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29
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33750681427
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note
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7 as solvent in 5 mm NMR tubes. Proton and carbon spectra were referenced internally to the TMS signal at 0.00 ppm. All spectra were measured by using the standard pulse programs installed by Bruker.
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30
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33750723240
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note
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2 (312.45): C, 65.35; H, 5.16; N, 8.97; S, 20.53. Found: C, 65.11; H, 5.27; N, 8.92; S, 20.66.
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31
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33750742879
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note
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3S (279.36): C, 68.79; H, 4.69; N, 15.04; S, 11.48. Found: C, 68.62; H, 4.91; N, 15.10; S, 11.62.
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32
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33750728683
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note
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3S (281.38): C, 68.30; H, 5.37; N, 14.93; S, 11.40. Found: C, 68.11; H, 5.53; N, 14.91; S, 11.67.
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34
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33750735835
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note
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Cervix adenocarcinoma (HeLa), breast adenocarcinoma (MCF7) and squamous skin carcinoma (A431) cells were maintained in minimal essential medium supplemented with 10% foetal bovine serum, antibiotic-antimycotic agents and non-essential amino acids. Briefly, human cancer cells were seeded onto a 96-well microplate. On the second day of the procedure, the original medium was removed and 200 μL new medium containing the test substances was added. The tested compounds were dissolved in DMSO, in a final concentration never exceeding 0.1%, which has no substantial effect on cell growth. After an incubation period of 72 h, living cells were assayed by the addition of 20 μL of 5 mg/mL MTT solution. MTT was converted by intact mitochondrial reductase and precipitated as blue crystals during a 4 h contact period. The medium was then removed and the precipitated crystals were dissolved in 100 mL DMSO during a 60 min period of shaking. Finally, the reduced MTT was assayed at 545 nm by using a microplate reader. All in vitro experiments were carried out on two microplates with at least 5 parallel wells.
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