Trans resveratrol

Wine alcohol 65% v/v was obtained from winery Ouro Verde-Miolo Wine Group (Casa Nova, BA, Brazil). Folin-Ciocalteu reagent, ethanol, potassium persulfate, phosphoric acid, and potassium phosphate monobasic were purchased from Merck (Darmstadt, Germany). Trolox (6-hydroxy-2,5,7,8-tetramethylchromate-2-carboxylic acid) and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals and 2,2-azino-bis (3-ethylbenzthiazoline-6 sulfonic acid) (ABTS) were supplied by Sigma-Aldrich (St. Louis, MO, USA). Methanol was from J.T. Baker (Phillipsburg, NJ, USA). Ultrapure water was obtained with a Marte Científica purification system (São Paulo, SP, Brazil). External standards of gallic acid, syringic acid, benzoic acid, p-coumaric acid, o-coumaric acid, cinnamic acid, procyanidin B₁, catechin, procyanidin B₂, and caffeic acid came from Sigma-Aldrich (St. Louis, MO, USA). Epigallocatechin gallate, epicatechin, epicatechin gallate, procyanidin A₂, quercetin 3-glucoside, rutin, kaempferol 3-glucoside, myricetin, and isorhamnetin 3-glucoside, quercetin-3-O-glucopyranoside came from Extrasynthese (Genay, France). The trans-Resveratrol was obtained from Cayman Chemical Company (Michigan, EUA, Ann Arbor, MI, USA).

We greased fly medium by paste of hydrolyzed yeast containing one of the substances. Control untreated animals were fed by yeast past without substances. To make the hydrolyz at yeast were boiled in water bath for 30 minutes. To prepare the 100 ml of paste 50 g of dry yeast per 60 mL of water were used.
Flies were treated throughout their whole lives with the substances as follows: aspirin (2-(acetyloxy)-benzoic acid), valeryl salicylate (2-[(1-oxopentyl)oxy]-benzoicacid), trans-resveratrol ((E)-5-[2-(4-hydroxyphenyl)ethenyl]-1,3-benzenediol), SC-560 (5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-(3fluoromethyl)-1H-pyrazole), APHS (2-(2-heptynylthio)-phenol-acetate), NS-398 (N-[2-(cyclohexyloxy)-4-nitrophenyl]-methanesolfonamide), SC-58125 (5-(4-fluorophenyl)-1-[4-(methylsulfonyl)-phenyl]-3-(trifluoromethyl)-1H-pirazole), valdecoxib (4-(5-methyl-3-phenyl-4-isoxazolyl)-benzenesulfonamide), CAY10404 (3-(4-methylsulphonylphenyl)-4-phenyl-5-trifluoromethylisoxazole), licofelone (6-(4-chlorophenyl)-2, 3-dihydro-2,2-dimethyl-7-phenyl-1H-pyrrolizine-5-acetic acid) at the concentrations of 0.05, 0.5, 1 μM (Cayman Chemical, USA).

Myricetin, 3,4-dihydroxy phenyl ethanol, homovanillic acid sulfate, equol, genistein, phloretin, kaempferol, enterolactone, hesperetin, isorhamnetin, protocatechuic acid, trans-resveratrol, roseoflavin, pyrogallol, isoferulic acid, urolithin A, phloroglucinol, (R)-γ-valerolactone and freight were purchased from Cayman company (Ann Arbor, MI, USA). Other polyphenol standards were purified in-house through chromatographic methods with mass spectrometry/NMR confirmation. Authentic standards corresponding to the measured metabolites were purchased from Sigma-Aldrich (Saint Louis, MO, USA) or IROA Technologies (Boston, MA, USA). The stable isotope-labeled amino acid mix (20 AA U-13C, 97–99%; U-15N, 97–99%) was purchased from Cambridge Isotope Laboratories (Tewksbury, MA, USA). HPLC-MS-grade acetonitrile, ammonium acetate, and acetic acid were all purchased from Fisher Scientific (Pittsburgh, PA, USA).

MDA-MB-231-luc-D3H2LN cells (Xenogen) and MCF7 cells (American Type Culture Collection) were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum and 1% antibiotic-antimycotic (Thermo Fisher Scientific) at 37° C in 5% CO₂. Trans-resveratrol (Cayman Chemical) was used at concentrations ranging from 12.5–50 mM.

Among 56 phenolic compounds, isoflavonoids, phenylpropanoids, and stilbenoid standards were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA), along with dimethyl sulfoxide (99.9%). The anthocyanin standards were purchased from Extrasynthese Impasse Jacquard, (Rue Jacquard, Genay, France). Biochanin A, daidzein, daidzin, formononetin, genistein, genistin, and glycitin were obtained from LC Laboratories (New Boston St. Woburn, MA, USA), while glycitein was purchased from Wako Pure Chemical Industries, Ltd. (Chuo-ku, Osaka, Japan), and polydatin (Piceid) and trans-resveratrol was obtained from Cayman Chemicals (Ann Arbor, MI, USA). HPLC grade methanol, absolute ethanol, and acetonitrile were purchased from Fisher Scientific (Seongnam-Si, Gyeonggi-Do, Korea), while formic acid (>98%) was obtained from Junsei Chemical Co., Ltd. (Chuo-ku, Tokyo, Japan). A 0.1 N standard solution of hydrochloric acid was purchased from Dae-Jung Chemicals & Metals Co., Ltd. (Shiheung-city, Gyeonggi-do, Korea), and deionized water with a resistivity of 18.2 MΩ was obtained from a PURELAB Option-Q System (Lane End, High Wycombe HP14 3BY, UK).

External standards for 1-kestose, nystose, raffinose, glucose, fructose, rhamnose, maltose and for citric tartaric, lactic and acetic acids were purchased from Sigma-Aldrich (St. Louis, MO, USA). External standards for formic and succinic acids were purchased from Merck (Darmstadt, HE, Germany). External standards for procyanidin A2, epigallocatechin gallate, epicatechin gallate, kaempferol 3-glucoside, quercetin 3-rutinoside (rutin), quercetin 3-glucoside, myricetin, came from Extrasynthese (Genay, France). cis-Resveratrol and trans-resveratrol were obtained from Cayman Chemical Company (Ann Arbor, MI, USA). External standards of the phenolics: gallic acid, p-coumaric acid, chlorogenic acid, syringic acid, trans-caftaric acid, caffeic acid, hesperidin, naringenin, procyanidin B1, catechin, epicatechin and procyanidin B2, were purchased from Sigma-Aldrich. Acetonitrile, methanol, potassium persulfate, gallic acid and ethyl alcohol were obtained from Merck. The ultrapure water was obtained using a Marte Científica purification system (São Paulo, SP, Brazil). The chromogen 3,3′diaminobenzidine tetrachloride was purchased from Vector Laboratories (Burlingame, CA, USA). All antibodies used in this study were purchased from Abcam (Cambridge, MA, UK). All chemical reagents used in the experiments were of analytical grade.