Ambient fine and coarse particles in Japan affect nasal and bronchial epithelial cells differently and elicit varying immune response.

Ambient particulate matter (PM) epidemiologically exacerbates respiratory and immune health, including allergic rhinitis (AR) and bronchial asthma (BA). Although fine and coarse particles can affect respiratory tract, the differences in their effects on the upper and lower respiratory tract and immune system, their underlying mechanism, and the components responsible for the adverse health effects have not been yet completely elucidated. In this study, ambient fine and coarse particles were collected at three different locations in Japan by cyclone technique. Both particles collected at all locations decreased the viability of nasal epithelial cells and antigen presenting cells (APCs), increased the production of IL-6, IL-8, and IL-1ß from bronchial epithelial cells and APCs, and induced expression of dendritic and epithelial cell (DEC) 205 on APCs. Differences in inflammatory responses, but not in cytotoxicity, were shown between both particles, and among three locations. Some components such as Ti, Co, Zn, Pb, As, OC (organic carbon) and EC (elemental carbon) showed significant correlations to inflammatory responses or cytotoxicity. These results suggest that ambient fine and coarse particles differently affect nasal and bronchial epithelial cells and immune response, which may depend on particles size diameter, chemical composition and source related particles types.

Nitro-PAH exposures of occupationally-exposed traffic workers and associated urinary 1-nitropyrene metabolite concentrations.

The assessment of occupational exposure to diesel exhaust (DE) is important from an epidemiological perspective. Urinary biomarkers of exposure have been proposed as a novel approach for measuring exposure to DE. In this study, we measured the concentrations of two urinary metabolites of 1-nitropyrene (1NP), a nitrated polycyclic aromatic hydrocarbon that has been suggested as a molecular marker of diesel particulate matter. These two metabolites, 6-hydroxy-1-nitropyrene and 8-hydroxy-1-nitropyrene, were determined in urine samples (10mL) from a small group of workers who were occupationally-exposed to vehicle exhaust in Trujillo, Peru, before and after their workshifts. Workshift exposures to 1NP, as well as PM2.5, 2-nitropyrene and 2-nitrofluoranthene, were also measured. Exposures to 1NP were similar in all studied workers, averaging 105±57.9pg/m3 (±standard deviation). Median urinary concentrations of the average of the pre- and post-exposure samples for 6-hydroxy-1-nitropyrene and 8-hydroxy-1-nitropyrene, were found to be 3.9 and 2.3pgmetabolite/mg creatinine, respectively in the group of occupationally-exposed subjects (n=17) studied. A direct relationship between workshift exposure to 1NP and urinary 1NP metabolites concentrations was not observed. However, the 1NP exposures and the creatinine-corrected urinary concentrations of the hydroxynitropyrene metabolites in these Peruvian traffic workers were similar to occupationally-exposed taxi drivers in Shenyang, China, and were higher than biomarker levels in office workers from Trujillo without occupational exposure to vehicle exhaust. This study provides further evidence that urinary metabolites of 1NP are associated with exposure to DE and may serve as a useful exposure biomarker.

Simultaneous determination of polycyclic aromatic hydrocarbon quinones by gas chromatography-tandem mass spectrometry, following a one-pot reductive trimethylsilyl derivatization.

We developed a sensitive and selective method to simultaneously analyze 37 polycyclic aromatic hydrocarbon quinones (PAHQs) with GC-MS/MS and applied the method to the analysis of standard atmospheric particulate matter samples. PAHQs were reduced with zinc granules and dithiothreitol (DTT) and the reductants were immediately converted to their silylated derivatives in a test tube. Two trimethylsilyl (TMS) groups were introduced into PAHQs through the one-pot reductive TMS derivatization. The PAHQs were derivatized with a mixed silylation reagent (BSA+TMCS+TMSI; (3:2:3)), which is one of the combinations of TMS-derivatization reagents with the highest reactivity. The derivatives produced different fragmentation between o-PAHQs and p-PAHQs. Therefore, isomers that have the same molecular weight are difficult to separate on a column were separated by the selected reaction monitoring (SRM) mode using the characteristic fragmentations, allowing separation and detection of all PAHQ derivatives in less than 30min. The instrumental detection limit (IDL) of each PAHQ was 1.2-29fg/injection and the method quantification limit (MQL) was 0.8-78µg/kg sample. For quantification, six deuterated PAHQs were used as internal standards to achieve high analytical precision. We applied the developed method to four standard atmospheric particulate matter samples. Results showed that out of 37 PAHQs, 33 compounds were identified and quantified. Moreover, from the 33 PAHQs, 14 were detected for the first time. Similar values were observed for the concentrations of PAHQs that have been quantified in previous reports. This method has the highest practicality in monitoring PAHQs in atmosphere, combustion exhaust gas, and toxicity evaluation. Thus, the method has the potential to become a standard analytical method for such applications.

Polycyclic Aromatic Hydrocarbons in Surface Water of the Southeastern Japan Sea.

Surface water samples were collected at 15 sampling sites in the southeastern Japan Sea along the Japanese Archipelago for analysis of polycyclic aromatic hydrocarbons (PAHs). Water samples were fractionated by filtration through a glass fiber membrane (pore size 0.5 µm) and analyzed by high-performance liquid chromatography with fluorescence detection. Thirteen PAHs having 3 to 6 rings were found in the dissolved phase (DP) and 12 were found in the particulate phase (PP). The total (DP+PP) PAH concentration ranged from 6.83 to 13.81 ng/L with the mean±standard deviation (S.D.) concentration of 9.36±1.92 ng/L. The mean±S.D. PAH concentration in the DP and PP was 5.99±1.80 and 3.38±0.65 ng/L, respectively. Three-ring PAHs predominated in the DP, while the proportion of 4-ring PAHs was higher in the PP. The mean total PAH concentration in the southeastern Japan Sea was higher than the concentration in the northwestern Japan Sea (8.5 ng/L). The Tsushima Current, which originates from the East China Sea with higher PAH concentration, is considered to be responsible for this higher concentration.

Mineral dust aerosols promote the formation of toxic nitropolycyclic aromatic compounds.

Atmospheric nitrated polycyclic aromatic hydrocarbons (NPAHs), which have been shown to have adverse health effects such as carcinogenicity, are formed in part through nitration reactions of their parent polycyclic aromatic hydrocarbons (PAHs) in the atmosphere. However, little is known about heterogeneous nitration rates of PAHs by gaseous NO2 on natural mineral substrates, such as desert dust aerosols. Herein by employing kinetic experiments using a flow reactor and surface analysis by Fourier transform infrared spectroscopy with pyridine adsorption, we demonstrate that the reaction is accelerated on acidic surfaces of mineral dust, particularly on those of clay minerals. In support of this finding, we show that levels of ambient particle-associated NPAHs in Beijing, China, significantly increased during heavy dust storms. These results suggest that mineral dust surface reactions are an unrecognized source of toxic organic chemicals in the atmosphere and that they enhance the toxicity of mineral dust aerosols in urban environments.

Function of rayon fibers with metallophthalocyanine derivatives: potential of low-molecular weight polycyclic aromatic hydrocarbon removal and Bacillus sp. removal.

Polycyclic aromatic hydrocarbons (PAHs) are known as carcinogenic and/or mutagenic substances, and are present at high concentration in polluted environments. It has recently been reported that spore-forming bacteria (e.g., Bacillus spp.) can be transported long distances alive in the atmosphere, which raises the possibility that some of the transported bacteria could have adverse effects on human health. There is thus a need for filters that can remove gaseous PAHs from the air that people breathe and that can inhibit bacterial growth on the filters. We focused on metallophthalocyanine derivatives (M-Pc) which are known to adsorb PAHs as well as to inhibit the growth of bacteria as a potential filtering agent. In this study, we developed different types of M-Pc-supported rayon fibers by changing central metals, functional groups, concentrations of M-Pc and rayon types, and evaluated their removal effects by measuring adsorption rates of 3- and 4-ring PAHs with a HPLC and growth curves of Bacillus sp. with a spectrophotometer. The results showed that both the effects depended on functional groups and concentrations of M-Pc, and rayon types. The most effective combination was observed in Fe-Pc with sulfo group supported on cationized rayon fiber at the concentration of 2 to 3.3 wt%. Central metal species of M-Pc were influenced only on the antibacterial properties. This fiber would be applicable to filtering agents and textiles.

Comparing the role of silica particle size with mineral fiber geometry in the release of superoxide from rat alveolar macrophages.

Particulate air pollutants and mineral fibers activate inflammatory cells to release oxidants, which contribute to inflammation and injury in the lower respiratory tract. Our aim was to compare the role of silica particle size with mineral fiber length and width in the ability to induce superoxide release from rat alveolar macrophages. We estimated the ability of four types of silica particle samples, with different mode diameter, and three types of mineral fiber samples, with different geometric mean lengths and widths, to induce lucigenin-dependent chemiluminescence (CL) from the cells per number of dust particles (i.e., silica particles and mineral fibers). A close positive correlation was observed between dust size and the ability to induce CL in silica as well as mineral fiber samples. Moreover, the ability of silica samples to induce CL was weaker than that of long mineral fiber sample. This ability increased at a larger rate in small silica particle and thin mineral fiber samples than in large silica particle and thick mineral fiber samples at the initial stage of administration. These results suggest that the kinetics of the induction superoxide release from macrophages is similar between silica particles and mineral fibers; moreover, this depends on silica particle size and mineral fiber geometry. Finally, large silica particles were more active than small ones.

Factors affecting atmospheric 1-, 2-nitropyrenes and 2-nitrofluoranthene in winter at Noto peninsula, a remote background site, Japan.

Airborne particulates were collected at a background site (Wajima Air Monitoring Station; WAMS) on the Noto Peninsula, Japan from January 2006 to December 2007. 1-, 2-nitropyrenes (1-, 2-NPs) and 2-nitrofluoranthene (2-NFR), in the particulates were determined with a sensitive HPLC method with chemiluminescence detection. The average concentrations were higher in winter than in summer. A meteorological analysis indicated that the air samples collected in winter were transported mainly from Northeast China over the Japan Sea. Both the concentration ratios of 2-NFR to 1-NP and 1-NP to pyrene were similar to those in Shenyang in Northeast China which located along the air transportation route to WAMS, but not in Kanazawa which near WAMS. These results strongly suggest that most of the atmospheric 1-, 2-NPs and 2-NFR at WAMS in winter were long range transported from Northeast China.

Identification and quantification of in vivo metabolites of 9,10-phenanthrenequinone in human urine associated with producing reactive oxygen species.

Polycyclic aromatic hydrocarbon quinones (PAHQs) are components in airborne particulate matter (PM) and generate reactive oxygen species (ROS) in a redox cycling process. 9,10-Phenanthrenequinone (9,10-PQ) is a PAHQ found in diesel exhaust particulates and PM. When inhaled, it produces much more ROS than other PAHQs. We hypothesized that urinary metabolites of 9,10-PQ could serve as biomarkers of PAHQ exposure. Here, we describe methods for pretreating urine samples and analyzing 9,10-PQ metabolites by liquid chromatography with tandem mass spectrometry (LC-MS/MS). In urine from rats intraperitoneally injected with 9,10-PQ, the monoglucuronide of 9,10-dihydroxyphenanthrene (9,10-PQHG) was found to be a major metabolite of 9,10-PQ. 9,10-PQHG was also identified in the urine of a nonoccupationally exposed human by its retention time and MS/MS spectra. Furthermore, the urine contained hardly any free (unmetabolized) 9,10-PQ, but treating it with hydrolytic enzymes released 9,10-PQ from conjugated metabolites such as 9,10-PQHG. The concentrations of 9,10-PQHG in urine samples from nonoccupationally exposed subjects who lived in a suburban area were 2.04-19.08 nmol/mol creatinine. This study is the first to demonstrate the presence of 9,10-PQHG in human urine. Determination of urinary 9,10-PQHG should be useful for determining 9,10-PQ exposure.

Determination of selected nitropolycyclic aromatic hydrocarbons in water samples.

A new method for the analysis of selected nitropolycyclic aromatic hydrocarbons (NPAHs) at ultra-trace levels in water samples is proposed. Particulate NPAHs were collected on a GC glass fiber filter. Soluble NPAHs were collected on a C18 Empore disk. After simple clean-up and concentration of NPAHs in both phases, the NPAHs were analyzed using HPLC equipped with clean-up, reducer, concentration and chemiluminescence detection units. The proposed method showed good linear calibration curves with correlation coefficients (r(2)) ranging from 0.9954 to 0.9998. The limits of detection ranged from 0.013 to 0.15 nmol/L (soluble NPAHs) and from 0.01 to 0.13 nmol/L (particulate NPAHs) for 5 NPAHs [(9-nitroanthracene, 1-nitropyrene, 6-nitrochrysene (6-NC), 7-nitrobenz[a]anthracene and 6-nitrobenzo[a]pyrene] having 3 to 5 rings analyzed, when 1.5 L of water sample was used. The repeatability for soluble NPAHs ranged between 4.2 and 6.8% (n=5), while for particulate NPAHs, it varied between 2.2 and 2.8% (n=5). The proposed method successfully determined the above NPAHs expect 6-NC in river water samples, suggesting that this method is suitable for the ultra-trace analysis of NPAHs in water samples.

Biological effects of polycyclic aromatic hydrocarbon derivatives.

Polycyclic aromatic hydrocarbons (PAHs) are included in various environmental pollutants such as airborne particles and have been reported to induce a variety of toxic effects. On the other hand, PAH derivatives are generated from PAHs both through chemical reaction in the atmosphere and metabolism in the body.PAH derivatives have become known for their specific toxicities such as estrogenic/antiestrogenic activities and oxidative stress, and correlations between the toxicities and structures of PAH derivatives have been shown in recent studies. These studies are indispensable for demonstrating the health effects of PAH derivatives, since they would contribute to the comprehensive toxicity prediction of many kinds of PAH derivatives.

Polycyclic aromatic hydrocarbons and nitropolycyclic aromatic hydrocarbons in particulates emitted by motorcycles.

We determined eleven PAHs and four NPAHs in particulates and regulated pollutants (CO, CO2, HC, NOx, PM) exhausted from motorcycles to figure out the characteristics of motorcycle exhausts. Fluoranthene and pyrene accounted for more than 50% of the total detected PAHs. Among four detected NPAHs, 6-nitrochrysene and 7-nitrobenz[a]anthracene were the predominant NPAHs and were highly correlated relationship with their parent PAHs (R = 0.93 and 0.97, respectively). The PM and HC emissions tended to be close to the PAH emissions. NOx and NPAHs were negatively correlated. Despite their small engine size, motorcycles emitted much more PM and PAHs, showed stronger PAH-related carcinogenicity and indirect-acting mutagenicity, but weaker NPAH-related direct-acting mutagenic potency than automobiles. This is the first study to analyze both PAHs and NPAHs emitted by motorcycles, which could provide useful information to design the emission regulations and standards for motorcycles such as PM.

Prediction of rate constants for the gas phase reactions of triphenylene with OH and NO3 radicals using a relative rate method in CCl4 liquid phase-system.

The kinetics of CCl(4) liquid-phase reactions of ten kinds of polycyclic aromatic compounds (PACs) including triphenylene (TP) with NO(3) radicals have been investigated at 273K by a relative rate method using naphthalene (NA) as a reference compound. The obtained relative reaction rates of the tested PACs to NA in CCl(4) were as follows: 2.57±0.24 (acenaphthene), 2.11±0.30 (2,3-dimethylnaphthalene), 1.21±0.13 (fluoranthene), 0.56±0.07 (fluorene), 1.85±0.19 (1-methylnaphthalene), 1.77±0.12 (2-methylnaphthalene), 0.11±0.03 (1-nitronaphthalene), 1.59±0.23 (phenanthrene), 2.40±0.29 (pyrene), 0.22±0.04 (TP). TP is a semi-volatile PAC with four aromatic rings and it is chemically changed into mutagenic 2-nitrotriphenylene (2-NTP) via the gas-phase OH or NO(3) radical-initiated reactions. On the basis of the relative reactivity of the PACs in the CCl(4) liquid phase-system, the rate constants of the gas-phase reactions of TP with OH and NO(3) radicals at 298 K were predicted to be (8.6±1.2)×10(-12) cm(3) molecule(-1) s(-1) and (6.6±1.5)×10(-29)[NO(2)] cm(3) molecule(-1) s(-1), respectively. Based on the ambient concentrations of TP and 2-NTP and the obtained rate constant for the reaction of TP with OH radicals, the atmospheric loss rate of 2-NTP was also evaluated.

Evaluation of urinary metabolites of 1-nitropyrene as biomarkers for exposure to diesel exhaust in taxi drivers of Shenyang, China.

Diesel exhaust (DE) is a significant contributor to the toxicity associated with particulate matter (PM). 1-Nitropyrene (1-NP) has been used as a molecular marker for DE, and the urinary metabolites of 1-NP have been proposed as biomarkers for exposure to DE. In this study, several urinary 1-NP metabolites were evaluated for their utility as markers of short-term exposures to DE. The study population was a cohort of 24 taxi drivers from Shenyang, China, who submitted urine samples collected before, after, and the next morning following their workshifts. The urinary metabolites studied were isomers of hydroxy-1-nitropyrene (3-, 6-, 8- OHNPs) and hydroxy-N-acetyl-1-aminopyrene (3-,6-, 8-OHNAAPs). Exposure to DE was estimated based on exposure to 1-NP in air samples collected during and after the driver's workshift; 6- and 8-OHNP, and 8-OHNAAP were consistently detected in the drivers' urine. Concentrations of the metabolites in the taxi drivers' urine were greater than metabolite levels previously reported in non-occupationally exposed subjects; however no associations were observed between subject-specific exposures to 1-NP and urinary metabolites measured at the end of the workshift or in the next morning void. Significant autocorrelation was observed in metabolite levels in successive urine samples, from which half-lives for urinary elimination of ~10-12 h were estimated. These observations suggest that, in an occupational setting, urinary 1-NP metabolites may be more suitable as markers of ongoing exposure (timescales of several days) rather than indicators of acute exposure associated with single workshifts.

Personal and atmospheric concentrations of ozone in southeastern Hyogo prefecture, Japan.

Twenty-one data sets composed of readings collected by atmospheric ozone monitors worn by individuals on their clothing and installed outside their home or office were collected using Ogawa passive ozone samplers in southeastern Hyogo prefecture, Japan from September 12 to 13, 2011. The concentrations of personal and outdoor ozone ranged from not detectable to 23.2 ppb and from 4.7 to 38.3 ppb, respectively. The mean concentration of personal exposure to ozone was 3.7 ppb and was significantly lower than that of outdoor ozone (18.5 ppb). This suggests that the concentrations of outdoor ozone affect personal ozone exposure. However, in this study, we found no correlation between the concentrations of personal ozone and the total time spent outdoors or the time of day the individual was outside. In contrast, the mean concentrations of outdoor ozone were similar to those of ozone measured at the 12 nearest Ambient Monitoring Stations (AMSs). However, when the AMS was situated near a main road, the regional ozone levels were underestimated.

On-line concentration and fluorescence determination HPLC for polycyclic aromatic hydrocarbons in seawater samples and its application to Japan Sea.

An on­line concentration and fluorescence determination HPLC for polycyclic aromatic hydrocarbons (PAHs) in seawater was proposed. An online concentration column packed with octadecyl polyvinyl alcohol polymer, a pump and a column switching valve were introduced in the conventional HPLC with a fluorescence detector. Only 1.0-100 mL seawater sample was introduced into the concentration column at 1.0 mL min(-1) without any other pretreatment except filtration. Then the trapped PAHs totally flew into the separation column and eluted separately to be detected fluorogenically. The proposed method had good linearity with correlation coefficients (r) ranged from 0.951 to 0.998, and limits of detection ranged from 0.002 to 0.50 ng L(-1) for 15 PAHs as 100 mL seawater was loaded. The sensitivity of the method was 10 to 100 times higher than those reported by other works. The proposed method was applied to the determination of PAHs in the seawater samples collected in the Japan Sea with satisfactory results and to check the present benzo[a]pyrene concentration at the beaches in Noto peninsula, Japan polluted with C-heavy oil spilled from the tanker in 1997.

Analysis of 8-hydroxy-2'-deoxyguanosine in human urine using hydrophilic interaction chromatography with tandem mass spectrometry.

Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) is a widely used noninvasive biomarker of oxidative stress. A selective, sensitive and rapid method for determining 8-OHdG in human urine was developed using hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS/MS) with electrospray ionization. 8-OHdG and isotopically labeled 8-OHdG (internal standard) were separated on a HILIC column with a mobile phase of 10 mM ammonium acetate: acetonitrile (1:9, v/v) within 10 min and detected by using a positive electrospray ionization interface under the selected reaction monitoring mode. The detection limits of 8-OHdG (corresponding to a signal-to-noise ratio of 3) for the HILIC-MS/MS system and the conventional method using a reversed-phase column with MS/MS were 1.0 and 26.0 fmol/injection, respectively. The proposed method makes it possible to monitor the basal level of urinary 8-OHdG from non-exposed healthy subjects and can be used for large-scale human studies.

[How do transport and metabolism affect the biological effects of polycyclic aromatic hydrocarbons?].

Polycyclic aromatic hydrocarbons (PAHs), some of which are carcinogenic/mutagenic, are generated by combustion of fossil fuels and also released through tanker or oilfield accident to cause a large scale environmental pollution. PAHs concentration in China is especially high in East Asia because of many kinds of generation sources such as coal heating systems, vehicles and factories without exhaust gas/particulate treatment systems. So, the atmospheric pollution caused by PAHs in China has been seriously concerned from the view point of health effects. Like yellow sand and sulfur oxide, PAHs exhausted in China are also transported to Japan. Additionally, strongly mutagenic nitrated PAHs (NPAHs), estrogenic/antiestrogenic PAH hydroxides (PAHOHs) and reactive oxygen species-producing PAH quinones (PAHQs) are formed from PAHs by the chemical reaction during the transport. Furthermore these PAHOHs and PAHQs are produced by the metabolism in animal body. In the biological activities caused by the above PAH derivatives, the structure-activity relationship was observed. In this review, our recent results on the generation of PAH derivatives by atmospheric transport and metabolism are reported. Also, the existing condition of PAHs as atmospheric pollutants is considered.

Determination of benzo[a]pyrene-7,10-quinone in airborne particulates by using a chemiluminescence reaction of hydrogen peroxide and hydrosulfite.

An ultraweak chemiluminescence (CL) was observed when sodium hydrosulfite (NaHSO(3)) reacts with hydrogen peroxide (H(2)O(2)) and was enhanced 70 times by adding 10 pmol benzo[a]pyrene-7,10-quinone (7,10-BaPQ). The CL reaction is fast, and it reached maximum intensity in 0.1 s, and then decayed to baseline in 3 s. Mechanism of NaHSO(3)-7,10-BaPQ-H(2)O(2) system were investigated by CL spectrum, radical scavengers and electron spin resonance (ESR). Hydroxyl radical ((•)OH), super oxide anion radical ((•)O(2)(-)), and sulfite radical ((•)SO(3)(-)) were generated in the NaHSO(3)-7, 10-BaPQ-H(2)O(2) system. Reduction of 7,10-BaPQ by (•)O(2)(-) radical gave excited semiquinone, which showed strong CL emission when decayed to its ground state. Maximum CL emission wavelength was located at 440 nm, which may belong to the excited semiquinone. This CL system was developed as post column detection of high performance liquid chromatography for the determination of 7,10-BaPQ. Linearity ranged from 50 fmol to 20 pmol (R(2) = 0.9995) with limit of detection of 30 fmol (S/N = 3). The proposed method was used to determine 7,10-BaPQ in airborne particulates. Average atmospheric concentrations of 7,10-BaPQ in Kanazawa in December 2010 and Wajima in October 2007 were 2.0 and 1.6 pg/m(3), respectively.

Hydrogen peroxide-sodium hydrosulfite chemiluminescence system combined with high-performance liquid chromatography for determination of 1-hydroxypyrene in airborne particulates.

In this research, a highly sensitive chemiluminescence method based on a sodium hydrosulfite (NaHSO(3))-hydrogen peroxide (H(2)O(2)) reaction for the determination of 1-hydroxypyrene (1-OHP) was developed. The response of this system was linear in the range from 0.5 to 50 pmol (R(2)=0.9983). The limit of detection for 1-OHP was 100 fmol (S/N=3). 1-OHP in airborne particulates was well separated from interfering compounds using an ODS column with 75% methanol as the mobile phase in isocratic mode. The proposed method was successfully applied to determine the 1-OHP in airborne particulates collected in Kanazawa, Japan. The average concentration of 1-OHP in the atmosphere was 2.0 pg/m(3) (9.2 fmol/m(3)).