Advertisement

Environmental Science and Pollution Research

, Volume 24, Issue 1, pp 353–362 | Cite as

Association between inflammation processes, DNA damage, and exposure to environmental pollutants

  • Anelise Barth
  • Natália Brucker
  • Angela M. Moro
  • Sabrina Nascimento
  • Gabriela Goethel
  • Caroline Souto
  • Rafael Fracasso
  • Elisa Sauer
  • Louise Altknecht
  • Bárbara da Costa
  • Marta Duarte
  • Camila B. Menezes
  • Tiana Tasca
  • Marcelo D. Arbo
  • Solange Cristina GarciaEmail author
Research Article

Abstract

Environmental exposure to pollutants, especially polycyclic aromatic hydrocarbons (PAHs), could lead to carcinogenesis development. However, there is a gap on the mechanisms involved in this effect. Therefore, the aim of this study was to investigate the potential relationship between exposure to environmental air pollution and inflammation process in DNA damage in taxi drivers. This study included 45 taxi drivers and 40 controls; non-smokers composed both groups. Biological monitoring was performed through quantification of urinary 1-hydroxypyrene (1-OHP). ICAM-1 (CD54) expression, NTPDase activity, inflammatory cytokine (IL-1β, IL-6, IL-10, TNF-α and IFN-γ) levels, and comet and micronucleus assays were evaluated. The results demonstrated that 1-OHP levels, ICAM-1 expression, NTPDase activity, and DNA damage biomarkers (% tail DNA and micronucleus frequency) were increased in taxi drivers compared to the control group (p < 0.01). Moreover, significant associations were found between 1-OHP levels and ICAM-1 expression, % tail DNA, and micronucleus frequency (p < 0.05). Besides, pro-inflammatory cytokine levels were positively correlated to % tail DNA and micronucleus frequency (p < 0.001). Our findings suggest an important association between environmental exposure to air pollution with increase of ICAM-1 expression and NTPDase activity in taxi drivers. Additionally, the multiple regression linear-analysis demonstrated association between IL-6 and DNA damage. Thus, the present study has provided important evidence that, in addition to environmental exposure to air pollutants, the inflammation process may contribute to DNA damage.

Keywords

Taxi drivers Inflammation ICAM-1 expression NTPDases activity DNA damage 

Notes

Acknowledgments

This work was supported by Fundação de Amparo à Pesquisa do Rio Grande do Sul (FAPERGS)/PPSUS-RS; Conselho Nacional de Pesquisa (CNPq) and Propesq, grant to S.C. Garcia. The authors are grateful for the careful English correction by J. Nardi. S.C. Garcia is a recipient of CNPq research fellowship.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

References

  1. Abe T, Harashima A, Xia T, Konno H, Konno K, Morales A, Ahn J, Gutman D, Barber GN (2013) STING recognition of cytoplasmic DNA instigates cellular defense. Mol Cell 50:5–15CrossRefGoogle Scholar
  2. Alkurdi F, Karabet F, Dimashki M (2014) Characterization and concentrations of polycyclic aromatic hydrocarbons in emissions from different heating systems in Damascus, Syria. Environ Sci Pollut Res 21(8):5747–5759CrossRefGoogle Scholar
  3. Bagatini MD, Martins CC, Battisti V, Spanevello RM, Gasparetto D, Rosa CS, Gonçalves JF, Schetinger MR, dos Santos RB, Morsch VM (2008) Hydrolysis of adenine nucleotides in platelets from patients with acute myocardial infarction. Clin Biochem 41:1181–1185CrossRefGoogle Scholar
  4. Banauch GI, Hall C, Weiden M, Cohen HW, Aldrich TK, Christodoulou V, Arcentales N, Kelly KJ, Prezant DJ (2006) Pulmonary function after exposure to the World Trade Center collapse in the New York fire department. Am Respir Crit Care Med 174(3):312–319CrossRefGoogle Scholar
  5. Becker LV, Rosa CS, Souza Vdo C, Bagatini MD, Casali EA, Leal CA, da Silva JC, Moretto MB, Pinheiro Fde V, Morsch VM, Schetinger MR, Leal DB (2010) Leal activities of enzymes that hydrolyze adenine nucleotides in platelets from patients with rheumatoid arthritis. Clin Biochem 43:1096–1100CrossRefGoogle Scholar
  6. Bind MA, Baccarelli A, Zanobetti A, Tarantini L, Suh H, Vokonas P, Schwartz J (2012) Air pollution and markers of coagulation, inflammation, and endothelial function. Epidemiology 23(2):332–340CrossRefGoogle Scholar
  7. Birk AV, Broekman J, Gladek EM, Robertson HD, Drosopoulos JH, Marcus AJ, Szeto HH (2002) Role of extracellular ATP metabolism in regulation of platelet reactivity. J Lab Clin Med 140:166–175CrossRefGoogle Scholar
  8. Bjelland S, Seeberg E (2003) Mutagenicity, toxicity and repair of DNA base damage induced by oxidation. Mutat Res 531:37–80CrossRefGoogle Scholar
  9. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254Google Scholar
  10. Brook RD, Rajagopalan S, Brook JR, Bhatnagar A, Diez-Roux AV, Holguin F, Hong Y, Luepker RV, Mittleman MA, Peters A, Siscovick D, Smith SC, Whitsel L, Kaufman JD (2010) Particulate matter air pollution and cardiovascular disease: an update to the scientific statement from the American Heart Association. Circulation 121:2331–2378CrossRefGoogle Scholar
  11. Brucker N, Charão MF, Moro AM, Ferrari P, Bubols G, Sauer E, Fracasso R, Durgante J, Thiesen FV, Duarte MM, Gioda A, Castro I, Saldiva PH, Garcia SC (2014) Atherosclerotic process in taxi drivers occupationally exposed to air pollution and co-morbidities. Environ Res 131:31–38CrossRefGoogle Scholar
  12. Brucker N, Moro AM, Charão MF, Durgante J, Freitas F, Baierle M, Nascimento S, Gauer B, Bulcão RP, Bubols GB, Ferrari PD, Thiesen FV, Gioda A, Duarte MM, de Castro I, Saldiva PH, Garcia SC (2013) Biomarkers of occupational exposure to air pollution, inflammation and oxidative damage in taxi drivers. Sci Total Environ 1(463–464):884–893CrossRefGoogle Scholar
  13. Brunekreef B, Holgate ST (2002) Air pollution and health. Lancet 360:1233–1242CrossRefGoogle Scholar
  14. Burns AR, Smith CW, Walker DC (2003) Unique structural features that influence neutrophil emigration into the lung. Physiol Rev 83(2):309–336CrossRefGoogle Scholar
  15. Chan KM, Delfert D, Junger KD (1986) A direct colorimetric assay for Ca2+-stimulated ATPase activity. Anal Biochem 157(2):375–380CrossRefGoogle Scholar
  16. Chen E, Schreier HMC, Strunk R, Brauer M (2008) Chronic traffic-related air pollution and stress interact to predict biologic and clinical outcomes in asthma. Environ Health Perspect 116:970–975CrossRefGoogle Scholar
  17. Chetiyanukornkul T, Toriba A, Kameda T, Tang N, Hayakawa A (2006) Simultaneous determination of urinary hydroxylated metabolites of naphthalene, fluorene, phenanthrene, fluoranthene and pyrene as multiple biomarkers of exposure to polycyclic aromatic hydrocarbons. Anal Bioanal Chem 386(3):712–718CrossRefGoogle Scholar
  18. Chuang CY, Chang CC (2007) Urinary 1-hydroxypyrene level relative to vehicle exhaust exposure mediated by metabolic enzyme polymorphisms. J Occup Health 49(2):140–151CrossRefGoogle Scholar
  19. Ciarrocca M, Rosati MB, Tomei F, Capozzella A, Andreozzi G, Bacaloni A, Casale T, Andrè GC, Fioravanti M, Cuartas MF, Caciari T (2014) Is urinary 1-hydroxypyrene a valid biomarker for exposure to air pollution in outdoor workers? A meta-analysis. J Expo Sci Environ Epid 24:17–26CrossRefGoogle Scholar
  20. Collins AR, Dusinska M, Gedik CM, Stetina R (1996) Oxidative damage to DNA: do we have a reliable biomarker? Environ Health Perspect 104(Suppl 3):465–469CrossRefGoogle Scholar
  21. Collins AR, Duthie SJ, Dobson VL (1993) Direct enzymic detection of endogenous oxidative base damage in human lymphocyte DNA. Carcinogenesis 14:1733–1735CrossRefGoogle Scholar
  22. Curtis L, Rea W, Smith-Willis P, Fenyves E, Pan Y (2006) Adverse health effects of outdoor air pollutants. Environ Int 32:815–830CrossRefGoogle Scholar
  23. da Silva J, de Freitas TR, Heuser V, Marinho JR, Erdtmann B (2000) Genotoxicity biomonitoring in coal regions using wild rodent Ctenomys torquatus by comet assay and micronucleus test. Environ Mol Mutagen 35:270–278CrossRefGoogle Scholar
  24. DeMarini DM (2013) Genotoxicity biomarkers associated with exposure to traffic and near-road atmospheres: a review. Mutagenesis 28(5):485–505CrossRefGoogle Scholar
  25. DETRAN/RS. Departamento de trânsito do estado do Rio Grande do Sul. Frota do RS. http://www.detran.rs.gov.br/conteudo/27453/frota-do-rs Accessed 27 January 2015.Google Scholar
  26. Dizdaroglu M, Jaruga P, Birincioglu M, Rodriguez H (2002) Free radical-induced damage to DNA: mechanisms and measurement. Free Radic Biol Med 32:1102–1115CrossRefGoogle Scholar
  27. Esen F, Tasdemir Y, Vardar N (2008) Atmospheric concentrations of PAHs, their possible sources and gas-to-particle partitioning at a residential site of Bursa, Turkey. Atmos Res 88(3–4):243–255CrossRefGoogle Scholar
  28. Fenech M, Kirsch-Volders M, Natarajan AT, Surralles J, Crott JW, Parry J, Norppa H, Eastmond DA, Tucker JD, Thomas P (2011) Molecular mechanisms of micronucleus, nucleoplasmic bridge and nuclear bud formation in mammalian and human cells. Mutagenesis 26:125–132CrossRefGoogle Scholar
  29. Freire C, Abril A, Fernández MF, Ramos R, Estarlich M, Manrique A, Aguirre A, Ibarluzea J, Olea N (2009) Urinary 1-hydroxypyrene and PAH exposure in 4-year-old Spanish children. Sci Total Environ 407:1562–1569CrossRefGoogle Scholar
  30. Gayle RB III, Malliszewski CR, Gimpel SD, Schoenborn MA, Caspary RG, Richards C, Brasel K, Price V, Drosopoulos JH, Islam N, Alyonycheva TN, Broekman MJ, Marcus AJ (1998) Inhibition of platelet unction by recombinant soluble ecto-ADPase/CD39. J Clin Invest 101(9):1851–1859CrossRefGoogle Scholar
  31. Hansen AM, Mathiesen L, Pedersen M, Knudsen LE (2008) Urinary 1-hydroxypyrene (1-HOP) in environmental and occupational studies—a review. Int J Hyg Environ Health 211:471–503CrossRefGoogle Scholar
  32. Hansen AM, Wallin H, Binderup ML, Dybdahl M, Autrup H, Loft S, Knudsen LE (2004) Urinary 1-hydroxypyrene and mutagenicity in bus drivers and mail carriers exposed to urban air pollution in Denmark. Mutat Res 557(1):7–17CrossRefGoogle Scholar
  33. Hayashi M, MacGregor JT, Gatehouse DG, Ilse-Dore A, Blakey DH, Dertinger SD (2000) In vivo rodent erythrocyte micronucleus assay. Some aspects of protocol design including repeated treatments integration with toxicity testing, and automated scoring. Environ Mol Mutagen 35(3):234–252CrossRefGoogle Scholar
  34. Health Effects Institute (HEI) (2010) Traffic-related air pollution: a critical review of the literature on emissions, exposure, and health effects, special report 17. Health Effects Institute, Boston, MA, USAGoogle Scholar
  35. Hellman B, Vaghef H, Boström B (1995) The concepts of tail moment and tail inertia in the single cell gel electrophoresis assay. Mutat Res 336(2):123–131CrossRefGoogle Scholar
  36. Hirano S, Furuyama A, Koike E, Kobayashi T (2003) Oxidative-stress potency of organic extracts of diesel exhaust and urban fine particles in rat heart microvessel endothelial cells. Toxicology 187(2–3):161–170CrossRefGoogle Scholar
  37. International Agency for Research on Cancer (IARC) (2013) Outdoor air pollution a leading environmental cause of cancer deaths.Google Scholar
  38. Ishikawa H, Ma Z, Barber GN (2009) TING regulates intracellular DNA mediated, type I interferon-dependent innate immunity. Nature 461:788–792CrossRefGoogle Scholar
  39. Kamal A, Cincinelli A, Martellini T, Palchetti I, Bettazzi F, Malik RN (2016) Health and carcinogenic risk evaluation for cohorts exposed to PAHs in petrochemical workplaces in Rawalpindi city (Pakistan). Int J Environ Health Res 26(1):37–57CrossRefGoogle Scholar
  40. Krieglstein CF, Granger DN (2001) Adhesion molecules and their role in vascular disease. Am J Hypertens 14:44S–54SCrossRefGoogle Scholar
  41. Krishna G, Hayashi M (2000) In vivo rodent micronucleus assay: protocol, conduct and data interpretation. Mutat Res 455:155–166CrossRefGoogle Scholar
  42. Lodovici M, Bigagli E (2011) Oxidative stress and air pollution exposure. J Toxicol 48:7074Google Scholar
  43. Lunkes GI, Lunkes D, Stefanello F, Morsch A, Morsch VM, Mazzanti CM, Schetinger MR (2003) Enzymes that hydrolyze adenine nucleotides in diabetes and associated pathologies. Thromb Res 109:189–194CrossRefGoogle Scholar
  44. Maldonado PA, Corrêa MC, Becker LV, Flores C, Moretto MB, Morsch V, Schetinger MRC (2008) Ectonucleotide pyrophosphatase/phosphodiesterase (E-NPP) and adenosine deaminase (ADA) activities in patients with uterine cervix neoplasia. Clin Biochem 41:400–406CrossRefGoogle Scholar
  45. Marcus AJ, Broekman MJ, Drosopoulos JFH, Islam N, Pisnky DJ, Sesti C, Levi R (2003) Heterologous cell–cell interactions: thromboregulation, cerebroprotection and cardioprotection by CD39 (NTPDase-1. J Thromb Haemost 1:2497–2509CrossRefGoogle Scholar
  46. Massberg S, Brand K, Grüner S, Page S, Müller E, Müller I, Bergmeier W, Richter T, Lorenz M, Konrad I, Nieswandt B, Gawaz M (2002) A critical role of platelet adhesion in the initiation of atherosclerotic lesion formation. J Exp Med 196:887–896CrossRefGoogle Scholar
  47. Matullo G, Dunning AM, Guarrera S, Baynes C, Polidoro S, Garte S, Autrup H, Malaveille C, Peluso M, Airoldi L, Veglia F, Gormally E, Hoek G, Krzyzanowski M, Overvad K, Raaschou-Nielsen O, Clavel-Chapelon F, Linseisen J, Boeing H, Trichopoulou A, Palli D, Krogh V, Tumino R, Panico S, Bueno-De-Mesquita HB, Peeters PH, Lund E, Pera G, Martinez C, Dorronsoro M, Barricarte A, Tormo MJ, Quiros JR, Day NE, Key TJ, Saracci R, Kaaks R, Riboli E, Vineis P (2006) DNA repair polymorphisms and cancer risk in non-smokers in a cohort study. Carcinogenesis 27(5):997–1007CrossRefGoogle Scholar
  48. Mills NL, Donaldson K, Hadoke PW, Boon NA, MacNee W, Cassee FR, Sandström T, Blomberg A, Newby DE (2009) Adverse cardiovascular effects of air pollution. Nat Clin Pract Cardiovasc Med 6:36–44CrossRefGoogle Scholar
  49. Montiel-Davalos A, Alfaro-Moreno E, Lopez-Marure R (2007) PM2.5 and PM10induce the expression of adhesion molecules and the adhesion of monocytic cells to human umbilical vein endothelial cells. Inhal Toxicol 19(Suppl. 1):91–98CrossRefGoogle Scholar
  50. Moore KW, de Waal MR, Coffman RL, O’Garra A (2001) Interleukin-10 and the interleukin-10 receptor. Annu Rev Immunol 19:683–765CrossRefGoogle Scholar
  51. Moro AM, Charão MF, Brucker N, Durgante J, Baierle M, Bubols G, Goethel G, Fracasso R, Nascimento S, Bulcão R, Gauer B, Barth A, Bochi G, Moresco R, Gioda A, Salvador M, Farsky S, Garcia SC (2013) Genotoxicity and oxidative stress in gasoline station attendants. Mutat Res 754(1–2):63–70CrossRefGoogle Scholar
  52. Mosser DM, Zhang X (2008) Interleukin-10: new perspectives on an old cytokine. Immunol Rev 226:205–218CrossRefGoogle Scholar
  53. Muñoz B, Albores A (2011) DNA Damage Caused by Polycyclic Aromatic Hydrocarbons: Mechanisms and Markers. In: Chen C, editor. Selected Topics in DNA Repair. ISBN: 978–953–307-606-5.Google Scholar
  54. Narizhneva NV, Razorenova OV, Podrez EA, Chen J, Chandrasekharan UM, DiCorleto PE, Plow EF, Topol EJ, Byzova TV (2005) Thrombo-spondin-1 up-regulates expression of cell adhesion molecules and promotes monocyte binding to endothelium. FASEB J 19:1158–1160Google Scholar
  55. Neophytou AM, Jaime EH, Jennifer MC, Thomas JS, Douglas WD, Brent AC, Eric G, Francine L (2013) Traffic-related exposures and biomarkers of systemic inflammation, endothelial activation and oxidative stress: a panel study in the US trucking industry. Environ Health 12:105CrossRefGoogle Scholar
  56. Petchpoung K, Kaojarern S, Yoovathaworn K, Sura T, Sirivarasai J (2011) The influence of metabolic gene polymorphisms on urinary 1-hydroxypyrene concentration in Thai bus drivers. Environ Toxicol Pharmacol 31(1):160–164CrossRefGoogle Scholar
  57. Pilla C, Emanuelli T, Frasetto SS, Battastini AMO, Dias RD, Sarkis JJF (1996) ATP diphosphohydrolase activity (apyrase, EC 3.6.1.5) in human blood platelets. Platelets 7:225–230CrossRefGoogle Scholar
  58. Reuter S, Subash CG, Madan MC, Bharat BA (2010) Oxidative stress, inflammation, and cancer: how are they linked? Free Radic Biol Med 49(11):1603–1616CrossRefGoogle Scholar
  59. Rex S, Freedman JE (2007) Inhibition of platelet function by the endothelium. Platelets 251–279.Google Scholar
  60. Rosenfeld ME (2013) Inflammation and atherosclerosis: direct versus indirect mechanisms. Curr Opin in Pharmacol 13:154–160CrossRefGoogle Scholar
  61. Rossner P Jr, Svecova V, Milcova A, Lnenickova Z, Solansky I, Sram RJ (2008) Seasonal variability of oxidative stress markers in city bus drivers. Part II. Oxidative damage to lipids and proteins. Mutat Res 642:21–27CrossRefGoogle Scholar
  62. Saitoh T, Fujita N, Hayashi T, Takahara K, Satoh T, Lee H, Matsunaga K, Kageyama S, Omori H, Noda T, Yamamoto N, Kawai T, Ishii K, Takeuchi O, Yoshimori T, Akira S (2009) Atg9acontrols dsDNA-driven dynamic translocation of STING and the innate immune response. Proc Natl Acad Sci U S A 106:20842–208426CrossRefGoogle Scholar
  63. Sancini A, Fioravanti M, Ciarrocca M, Palermo P, Fiaschetti M, Schifano MP, Tomei G, Tomei F (2010) Pulmonary nodules in workers exposed to urban stressor. Environ Res 110:519–525CrossRefGoogle Scholar
  64. Savi P, Herbert JM (2005) Clopidogrel and ticlopidine: P2Y12 adenosine diphosphate-receptor antagonists for the prevention of atherothrombosis. Semin Thromb Hemost 31(2):174–183CrossRefGoogle Scholar
  65. Schmatz R, Schetinger MRC, Spanevello RM, Mazzanti CM, Stefanello N, Maldonado PA, Gutierres J, Corrêa Mde C, Girotto E, Moretto MB, Morsch VM et al (2009) Effects of resveratrol on nucleotide degrading enzymes in streptozotocin-induced diabetic rats. Life Sci 84:345–350CrossRefGoogle Scholar
  66. Shoenfelt J, Mitkus RJ, Zeisler R, Spatz RO, Powell J, Fenton MJ, Squibb KA, Medvedev AE (2009) Involvement of TLR2 and TLR4 in inflammatory immune responses induced by fine and coarse ambient air particulate matter. J Leukoc Biol 86:303–312CrossRefGoogle Scholar
  67. Simon SI, Green CE (2005) Molecular mechanics and dynamics of leukocyte recruitment during inflammation. Annu Rev Biomed Eng 7:151–185CrossRefGoogle Scholar
  68. Tice R, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu JC, Sasaki YF (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206–221CrossRefGoogle Scholar
  69. Timea P, Csanad ZB (2014) Inflammation-induced DNA damage and damage-induced inflammation: a vicious cycle. Microbes Infect 16(10):822–832CrossRefGoogle Scholar
  70. Tuakuila J, Kabamba M, Mata H (2013) High human exposure to pyrene (polycyclic aromatic hydrocarbon) in Kinshasa, a capital of the Democratic Republic of Congo. Arch of Public Health 71:14CrossRefGoogle Scholar
  71. Tucker JD, Preston RJ (1996) Chromosome aberrations, micronuclei aneuploidy, sister chromatid exchanges, and cancer risk assessment. Mutat Res 365:147–159CrossRefGoogle Scholar
  72. Vitta JA, Keaney JF Jr (2002) Endothelial function: a barometer for cardiovascular risk? Circulation 106:640–642CrossRefGoogle Scholar
  73. Wang L, Lin L, Lai S (2009) Emissions of polycyclic aromatic hydrocarbons from fluidized and fixed bed incinerators disposing petrochemical industrial biological sludge. J Hazard Mater 168:438–444CrossRefGoogle Scholar
  74. Wold LE, Simkhovich BZ, Kleinman MT, Nordlie MA, Dow JS, Sioutas C, Kloner RA (2006) In vivo and in vitro models to test the hypothesis of particle-induced effects on cardiac function and arrhythmias. Cardiovasc Toxicol 6(1):69–78CrossRefGoogle Scholar
  75. Yang C, Chen A, Chen R, Qi Y, Ye J, Li S, Li W, Liang Z, Liang Q, Guo D, Kan H, Chen X (2014) Acute effect of ambient air pollution on heart failure in Guangzhou, China. Int J Cardiol 177(2):436–441CrossRefGoogle Scholar
  76. Zhai Q, Duan H, Wang Y, Huang C, Niu Y, Dai Y, Bin P, Liu Q, Chen W, Ma J, Zheng Y (2012) Genetic damage induced by organic extract of coke oven emissions on human bronchial epithelial cells. Toxicol in Vitro 26(5):752–758CrossRefGoogle Scholar
  77. Zimmermann H (2001) Ectonucleotidases: some recent developments and a note on nomenclature. Drug Develop Res 52:44–56CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Anelise Barth
    • 1
  • Natália Brucker
    • 1
  • Angela M. Moro
    • 1
  • Sabrina Nascimento
    • 1
    • 2
  • Gabriela Goethel
    • 1
    • 2
  • Caroline Souto
    • 1
  • Rafael Fracasso
    • 1
  • Elisa Sauer
    • 1
    • 2
  • Louise Altknecht
    • 1
  • Bárbara da Costa
    • 1
  • Marta Duarte
    • 3
  • Camila B. Menezes
    • 2
    • 4
  • Tiana Tasca
    • 2
    • 4
  • Marcelo D. Arbo
    • 1
    • 2
  • Solange Cristina Garcia
    • 1
    • 2
    • 5
    Email author
  1. 1.Laboratory of Toxicology (LATOX), Department of Analysis, Faculty of PharmacyFederal University of Rio Grande do SulPorto AlegreBrazil
  2. 2.Post-graduate Program in Pharmaceutical Sciences, Faculty of PharmacyFederal University of Rio Grande do SulPorto AlegreBrazil
  3. 3.Department of Health SciencesLutheran University of BrazilSanta MariaBrazil
  4. 4.Laboratory of Research in Parasitology, Department of Analysis, Faculty of PharmacyFederal University of Rio Grande do SulPorto AlegreBrazil
  5. 5.Institute of CardiologyUniversity Cardiology FoundationPorto AlegreBrazil

Personalised recommendations