Papers from the current funding period 

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Ahn KC, Ranganathan A, Bever CS, Hwang SH, Holland EB, Morisseau C, Pessah IN, Hammock BD, Gee SJ. Detection of the Antimicrobial Triclosan in Environmental Samples by Immunoassay. Environmental science & technology. 2016;50(7):3754-61. Epub 2016/03/05. doi: 10.1021/acs.est.5b05357. PubMed PMID: 26937944; PMCID: PMC4821808. (Project 5, Core B)

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Barnych B, Vasylieva N, Joseph T, Hulsizer S, Nguyen HM, Cajka T, Pessah I, Wulff H, Gee SJ, Hammock BD. Development of Tetramethylenedisulfotetramine (TETS) Hapten Library: Synthesis, Electrophysiological Studies, and Immune Response in Rabbits. Chemistry (Weinheim an der Bergstrasse, Germany). 2017;23(35):8466-72. Epub 2017/04/16. doi: 10.1002/chem.201700783. PubMed PMID: 28411375; PMCID: PMC5808876.

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Bever CS, Dong JX, Vasylieva N, Barnych B, Cui Y, Xu ZL, Hammock BD, Gee SJ. VHH antibodies: emerging reagents for the analysis of environmental chemicals. Analytical and bioanalytical chemistry. 2016;408(22):5985-6002. Epub 2016/05/23. doi: 10.1007/s00216-016-9585-x. PubMed PMID: 27209591; PMCID: PMC4983233.

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Bever CS, Rand AA, Nording M, Taft D, Kalanetra KM, Mills DA, Breck MA, Smilowitz JT, German JB, Hammock BD. Effects of triclosan in breast milk on the infant fecal microbiome. Chemosphere. 2018;203:467-73. Epub 2018/04/11. doi: 10.1016/j.chemosphere.2018.03.186. PubMed PMID: 29635158; PMCID: PMC5915298. (Core A)

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Cam M, Durieu E, Bodin M, Manousopoulou A, Koslowski S, Vasylieva N, Barnych B, Hammock BD, Bohl B, Koch P, Omori C, Yamamoto K, Hata S, Suzuki T, Karg F, Gizzi P, Erakovic Haber V, Bencetic Mihaljevic V, Tavcar B, Portelius E, Pannee J, Blennow K, Zetterberg H, Garbis SD, Auvray P, Gerber H, Fraering J, Fraering PC, Meijer L. Induction of Amyloid-β42 Production by Fipronil and Other Pyrazole Insecticides. Journal of Alzheimer's disease : JAD. 2018;62(4):1663-81. Epub 2018/03/06. doi: 10.3233/jad-170875. PubMed PMID: 29504531; PMCID: PMC7065497. (Project 5)

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Chen H, Ding Y, Yang Q, Barnych B, González-Sapienza G, Hammock BD, Wang M, Hua X. Fluorescent "Turn off-on" Small-Molecule-Monitoring Nanoplatform Based on Dendrimer-like Peptides as Competitors. ACS applied materials & interfaces. 2019;11(36):33380-9. Epub 2019/08/23. doi: 10.1021/acsami.9b13111. PubMed PMID: 31433617; PMCID: PMC7059760.

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Chen H, Yang Q, Ding Y, Vasylieva N, Bever CS, Hua X, Wang M, Hammock BD. Competitive and noncompetitive immunoassays for the detection of benzothiostrobin using magnetic nanoparticles and fluorescein isothiocyanate-labeled peptides. Analytical and bioanalytical chemistry. 2019;411(2):527-35. Epub 2018/11/28. doi: 10.1007/s00216-018-1478-8. PubMed PMID: 30478514; PMCID: PMC7263471.

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Chen ZJ, Liu XX, Xiao ZL, Fu HJ, Huang YP, Huang SY, Shen YD, He F, Yang XX, Hammock BD, Xu ZL. Production of a specific monoclonal antibody for 1-naphthol based on novel hapten strategy and development of an easy-to-use ELISA in urine samples. Ecotoxicology and environmental safety. 2020;196:110533. Epub 2020/04/05. doi: 10.1016/j.ecoenv.2020.110533. PubMed PMID: 32247241; PMCID: PMC7200204.

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Cui X, He Q, Shen D, Jiang Z, Chen Y, Zhao S, Hammock BD. Production and characterization of a single-chain variable fragment-alkaline phosphatase fusion protein for glycocholic acid detection in a one-step enzyme-linked immunosorbent assay. Analytical Methods. 2018;10(22):2629-35. doi: 10.1039/C8AY00848E.

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Cui X, Vasylieva N, Shen D, Barnych B, Yang J, He Q, Jiang Z, Zhao S, Hammock BD. Biotinylated single-chain variable fragment-based enzyme-linked immunosorbent assay for glycocholic acid. The Analyst. 2018;143(9):2057-65. Epub 2018/04/10. doi: 10.1039/c7an02024d. PubMed PMID: 29629470; PMCID: PMC6449042. (Core A)

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Cui X, Vasylieva N, Wu P, Barnych B, Yang J, Shen D, He Q, Gee SJ, Zhao S, Hammock BD. Development of an Indirect Competitive Enzyme-Linked Immunosorbent Assay for Glycocholic Acid Based on Chicken Single-Chain Variable Fragment Antibodies. Analytical chemistry. 2017;89(20):11091-7. Epub 2017/09/26. doi: 10.1021/acs.analchem.7b03190. PubMed PMID: 28942640; PMCID: PMC6459402. (Core A)

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Ding Y, Hua X, Chen H, Gonzalez-Sapienza G, Barnych B, Liu F, Wang M, Hammock BD. A dual signal immunochromatographic strip for the detection of imidaclothiz using a recombinant fluorescent-peptide tracer and gold nanoparticles. Sensors and Actuators B: Chemical. 2019;297:126714. doi: https://doi.org/10.1016/j.snb.2019.126714.

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Dong J, Li Z, Wang Y, Jin M, Shen Y, Xu Z, Abd El-Aty AM, Gee SJ, Hammock BD, Sun Y, Wang H. Generation of functional single-chain fragment variable from hybridoma and development of chemiluminescence enzyme immunoassay for determination of total malachite green in tilapia fish. Food chemistry. 2021;337:127780. Epub 2020/08/18. doi: 10.1016/j.foodchem.2020.127780. PubMed PMID: 32799164; PMCID: PMC7541715.

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Duflot T, Moreau-Grangé L, Roche C, Iacob M, Wils J, Rémy-Jouet I, Cailleux AF, Leuillier M, Renet S, Li D, Morisseau C, Lamoureux F, Richard V, Prévost G, Joannidès R, Bellien J. Altered bioavailability of epoxyeicosatrienoic acids is associated with conduit artery endothelial dysfunction in type 2 diabetic patients. Cardiovascular diabetology. 2019;18(1):35. Epub 2019/03/20. doi: 10.1186/s12933-019-0843-z. PubMed PMID: 30885203; PMCID: PMC6423843. (Project 5)

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El-Moghazy AY, Huo J, Amaly N, Vasylieva N, Hammock BD, Sun G. An Innovative Nanobody-Based Electrochemical Immunosensor Using Decorated Nylon Nanofibers for Point-of-Care Monitoring of Human Exposure to Pyrethroid Insecticides2020;12(5):6159-68. doi: 10.1021/acsami.9b16193. PubMed PMID: 31927905. (Project 2)

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Fu HJ, Wang Y, Xiao ZL, Wang H, Li ZF, Shen YD, Lei HT, Sun YM, Xu ZL, Hammock BD. A rapid and simple fluorescence enzyme-linked immunosorbent assay for tetrabromobisphenol A in soil samples based on a bifunctional fusion protein. Ecotoxicology and environmental safety. 2020;188:109904. Epub 2019/11/11. doi: 10.1016/j.ecoenv.2019.109904. PubMed PMID: 31704326; PMCID: PMC7198468.

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Gee SJ, Kennedy IR, Lee NA, Ohkawa H, Prapamontol T, Xu T. Immunoanalysis for environmental monitoring and human health. Analytical and bioanalytical chemistry. 2016;408(22):5959-61. Epub 2016/06/10. doi: 10.1007/s00216-016-9666-x. PubMed PMID: 27277933.

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He J, Tian J, Xu J, Wang K, Li J, Gee SJ, Hammock BD, Li QX, Xu T. Strong and oriented conjugation of nanobodies onto magnetosomes for the development of a rapid immunomagnetic assay for the environmental detection of tetrabromobisphenol-A. Analytical and bioanalytical chemistry. 2018;410(25):6633-42. Epub 2018/08/02. doi: 10.1007/s00216-018-1270-9. PubMed PMID: 30066195; PMCID: PMC7105167.

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Hongsibsong S, Prapamontol T, Dong J-X, Bever CS, Xu Z-L, Gee SJ, Hammock BD. Exposure of consumers and farmers to organophosphate and synthetic pyrethroid insecticides in Northern Thailand. Journal of Consumer Protection and Food Safety. 2019;14(1):17-23. doi: 10.1007/s00003-019-01207-7.

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Hongsibsong S, Prapamontol T, Xu T, Hammock BD, Wang H, Chen ZJ, Xu ZL. Monitoring of the Organophosphate Pesticide Chlorpyrifos in Vegetable Samples from Local Markets in Northern Thailand by Developed Immunoassay. International journal of environmental research and public health. 2020;17(13). Epub 2020/07/08. doi: 10.3390/ijerph17134723. PubMed PMID: 32630084; PMCID: PMC7369984.

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Huo J, Barnych B, Li Z, Wan D, Li D, Vasylieva N, Knezevic SZ, Osipitan OA, Scott JE, Zhang J, Hammock BD. Hapten Synthesis, Antibody Development, and a Highly Sensitive Indirect Competitive Chemiluminescent Enzyme Immunoassay for Detection of Dicamba. Journal of Agricultural and Food Chemistry. 2019. doi: 10.1021/acs.jafc.8b07134.

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Huo J, Li Z, Wan D, Li D, Qi M, Barnych B, Vasylieva N, Zhang J, Hammock BD. Development of a Highly Sensitive Direct Competitive Fluorescence Enzyme Immunoassay Based on a Nanobody-Alkaline Phosphatase Fusion Protein for Detection of 3-Phenoxybenzoic Acid in Urine. Journal of agricultural and food chemistry. 2018;66(43):11284-90. Epub 2018/10/09. doi: 10.1021/acs.jafc.8b04521. PubMed PMID: 30293433; PMCID: PMC6442738. (Core A)

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Joshi U, Pearson A, Evans JE, Langlois H, Saltiel N, Ojo J, Klimas N, Sullivan K, Keegan AP, Oberlin S, Darcey T, Cseresznye A, Raya B, Paris D, Hammock BD, Vasylieva N, Hongsibsong S, Stern LJ, Crawford F, Mullan M, Abdullah L. A permethrin metabolite is associated with adaptive immune responses in Gulf War Illness. Brain, behavior, and immunity. 2019;81:545-59. Epub 2019/07/22. doi: 10.1016/j.bbi.2019.07.015. PubMed PMID: 31325531; PMCID: PMC7155744. (Project 5)

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Kumar S, Vasylieva N, Singh V, Hammock BD, Singh SG. A Facile, Sensitive and Rapid Sensing Platform Based on CoZnO for Detection of Fipronil; an Environmental Toxin. Electroanalysis. 2020;32(9):2056-64. doi: 10.1002/elan.202000051, NIHMSID: 1655891.

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Lassabe G, Kramer K, Hammock BD, González-Sapienza G, González-Techera A. Noncompetitive Homogeneous Detection of Small Molecules Using Synthetic Nanopeptamer-Based Luminescent Oxygen Channeling. Analytical chemistry. 2018;90(10):6187-92. Epub 2018/04/26. doi: 10.1021/acs.analchem.8b00657. PubMed PMID: 29694028; PMCID: PMC5963685. (Project 5)

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Li D, Cui Y, Morisseau C, Gee SJ, Bever CS, Liu X, Wu J, Hammock BD, Ying Y. Nanobody Based Immunoassay for Human Soluble Epoxide Hydrolase Detection Using Polymeric Horseradish Peroxidase (PolyHRP) for Signal Enhancement: The Rediscovery of PolyHRP? Analytical chemistry. 2017;89(11):6248-56. Epub 2017/05/04. doi: 10.1021/acs.analchem.7b01247. PubMed PMID: 28460522; PMCID: PMC5611449. (Core B)

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Li D, Morisseau C, McReynolds CB, Duflot T, Bellien J, Nagra RM, Taha AY, Hammock BD. Development of Improved Double-Nanobody Sandwich ELISAs for Human Soluble Epoxide Hydrolase Detection in Peripheral Blood Mononuclear Cells of Diabetic Patients and the Prefrontal Cortex of Multiple Sclerosis Patients. Analytical chemistry. 2020;92(10):7334-42. doi: 10.1021/acs.analchem.0c01115. PubMed PMID: 32253910. (Project 5 and Core B)

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Li P, Yang C, Liu B, Wu Q, Wang Y, Dong S, Zhang H, Vasylieva N, Hammock BD, Zhang C. Sensitive Immunochromatographic Assay Using Highly Luminescent Quantum Dot Nanobeads as Tracer for the Detection of Cyproheptadine Hydrochloride in Animal-Derived Food. Frontiers in chemistry. 2020;8:575. Epub 2020/08/08. doi: 10.3389/fchem.2020.00575. PubMed PMID: 32760698; PMCID: PMC7372008.

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Liu X, Tang Z, Duan Z, He Z, Shu M, Wang X, Gee SJ, Hammock BD, Xu Y. Nanobody-based enzyme immunoassay for ochratoxin A in cereal with high resistance to matrix interference. Talanta. 2017;164:154-8. Epub 2017/01/22. doi: 10.1016/j.talanta.2016.11.039. PubMed PMID: 28107910; PMCID: PMC5862147.

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Liu Z, Wang K, Wu S, Wang Z, Ding G, Hao X, Li QX, Li J, Gee SJ, Hammock BD, Xu T. Development of an immunoassay for the detection of carbaryl in cereals based on a camelid variable heavy-chain antibody domain. Journal of the science of food and agriculture. 2019;99(9):4383-90. Epub 2019/03/10. doi: 10.1002/jsfa.9672. PubMed PMID: 30851058; PMCID: PMC7061733.

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Luo L, Wei XQ, Jia BZ, Yang J, Shen YD, Hammock BD, Dong JX, Wang H, Lei HT, Xu ZL. Modulating Linker Composition of Haptens Resulted in Improved Immunoassay for Histamine. Biomolecules. 2019;9(10). Epub 2019/10/17. doi: 10.3390/biom9100597. PubMed PMID: 31614550; PMCID: PMC6843301. (Core A)

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Mishra M, Tiwari S, Gunaseelan A, Li D, Hammock BD, Gomes AV. Improving the sensitivity of traditional Western blotting via Streptavidin containing Poly-horseradish peroxidase (PolyHRP). Electrophoresis. 2019;40(12-13):1731-9. Epub 2019/04/26. doi: 10.1002/elps.201900059. PubMed PMID: 31021001; PMCID: PMC7036291. (Project 4 and Core B)

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Mukhtar H, Ma L, Pang Q, Zhou Y, Wang X, Xu T, Hammock BD, Wang J. Cyclic peptide: a safe and effective alternative to synthetic aflatoxin B1-competitive antigens. Analytical and bioanalytical chemistry. 2019;411(17):3881-90. doi: 10.1007/s00216-019-01862-7.

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Qi M, Huo J, Li Z, He C, Li D, Wang Y, Vasylieva N, Zhang J, Hammock BD. On-spot quantitative analysis of dicamba in field waters using a lateral flow immunochromatographic strip with smartphone imaging. Analytical and bioanalytical chemistry. 2020;412(25):6995-7006. Epub 2020/08/02. doi: 10.1007/s00216-020-02833-z. PubMed PMID: 32737550.

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Qiu Y, Li P, Dong S, Zhang X, Yang Q, Wang Y, Ge J, Hammock BD, Zhang C, Liu X. Phage-Mediated Competitive Chemiluminescent Immunoassay for Detecting Cry1Ab Toxin by Using an Anti-Idiotypic Camel Nanobody. Journal of agricultural and food chemistry. 2018;66(4):950-6. Epub 2018/01/03. doi: 10.1021/acs.jafc.7b04923. PubMed PMID: 29293334; PMCID: PMC7314401.

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Ren W, Li Z, Xu Y, Wan D, Barnych B, Li Y, Tu Z, He Q, Fu J, Hammock BD. One-Step Ultrasensitive Bioluminescent Enzyme Immunoassay Based on Nanobody/Nanoluciferase Fusion for Detection of Aflatoxin B(1) in Cereal2019;67(18):5221-9. doi: 10.1021/acs.jafc.9b00688. PubMed PMID: 30883117.

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Ren W, Xu Y, Huang Z, Li Y, Tu Z, Zou L, He Q, Fu J, Liu S, Hammock BD. Single-chain variable fragment antibody-based immunochromatographic strip for rapid detection of fumonisin B(1) in maize samples. Food chemistry. 2020;319:126546. Epub 2020/03/18. doi: 10.1016/j.foodchem.2020.126546. PubMed PMID: 32179370.

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Riangrungroj P, Bever CS, Hammock BD, Polizzi KM. A label-free optical whole-cell Escherichia coli biosensor for the detection of pyrethroid insecticide exposure. Scientific reports. 2019;9(1):12466. Epub 2019/08/30. doi: 10.1038/s41598-019-48907-6. PubMed PMID: 31462650; PMCID: PMC6713742.

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Rossotti MA, Pirez M, Gonzalez-Techera A, Cui Y, Bever CS, Lee KS, Morisseau C, Leizagoyen C, Gee S, Hammock BD, González-Sapienza G. Method for Sorting and Pairwise Selection of Nanobodies for the Development of Highly Sensitive Sandwich Immunoassays. Analytical chemistry. 2015;87(23):11907-14. Epub 2015/11/07. doi: 10.1021/acs.analchem.5b03561. PubMed PMID: 26544909; PMCID: PMC4666776. (Project 5)

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Shu M, Xu Y, Dong J-x, Zhong C, Hammock BD, Wang W-j, Wu G-p. Development of a noncompetitive idiometric nanobodies phage immumoassay for the determination of fumonisin B1. Food and Agricultural Immunology. 2019;30(1):510-21. doi: 10.1080/09540105.2019.1604637.

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Späth J, Nording M, Lindberg R, Brodin T, Jansson S, Yang J, Wan D, Hammock BD, Fick J. Novel metabolomic method to assess the effect-based removal efficiency of advanced wastewater treatment techniques. Environmental Chemistry. 2020;17(1):1. doi: 10.1071/en19270. (Core A)

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Sun Z, Lv J, Liu X, Tang Z, Wang X, Xu Y, Hammock BD. Development of a Nanobody-AviTag Fusion Protein and Its Application in a Streptavidin-Biotin-Amplified Enzyme-Linked Immunosorbent Assay for Ochratoxin A in Cereal. Analytical chemistry. 2018;90(17):10628-34. Epub 2018/08/11. doi: 10.1021/acs.analchem.8b03085. PubMed PMID: 30092629; PMCID: PMC7307665.

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Tang Z, Liu X, Wang Y, Chen Q, Hammock BD, Xu Y. Nanobody-based fluorescence resonance energy transfer immunoassay for noncompetitive and simultaneous detection of ochratoxin a and ochratoxin B. Environmental pollution (Barking, Essex : 1987). 2019;251:238-45. Epub 2019/05/15. doi: 10.1016/j.envpol.2019.04.135. PubMed PMID: 31082608; PMCID: PMC7103568.

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Tang Z, Mo F, Liu A, Duan S, Yang X, Liang L, Hou X, Yin S, Jiang X, Vasylieva N, Dong J, Barnych B, Hammock BD, Lu X. A Nanobody Against Cytotoxic T-Lymphocyte Associated Antigen-4 Increases the Anti-Tumor Effects of Specific CD8 + T Cells. Journal of Biomedical Nanotechnology. 2019;15:2229-39. doi: 10.1166/jbn.2019.2859.

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Vasylieva N, Barnych B, Rand A, Inceoglu B, Gee SJ, Hammock BD. Sensitive Immunoassay for Detection and Quantification of the Neurotoxin, Tetramethylenedisulfotetramine. Analytical chemistry. 2017;89(10):5612-9. Epub 2017/04/12. doi: 10.1021/acs.analchem.7b00846. PubMed PMID: 28398746; PMCID: PMC5920647.

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Vasylieva N, Barnych B, Wan D, El-Sheikh EA, Nguyen HM, Wulff H, McMahen R, Strynar M, Gee SJ, Hammock BD. Hydroxy-fipronil is a new urinary biomarker of exposure to fipronil. Environment international. 2017;103:91-8. Epub 2017/03/28. doi: 10.1016/j.envint.2017.03.012. PubMed PMID: 28343720; PMCID: PMC5432128. (Core A)

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Vasylieva N, Kitamura S, Dong J, Barnych B, Hvorecny KL, Madden DR, Gee SJ, Wolan DW, Morisseau C, Hammock BD. Nanobody-based binding assay for the discovery of potent inhibitors of CFTR inhibitory factor (Cif). Analytica chimica acta. 2019;1057:106-13. Epub 2019/03/06. doi: 10.1016/j.aca.2018.12.060. PubMed PMID: 30832908; PMCID: PMC6426448. (Core B)

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Wagner KD, Kurobe T, Hammock BD, Lam CH, Wu G, Vasylieva N, Gee SJ, Hammock BD, Teh SJ. Developmental effects of fipronil on Japanese Medaka (Oryzias latipes) embryos. Chemosphere. 2017;166:511-20. Epub 2016/10/07. doi: 10.1016/j.chemosphere.2016.09.069. PubMed PMID: 27710886; PMCID: PMC5560107.

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Wang F, Li ZF, Yang YY, Wan DB, Vasylieva N, Zhang YQ, Cai J, Wang H, Shen YD, Xu ZL, Hammock BD. Chemiluminescent Enzyme Immunoassay and Bioluminescent Enzyme Immunoassay for Tenuazonic Acid Mycotoxin by Exploitation of Nanobody and Nanobody-Nanoluciferase Fusion. Analytical chemistry. 2020;92(17):11935-42. doi: 10.1021/acs.analchem.0c02338. PubMed PMID: 32702970.

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Wang K, Liu Z, Ding G, Li J, Vasylieva N, Li QX, Li D, Gee SJ, Hammock BD, Xu T. Development of a one-step immunoassay for triazophos using camel single-domain antibody-alkaline phosphatase fusion protein. Analytical and bioanalytical chemistry. 2019;411(6):1287-95. Epub 2019/02/02. doi: 10.1007/s00216-018-01563-7. PubMed PMID: 30706076; PMCID: PMC7153572.

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Wang K, Vasylieva N, Wan D, Eads DA, Yang J, Tretten T, Barnych B, Li J, Li QX, Gee SJ, Hammock BD, Xu T. Quantitative Detection of Fipronil and Fipronil-Sulfone in Sera of Black-Tailed Prairie Dogs and Rats after Oral Exposure to Fipronil by Camel Single-Domain Antibody-Based Immunoassays. Analytical chemistry. 2019;91(2):1532-40. Epub 2018/12/07. doi: 10.1021/acs.analchem.8b04653. PubMed PMID: 30521755; PMCID: PMC7144541. (Core A)

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Wang Y, Li Z, Barnych B, Huo J, Wan D, Vasylieva N, Xu J, Li P, Liu B, Zhang C, Hammock BD. Investigation of the Small Size of Nanobodies for a Sensitive Fluorescence Polarization Immunoassay for Small Molecules: 3-Phenoxybenzoic Acid, an Exposure Biomarker of Pyrethroid Insecticides as a Model. Journal of agricultural and food chemistry. 2019;67(41):11536-41. Epub 2019/10/08. doi: 10.1021/acs.jafc.9b04621. PubMed PMID: 31589045; PMCID: PMC7134064. (Core A)

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Wang Y, Xu J, Qiu Y, Li P, Liu B, Yang L, Barnych B, Hammock BD, Zhang C. Highly Specific Monoclonal Antibody and Sensitive Quantum Dot Beads-Based Fluorescence Immunochromatographic Test Strip for Tebuconazole Assay in Agricultural Products. Journal of agricultural and food chemistry. 2019;67(32):9096-103. Epub 2019/07/30. doi: 10.1021/acs.jafc.9b02832. PubMed PMID: 31356079; PMCID: PMC7069222.

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Wang, K., Liu, Z., Ding, G. et al. Development of a one-step immunoassay for triazophos using camel single-domain antibody–alkaline phosphatase fusion protein. Anal Bioanal Chem 411, 1287–1295 (2019). https://doi.org/10.1007/s00216-018-01563-7

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Yu Y, Tang P, Barnych B, Zhao C, Sun G, Ge M. Design and Synthesis of Core–Shell Carbon Polymer Dots with Highly Stable Fluorescence in Polymeric Materials. ACS Applied Nano Materials. 2019;2(10):6503-12. doi: 10.1021/acsanm.9b01446. NIHMSID: 1667599 (Project 2)

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