Chemical Oxidative Potential and Cellular Oxidative Stress from Open

Feb 15, 2019 - Particulate matter (PM) exposure is a leading global human health risk. In this study, water-soluble oxidative potential (OP) and intra...
0 downloads 0 Views 311KB Size
Subscriber access provided by UNIV OF NEW ENGLAND ARMIDALE

Ecotoxicology and Human Environmental Health

Chemical oxidative potential and cellular oxidative stress from open biomass burning aerosol Wing Y Tuet, Fobang Liu, Nilmara de Oliveira Alves, Shierly Fok, Paulo Artaxo, Perola C Vasconcellos, Julie Champion, and Nga Lee Ng Environ. Sci. Technol. Lett., Just Accepted Manuscript • DOI: 10.1021/acs.estlett.9b00060 • Publication Date (Web): 15 Feb 2019 Downloaded from http://pubs.acs.org on February 17, 2019

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 27

Environmental Science & Technology Letters

1

Chemical oxidative potential and cellular oxidative stress from open biomass burning

2

aerosol

3

Wing Y. Tuet1, Fobang Liu1, Nilmara de Oliveira Alves2, Shierly Fok1, Paulo Artaxo3, Pérola

4

Vasconcellos4, Julie A. Champion1, Nga L. Ng1,5*

5

1School

6

GA 30332, USA

7

2School

8

3Institute

of Physics, University of São Paulo, São Paulo, 05508, Brazil

9

4Institute

of Chemistry, University of São Paulo, São Paulo, 05508, Brazil

of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta,

of Medicine, University of São Paulo, São Paulo, 03178, Brazil

10

5School

11

30332, USA

12

Corresponding Author

13

*email:

of Earth and Atmospheric Sciences, Georgia Institute of Technology, Atlanta, GA

[email protected]

ACS Paragon Plus Environment

1

Environmental Science & Technology Letters

14

Keywords: oxidative potential, reactive oxygen/nitrogen species, biomass burning, particulate

15

matter

Page 2 of 27

ACS Paragon Plus Environment

2

Page 3 of 27

16

Environmental Science & Technology Letters

Abstract

17

Particulate matter (PM) exposure is a leading global human health risk. In this

18

study, water-soluble oxidative potential (OP) and intracellular reactive oxygen and

19

nitrogen species (ROS/RNS) production were measured for open biomass burning

20

aerosol collected from the Brazilian Amazon. Compared to ambient samples collected

21

from Atlanta and laboratory-generated secondary organic aerosol (SOA), biomass

22

burning aerosol had comparable OP and induced higher levels of ROS/RNS. Compared

23

to regressed OP ranges for biomass burning factors resolved using source apportionment

24

in prior studies, the samples investigated in this study spanned a wider OP range,

25

suggesting that concentration addition may not be applicable for OP measurements. The

26

discrepancy between ROS/RNS estimated using laboratory polycyclic aromatic

27

hydrocarbons (PAHs) solution mixtures and ROS/RNS measured for the water-soluble

28

hydrophobic fraction of Amazon filter samples further supports this conclusion. These

29

results have important implications as many previous studies are based on linear

30

regressions that assume concentration addition. Finally, a significant correlation was

31

observed between ROS/RNS and levoglucosan concentrations although exposure to

32

pure solutions of levoglucosan induced negligible ROS/RNS. These results demonstrate

33

that levoglucosan may be considered as predictors for ROS/RNS even though

34

concentration addition may not be an applicable mixture effect model.

ACS Paragon Plus Environment

3

Environmental Science & Technology Letters

35

Page 4 of 27

Introduction

36

Exposure to inhalable particulate matter (PM) is a leading cause of premature

37

death worldwide.1, 2 According to recent data from the World Health Organization (WHO),

38

an estimated 7 million premature deaths each year may be attributed to PM inhalation,

39

and 9 out of 10 individuals worldwide breathe air containing high levels of pollutants.3

40

Over the past few decades, multiple epidemiological studies have reported associations

41

between elevated PM concentrations and increased incidences of cardiopulmonary

42

morbidity and mortality.2, 4-9 The specific mechanism by which PM exposure results in

43

adverse health outcomes is still unclear. Nevertheless, toxicological studies have

44

suggested a possible mechanism whereby oxidative stress is induced through PM-

45

initiated oxidant production,10-13 and multiple assays14-17 have been developed to

46

measure the production of these oxidants.

47

Recently, several studies on the relative toxicities of different aerosol subtypes

48

resolved using various source apportionment methods reported a high oxidative potential

49

(OP) for biomass burning aerosol (BURN18 and biomass burning organic aerosol,

50

BBOA19). However, direct measures of biomass burning aerosol toxicity are scarce20, 21

51

or include measures that cannot be readily compared to the toxicity of other aerosol

52

subtypes.22-25 Furthermore, the BURN18 and BBOA19 factors were resolved assuming a

53

concentration addition model for oxidative potentials, which may not be applicable

54

according to recent studies.26-28

ACS Paragon Plus Environment

4

Page 5 of 27

Environmental Science & Technology Letters

55

The Brazilian Amazon presents an ideal environment for studying the toxicity of

56

biomass burning aerosol with minimal influence from industrial emissions or other

57

sources.29, 30 The region contains about 40% of the world’s remaining tropical rainforests

58

and has been severely affected by deforestation and open biomass burning emissions in

59

recent years.31 These activities represent a major source of inhalable PM,32 especially

60

during the dry season, and the increased PM has been shown to have a significant health

61

impact on the surrounding population (e.g., increased incidence of respiratory

62

diseases).22, 29, 33

63

In the present study, OP and reactive oxygen and nitrogen species (ROS/RNS)

64

production were measured for ambient PM samples collected in the southwestern region

65

of the Brazilian Amazon during dry and wet seasons. The dithiothreitol (DTT) assay was

66

utilized to measure OP, which may represent the concentration of redox-active species

67

present in the PM sample.34 A cellular assay involving murine alveolar macrophages was

68

employed to measure intracellular ROS/RNS produced as a result of PM exposure.16, 28,

69

35

70

composition were explored to identify and elucidate components associated with biomass

71

burning aerosol toxicity.

72

Methods

Correlations between aerosol toxicity measures (OP or ROS/RNS levels) and chemical

ACS Paragon Plus Environment

5

Environmental Science & Technology Letters

Page 6 of 27

73

PM collection and extraction. Ambient PM10 samples were collected from a site

74

(8.69⁰ S, 63.87⁰ W) located in the southwestern region of the Brazilian Amazon, about 5

75

km north of Porto Velho,36 during the dry (28 July 2011 – 10 October 2011) and wet (10

76

October 2011 – 15 March 2012) seasons. High-volume samplers with a flow rate of 1.3

77

m3 min-1 were used to collect particles onto pre-baked quartz filters. Concentrations of

78

organic carbon (OC) and elemental carbon (EC) were determined using thermal-optical

79

transmittance (TOT, Sunset Laboratory Inc.), while concentrations of monosaccharide

80

anhydrides (levoglucosan, mannosan, and galactosan) were measured using high-

81

performance anion-exchange chromatography coupled to electrospray ionization mass

82

spectrometry (HPAEC/ESI-MS).37 Concentrations of various polycyclic aromatic

83

hydrocarbons (PAHs) and their derivatives (fluorene, phenanthrene, anthracene,

84

luoranthene, pyrene, retene, benzo(a)anthracene, chrysene, benzo(b)fluoranthene,

85

benzo(k)fluoranthene,

86

dibenzo(a,h)anthracene, and benzo(g,h,i)perylene), oxygenated-PAHs (oxy-PAHs; 9-

87

fluorenone, 9,10-anthraquinone, 2-methylanthraquinone, and benzo(a)anthracene-7,12-

88

dione), and nitrogenated-PAHs (nitro-PAHs; 9-nitrophenanthrene, 3-nitrophenanthrene,

89

2-nitrophenanthrene, 2-nitroanthracene, 3-nitrofluoranthene, 2-nitrofluoranthene, 1-

90

nitropyrene,

91

nitrobenzo(a)pyrene) were determined using gas chromatography-mass spectrometry

92

(GC-MS, Agilent 7820A).37,

93

these samples can be found in Oliveira Alves et al. 37, 38

benzo(e)pyrene,

4-nitropyrene,

benzo(a)pyrene,

7-nitrobenzo(a)anthracene,

38

indenol(1,2,3-c,d)pyrene,

6-nitrochrysene,

and

6-

Mass concentrations and detailed chemical analysis of

ACS Paragon Plus Environment

6

Page 7 of 27

Environmental Science & Technology Letters

94

Collected samples were extracted following pre-established procedures for OP

95

determination39 and intracellular ROS/RNS measurement.16 Briefly, filters were

96

submerged in extraction media (DI water for OP and cell culture media (RPMI-1640) for

97

ROS/RNS) and sonicated for 30 min using an Ultrasonic Cleanser (VWR International).

98

Extracts were filtered using a 0.45 µm polytetrafluoroethylene (PTFE) syringe filter

99

(Fisherbrand™) to remove insoluble material post-sonication34 and extracts for cellular

100

exposure were supplemented with 10% fetal bovine serum (FBS).16

101

Polycyclic aromatic hydrocarbon solutions. Laboratory solutions of pure oxy- and

102

nitro-PAHs were prepared to final concentrations of 1 ng µL-1 (9-fluorenone), 2 ng µL-1 (1-

103

nitropyrene, 6-nitrochrysene), and 4 ng µL-1 (2-methylanthraquinone) and diluted 1000x

104

prior to cellular exposure. Concentrations were chosen to span the range observed for

105

these species in the collected samples.38

106

Oxidative potential. Intrinsic water-soluble OP as measured by DTT was measured

107

using a semi-automated DTT system.34 Briefly, the method consisted of three major

108

steps: (1) oxidation of DTT by redox-active species, (2) reaction of unoxidized DTT with

109

5,5-dithio-bis-(2-nitrobenzoic acid) (DTNB) to form 2-nitro-5-mercaptobenoic acid (TNB),

110

and (3) measurement of TNB at 412 nm using an online spectrophotometer (Ocean

111

Optics, Inc., Dunedin, USA).

112

Intracellular ROS/RNS measurement. Murine alveolar macrophages (MH-S,

113

ATCC®CRL-2019™), maintained in RPMI-1640 media supplemented with 10% FBS, 1%

ACS Paragon Plus Environment

7

Environmental Science & Technology Letters

Page 8 of 27

114

penicillin-streptomycin, and 50 µM β-mercaptoethanol at 37°C and 5% CO2, were

115

exposed to PM extracts or laboratory-prepared solutions of pure oxy- and nitro-PAHs for

116

24 hrs. Intracellular ROS/RNS levels were measured post-exposure following the

117

methodology described in Tuet et al.16 (cell density: 2 x 104 cells well-1; ROS/RNS probe:

118

carboxy-H2DCFDA, Molecular Probes C-400, 10 µM). A detailed description of ROS/RNS

119

measurement is provided in supporting information and have been described in Tuet et

120

al.16, 28, 35

121

Cellular

metabolic

activity.

MTT

(3-(4,5-dimethylthiazol-2-yl)-2,5-

122

diphenyltetrazolium bromide) was used to assess cellular metabolic activity post PM

123

exposure following manufacturer’s instruments (Biotium). No decrease of metabolic

124

activity was observed upon exposure to filter extract.

125

Statistical analysis. Linear regressions were evaluated using Pearson’s correlation

126

coefficient with the significance determined using multiple imputation40 and a 95%

127

confidence level.

128

Results and Discussion

129

Ambient and chamber context. Open biomass burning aerosol collected from the

130

Brazilian Amazon during both dry (intense biomass burning)37, 38 and wet (low to moderate

131

biomass burning)37,

132

manner (Fig. S1). Over the same dose range, Amazon aerosol generally induced higher

38

seasons induced ROS/RNS production in a dose-dependent

ACS Paragon Plus Environment

8

Page 9 of 27

Environmental Science & Technology Letters

133

ROS/RNS levels (AUC per µg of PM) than ambient samples collected around the Greater

134

Atlanta region16 and secondary organic aerosol (SOA) generated from a variety of

135

biogenic and anthropogenic precursors28,

136

during the dry season. The high ROS/RNS production observed for biomass burning

137

aerosol suggests that oxidative stress may be a likely mechanism by which biomass

138

burning aerosol exposure results in the adverse health endpoints reported in prior

139

studies.41, 42 In particular, de Oliveira Alves et al.22 showed that biomass burning aerosol

140

(the same samples used in this work) induced oxidative stress in human lung cells.

141

Furthermore, previous studies have shown that exposure to biomass burning aerosol was

142

associated with increased phlegm production in humans.42 A well-established link exists

143

between oxidative stress, characterized by increased oxidant production such as

144

ROS/RNS, and mucus hypersecretion,43-46 further supporting oxidative stress as a

145

possible mechanism for the health effects of biomass burning aerosol. Increased oxidant

146

production may also result in other cellular damages, including DNA/RNA damage,

147

protein oxidation, and lipid peroxidation.47, 48 A higher occurrence of DNA damage have

148

been found in a prior study23 from exposure to biomass burning aerosol collected from

149

the Brazilian Amazon. These results are in agreement with the ROS/RNS measurements

150

obtained in this study, where higher ROS/RNS levels were observed for samples

151

collected during the dry season (Fig. S2).

35

(Fig. 1), especially for aerosol collected

ACS Paragon Plus Environment

9

Environmental Science & Technology Letters

Page 10 of 27

152

In terms of OP, biomass burning aerosol had redox activities comparable to that

153

observed for ambient samples collected from multiple sites in Atlanta19, 39, 49 and chamber

154

SOA generated from different precursors50 (Fig.1). Additionally, higher DTT activities

155

were observed for samples collected during the dry season compared to the wet season

156

(Fig.S2). Previous studies using factor analysis and linear regression reported that BBOA

157

resolved from ambient high resolution time-of-flight aerosol mass spectrometer (AMS)

158

data was the most redox-active among different aerosol subtypes.19 The higher redox

159

activity observed during the dry season is therefore consistent with the more intense open

160

biomass burning that is characteristic of the dry season. It is also interesting to note that

161

many DTT activities measured for open biomass burning aerosol collected from the

162

Amazon region overlap the ranges in between two biomass burning aerosol factors

163

(BBOA and BURN) resolved using different source apportionment methods.18, 19 Note that

164

aerosol samples collected in this study were PM10 while PM2.5 samples were analyzed in

165

the BBOA and BURN studies.18,

166

accounted for ~80% mass concentration of PM10 during the dry season at the same

167

sampling site of this study.32 To our knowledge, the measurements acquired in this study

168

represent the first direct measures of aerosol oxidative potential during intense open

169

biomass burning from the Amazon region. It is promising that there exists some overlap

170

between direct measures and regressed values. However, it should be noted that the

171

direct measurements of open biomass burning OP span a range that extends lower than

172

that predicted by either regression study,18, 19 although components from other types of

19

However, a previous study showed that PM2.5

ACS Paragon Plus Environment

10

Page 11 of 27

Environmental Science & Technology Letters

173

aerosol might contribute to the lower range in this study. Additionally, this study confirms

174

our previous conclusion16, 35 that there is not one simple correlation between oxidative

175

potential and cellular ROS/RNS levels for different PM samples, while low DTT activity

176

will likely correspond to a low cellular response (Fig.1).

177

PM constituents associated with ROS/RNS production. PM-induced ROS/RNS

178

production (AUC per m3 of air) was significantly correlated with concentrations of various

179

monosaccharide anhydrides (levoglucosan, mannosan, and galactosan) (Fig. 2). These

180

monosaccharides are known tracers for biomass burning and ratios of different

181

monosaccharides may be used to infer the source of biomass burning.51 For wet season

182

samples, low but appreciable levels of ROS/RNS were induced despite negligible

183

amounts of all three monosaccharides. These results suggest that monosaccharide

184

concentrations may serve not only as a tracer for biomass burning, but also as a good

185

predictor for ROS/RNS produced as a result of exposure to biomass burning aerosol.

186

Note that exposure to pure levoglucosan solutions (prepared in the laboratory) induced

187

negligible ROS/RNS production over the range observed in the collected samples (Fig.

188

S3).

189

ROS/RNS levels were also significantly correlated with concentrations of OC and

190

EC (Fig. S4). Prior studies involving both cellular and acellular assays have highlighted

191

the importance of organic species in terms of aerosol toxicity.28,

192

correlations between ROS/RNS production and organic species (water-soluble organic

35, 52-55

For instance,

ACS Paragon Plus Environment

11

Environmental Science & Technology Letters

Page 12 of 27

193

carbon and brown carbon) have been previously reported using the same cellular assay

194

used in this study.16 Another study using a different cellular assay reported significant

195

correlations between the biomass burning fraction of water-soluble organic carbon and

196

ROS activity as well.54 Additionally, a meta-analysis performed using data from multiple

197

epidemiological studies found that the relative health risk per mass of EC was about an

198

order of magnitude greater than that of PM2.5.56 This indicates that EC was a tracer for

199

many chemical species co-emitted during biomass burning.

200

To gain further insight into the organic species associated with biomass burning

201

aerosol toxicity, correlations between the concentrations of multiple PAHs and ROS/RNS

202

levels were evaluated. The sum of all PAHs detected in the biomass burning samples

203

was not significantly correlated with ROS/RNS production (Fig. S5a). For samples

204

collected during the wet season, a low level of ROS/RNS was induced regardless of total

205

PAH concentration (blue markers in Fig. S5a). A larger range of ROS/RNS was observed

206

for samples collected during the dry season, although no significant correlation was

207

found. Similar relationships were observed for individual PAHs detected. Previous studies

208

have shown that oxy-PAHs are more redox active and induce the formation of more

209

ROS/RNS upon cellular exposure.35, 50, 57, 58 Correlations between oxy- and nitro-PAHs

210

and ROS/RNS, however, were also not significant for the majority of individual oxy- and

211

nitro-PAHs as well as the sum of oxy- and nitro-PAHs (Fig. S5). The lack of correlation

212

did not necessarily indicate a lack of response. As in laboratory experiments, exposure

ACS Paragon Plus Environment

12

Page 13 of 27

Environmental Science & Technology Letters

213

to pure solutions of four oxy- and nitro-PAHs (1-nitropyrene, 6-nitrochrysene, 9-

214

fluorenone, and 2-methylanthraquinone) induced considerable levels of ROS/RNS

215

production (Fig. S6). These specific compounds were chosen due to their demonstrated

216

toxicity, including DNA adduct formation, carcinogenicity, and cytotoxicity.58-62 In

217

particular, 9-fluorenone has been shown to induce ROS production61 and a significant

218

correlation between concentrations of 9-fluorenone and ROS/RNS was observed for the

219

biomass burning samples (Fig. S7). Together with the results obtained for levoglucosan,

220

these results suggest that there is not a simple relationship between correlation and

221

causation, and caution must be exercised when interpreting correlation results.

222

Compared to the Pearson correlation, it is suggested to use non-parameter methods in

223

statistical analysis. It should however be noted that ambient samples contain other

224

compounds that were not identified in this study. These compounds may induce

225

ROS/RNS production, which may potentially affect correlation results.

226

To gauge the relative contribution of oxy- and nitro-PAHs to ROS/RNS production

227

and to evaluate the applicability of concentration addition as an effect model, an estimated

228

ROS/RNS level was calculated using individual dose-response curves (Fig. S6) and

229

concentrations for the four oxy- and nitro-PAHs (Fig. 3, aqua bars). ROS/RNS produced

230

as a result of exposure to the water-soluble hydrophobic fraction of biomass burning

231

aerosol collected from the Brazilian Amazon, obtained by passing the water-soluble

232

extract through a C-18 column,63 was also measured for comparison (Fig. 3, pink bars).

ACS Paragon Plus Environment

13

Environmental Science & Technology Letters

Page 14 of 27

233

For the eight samples investigated, the ROS/RNS estimated using oxy- and nitro-PAHs

234

was lower than that measured for the water-soluble hydrophobic fraction for six samples

235

and higher for two samples. These results directly demonstrate that concentration

236

addition is not an accurate effect model for estimating cellular responses induced as a

237

result of aerosol exposure, in agreement with recent findings regarding the non-additivity

238

of redox activities.26-28

239

Implications. Results from this study represent the first direct measurements for

240

OP and ROS/RNS of open biomass burning aerosol from the Amazon region. The

241

perspective gained on the relative toxicity of biomass burning aerosol in the context of

242

other aerosol subtypes as well as the information obtained from the differences observed

243

between samples collected during dry and wet seasons can guide future studies to focus

244

more on biomass burning emissions due to its high relative toxicity. Furthermore, the

245

discrepancies between OP measured in this study and biomass burning factors resolved

246

from source apportionment methods (BURN and BBOA) and between ROS/RNS

247

estimated using oxy- and nitro-PAHs and ROS/RNS measured demonstrates that

248

concentration addition may not be an applicable mixture effect model for both OP and

249

ROS/RNS measurements. These results have important implications as many previous

250

studies are based on linear regressions that assume concentration addition.18, 19, 64

251

ACS Paragon Plus Environment

14

Page 15 of 27

Environmental Science & Technology Letters

252

Supporting Information

253

The supporting material consists of a detailed description of ROS/RNS measurement

254

and seven figures.

255 256

AUTHOR INFORMATION

257

Corresponding author

258

Nga Lee Ng: Phone: (404) 385 2148; Email: [email protected]

259 260

Notes

261

The authors declare no competing financial interest.

262 263

ACKNOWLEDGMENT

264

This work was supported by the Health Effects Institute under research agreement No.

265

4943-RFA13-2/14-4. W. Y. Tuet acknowledges support by the National Science

266

Foundation Graduate Research Fellowship under Grant No. DGE-1650044. P. Artaxo

267

acknowledges support from FAPESP under Grants 2013/05014-0 and 2017/17047-0 and

268

CNPq. The authors thank Fernando Morais and Fábio Jorge for assisting with sample

269

collection in the Amazon and acknowledge support from the LBA Central office at INPA,

270

Manaus. The authors also thank Rodney J. Weber for use of the DTT assay setup.

ACS Paragon Plus Environment

15

Environmental Science & Technology Letters

271

ABBREVIATIONS

272

PM: particulate matter; ROS/RNS: reactive oxygen and nitrogen species; DTT:

273

dithiothreitol; OP: oxidative potential

Page 16 of 27

ACS Paragon Plus Environment

16

Page 17 of 27

Environmental Science & Technology Letters

274

REFERENCES

275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317

1. Gakidou, E.; Afshin, A.; Abajobir, A. A.; Abate, K. H.; Abbafati, C.; Abbas, K. M.; AbdAllah, F.; Abdulle, A. M.; Abera, S. F.; Aboyans, V., et al., Global, regional, and national comparative risk assessment of 84 behavioural, environmental and occupational, and metabolic risks or clusters of risks, 1990–2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet 2017, 390 (10100), 1345-1422. 2. Anderson, J. O.; Thundiyil, J. G.; Stolbach, A., Clearing the air: a review of the effects of particulate matter air pollution on human health. J. Med. Toxicol. 2012, 8 (2), 166-175. 3. WHO, 9 out of 10 people worldwide breathe polluted air. 2018, 39 (6), 641-643. 4. Li, N.; Xia, T.; Nel, A. E., The role of oxidative stress in ambient particulate matterinduced lung diseases and its implications in the toxicity of engineered nanoparticles. Free. Radic. Biol. Med. 2008, 44 (9), 1689-1699. 5. Pope III, C. A.; Dockery, D. W., Health effects of fine particulate air pollution: lines that connect. J. Air. Waste. Manag. Assoc. 2006, 56 (6), 709-742. 6. Brunekreef, B.; Holgate, S. T., Air pollution and health. Lancet 2002, 360 (9341), 12331242. 7. Dockery, D. W.; Pope, C. A.; Xu, X.; Spengler, J. D.; Ware, J. H.; Fay, M. E.; Ferris Jr, B. G.; Speizer, F. E., An association between air pollution and mortality in six US cities. N. Engl. J. Med. 1993, 329 (24), 1753-1759. 8. Hoek, G.; Krishnan, R. M.; Beelen, R.; Peters, A.; Ostro, B.; Brunekreef, B.; Kaufman, J. D., Long-term air pollution exposure and cardio-respiratory mortality: a review. Environ. health. 2013, 12 (1), 43. 9. Pope III, C. A.; Burnett, R. T.; Thun, M. J.; Calle, E. E.; Krewski, D.; Ito, K.; Thurston, G. D., Lung cancer, cardiopulmonary mortality, and long-term exposure to fine particulate air pollution. Jama-Journal of the American Medical Association 2002, 287 (9), 1132-1141. 10. Li, N.; Hao, M.; Phalen, R. F.; Hinds, W. C.; Nel, A. E., Particulate air pollutants and asthma: a paradigm for the role of oxidative stress in PM-induced adverse health effects. Clin. Immunol. 2003, 109 (3), 250-265. 11. Tao, F.; Gonzalez-Flecha, B.; Kobzik, L., Reactive oxygen species in pulmonary inflammation by ambient particulates. Free. Radic. Biol. Med. 2003, 35 (4), 327-340. 12. Castro, L.; Freeman, B. A., Reactive oxygen species in human health and disease. Nutrition 2001, 17 (2), 161-165. 13. Gurgueira, S. A.; Lawrence, J.; Coull, B.; Murthy, G. K.; González-Flecha, B., Rapid increases in the steady-state concentration of reactive oxygen species in the lungs and heart after particulate air pollution inhalation. Environ. Health Perspect. 2002, 110 (8), 749. 14. Kumagai, Y.; Koide, S.; Taguchi, K.; Endo, A.; Nakai, Y.; Yoshikawa, T.; Shimojo, N., Oxidation of proximal protein sulfhydryls by phenanthraquinone, a component of diesel exhaust particles. Chem. Res. Toxicol. 2002, 15 (4), 483-489. 15. Landreman, A. P.; Shafer, M. M.; Hemming, J. C.; Hannigan, M. P.; Schauer, J. J., A macrophage-based method for the assessment of the reactive oxygen species (ROS) activity of atmospheric particulate matter (PM) and application to routine (daily-24 h) aerosol monitoring studies. Aerosol. Sci. Technol. 2008, 42 (11), 946-957. 16. Tuet, W. Y.; Fok, S.; Verma, V.; Rodriguez, M. S. T.; Grosberg, A.; Champion, J. A.; Ng, N. L., Dose-dependent intracellular reactive oxygen and nitrogen species (ROS/RNS)

ACS Paragon Plus Environment

17

Environmental Science & Technology Letters

318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361

Page 18 of 27

production from particulate matter exposure: comparison to oxidative potential and chemical composition. Atmos. Environ. 2016, 144, 335-344. 17. Cho, A. K.; Sioutas, C.; Miguel, A. H.; Kumagai, Y.; Schmitz, D. A.; Singh, M.; Eiguren-Fernandez, A.; Froines, J. R., Redox activity of airborne particulate matter at different sites in the Los Angeles Basin. Environ. Res. 2005, 99 (1), 40-47. 18. Bates, J. T.; Weber, R. J.; Abrams, J.; Verma, V.; Fang, T.; Klein, M.; Strickland, M. J.; Sarnat, S. E.; Chang, H. H.; Mulholland, J. A., Reactive oxygen species generation linked to sources of atmospheric particulate matter and cardiorespiratory effects. Environ. Sci. Technol. 2015, 49 (22), 13605-13612. 19. Verma, V.; Fang, T.; Xu, L.; Peltier, R. E.; Russell, A. G.; Ng, N. L.; Weber, R. J., Organic aerosols associated with the generation of reactive oxygen species (ROS) by watersoluble PM2.5. Environ. Sci. Technol. 2015, 49 (7), 4646-4656. 20. Dou, J.; Lin, P.; Kuang, B.-Y.; Yu, J. Z., Reactive oxygen species production mediated by humic-like substances in atmospheric aerosols: enhancement effects by pyridine, imidazole, and their derivatives. Environ. Sci. Technol. 2015, 49 (11), 6457-6465. 21. Park, M.; Joo, H. S.; Lee, K.; Jang, M.; Kim, S. D.; Kim, I.; Borlaza, L. J. S.; Lim, H.; Shin, H.; Chung, K. H., Differential toxicities of fine particulate matters from various sources. Sci. Rep. 2018, 8 (1), 17007. 22. de Oliveira Alves, N.; Vessoni, A. T.; Quinet, A.; Fortunato, R. S.; Kajitani, G. S.; Peixoto, M. S.; de Souza Hacon, S.; Artaxo, P.; Saldiva, P.; Menck, C. F. M., Biomass burning in the Amazon region causes DNA damage and cell death in human lung cells. Sci. Rep. 2017, 7 (1), 10937. 23. de Oliveira Alves, N.; de Souza Hacon, S.; de Oliveira Galvão, M. F.; Peixotoc, M. S.; Artaxo, P.; de Castro Vasconcellos, P.; de Medeiros, S. R. B., Genetic damage of organic matter in the Brazilian Amazon: A comparative study between intense and moderate biomass burning. Environ. Res. 2014, 130, 51-58. 24. Kim, Y. H.; Warren, S. H.; Krantz, Q. T.; King, C.; Jaskot, R.; Preston, W. T.; George, B. J.; Hays, M. D.; Landis, M. S.; Higuchi, M., Mutagenicity and lung toxicity of smoldering vs. flaming emissions from various biomass fuels: implications for health effects from wildland fires. Environ. Health Perspect. (Online) 2018, 126 (1). 25. Naeher, L. P.; Brauer, M.; Lipsett, M.; Zelikoff, J. T.; Simpson, C. D.; Koenig, J. Q.; Smith, K. R., Woodsmoke health effects: a review. Inhal. Toxicol. 2007, 19 (1), 67-106. 26. Yu, H.; Wei, J.; Cheng, Y.; Subedi, K.; Verma, V., Synergistic and antagonistic interactions among the particulate matter components in generating reactive oxygen species based on the dithiothreitol assay. Environ. Sci. Technol. 2018, 52 (4), 2261-2270. 27. Wang, S.; Ye, J.; Soong, R.; Wu, B.; Yu, L.; Simpson, A. J.; Chan, A. W., Relationship between chemical composition and oxidative potential of secondary organic aerosol from polycyclic aromatic hydrocarbons. Atmos. Chem. Phys. 2018, 18 (6), 3987-4003. 28. Tuet, W. Y.; Chen, Y.; Fok, S.; Gao, D.; Weber, R. J.; Champion, J. A.; Ng, N. L., Chemical and cellular oxidant production induced by naphthalene secondary organic aerosol (SOA): effect of redox-active metals and photochemical aging. Sci. Rep. 2017, 7 (1), 15157. 29. Reddington, C.; Butt, E.; Ridley, D.; Artaxo, P.; Morgan, W.; Coe, H.; Spracklen, D., Air quality and human health improvements from reductions in deforestation-related fire in Brazil. Nat. Geosci. 2015, 8 (10), 768.

ACS Paragon Plus Environment

18

Page 19 of 27

362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407

Environmental Science & Technology Letters

30. Marle, M. J.; Field, R. D.; Werf, G. R.; Estrada de Wagt, I. A.; Houghton, R. A.; Rizzo, L. V.; Artaxo, P.; Tsigaridis, K., Fire and deforestation dynamics in Amazonia (1973–2014). Global. Biogeochem. Cy. 2017, 31 (1), 24-38. 31. Laurance, W. F.; Cochrane, M. A.; Bergen, S.; Fearnside, P. M.; Delamônica, P.; Barber, C.; D'Angelo, S.; Fernandes, T., The Future of the Brazilian Amazon. Science 2001, 291 (5503), 438-439. 32. Artaxo, P.; Rizzo, L. V.; Brito, J. F.; Barbosa, H. M.; Arana, A.; Sena, E. T.; Cirino, G. G.; Bastos, W.; Martin, S. T.; Andreae, M. O., Atmospheric aerosols in Amazonia and land use change: from natural biogenic to biomass burning conditions. Faraday. Discuss. 2013, 165, 203235. 33. Jacobson, L. d. S. V.; de Souza Hacon, S.; de Castro, H. A.; Ignotti, E.; Artaxo, P.; Saldiva, P. H. N.; de Leon, A. C. M. P., Acute effects of particulate matter and black carbon from seasonal fires on peak expiratory flow of schoolchildren in the Brazilian Amazon. PloS one 2014, 9 (8), e104177. 34. Fang, T.; Verma, V.; Guo, H.; King, L.; Edgerton, E.; Weber, R., A semi-automated system for quantifying the oxidative potential of ambient particles in aqueous extracts using the dithiothreitol (DTT) assay: results from the Southeastern Center for Air Pollution and Epidemiology (SCAPE). Atmos. Meas. Tech. 2015, 8 (1), 471-482. 35. Tuet, W. Y.; Chen, Y.; Fok, S.; Champion, J. A.; Ng, N. L., Inflammatory responses to secondary organic aerosols (SOA) generated from biogenic and anthropogenic precursors. Atmos. Chem. Phys. 2017, 17 (18), 11423-11440. 36. Brito, J.; Rizzo, L. V.; Morgan, W. T.; Coe, H.; Johnson, B.; Haywood, J.; Longo, K.; Freitas, S.; Andreae, M. O.; Artaxo, P., Ground-based aerosol characterization during the South American Biomass Burning Analysis (SAMBBA) field experiment. Atmos. Chem. Phys. 2014, 14 (22), 12069-12083. 37. de Oliveira Alves, N.; Brito, J.; Caumo, S.; Arana, A.; de Souza Hacon, S.; Artaxo, P.; Hillamo, R.; Teinilä, K.; de Medeiros, S. R. B.; de Castro Vasconcellos, P., Biomass burning in the Amazon region: Aerosol source apportionment and associated health risk assessment. Atmos. Environ. 2015, 120, 277-285. 38. de Oliveira Galvão, M. F.; de Oliveira Alves, N.; Ferreira, P. A.; Caumo, S.; de Castro Vasconcellos, P.; Artaxo, P.; de Souza Hacon, S.; Roubicek, D. A.; de Medeiros, S. R. B., Biomass burning particles in the Brazilian Amazon region: Mutagenic effects of nitro and oxyPAHs and assessment of health risks. Environ. Pollut. 2018, 233, 960-970. 39. Fang, T.; Guo, H.; Verma, V.; Peltier, R. E.; Weber, R. J., PM2.5 water-soluble elements in the southeastern United States: automated analytical method development, spatiotemporal distributions, source apportionment, and implications for heath studies. Atmos. Chem. Phys. 2015, 15 (20), 11667-11682. 40. Pan, Q.; Shimizu, I. In Imputation Variance Estimation by Multiple Imputation Method for the National Hospital Discharge Survey, JSM Proceedings, 2009; 2009; pp 1106-1114. 41. Arbex, M. A.; Martins, L. C.; de Oliveira, R. C.; Pereira, L. A. A.; Arbex, F. F.; Cançado, J. E. D.; Saldiva, P. H. N.; Braga, A. L. F., Air pollution from biomass burning and asthma hospital admissions in a sugar cane plantation area in Brazil. J. Epidemiol. Community. Health. 2007, 61 (5), 395-400. 42. Regalado, J.; Pérez-Padilla, R.; Sansores, R.; Paramo Ramirez, J. I.; Brauer, M.; Paré, P.; Vedal, S., The effect of biomass burning on respiratory symptoms and lung function in rural Mexican women. Am. J. Respir. Crit. Care. Med. 2006, 174 (8), 901-905.

ACS Paragon Plus Environment

19

Environmental Science & Technology Letters

408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451

Page 20 of 27

43. MacNee, W.; Rahman, I., Is oxidative stress central to the pathogenesis of chronic obstructive pulmonary disease? Trends. Mol. Med. 2001, 7 (2), 55-62. 44. McGuinness, A. J. A.; Sapey, E., Oxidative stress in COPD: sources, markers, and potential mechanisms. J. Clin. Med. 2017, 6 (2), 21. 45. Santus, P.; Corsico, A.; Solidoro, P.; Braido, F.; Di Marco, F.; Scichilone, N., Oxidative stress and respiratory system: pharmacological and clinical reappraisal of N-acetylcysteine. COPD 2014, 11 (6), 705-717. 46. Repine, J. E.; Bast, A.; Lankhorst, I.; Group, O. S. S., Oxidative stress in chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care. Med. 1997, 156 (2), 341-357. 47. Wiseman, H.; Halliwell, B., Damage to DNA by reactive oxygen and nitrogen species: role in inflammatory disease and progression to cancer. Biochem. J. 1996, 313 (Pt 1), 17-29. 48. Hensley, K.; Robinson, K. A.; Gabbita, S. P.; Salsman, S.; Floyd, R. A., Reactive oxygen species, cell signaling, and cell injury. Free. Radic. Biol. Med. 2000, 28 (10), 1456-1462. 49. Verma, V.; Fang, T.; Guo, H.; King, L.; Bates, J.; Peltier, R.; Edgerton, E.; Russell, A.; Weber, R., Reactive oxygen species associated with water-soluble PM2.5 in the southeastern United States: spatiotemporal trends and source apportionment. Atmos. Chem. Phys. 2014, 14 (23), 12915-12930. 50. Tuet, W. Y.; Chen, Y.; Xu, L.; Fok, S.; Gao, D.; Weber, R. J.; Ng, N. L., Chemical oxidative potential of secondary organic aerosol (SOA) generated from the photooxidation of biogenic and anthropogenic volatile organic compounds. Atmos. Chem. Phys. 2017, 17 (2), 839853. 51. Simoneit, B. R., Biomass burning—a review of organic tracers for smoke from incomplete combustion. Appl. Geochem. 2002, 17 (3), 129-162. 52. Li, N.; Sioutas, C.; Cho, A.; Schmitz, D.; Misra, C.; Sempf, J.; Wang, M.; Oberley, T.; Froines, J.; Nel, A., Ultrafine particulate pollutants induce oxidative stress and mitochondrial damage. Environ. Health Perspect. 2003, 111 (4), 455. 53. Kleinman, M. T.; Hamade, A.; Meacher, D.; Oldham, M.; Sioutas, C.; Chakrabarti, B.; Stram, D.; Froines, J. R.; Cho, A. K., Inhalation of concentrated ambient particulate matter near a heavily trafficked road stimulates antigen-induced airway responses in mice. J. Air. Waste. Manag. Assoc. 2005, 55 (9), 1277-1288. 54. Hamad, S. H.; Shafer, M. M.; Kadhim, A. K. H.; Al-Omran, S. M.; Schauer, J. J., Seasonal trends in the composition and ROS activity of fine particulate matter in Baghdad, Iraq. Atmos. Environ. 2015, 100, 102-110. 55. Verma, V.; Wang, Y.; El-Afifi, R.; Fang, T.; Rowland, J.; Russell, A. G.; Weber, R. J., Fractionating ambient humic-like substances (HULIS) for their reactive oxygen species activity – Assessing the importance of quinones and atmospheric aging. Atmos. Environ. 2015, 120, 351359. 56. Janssen, N. A.; Hoek, G.; Simic-Lawson, M.; Fischer, P.; Van Bree, L.; Ten Brink, H.; Keuken, M.; Atkinson, R. W.; Anderson, H. R.; Brunekreef, B., Black carbon as an additional indicator of the adverse health effects of airborne particles compared with PM10 and PM2.5. Environ. Health Perspect. 2011, 119 (12), 1691-1699. 57. Charrier, J. G.; Anastasio, C., On dithiothreitol (DTT) as a measure of oxidative potential for ambient particles: evidence for the importance of soluble transition metals. Atmos. Chem. Phys. 2012, 12 (19), 9321-9333.

ACS Paragon Plus Environment

20

Page 21 of 27

452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470 471 472

Environmental Science & Technology Letters

58. Doroshow, J. H., Role of hydrogen peroxide and hydroxyl radical formation in the killing of Ehrlich tumor cells by anticancer quinones. Proc. Natl. Acad. Sci. U. S. A. 1986, 83 (12), 4514-4518. 59. Stanton, C. A.; Chow, F. L.; Phillips, D. H.; Grover, P. L.; Garner, R. C.; Martin, C. N., Evidence for N-(deoxyguanosin-8-yl)-1-aminopyrene as a major DNA adduct in female rats treated with 1-nitropyrene. Carcinogenesis 1985, 6 (4), 535-538. 60. Busby, J. W. F.; Garner, R. C.; Chow, F. L.; Martin, C. N.; Stevens, E. K.; Newberne, P. M.; Wogan, G. N., 6-Nitrochrysene is a potent tumorigen in newborn mice. Carcinogenesis 1985, 6 (5), 801-803. 61. Atsumi, T.; Ishihara, M.; Kadoma, Y.; Tonosaki, K.; Fujisawa, S., Comparative radical production and cytotoxicity induced by camphorquinone and 9‐fluorenone against human pulp fibroblasts. J. Oral. Rehabil. 2004, 31 (12), 1155-1164. 62. Lin, T.-S.; Tiecher, B. A.; Sartorelli, A. C., 2-Methylanthraquinone derivatives as potential bioreductive alkylating agents. J. Med. Chem. 1980, 23 (11), 1237-1242. 63. Verma, V.; Rico-Martinez, R.; Kotra, N.; King, L.; Liu, J.; Snell, T. W.; Weber, R. J., Contribution of water-soluble and insoluble components and their hydrophobic/hydrophilic subfractions to the reactive oxygen species-generating potential of fine ambient aerosols. Environ. Sci. Technol. 2012, 46 (20), 11384-11392. 64. Verma, V.; Shafer, M. M.; Schauer, J. J.; Sioutas, C., Contribution of transition metals in the reactive oxygen species activity of PM emissions from retrofitted heavy-duty vehicles. Atmos. Environ. 2010, 44 (39), 5165-5173.

473

ACS Paragon Plus Environment

21

Environmental Science & Technology Letters

Dry season Wet season Ambient samples (Tuet et al., 2016) Chamber SOA (Tuet el al., 2017) BURN (Bates et al. 2015) BBOA (Verma et al. 2015)

1.4

ROS/RNS (per μg of PM)

Page 22 of 27

1.2 1.0 0.8 0.6 0.4 0.2 0.0 0.00

0.05

0.10

0.15 -1

474

0.20

-1

DTT activity (nmol min μg )

475

Figure 1. ROS/RNS production and DTT activities for biomass burning aerosol collected

476

from the Brazilian Amazon, ambient samples collected from around the greater Atlanta

477

area,16 and chamber SOA generated from a variety of biogenic (isoprene, α-pinene, β-

478

caryophyllene) and anthropogenic (pentadecane, m-xylene, naphthalene) precursors

479

under different formation conditions.35, 50 Biomass burning samples are colored based on

480

season (dry in red and wet in blue). All samples were analyzed following the methology

481

outlined in Fang et al.34 and Tuet et al.16 Biomass burning factors obtained from source

482

apportionment (BURN18 and BBOA19) and their respective DTT activities estimated using

483

multiple linear regression are shown as shaded regions for comparison. BURN was

484

derived from offline chemical analysis and chemical mass balance (CMB) model.18 BBOA

ACS Paragon Plus Environment

22

Page 23 of 27

Environmental Science & Technology Letters

485

was derived from high-resolution aerosol mass spectrometer (AMS) measurements and

486

positive matrix factorization (PMF) analysis.19

487

ACS Paragon Plus Environment

23

50 40

Page 24 of 27

Dry season Wet season

3

A ROS/RNS (per m of air)

Environmental Science & Technology Letters

30

R = 0.89**

20 10 0 0

1000

2000

3000

4000

-3

Levoglucosan (ng m ) 50 40

3

ROS/RNS (per m of air)

B

R = 0.90**

30 20 10 0 0

50

100

150

200

250

-3

Galactosan (ng m ) 50 40

3

ROS/RNS (per m of air)

C

30

R = 0.89**

20 10 0 0

488

50

100

150

200

250

300

350

-3

Mannosan (ng m )

489

Figure 2. Correlations between ROS/RNS levels and concentrations of three

490

monosaccharide anhydrides, i.e., levoglucosan (R=0.89**), galactosan (R=0.90**), and

491

mannosan (R=0.89**). Pearson’s correlation coefficients for dry and wet season

ACS Paragon Plus Environment

24

Page 25 of 27

Environmental Science & Technology Letters

492

separately were: Rdry=0.88** and Rwet=-0.07 for levoglucosan, Rdry=0.91** and Rwet=0.33

493

for galactosan, and Rdry=0.87** and Rwet=0.25 for mannosan. ** indicates significance of p

494

< 0.01.

ACS Paragon Plus Environment

25

Environmental Science & Technology Letters

ROS/RNS (per µg of PM)

20

Page 26 of 27

Amazon samples Laboratory prepared oxy- and nitro-PAHs

15

10

5

0 4 495

5

6

17

21

28

32

45

Sample ID

496

Figure 3. ROS/RNS estimated from four oxy- and nitro-PAHs (1-nitropyrene, 6-nitrochrysene, 9-

497

fluorenone, and 2-methylanthraquinone) (aqua bars) prepared in the laboratory and ROS/RNS

498

measured for the water-soluble hydrophobic fraction of biomass burning aerosol samples collected

499

from the Brazilian Amazon (pink bars). Estimations were obtained using dose-response

500

relationships for individual oxy- and nitro-PAHs (Fig. S7) and concentrations of oxy- and nitro-

501

PAHs assuming concentration addition as the mixture model. The water-soluble hydrophobic

502

fraction was obtained by passing the water-soluble extract through a C-18 column. The sample ID

503

is the numbering of collected filter samples. Samples 4, 5, 6, 17 and 21 were collected in dry season

504

and samples 28, 32 and 45 were collected in wet season.

505

ACS Paragon Plus Environment

26

Page 27 of 27

506

Environmental Science & Technology Letters

For Table of Contents Use Only

507

508

ACS Paragon Plus Environment

27