Resveratrol assay

An extensive mutagenesis protocol was used to probe the contribution of these observed changes in the STS fold towards cyclization specificity in CHS. Initially, an 18X mutant of alfalfa CHS was created to probe the mechanistic relevance of the structural differences observed in STS. Introduction of the pine STS primary sequence, consisting of 18 amino acid changes in areas 1-3, into alfalfa CHS results in an enzyme with similar kinetic efficiency to wild-type CHS. However, the mutant now produces resveratrol as the major product by using coumaroyl-CoA and malonyl-CoA in assays (Fig. 12.9B). Further deconvolution of the necessary... 

J. W. Hirschelman, W. H. et ah Increasing the throughput and productivity of Caco-2 cell permeability assays using liquid chromatography-mass spectrometry application to resveratrol absorption and metabolism. Comb Chem High Throughput Screen 2003, 6, 757-767. 

The pharmacokinetic dispositions of other stilbenes that are structurally similar to resveratrol and have pharmacological activity across many anticancer, anti-inflammatory, and antioxidant assays have been studied. The pharmacokinetics was characterized in male Sprague-Dawley rats after single intravenous... 

Soleas GJ, Yan J, Goldberg DM. 2001b. Ultrasensitive assay for three polyphenols (catechin, quercetin and resveratrol) and their conjugates in biological fluids utilizing gas chromatography with mass selective detection. J Chromatogr B Biomed Sci Appl 757 161-172. 

Some further comments include the fact that resveratrol has been extensively studied at the University of Illinois at Chicago, an institution that is said to have the broadest-based chemoprevention drug discovery program in the world. (Information about resveratrol is also provided by Hoffman, 1999a, p. 214). Some 2500 natural substances have been tested under the direction of pharmacy professor John Pezzuto (who is cited elsewhere as coauthor with Mathew Suffness of a chapter in Methods in Plant Biochemistry, Vol. 6, Assays for Bioactivity, pp. 72ff, 116). This anticancer substance (resveratrol) has been identified in over 70 plant species, notably in red-grape skins (and in red wine), but also in mulberries and peanuts. 

The quality of data was such that even differences in activity between identical compounds supplied by different vendors sufficient to cause them to be misclassified using less rigorous assays could be noted. For example, resveratrol would be identified as active at 2.3 i.M in one sample but inactive in the other, when a 50% threshold is used. The authors suggest that qHTS shifts the analysis of screening data from a statistical to a pharmacological... 

Finally, we have assayed the in vivo antidepressant effect of a natural phenolic compound, frans-resveratrol, and an oxoisoaporphine derivative, 5-methoxy-7/f-dibenzo[Je,/i]quinolin-7-one, 0X0 4 (Figime 12). The antidepressant-like effect of trans-resveratrol and 0X0 4 was evaluated through a Forced Swimming Test (FST), and they were compared with reference antidepressants imipramine, desipramine, nomifensine, bupropion, nisoxetine, citalopram and moclobemide. In this sense, we evaluate 0X0 4 at 15, 30 and 45 min. affording the mayor reduction at 30 min after the administration. 

Pharmacometrics of stilbenes and its relationship with clinical studies were recently reviewed [9]. The aim of the research [31] was to study the sulfation of resveratrol in the human liver and duodenum. A simple and reproducible radiometric assay for resveratrol sulfation was developed. It employed 3 -phosphoadenosine-5 -phospho-sulfate-[35S] as the sulfate donor, and the rate of resveratrol sulfation (pmol/min/mg cytosolic protein) were found to be 90 (liver) and 74 (duodenum). Resveratrol sulfotransferase followed Michaelis-Menten kinetics, and (fiM) was 0.63 (liver) and 0.50 (duodenum) and (pmol/min/mg cytosolic protein) were 125 (liver) and... 

To explore the contribution of resveratrol to the proliferation and apoptosis of lens epithelial cells, the thiazolyl tetrazolium (MTT) assay was conducted [47]. The stilbene effects on the proliferation of lens epithelial cells cultured with different concentrations of this compound were measured. HE staining, transmission electron microscopy (TEM), TUNEL fluorescence staining, and the aimexin V assay... 

Figure 7.4 Resveratrol (Res) induces loss of cell viability in IM-S and IM-R cells. IM-S (a) and IM-R (b) K562 cells (lO /ml) were incubated for 48 h (filled bars) or 72 h (open bars) at 37°C with increasing concentrations of resveratrol in 100)tl of RPMI 1640 medium containing 5% FCS in 96-well plates. Cell viability was measured by the XTT assay. Results are mean SD of four different determinations. Error bars = 95% confidence intervals. Resveratrol in the l-lOOpM range or 1 pM imatinib (Ima) was added to IM-S (c) or IM-R (d) CML cell lines growing in semisolid methyl...

The aim of the research [44] was to study the sulfation of resveratrol in the human liver and duodenum A simple and reproducible radiometric assay for resveratrol sulfation was developed. It employed 3 -phosphoadenosine-5 -phosphosulfate-[35S]... 

---