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6 protocols using diallyl disulphide

1

Antioxidant Pathway Regulation Protocol

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Diphenyl dichloromethane, diallyl disulphide, hydrogen peroxide, 4-nitrophenol, methanesulfonic acid, tetramethoxypropane, dinitrophenylhydrazine, 1-methyl-2-phenylindole 4-nitrophenyl-N-acetyl-β-d-glucosaminide, acetonitrile, methanol, and guanidine were purchased from Sigma (St. Louis, MO, USA). The anti-angiotensin II type 1 receptor antibody (ab124734) and anti-angiotensin II type 2 receptor antibody (ab92445) were purchased from Abcam (Cambridge, MA, USA). The β-actin antibody GTX109639, eNOS antibody GTX54637, KEAP1 antibody GTX60660, and NRF2 antibody GTX103322 were purchased from GeneTex Inc. (Irvine, CA, USA). The nephrin antibody ALX-810-016-R100 was purchased from Enzo Life Sciences Inc. (Farmingdale, NY, USA). The catalase antibody sc-34281, heme oxygenase 1 antibody sc-1797, and superoxide dismutase antibodysc-8637 were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). All other chemicals used herein were of the highest analytical grade available.
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2

Comprehensive Analytical Protocol for Natural and Synthetic Compounds

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A mixture of C7–C30 normal alkanes in n-hexane (HPLC grade) was purchased from Beijing Huawei Ruike Chemical Co., Ltd. (Beijing, China); 6-undecanone (AR) was obtained from Tokyo Huacheng Industrial Pearl Club; sec-butylisothiocyanate-(96%), allyl isothiocyanate (95%), 3-(methylthio)propyl isothiocyanate(98%), benzyl isothiocyanate(97%), phenylethyl isothiocyanate(96%), allyl mercaptan(97%), diallyl sulfide(95%), diallyl disulphide(98%), carbon disulfide(97%), allyl chloride(96%), hexanoic acid(95%), 3-methylphenol(98%), and propylene glycol (99%) were purchased from Sigma-Aldrich Chemical Co. (Saint Louis, USA). Sudan Ⅲ and liquid paraffin were obtained from Beijing Chemical Reagent Co., Ltd. (Beijing, China).
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3

Biochemical Reagents Acquisition Protocol

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Angiotensin II (Ang II), dichloromethane, diallyl disulphide, hydrogen peroxide, 4-nitrophenol, tetramethoxypropane, dinitrophenylhydrazine, methanesulfonic acid, and 1-methyl-2-phenylindole were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other chemicals used were of the highest purity available.
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4

Quantification of Volatile Compounds

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All 23 standard compounds used for the quantification analysis were purchased from Sigma-Aldrich (purity 99.0%, all chromatographic grade), including butanoic acid, acetic acid, thujone, methyl eugenol, p-cymene, α-pinene, diallyl disulphide, α-terpineol, β-caryophyllene, β-myrcene, 1,8-cineole, D-limonene, linalool, methional, nonanal, phenylethanal, ethyl caprylate, 4-methyl-5-thiazoleethanol, (E)-2-octenal, (E)-2-heptenal, humulene, acetophenone, and linalyl acetate. The internal standard (2,4,5-trimethylthiazole) and n-alkanes (C8–C30) used to calculate the retention index (RI) were of chromatographic grade and purchased from Sigma-Aldrich (St, Louis, MO, USA). Diethyl ether and n-pentane for dilution and preparation of sample were obtained from Thermo Fisher Scientific Inc. (Waltham, MA, USA). High purity helium (99.999% purity) and nitrogen (99.99% purity) were purchased and produced by Beijing Tianlirenhe Trading Co., Ltd. (Beijing, China).
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5

Analytical Grade Reagent Analysis

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All reagents were of analytical grade. Dichloromethane of chromatographic grade and diallyl disulphide (DADS) were purchased from Sigma-Aldrich (St Louis, MO). Dimethyl trisulphide (DMTS) and dimethyl disulphide (DMDS) (purity 98%) were supplied by Natural Advantage (Oakdale, LA).
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6

Behavioral Bioassays of Volatile Compounds

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Garlic oil, A. sativum, was purchased from Elixarome Ltd; rosemary, R. officinalis, and lavender, La. angustifolia, essential oils were provided by Botanix Ltd, and bog myrtle oil, Myrica gale, was purchased from Amphora Aromatics Ltd. Diallyl sulphide, diallyl disulphide, allyl, propyl, butyl and 2-phenylethyl isothiocyanate were purchased from Sigma-Aldrich. 4-Pentenyl isothiocyanate was synthesized from 4-pentenylamine hydrochloride as described previously (Dawson et al. 1993) . Solutions of extracts and individual compounds in absolute ethanol were prepared (0.1, 0.05, 0.01 and 0.001 mg mL 1 ) and diluted to 1 lL L 1 in ASW (Ingvarsdottir et al. 2002b) to give a final concentration of 100, 50, 10 and 1 parts per trillion, respectively, for use in Y-tube behavioural bioassays.
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