EpiQuik DNA Methyltransferase 1 kit and a 3B Activity/Inhibitor Screening Assay kit (Epigentek, Brooklyn, NY) were used to access inhibition of DNMT1 and DNMT3B, respectively. Ellagic acid, urolithin A, urolithin B, cyanidin-glucoside, cyanidin-rutinoside, protocatechuic acid, fructo-oligosaccharides, galacto-oligosaccharides, xylo-oligosaccharides, butyric acid, acetic acid, propionic acid, and valenic acid were purchased from SigmaAldrich (St. Louis, MO). Each compound was dissolved in dimethyl sulfoxide to prepare stock solution, then diluted to 1 nM, 100 nM, and 10 μM and tested in DNMT1 and DNMT3B inhibition assays. Each compound was tested in both assays in three independent runs.
Fructo oligosaccharides
Manufactured by Merck Group
Sourced in Canada, United States
Fructo-oligosaccharides are a type of carbohydrate that naturally occur in various plants. They are used as a food ingredient and in the production of dietary supplements. Fructo-oligosaccharides have a function as a prebiotic, which means they promote the growth of beneficial gut bacteria.
Automatically generated - may contain errors
Lab products found in correlation
Valenic acid, by Merck Group (2 mentions)
Propionic acid, by Merck Group (2 mentions)
Butyric acid, by Merck Group (2 mentions)
Epiquik dna methyltransferase 1 kit, by Epigentek (2 mentions)
A 3b activity inhibitor screening assay kit, by Epigentek (2 mentions)
Cyanidin glucoside, by Merck Group (2 mentions)
Cyanidin rutinoside, by Merck Group (2 mentions)
Galacto oligosaccharides, by Merck Group (2 mentions)
Xylo oligosaccharides, by Merck Group (2 mentions)
Urolithin a, by Merck Group (2 mentions)
Urolithin b, by Merck Group (2 mentions)
Acetic acid, by Merck Group (2 mentions)
Protocatechuic acid, by Merck Group (2 mentions)
Ellagic acid, by Merck Group (2 mentions)
Inulin, by Merck Group (1 mentions)
Gum arabic, by Merck Group (1 mentions)
4 protocols using fructo oligosaccharides
1
Screening Natural Compounds for DNMT Inhibition
2
Optimizing Bee Prebiotic and Probiotic Intake
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The prebiotics inulin, fructooligosaccharides, and gum arabic were obtained from Sigma-Aldrich (Oakville, ON, Canada). The Vetafarm Probotic was obtained from Vetafarm (Wagga Wagga, NSW, Australia). The probiotics Protexin Concentrate single-strain and Protexin Concentrate multi-strain were obtained from Probiotics International Limited (Lopen, Somerset, UK). Because the prebiotic compounds had not previously been tested on bees, doses were determined by looking at studies on other organisms, and adjusting the dose based on an average body weight of 100 mg per bee or average feed consumption over 16 days. For the probiotics, the feeding guide on the packaging (Vetafarm Probotic) or the manufacturer’s website (Protexin Concentrate single-strain and Protexin Concentrate multi-strain) was used. The appropriate dose for a bee was calculated using an average body weight of 100 mg per bee. The doses used are shown in Table 1 .
3
Screening Natural Compounds for DNMT Inhibition
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EpiQuik DNA Methyltransferase 1 kit and a 3B Activity/Inhibitor Screening Assay kit (Epigentek, Brooklyn, NY) were used to access inhibition of DNMT1 and DNMT3B, respectively. Ellagic acid, urolithin A, urolithin B, cyanidin-glucoside, cyanidin-rutinoside, protocatechuic acid, fructo-oligosaccharides, galacto-oligosaccharides, xylo-oligosaccharides, butyric acid, acetic acid, propionic acid, and valenic acid were purchased from SigmaAldrich (St. Louis, MO). Each compound was dissolved in dimethyl sulfoxide to prepare stock solution, then diluted to 1 nM, 100 nM, and 10 μM and tested in DNMT1 and DNMT3B inhibition assays. Each compound was tested in both assays in three independent runs.
4
Prebiotic Screening of Fermented Hemp Biomass
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The best performing time point, selected based on the best growth and pH reduction, was used to choose the FBH samples to screen for prebiotic activity, that was calculated with the related formula from two independent experiments and triplicates as previously described (Fissore, Santo Domingo, Gerschenson & Giannuzzi, 2015; (link)Huebner, Wehling, Parkhurst & Hutkins, 2008) , including qPCR quantifications (Nissen, di Carlo & Gianotti, 2020) (link). FBH samples were filtered (Minisart® Syringe Filter 0.22 μm, Sartorius, Gottingen, Germany) and then all samples including BH and TBH controls were freeze dried using a Savant freeze-dryer Lyolab 3000 apparatus (Thermo Fisher Scientific, USA), in order to add a 1g/dL of product to 10 mL of culture media.
FOS from chicory (Sigma, USA) was used as prebiotic positive control, and FH (commercial hemp seed flour) sample was used as an additional control, along with prebiotic positive control fructooligosaccharides (FOS) from chicory (Sigma, USA). The media employed as control to calculate the prebiotic scores were instead added with 1g/dL of glucose. The bacterial type strains Lp. plantarumLb. plantarum 98b, B. bifidum NCIMB 700795, and E. coli ATCC 25922 were used at final concentration of 6 Log10 CFU/mL (Fissore, Santo Domingo, Gerschenson & Giannuzzi, 2015; (link)Nissen, di Carlo & Gianotti, 2020) (link).
FOS from chicory (Sigma, USA) was used as prebiotic positive control, and FH (commercial hemp seed flour) sample was used as an additional control, along with prebiotic positive control fructooligosaccharides (FOS) from chicory (Sigma, USA). The media employed as control to calculate the prebiotic scores were instead added with 1g/dL of glucose. The bacterial type strains Lp. plantarumLb. plantarum 98b, B. bifidum NCIMB 700795, and E. coli ATCC 25922 were used at final concentration of 6 Log10 CFU/mL (Fissore, Santo Domingo, Gerschenson & Giannuzzi, 2015; (link)Nissen, di Carlo & Gianotti, 2020) (link).
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