Arsenic alters transcriptional responses to Pseudomonas aeruginosa infection and decreases antimicrobial defense of human airway epithelial cells

Toxicology and Applied Pharmacology

Volumn 331, Issue , 2017, Pages 154-163

  Goodale, Britton C ^a   Rayack, Erica J ^a   Stanton, Bruce A ^a  

^a DARTMOUTH MEDICAL SCHOOL   (United States)

Author keywords

Arsenic; Immunity; Lung; Lysozyme; RNA Seq

Indexed keywords

ARSENIC; CYSTEINE RICH SECRETORY PROTEIN 3; FERRETIN HEAVY CHAIN 1; GLUCURONOSYLTRANSFERASE 1A8; HEME OXYGENASE 1; HLA DQA1 ANTIGEN; INDUCIBLE NITRIC OXIDE SYNTHASE; INTERLEUKIN 19; INTERLEUKIN 6; LACTATE DEHYDROGENASE; LYSOZYME; PEPTIDES AND PROTEINS; PLA2G7 PROTEIN; SECRETOGLOBULIN; SLC7A11 PROTEIN; STATHERIN; TRANSCRIPTION FACTOR NRF2; TRANSCRIPTION FACTOR SOX4; TUMOR NECROSIS FACTOR; UNCLASSIFIED DRUG;

AIRWAY EPITHELIUM CELL; ANTIBACTERIAL ACTIVITY; ANTIMICROBIAL ACTIVITY; ARTICLE; BACTERICIDAL ACTIVITY; BACTERIOLYSIS; BIOLOGICAL ACTIVITY; CELL COUNT; CELL CULTURE; CELL DEATH; CONTROLLED STUDY; CYTOTOXICITY; ENZYME ACTIVITY; ENZYME LINKED IMMUNOSORBENT ASSAY; GENE EXPRESSION ASSAY; GENE EXPRESSION REGULATION; GENE ONTOLOGY; HOST RESISTANCE; HUMAN; HUMAN CELL; IMMUNE RESPONSE; LUNG EPITHELIUM; MICROCOCCUS LUTEUS; NONHUMAN; PATHOGENESIS; PH; PSEUDOMONAS AERUGINOSA; PSEUDOMONAS INFECTION; RESPIRATORY EPITHELIUM; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; RNA ISOLATION; RNA SEQUENCE; SIGNAL TRANSDUCTION; ADULT; AGED; CYTOLOGY; DOSE RESPONSE; DRUG EFFECTS; FEMALE; GENE REGULATORY NETWORK; GENETIC TRANSCRIPTION; GENETICS; IMMUNOLOGY; INNATE IMMUNITY; MALE; MIDDLE AGED; PHYSIOLOGY; RESPIRATORY MUCOSA; YOUNG ADULT;

ADULT; AGED; ARSENIC; DOSE-RESPONSE RELATIONSHIP, DRUG; FEMALE; GENE REGULATORY NETWORKS; HUMANS; IMMUNITY, INNATE; MALE; MIDDLE AGED; PSEUDOMONAS AERUGINOSA; PSEUDOMONAS INFECTIONS; RESPIRATORY MUCOSA; TRANSCRIPTION, GENETIC; YOUNG ADULT;

EID: 85021136955     PISSN: 0041008X     EISSN: 10960333     Source Type: Journal    
DOI: 10.1016/j.taap.2017.06.010     Document Type: Article

References (70)

1. Aksamit, T.R., O'Donnell, A.E., Barker, A., Olivier, K.N., Winthrop, K.L., Daniels, M.L.A., Johnson, M., Eden, E., Griffith, D., Knowles, M., Metersky, M., Salathe, M., Thomashow, B., Tino, G., Turino, G., Carretta, B., Daley, C.L., Adult patients with bronchiectasis: a first look at the US bronchiectasis Research Registry. Chest 151 (2017), 982–992, 10.1016/j.chest.2016.10.055.
2. Andrew, A.S., Bernardo, V., Warnke, L.A., Davey, J.C., Hampton, T., Mason, R.A., Thorpe, J.E., Ihnat, M.A., Hamilton, J.W., Exposure to arsenic at levels found in U.S. drinking water modifies expression in the mouse lung. Toxicol. Sci. 100 (2007), 75–87, 10.1093/toxsci/kfm200.
3. ATSDR, Toxicological Profile for Arsenic. 2007, US Department of Health and Human Services, Agency for Toxic Substances and Disease Registry.
4. Bailey, K.A., Smith, A.H., Tokar, E.J., Graziano, J.H., Kim, K.-W., Navasumrit, P., Ruchirawat, M., Thiantanawat, A., Suk, W.A., Fry, R.C., Mechanisms underlying latent disease risk associated with early-life arsenic exposure: current Research trends and scientific gaps. Environ. Health Perspect. 124 (2016), 170–175, 10.1289/ehp.1409360.
5. Bomberger, J.M., Coutermarsh, B.A., Barnaby, R.L., Stanton, B.A., Arsenic promotes ubiquitinylation and lysosomal degradation of cystic fibrosis transmembrane conductance regulator (CFTR) chloride channels in human airway epithelial cells. J. Biol. Chem. 287 (2012), 17130–17139, 10.1074/jbc.M111.338855.
6. Carignan, C.C., Cottingham, K.L., Jackson, B.P., Farzan, S.F., Gandolfi, A.J., Punshon, T., Folt, C.L., Karagas, M.R., Estimated exposure to arsenic in breastfed and formula-fed infants in a United States cohort. Environ. Health Perspect., 2015, 10.1289/ehp.1408789.
7. Carlin, D.J., Naujokas, M.F., Bradham, K.D., Cowden, J., Heacock, M., Henry, H.F., Lee, J.S., Thomas, D.J., Thompson, C., Tokar, E.J., Waalkes, M.P., Birnbaum, L.S., Suk, W.A., Arsenic and environmental health: state of the science and future Research opportunities. Environ. Health Perspect. 124 (2016), 890–899, 10.1289/ehp.1510209.
8. Cole, A.M., Thapa, D.R., Gabayan, V., Liao, H.-I., Liu, L., Ganz, T., Decreased clearance of Pseudomonas aeruginosa from airways of mice deficient in lysozyme M. J. Leukoc. Biol. 78 (2005), 1081–1085, 10.1189/jlb.0205073.
9. Dajani, R., Zhang, Y., Taft, P.J., Travis, S.M., Starner, T.D., Olsen, A., Zabner, J., Welsh, M.J., Engelhardt, J.F., Lysozyme secretion by submucosal glands protects the airway from bacterial infection. Am. J. Respir. Cell Mol. Biol. 32 (2005), 548–552, 10.1165/rcmb.2005-0059OC.
10. Efremenko, A.Y., Seagrave, J., Clewell, H.J., Van Landingham, C., Gentry, P.R., Yager, J.W., Evaluation of gene expression changes in human primary lung epithelial cells following 24-hr exposures to inorganic arsenic and its methylated metabolites and to arsenic trioxide. Environ. Mol. Mutagen, 2015, 10.1002/em.21937 (n/a-n/a).
11. Escudero-Lourdes, C., Medeiros, M., Cardenas-Gonzalez, M., Wnek, S., Gandolfi, J., Low level exposure to monomethyl arsonous acid-induced the over-production of inflammation-related cytokines and the activation of cell signals associated with tumor progression in an urothelial cell model. Toxicol. Appl. Pharmacol. 244 (2010), 162–173, 10.1016/j.taap.2009.12.029.
12. Farzan, S.F., Korrick, S., Li, Z., Enelow, R., Gandolfi, A.J., Madan, J., Nadeau, K., Karagas, M.R., In utero arsenic exposure and infant infection in a United States cohort: a prospective study. Environ. Res. 126 (2013), 24–30, 10.1016/j.envres.2013.05.001.
13. Farzan, S.F., Li, Z., Korrick, S.A., Spiegelman, D., Enelow, R., Nadeau, K., Baker, E., Karagas, M.R., Infant infections and respiratory symptoms in relation to in utero arsenic exposure in a U.S. cohort. Environ. Health Perspect. 124 (2016), 840–847, 10.1289/ehp.1409282.
14. Fleming, A., On a remarkable bacteriolytic element found in tissues and secretions. Proc. R. Soc. Lond. Ser. B Contain. Pap. Biol. Character 93 (1922), 306–317.
15. Fry, R.C., Navasumrit, P., Valiathan, C., Svensson, J.P., Hogan, B.J., Luo, M., Bhattacharya, S., Kandjanapa, K., Soontararuks, S., Nookabkaew, S., Mahidol, C., Ruchirawat, M., Samson, L.D., Activation of inflammation/NF-?B signaling in infants born to arsenic-exposed mothers. PLoS Genet., 3, 2007, 10.1371/journal.pgen.0030207.
16. Fulcher, M.L., Randell, S.H., Human nasal and tracheo-bronchial respiratory epithelial cell culture. Randell, S.H., Fulcher, M.L., (eds.) Epithelial Cell Culture Protocols, 2013, Humana Press, 109–121.
17. Ganz, T., Antimicrobial polypeptides. J. Leukoc. Biol. 75 (2004), 34–38, 10.1189/jlb.0403150.
18. Gonzalez, H.O., Hu, J., Gaworecki, K.M., Roling, J.A., Baldwin, W.S., Gardea-Torresdey, J.L., Bain, L.J., Dose-responsive gene expression changes in juvenile and adult mummichogs (Fundulus heteroclitus) after arsenic exposure. Mar. Environ. Res. 70 (2010), 133–141, 10.1016/j.marenvres.2010.04.003.
19. Hall, M., Gamble, M., Slavkovich, V., Liu, X., Levy, D., Cheng, Z., van Geen, A., Yunus, M., Rahman, M., Pilsner, J.R., Graziano, J., Determinants of arsenic metabolism: blood arsenic metabolites, plasma folate, cobalamin, and homocysteine concentrations in maternal-newborn pairs. Environ. Health Perspect. 115 (2007), 1503–1509, 10.1289/ehp.9906.
20. Iacono, V.J., MacKay, B.J., DiRienzo, S., Pollock, J.J., Selective antibacterial properties of lysozyme for oral microorganisms. Infect. Immun. 29 (1980), 623–632.
21. Jackson, B.P., Taylor, V.F., Karagas, M.R., Punshon, T., Cottingham, K.L., Arsenic, organic foods, and brown rice syrup. Environ. Health Perspect. 120 (2012), 623–626, 10.1289/ehp.1104619.
22. Jara, E.A., Winter, C.K., Dietary exposure to total and inorganic arsenic in the United States, 2006–2008. Int. J. Food Contam., 1, 2014, 3, 10.1186/s40550-014-0003-x.
23. Kai, H., Hisatsune, A., Chihara, T., Uto, A., Kokusho, A., Miyata, T., Basbaum, C., Myeloid ELF-1-like factor up-regulates lysozyme transcription in epithelial cells. J. Biol. Chem. 274 (1999), 20098–20102, 10.1074/jbc.274.29.20098.
24. Konstan, M.W., Chen, P.W., Sherman, J.M., Thomassen, M.J., Wood, R.E., Boat, T.F., Human lung lysozyme: sources and properties. Am. Rev. Respir. Dis. 123 (1981), 120–124, 10.1164/arrd.1981.123.1.120.
25. Kozul, C.D., Ely, K.H., Enelow, R.I., Hamilton, J.W., Low-dose arsenic compromises the immune response to influenza a infection in vivo. Environ. Health Perspect. 117 (2009), 1441–1447, 10.1289/ehp.0900911.
26. Kozul, C.D., Hampton, T.H., Davey, J.C., Gosse, J.A., Nomikos, A.P., Eisenhauer, P.L., Weiss, D.J., Thorpe, J.E., Ihnat, M.A., Hamilton, J.W., Chronic exposure to arsenic in the drinking water alters the expression of immune response genes in mouse lung. Environ. Health Perspect. 117 (2009), 1108–1115, 10.1289/ehp.0800199.
27. Kurzius-Spencer, M., Burgess, J.L., Harris, R.B., Hartz, V., Roberge, J., Huang, S., Hsu, C.-H., O'Rourke, M.K., Contribution of diet to aggregate arsenic exposures—an analysis across populations. J. Expo. Sci. Environ. Epidemiol., 2013, 10.1038/jes.2013.37.
28. Lantz, R.C., Chau, B., Sarihan, P., Witten, M.L., Pivniouk, V.I., Chen, G.J., In utero and postnatal exposure to arsenic alters pulmonary structure and function. Toxicol. Appl. Pharmacol. 235 (2009), 105–113, 10.1016/j.taap.2008.11.012.
29. Lau, A., Whitman, S.A., Jaramillo, M.C., Zhang, D.D., Arsenic-mediated activation of the Nrf2-Keap1 antioxidant pathway. J. Biochem. Mol. Toxicol. 27 (2013), 99–105, 10.1002/jbt.21463.
30. Lau, A., Zheng, Y., Tao, S., Wang, H., Whitman, S.A., White, E., Zhang, D.D., Arsenic inhibits autophagic flux, activating the Nrf2-Keap1 pathway in a p62-dependent manner. Mol. Cell. Biol. 33 (2013), 2436–2446, 10.1128/MCB.01748-12.
31. Lavoie, E.G., Wangdi, T., Kazmierczak, B.I., Innate immune responses to Pseudomonas aeruginosa infection. Microbes Infect. 13 (2011), 1133–1145, 10.1016/j.micinf.2011.07.011.
32. Lee, Y.C., Yang, D., Determination of lysozyme activities in a microplate format. Anal. Biochem. 310 (2002), 223–224, 10.1016/S0003-2697(02)00320-2.
33. Lefevre, P., Witham, J., Lacroix, C.E., Cockerill, P.N., Bonifer, C., The LPS-induced transcriptional upregulation of the chicken lysozyme locus involves CTCF eviction and noncoding RNA transcription. Mol. Cell 32 (2008), 129–139, 10.1016/j.molcel.2008.07.023.
34. Liao, C.-M., Chio, C.-P., Cheng, Y.-H., Hsieh, N.-H., Chen, W.-Y., Chen, S.-C., Quantitative links between arsenic exposure and influenza a (H1N1) infection-associated lung function exacerbations risk. Risk Anal. 31 (2011), 1281–1294, 10.1111/j.1539-6924.2010.01575.x.
35. Mandal, B.K., Suzuki, K.T., Arsenic round the world: a review. Talanta 58 (2002), 201–235, 10.1016/S0039-9140(02)00268-0.
36. McCarthy, D.J., Chen, Y., Smyth, G.K., Differential expression analysis of multifactor RNA-Seq experiments with respect to biological variation. Nucleic Acids Res. 40 (2012), 4288–4297, 10.1093/nar/gks042.
37. Millar, I.T., Heaney, H., Heinekey, D.M., Fernelius, W.C., Maier, L., Rochow, E.G., Methyldiiodoarsine. Rochow, E.G., (eds.) Inorganic Syntheses, 1960, John Wiley & Sons, Inc., 113–115, 10.1002/9780470132371.ch36.
38. Moreau-Marquis, S., Redelman, C.V., Stanton, B.A., Anderson, G.G., Co-culture models of Pseudomonas aeruginosa biofilms grown on live human airway cells. J. Vis. Exp., 2010, 10.3791/2186.
39. Murphy, T.F., Brauer, A.L., Eschberger, K., Lobbins, P., Grove, L., Cai, X., Sethi, S., Pseudomonas aeruginosa in chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 177 (2008), 853–860, 10.1164/rccm.200709-1413OC.
40. Nachman, K.E., Ginsberg, G.L., Miller, M.D., Murray, C.J., Nigra, A.E., Pendergrast, C.B., Mitigating dietary arsenic exposure: current status in the United States and recommendations for an improved path forward. Sci. Total Environ. 581–582 (2017), 221–236, 10.1016/j.scitotenv.2016.12.112.
41. Nadeau, K.C., Li, Z., Farzan, S., Koestler, D., Robbins, D., Fei, D.L., Malipatlolla, M., Maecker, H., Enelow, R., Korrick, S., Karagas, M.R., In utero arsenic exposure and fetal immune repertoire in a US pregnancy cohort. Clin. Immunol. 155 (2014), 188–197, 10.1016/j.clim.2014.09.004.
42. Nash, J.A., Ballard, T.N.S., Weaver, T.E., Akinbi, H.T., The peptidoglycan-degrading property of lysozyme is not required for bactericidal activity in vivo. J. Immunol. 177 (2006), 519–526, 10.4049/jimmunol.177.1.519.
43. Naujokas, M.F., Anderson, B., Ahsan, H., Aposhian, H.V., Graziano, J.H., Thompson, C., Suk, W.A., The broad scope of health effects from chronic arsenic exposure: update on a worldwide public health problem. Environ. Health Perspect. 121 (2013), 295–302, 10.1289/ehp.1205875.
44. Nayak, A.S., Lage, C.R., Kim, C.H., Effects of low concentrations of arsenic on the innate immune system of the zebrafish (Danio rerio). Toxicol. Sci. 98 (2007), 118–124, 10.1093/toxsci/kfm072.
45. Notch, E.G., Goodale, B.C., Barnaby, R., Coutermarsh, B., Berwin, B., Taylor, V.F., Jackson, B.P., Stanton, B.A., Monomethylarsonous acid (MMAIII) has an adverse effect on the innate immune response of human bronchial epithelial cells to Pseudomonas aeruginosa. PLoS One, 10, 2015, 10.1371/journal.pone.0142392.
46. Olivas-Calderón, E., Recio-Vega, R., Jay Gandolfi, A., Clark Lantz, R., González-Cortes, T., Alba, C.G.-D., Froines, J.R., Espinosa-Fematt, J.A., Lung inflammation biomarkers and lung function in children chronically exposed to arsenic. Toxicol. Appl. Pharmacol., 2015, 10.1016/j.taap.2015.06.001.
47. Panarelli, M., Holloway, C.D., Mulatero, P., Fraser, R., Kenyon, C.J., Inhibition of lysozyme synthesis by dexamethasone in human mononuclear leukocytes: an index of glucocorticoid sensitivity. J. Clin. Endocrinol. Metab. 78 (1994), 872–877, 10.1210/jcem.78.4.8157714.
48. Parvez, F., Chen, Y., Yunus, M., Olopade, C., Segers, S., Slavkovich, V., Argos, M., Hasan, R., Ahmed, A., Islam, T., Akter, M.M., Graziano, J.H., Ahsan, H., Arsenic exposure and impaired lung function. Findings from a large population-based prospective cohort study. Am. J. Respir. Crit. Care Med. 188 (2013), 813–819, 10.1164/rccm.201212-2282OC.
49. R Core Team, R: A Language and Environment for Statistical Computing. 2014.
50. Ramsey, K.A., Bosco, A., McKenna, K.L., Carter, K.W., Elliot, J.G., Berry, L.J., Sly, P.D., Larcombe, A.N., Zosky, G.R., In utero exposure to arsenic alters lung development and genes related to immune and mucociliary function in mice. Environ. Health Perspect., 121, 2013, 244, 10.1289/ehp.1205590 (Online Res. Triangle Park).
51. Robinson, M.D., McCarthy, D.J., Smyth, G.K., edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26 (2010), 139–140, 10.1093/bioinformatics/btp616.
52. Ryu, J.-H., Kim, C.-H., Yoon, J.-H., Innate immune responses of the airway epithelium. Mol. Cells 30 (2010), 173–183, 10.1007/s10059-010-0146-4.
53. Sanders, A.P., Messier, K.P., Shehee, M., Rudo, K., Serre, M.L., Fry, R.C., Arsenic in North Carolina: public health implications. Environ. Int. 38 (2012), 10–16, 10.1016/j.envint.2011.08.005.
54. Sherwood, C.L., Lantz, R.C., Boitano, S., Chronic arsenic exposure in nanomolar concentrations compromises wound response and intercellular signaling in airway epithelial cells. Toxicol. Sci. 132 (2013), 222–234, 10.1093/toxsci/kfs331.
55. Shi, L., Reid, L.H., Jones, W.D., Shippy, R., Warrington, J.A., Baker, S.C., Collins, P.J., Longueville, F. de, Kawasaki, E.S., Lee, K.Y., Luo, Y., Sun, Y.A., Willey, J.C., Setterquist, R.A., Fischer, G.M., Tong, W., Dragan, Y.P., Dix, D.J., Frueh, F.W., Goodsaid, F.M., Herman, D., Jensen, R.V., Johnson, C.D., Lobenhofer, E.K., Puri, R.K., Scherf, U., Thierry-Mieg, J., Wang, C., Wilson, M., Wolber, P.K., Zhang, L., Amur, S., Bao, W., Barbacioru, C.C., Lucas, A.B., Bertholet, V., Boysen, C., Bromley, B., Brown, D., Brunner, A., Canales, R., Cao, X.M., Cebula, T.A., Chen, J.J., Cheng, J., Chu, T.-M., Chudin, E., Corson, J., Corton, J.C., Croner, L.J., Davies, C., Davison, T.S., Delenstarr, G., Deng, X., Dorris, D., Eklund, A.C., Fan, X., Fang, H., Fulmer-Smentek, S., Fuscoe, J.C., Gallagher, K., Ge, W., Guo, L., Guo, X., Hager, J., Haje, P.K., Han, J., Han, T., Harbottle, H.C., Harris, S.C., Hatchwell, E., Hauser, C.A., Hester, S., Hong, H., Hurban, P., Jackson, S.A., Ji, H., Knight, C.R., Kuo, W.P., LeClerc, J.E., Levy, S., Li, Q.-Z., Liu, C., Liu, Y., Lombardi, M.J., Ma, Y., Magnuson, S.R., Maqsodi, B., McDaniel, T., Mei, N., Myklebost, O., Ning, B., Novoradovskaya, N., Orr, M.S., Osborn, T.W., Papallo, A., Patterson, T.A., Perkins, R.G., Peters, E.H., Peterson, R., Philips, K.L., Pine, P.S., Pusztai, L., Qian, F., Ren, H., Rosen, M., Rosenzweig, B.A., Samaha, R.R., Schena, M., Schroth, G.P., Shchegrova, S., Smith, D.D., Staedtler, F., Su, Z., Sun, H., Szallasi, Z., Tezak, Z., Thierry-Mieg, D., Thompson, K.L., Tikhonova, I., Turpaz, Y., Vallanat, B., Van, C., Walker, S.J., Wang, S.J., Wang, Y., Wolfinger, R., Wong, A., Wu, J., Xiao, C., Xie, Q., Xu, J., Yang, W., Zhang, L., Zhong, S., Zong, Y., Slikker, W., The MicroArray Quality Control (MAQC) project shows inter- and intraplatform reproducibility of gene expression measurements. Nat. Biotechnol. 24 (2006), 1151–1161, 10.1038/nbt1239.
56. Shugar, D., The measurement of lysozyme activity and the ultra-violet inactivation of lysozyme. Biochim. Biophys. Acta 8 (1952), 302–309, 10.1016/0006-3002(52)90045-0.
57. Smith, A.H., Marshall, G., Yuan, Y., Ferreccio, C., Liaw, J., Ehrenstein, O. von, Steinmaus, C., Bates, M.N., Selvin, S., Increased mortality from lung cancer and bronchiectasis in young adults after exposure to arsenic in utero and in early childhood. Environ. Health Perspect. 114 (2006), 1293–1296.
58. Smith, A.H., Marshall, G., Yuan, Y., Liaw, J., Ferreccio, C., Steinmaus, C., Evidence from Chile that arsenic in drinking water may increase mortality from pulmonary tuberculosis. Am. J. Epidemiol. 173 (2011), 414–420, 10.1093/aje/kwq383.
59. Smith, A.H., Yunus, M., Khan, A.F., Ercumen, A., Yuan, Y., Smith, M.H., Liaw, J., Balmes, J., von Ehrenstein, O., Raqib, R., Kalman, D., Alam, D.S., Streatfield, P.K., Steinmaus, C., Chronic respiratory symptoms in children following in utero and early life exposure to arsenic in drinking water in Bangladesh. Int. J. Epidemiol. 42 (2013), 1077–1086, 10.1093/ije/dyt120.
60. Suico, M.A., Koga, T., Shuto, T., Hisatsune, A., Lu, Z., Basbaum, C., Okiyoneda, T., Kai, H., Sp1 is involved in the transcriptional activation of lysozyme in epithelial cells. Biochem. Biophys. Res. Commun. 324 (2004), 1302–1308, 10.1016/j.bbrc.2004.09.195.
61. Tarran, R., Regulation of airway surface liquid volume and mucus transport by active ion transport. Proc. Am. Thorac. Soc. 1 (2004), 42–46, 10.1513/pats.2306014.
62. Thompson, A.B., Bohling, T., Payvandi, F., Rennard, S.I., Lower respiratory tract lactoferrin and lysozyme arise primarily in the airways and are elevated in association with chronic bronchitis. J. Lab. Clin. Med. 115 (1990), 148–158.
63. Travis, S.M., Conway, B.-A.D., Zabner, J., Smith, J.J., Anderson, N.N., Singh, P.K., Peter Greenberg, E., Welsh, M.J., Activity of abundant antimicrobials of the human airway. Am. J. Respir. Cell Mol. Biol. 20 (1999), 872–879, 10.1165/ajrcmb.20.5.3572.
64. Tsai, C.-M., Wong, K.-S., Lee, W.-J., Hsieh, K.-S., Hung, P.-L., Niu, C.-K., Yu, H.-R., Diagnostic value of bronchoalveolar lavage in children with nonresponding community-acquired pneumonia. Pediatr. Neonatol., 2017, 10.1016/j.pedneo.2016.09.004.
65. Vahter, M., Mechanisms of arsenic biotransformation. Toxicology 181–182 (2002), 211–217, 10.1016/S0300-483X(02)00285-8.
66. Vervoort, S.J., van Boxtel, R., Coffer, P.J., The role of SRY-related HMG box transcription factor 4 (SOX4) in tumorigenesis and metastasis: friend or foe?. Oncogene 32 (2013), 3397–3409, 10.1038/onc.2012.506.
67. Wickham, H., ggplot2 - Elegant Graphics for Data Analysis. 2009, Springer.
68. World Health Organization, Guidelines for Drinking-Water Quality: First Addendum to Third Edition. 2006.
69. Xie, H., Huang, S., Martin, S., Wise, J.P. Sr., Arsenic is cytotoxic and genotoxic to primary human lung cells. Mutat. Res. Toxicol. Environ. Mutagen. 760 (2014), 33–41, 10.1016/j.mrgentox.2013.11.001.
70. Young, M.D., Wakefield, M.J., Smyth, G.K., Oshlack, A., Gene ontology analysis for RNA-seq: accounting for selection bias. Genome Biol., 11, 2010, R14, 10.1186/gb-2010-11-2-r14.