Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Comparison of AOP1 EC 50 s evaluated in Caco-2 and HepG2 cells for the 12 natural extracts selected for the intestinal barrier transport study. Extracts were selected according to the best EC 50 s obtained with AOP1 assay. In order to increase the diversity of tested extracts, some Vitis vinifera extracts were removed from the list because of redundancy with each other. ND = not determined (no full effect observed).

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques:

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Indirect antioxidant ARE/Nrf2 assay data obtained for Syzygium aromaticum extract (given as an example) in HepG2/Nrf2 cells. All other extracts were subjected to the same protocol and data analyses. ( A ) Assay of increasing concentrations showing the typical bell-shaped profile (see text for explanation). Sulforaphane 25 µM was used as positive control; ( B ) dose–response and sigmoid fit curve used for Efficacy Concentration (EC) estimations.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques: Positive Control, Concentration Assay

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: ARE/Nrf2 EC 50 s established in HepG2 cells for the 12 natural extracts selected for the Caco-2 intestinal barrier transport study. ND = not determined; R 2 = determination coefficient.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques:

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Effect of post-intestinal barrier transport compartments on ARE/NRF2 pathway activation in HepG2 cells. Results are expressed as fold increase compared to constitutive medium ARE/Nrf2 gene expression. Values represent means of quadruplicates obtained from two independent intestinal barrier transport experiments. Sulforaphane 25 µM data is provided as positive control; *** p < 0.005.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques: Activation Assay, Expressing, Positive Control

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Effect of combinations of post-intestinal barrier compartments on ARE/NRF2 pathway in HepG2 cells. Results are expressed as ARE/Nrf2 gene expression fold increase compared to constitutive medium. Values represent means of pentaplicates for all 50/50 combinations and of triplicates for all others. All data were normalized to constitutive medium values; *** p < 0.001; ** p < 0.01.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques: Expressing

Journal: Antioxidants

Article Title: Cell-Based Antioxidant Properties and Synergistic Effects of Natural Plant and Algal Extracts Pre and Post Intestinal Barrier Transport

doi: 10.3390/antiox11030565

Figure Lengend Snippet: Synergistic effects on ARE/Nrf2 gene expression of post-intestinal barrier compartment combinations. Combin. FI = gene expression fold increase (FI) measure for the combination; S indep. FI = sum of the increase in gene expression fold increase (FI) measures of each of the two extracts taken independently as depicted in Figure 6 ; Var. (%) = variation in percentage of the two values. A positive percentage value indicates a synergistic effect.

Article Snippet: The HepG2/Nrf2 cells were treated with conditions for 17 h; then, they were treated with a mix (BPS Bioscience, USA) comprising cell lysis solution and luciferin (substrate of luciferase) for 40 min. Luminescence was read on a Varioskan Flash Spectral Scanning Multimode Reader.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Glucose-coated superparamagnetic iron oxide nanoparticles prepared by metal vapor synthesis can target GLUT1 overexpressing tumors: In vitro tests and in vivo preliminary assessment

doi: 10.1371/journal.pone.0269603

Figure Lengend Snippet: GLUT1 levels estimated by ELISA in PSN-1, BCPAP and HEK293 cells (at the top). Uptake studies: PSN-1 and BCPAP cells were incubated with 0.1 mg/mL of Glc-SPIONs for 30 min, 1, 3, 6, and 12 h. The Fe cellular content was estimated by GF-AAS analysis. Uptake studies after treating the cells with GLUT1 inhibitors: PSN1 and BCPAP cells were pre-incubated for 1 h with anti-GLUT1 polyclonal antibody, WZB117, Fasentin, BAY-876, and STF-31. Subsequently, cells were treated for 3 or 6 h with 0.1 mg/mL of Glc-SPIONs.

Article Snippet: Expression levels in cancer cells were evaluated by means of the GLUT1 Colorimetric Cell-Based ELISA kit (Boster Biological Technology, Pleasanton CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Incubation

Journal: Molecular Nutrition & Food Research

Article Title: Substrate‐Driven Differences in Tryptophan Catabolism by Gut Microbiota and Aryl Hydrocarbon Receptor Activation

doi: 10.1002/mnfr.202100092

Figure Lengend Snippet: Quantification of the AhR activity of supernatants collected during fermentation using the cell reporter assay. HepG2‐Lucia AhR reporter cells were incubated for 48 h with vehicle (medium or 1% v/v DMSO) and fermented samples. Trp, fermented with tryptophan; SP, fermented with isolated soybean protein; SC, fermented with single soybean cells; CC, fermented with clustered soybean cells. Results are normalized by control samples fermented without supplied substrates and expressed as mean ± SEM ( n = 3). Bars with different lowercase letters indicate treatments that are significantly different ( p < 0.05, two‐way ANOVA followed by a Tukey post‐hoc test).

Article Snippet: [ ] In brief, HepG2‐Lucia AhR reporter cells (InvivoGen, San Diego, CA) were grown and maintained according to the manufactures instructions.

Techniques: Activity Assay, Reporter Assay, Incubation, Isolation

Journal: Molecular Nutrition & Food Research

Article Title: Substrate‐Driven Differences in Tryptophan Catabolism by Gut Microbiota and Aryl Hydrocarbon Receptor Activation

doi: 10.1002/mnfr.202100092

Figure Lengend Snippet: Quantification of the AhR activity of SCFAs and tryptophan‐derived catabolites using the cell reporter assay. HepG2‐Lucia AhR reporter cells were incubated for 48 h with vehicle (medium or 1% v/v DMSO) and increasing concentrations of SCFAs or tryptophan‐derived catabolites, in the absence (white bars) or the presence (gray bars) of β‐NAPH (β‐naphthoflavone; 5 µmol L −1 ). Results are expressed as mean ± SEM ( n = 3). Bars with different lowercase or uppercase letters indicate treatments that are significantly different ( p < 0.05, one‐way ANOVA followed by a Tukey post‐hoc test).

Article Snippet: [ ] In brief, HepG2‐Lucia AhR reporter cells (InvivoGen, San Diego, CA) were grown and maintained according to the manufactures instructions.

Techniques: Activity Assay, Derivative Assay, Reporter Assay, Incubation

Journal: PLoS ONE

Article Title: Construction of the experimental rat model of gestational diabetes

doi: 10.1371/journal.pone.0273703

Figure Lengend Snippet: (A) Effects on the protein expression levels of GLUT1 and GLUT3 and the internal reference β-actin as determined by Western blot analysis, (B) Comparison of the protein expression levels of GLUT1 and GLUT3. * P < 0.05 vs. the NC group; # P < 0.05 vs. the HFHS group.

Article Snippet: After blocking with 5% skimmed milk in PBS-Tween, the membranes were probed with anti-glucose transporter 1 (GLUT1) or anti-glucose transporter 1 (GLUT3) primary antibody (1:500, Boeter) at 4°C overnight and conjugated with secondary antibodies at room temperature for 45 min. Antibodies against GLUT1 and GLUT3 were obtained from BOSTER Biological Technology Co., Ltd.

Techniques: Expressing, Western Blot, Comparison

Journal: Journal of Diabetes Research

Article Title: Distinct Effects of Carrageenan and High-Fat Consumption on the Mechanisms of Insulin Resistance in Nonobese and Obese Models of Type 2 Diabetes

doi: 10.1155/2019/9582714

Figure Lengend Snippet: Triglycerides and diacylglycerol are increased by HFD. (a–d) Oil Red O staining showed a marked increase in fat droplets in mouse hepatic tissue following high fat (c) and high fat with carrageenan (d) with a little sign of fat droplets in the carrageenan-treated (b) or control hepatic tissue (a). Scale bar = 50 μ m. (e) Triglycerides were measured in the hepatic and pancreatic tissues of the study mice and increased following HFD but not carrageenan ( p < 0.001, n = 12). (f) DAG levels in the hepatic and pancreatic tissues increased following HFD in the mice but not following carrageenan exposure ( p < 0.001, n = 12). (g) Palmitic acid increased DAG in the HepG2 cells ( p < 0.001, n = 3). CGN = carrageenan; DAG = diacylglycerol; PA = palmitic acid.

Article Snippet: Diacylglycerol (DAG) concentration was determined in tissue homogenates and in HepG2 cell extracts by a competitive enzyme immunoassay, as per directions (Aviva Systems Biology, San Diego, CA).

Techniques: Staining, Control

Journal: Journal of Diabetes Research

Article Title: Distinct Effects of Carrageenan and High-Fat Consumption on the Mechanisms of Insulin Resistance in Nonobese and Obese Models of Type 2 Diabetes

doi: 10.1155/2019/9582714

Figure Lengend Snippet: Serum galectin-3 increased following carrageenan or HFD. (a) Serum galectin-3 increased following carrageenan or HFD or their combination ( p < 0.001, n = 18). (b) Galectin-3 binding with the insulin receptor increased in the liver and muscle membrane preparations of the treated mice, with the greatest effect following the combined exposure ( p < 0.001, n = 12). (c) In HepG2 cells, the insulin-induced increase in phospho(Tyr)-IRS1 was significantly inhibited by administration of exogenous recombinant human galectin-3 ( p < 0.001, n = 3). (d) The insulin-induced glucose uptake in the HepG2 cells was blocked by administration of exogenous recombinant human galectin-3 ( p < 0.001, n = 3). (e) The Pearson correlation r between the glucose uptake and the phospho(Tyr)-IRS-1 in the HepG2 cells was 0.985. (f) Activity of the enzyme arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase), which removes 4-sulfate groups from the nonreducing end of chondroitin 4-sulfate and dermatan sulfate, was inhibited by exposure to carrageenan, but not by the HFD, in the liver and pancreas of the treated mice ( p < 0.001, n = 12). ARSB = arylsulfatase B; CGN = carrageenan; IRS = insulin receptor substrate.

Article Snippet: Diacylglycerol (DAG) concentration was determined in tissue homogenates and in HepG2 cell extracts by a competitive enzyme immunoassay, as per directions (Aviva Systems Biology, San Diego, CA).

Techniques: Binding Assay, Membrane, Recombinant, Activity Assay