High-Throughput Assay of Oxygen Radical Absorbance Capacity (ORAC) Using a Multichannel Liquid Handling System Coupled with a Microplate Fluorescence Reader in 96-Well Format

“…The ORAC values of LCS were evaluated according to the method of Huang et al (2002). For calibration, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox, Sigma-Aldrich; 6.25-50 mM) was used as a standard and was prepared fresh by diluting the stock solution (2 mM).…”

Section: Methodsmentioning

confidence: 99%

Probiotic attributes, antioxidant, anti-inflammatory and neuromodulatory effects of Enterococcus faecium CFR 3003: in vitro and in vivo evidence

Divyashri

Krishna

Muralidhara

et al. 2015

Journal of Medical Microbiology

View full text Add to dashboard Cite

Accumulating evidence suggests that probiotic bacteria play a vital role in modulating various aspects integral to the health and well-being of humans. In the present study, probiotic attributes and the antioxidant, anti-inflammatory and neuromodulatory potential of Enterococcus faecium CFR 3003 were investigated by employing suitable model systems. E. faecium exhibited robust resistance to gastrointestinal stress conditions as it could withstand acid stress at pH 1.5, 2 and 3. The bacterium also survived at a bile salt concentration of 0.45 %, and better tolerance was observed towards pepsin and trypsin. E. faecium produced lactic acid as a major metabolic product, followed by butyric acid. Lyophilized cell-free supernatant (LCS) of E. faecium exhibited significant antioxidant capacity evaluated against 1,1-diphenyl-2-picryl-hydrazyl, ascorbate autooxidation, oxygen radical absorbance and reducing power. Interestingly, E. faecium, Lactobacillus rhamnosus GG MTCC 1408 and LCS showed a significant anti-inflammatory effect by negatively modulating TNF-a production and upregulating IL-10 levels in LPS-stimulated macrophage cell lines. In an in vivo mice model, the propensity of probiotic supplements to modulate endogenous oxidative markers and redox status in brain regions was assessed. Young mice provided with oral supplements (daily for 28 days) of E. faecium and L. rhamnosus exhibited diminished oxidative markers in the brain and enhanced activities of antioxidant enzymes with a concomitant increase in c-aminobutyric acid and dopamine levels. Collectively, our findings clearly suggest the propensity of these bacteria to protect against tissue damage mediated through free radicals and inflammatory cytokines. Although the underlying molecular mechanisms need further studies, it is tempting to speculate that probiotics confer a neuroprotective advantage in vivo against oxidative damage-mediated neurodegenerative conditions.

show abstract

“…The ORAC assay was performed essentially as described by Huang et al (2002). A volume of 150 µl of working solution of sodium fluorescein (4 x 10-3 mM) was added to 25 µl samples, in a 12 well-microplate.…”

Section: Oxygen Radical Absorbance Capacity (Orac) Methodsmentioning

confidence: 99%

HPLC Fingerprint of Bioactive Compounds and Antioxidant Activities of Viscum album from Different Host Trees

Vicaş

Rugină

Leopold

et al. 2011

Not Bot Hort Agrobot Cluj

View full text Add to dashboard Cite

Viscum album L. is a medicinal plants used for many years as a remedy in the traditional medicine and in complementary cancer therapies. We investigated the influence of some host trees: Acer campestre (VAA), Fraxinus excelsior (VAF), Populus nigra (VAP), Malus domestica (VAM), and Robinia pseudoacacia (VAR), on the chemical composition and antioxidant activity of leaves and stems from V. album, as well the influence of the solvent (water and ethanol) used for extraction on biological activity. HPLC with photodiode array detector analysis of bioactive compounds from leaves and stems of mistletoes (V. album) hosted by five differents trees was performed. Antioxidant activities, determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH), Oxygen Radical Absorbance Capacity (ORAC), (Trolox-equivalent antioxidant capacity (TEAC) methods were compared with the quantification of total phenolics using the Folin -Ciocalteu reagent. V. album hosted by Fraxinus excelsior (VAF), recorded the highest level of total phenolic acids (108.64 µg/g dry matter) while the mistletoe hosted by Malus domestica (VAM) had the lowest level of total polyphenols (39.37 µg/g dry matter). In general, higher antioxidant activity was detected in the alcoholic than in the aqueous extracts. The values obtained for total phenolics in both, aqueous and ethanol extracts, decreased in the order: VAR > VAF > VAP > VAA > VAM. The mistletoe stem extracts contained lower levels of phenolics, as compared to leaves, in both solvents. Our results suggest that mistletoe extracts possess significant antioxidant activities that may be due to their bioactive compounds content, suggesting that this plant may be an important source of natural products with chemopreventive and chemotherapeutic activities.

show abstract

High-Throughput Assay of Oxygen Radical Absorbance Capacity (ORAC) Using a Multichannel Liquid Handling System Coupled with a Microplate Fluorescence Reader in 96-Well Format

Cited by 1,353 publications

References 19 publications

Optimization of microwave-assisted extraction of hydrophilic and lipophilic antioxidants from a surplus tomato crop by response surface methodology

Optimization of microwave-assisted extraction of hydrophilic and lipophilic antioxidants from a surplus tomato crop by response surface methodology

Probiotic attributes, antioxidant, anti-inflammatory and neuromodulatory effects of Enterococcus faecium CFR 3003: in vitro and in vivo evidence

HPLC Fingerprint of Bioactive Compounds and Antioxidant Activities of Viscum album from Different Host Trees

Contact Info

Product

Resources

About