovascular risk factor. However, few longitudinal studies have examined the effects of air pollution on kidney function. We evaluated associations between long-term air pollution exposure and estimated glomerular filtration rate (eGFR) using data from a cohort of the Electricity Generating Authority of Thailand (EGAT) study in Bangkok Metropolitan Region, Thailand. METHODS: This longitudinal study included 1839 subjects (aged 52-71 years in 2002) from the EGAT1 cohort study during 2002-2012. eGFR, based on creatinine, was measured in 2002, 2007, and 2012. Annual mean concentrations of air pollutants (i.e., particulate matter with an aerodynamic diameter ≤10 µm (PM10), ozone (O3), nitrogen dioxide (NO2), sulfur dioxide (SO2), and carbon monoxide (CO)) prior to a measurement of creatinine were assessed with the ordinary kriging method. Mixed-effect linear regression models were used to assess associations between air pollutants and eGFR, while controlling for potential covariates. eGFR values are expressed as percent change per interquartile range (IQR) increments of each pollutant. RESULTS: Lower eGFR was associated with higher concentrations of PM10 (-1.99%, 95% confidence interval (CI): -3.33, -0.63), SO2 (-4.89%, 95%CI: -6.69, -3.07), and CO (-0.97%, 95%CI: -1.96, 0.03). However, after adjusting for temperature, relative humidity, PM10, and SO2, no significant association was observed between CO and eGFR. CONCLUSIONS: Our findings support the hypothesis that long-term exposure to high concentrations of PM10 and SO2 is associated with the progression of kidney dysfunction in subjects of the EGAT cohort study.

Investigation of association between smoke haze and under-five mortality in Malaysia, accounting for time lag, duration and intensity.

BACKGROUND: Studies on the association between smoke haze (hereafter 'haze') and adverse health effects have increased in recent years due to extreme weather conditions and the increased occurrence of vegetation fires. The possible adverse health effects on under-five children (U5Y) is especially worrying due to their vulnerable condition. Despite continuous repetition of serious haze occurrence in Southeast Asia, epidemiological studies in this region remained scarce. Furthermore, no study had examined the association accounting for three important aspects (time lag, duration and intensity) concurrently. OBJECTIVE: This study aimed to examine the association between haze and U5Y mortality in Malaysia, considering time lag, duration and intensity of exposure. METHODS: We performed a time-stratified case-crossover study using a generalized additive model to examine the U5Y mortality related to haze in 12 districts in Malaysia, spanning from 2014 to 2016. A 'haze day' was characterized by intensity [based on concentrations of particulate matter (PM)] and duration (continuity of haze occurrence, up to 3 days). RESULTS: We observed the highest but non-significant odds ratios (ORs) of U5Y mortality at lag 4 of Intensity-3. Lag patterns revealed the possibility of higher acuteness at prolonged and intensified haze. Stratifying the districts by the 95th-percentile of PM distribution, the 'low' category demonstrated marginal positive association at Intensity-2 Duration-3 [OR: 1.210 (95% confidence interval: 1.000, 1.464)]. CONCLUSIONS: We found a null association between haze and U5Y mortality. The different lag patterns of the association observed over different duration and intensity suggest consideration of these aspects in future studies.

Application of three-dimensional Raman imaging to determination of the relationship between cellular localization of diesel exhaust particles and the toxicity.

A diesel exhaust particle (DEP) is a type of particulate matter that is easily produced from combustion in a diesel power engine. It has been reported that DEPs can cause short- and long-term health problems. This is because DEPs are complex mixtures that are highly inhalable through the airways due to their small particle size. However, the relationship between intracellular localization of DEPs after their deposition in the lungs and the subsequent biological responses remains to be clarified. This is due to difficulties in distinguishing particles that are inside the cells from those that are outside. In this study, A549 human lung epithelial cells were exposed to DEPs at concentrations of 0, 25, 75, or 200 µg/mL for different periods, after that particles in the A549 cells were analyzed by three-dimensional (3D) images obtained from a Raman microscope. The cytotoxic effects of DEPs on the A549 cells were investigated by measuring cell viability, the levels of intracellular reactive oxygen species (ROS) and cell death. The Raman microscopy revealed that the particles invaded the A549 cells, and at a concentration of 200 µg/mL, they markedly decreased cell viability, increased intracellular ROS production, triggered late apoptosis/necrosis and induced nuclear damage. These results suggest that intracellular DEPs exposed at a high concentration may be highly toxic and can impair the viability of A549 cells. Furthermore, the 3D images from the Raman microscopy can be used to evaluate intracellular particle dynamics.

Biological Response-Enhancing Activity with Antigens in A549 Cells Exposed to Representative Polycyclic Aromatic Hydrocarbons.

The question of what kinds of airborne particles, including diesel exhaust particles and their adherent chemical constituents, exacerbate the activity of allergic and inflammatory respiratory diseases has not been elucidated in detail. Therefore, chemicals that have amplifying actions on Dermatophagoides farinae (Df) body extract-induced IL-8, the inflammatory cytokines of the innate immune system, were comprehensively examined using commonly used human alveolar epithelial cells, A549, as simple screening for 17 polycyclic aromatic hydrocarbons (PAHs), which are representative organic constituents in atmospheric samples. The significant amplifying actions of two PAHs, dibenzo[a,l]pyrene (DB[a,l]P) at 50 nM and dibenzo[a,i]pyrene (DB[a,i]P) at 2 µM for 48 h, for IL-8 protein release induced by mite antigens in epithelial cells were observed for the first time. In contrast, the enhancement of IL-8 was not observed in protein levels for these PAHs without the antigens. Meanwhile, the significant synergistic amplifying effect of DB[a,l]P at 50 nM on proinflammatory actions was measured in gene expression (i.e., IL-8, IL-6, ICAM-1, and TNF-α) levels in the experimental setting; for the results, the induction of TNF-α may have been the essential factor that enhanced the amplifying activity of DB[a,l]P for IL-8 gene expression and protein release. Examining the exacerbating effect on allergic pathophysiological states for DB[a,l]P is planned for further study.

Impact of dietary exposure to low-dose tris(1,3-dichloro-2-propyl)phosphate in allergic asthmatic mice.

OBJECTIVE: Tris(1,3-dichloro-2-propyl)phosphate (TDCIPP) is an organophosphorus flame retardant that is an alternative to brominated flame retardants. Although TDCIPP can adversely affect human health, information about its effects on immune and allergic responses is scarce. We aimed to investigate the effects of dietary exposure to TDCIPP using less than the human tolerable daily intake (TDI) in allergic asthmatic mice. METHODS: Male C3H/HeJSlc mice were fed a chow diet containing TDCIPP equivalent to 0.02 µg/kg/day (low; L), 0.2 µg/kg/day (medium; M), or 2 µg/kg/day (high; H) and were intratracheally administered ovalbumin (OVA, 1 µg/animal) every 2 weeks from 5 to 11 weeks of age. RESULTS: In OVA-treated mice, TDCIPP-H exposure tended to enhance pulmonary inflammation compared with vehicle exposure. TDCIPP dose-dependently decreased mRNA level of G protein-coupled estrogen receptor (GPER) in the lungs with or without OVA. OVA + TDCIPP-H treatment tended to increase the total cell number and promoted CD4+ cell activation compared with OVA alone treatment in mediastinal lymph nodes. In splenocytes, an increase in the fraction of Breg cells, but not of total B and T cells, and an increase in IL-5 in cell culture supernatants following OVA re-stimulation in OVA + TDCIPP-H-treated mice was observed compared with OVA-alone-treated mice. Moreover, OVA + TDCIPP-H exposure decreased Gr-1 expression in bone marrow (BM) cells. DISCUSSION: These results suggested that dietary exposure to TDCIPP at TDI level slightly enhances allergic diseases, such as allergic asthma, via GPER regulation at inflamed sites and secondary lymphoid tissue and BM cell alternations.