Journal: Nutrients

Article Title: Distinct Postprandial Bile Acids Responses to a High-Calorie Diet in Men Volunteers Underscore Metabolically Healthy and Unhealthy Phenotypes

doi: 10.3390/nu12113545

Figure Lengend Snippet: Interindividual variability of maximal absolute postprandial change from fasting sate in circulating BA levels. ( A ) Primary BAs. ( B ) Secondary and Tertiary BAs. ( C ) Sulfo-conjugated BAs. ( D ) Total BAs, unconjugated, conjugated BAs, and BA synthesis marker 7α-hydroxy-4-cholesten-3-one, C4.

Article Snippet: The 7α-hydroxy-4-cholesten-3-one (C4) and 7α-hydroxy-4-cholesten-3-one-D7 (C4-D7) were obtained from Avanti Polar Lipid.

Techniques: Marker

Journal: Biochemistry

Article Title: Sterol Structure Strongly Modulates Membrane-Islet Amyloid Polypeptide Interactions.

doi: 10.1021/acs.biochem.7b01190

Figure Lengend Snippet: Figure 3. Effect of different sterols on membrane leakage. The time course of carboxyfluorescein leakage experiments are displayed for vesicles containing 100 mol % POPC (red), 80 mol % POPC, and 20 mol % sterol: cholesterol (blue), cholestanol (cyan), lathosterol (dark cyan), 7-dehydrocholesterol (yellow), epicholesterol (green), pregne- nolone (pink), cholestenone (dark red), coprostanol (purple), and 5α- cholestan-3-one (gray). Experiments were conducted in 20 mM Tris· HCl, 100 mM NaCl, pH 7.4 at 25 °C with 400 μM lipid and 20 μM hIAPP.

Article Snippet: 5αCholestan-3-one, cholestenone, coprostanol, and epicholesterol were obtained from steraloids.

Techniques: Membrane

Journal: Biochemistry

Article Title: Sterol Structure Strongly Modulates Membrane-Islet Amyloid Polypeptide Interactions.

doi: 10.1021/acs.biochem.7b01190

Figure Lengend Snippet: Figure 5. Effect of different sterols on amyloid formation. Vesicles were prepared with different sterols and the zwitterionic lipid POPC. The results of thioflavin-T assays are displayed. Data are plotted for hIAPP in solution (black), LUVs containing 100 mol % POPC (red), and LUVs containing 80 mol % POPC and 20 mol % of the following sterols: cholesterol (blue), cholestanol (cyan), lathosterol (dark cyan), 7-dehydrocholesterol (yellow), epicholesterol (green), pregnenolone (pink), cholestenone (dark red), coprostanol (purple), and 5α- cholestan-3-one (gray). Experiments were conducted in 20 mM Tris· HCl, 100 mM NaCl, pH 7.4 at 25 °C with 400 μM lipid and 20 μM hIAPP.

Article Snippet: 5αCholestan-3-one, cholestenone, coprostanol, and epicholesterol were obtained from steraloids.

Techniques:

Journal: Nutrients

Article Title: Distinct Postprandial Bile Acids Responses to a High-Calorie Diet in Men Volunteers Underscore Metabolically Healthy and Unhealthy Phenotypes

doi: 10.3390/nu12113545

Figure Lengend Snippet: Interindividual variability of maximal absolute postprandial change from fasting sate in circulating BA levels. ( A ) Primary BAs. ( B ) Secondary and Tertiary BAs. ( C ) Sulfo-conjugated BAs. ( D ) Total BAs, unconjugated, conjugated BAs, and BA synthesis marker 7α-hydroxy-4-cholesten-3-one, C4.

Article Snippet: The 7α-hydroxy-4-cholesten-3-one (C4) and 7α-hydroxy-4-cholesten-3-one-D7 (C4-D7) were obtained from Avanti Polar Lipid.

Techniques: Marker

Journal:

Article Title: Identification of bile acid precursors as endogenous ligands for the nuclear xenobiotic pregnane X receptor

doi: 10.1073/pnas.0237082100

Figure Lengend Snippet: Hepatic mRNA expression levels of several enzymes involved in bile acid biosynthesis are altered in the Cyp27a1−/− mouse. Relative hepatic RNA levels were determined by Northern analysis from male, mixed-strain, wild-type and Cyp27a1−/− mice fed a standard rodent diet. The values shown in boxes represent the mean ± SEM of the fold change in mRNA levels (n = 6 mice per genotype). Bile acid intermediates that would be predicted to accumulate upstream of CYP3A11 because of these changes in enzyme levels include the following: 1, 7α-hydroxycholesterol (5-cholesten-3β,7α-diol); 2, 7α-hydroxy-4-cholesten-3-one (4-cholesten-7α-ol-3-one); 3, 5β-cholestan-3α,7α,12α-triol; 4, 4-cholesten-3-one; and 5, cholestanol (5α-cholestan-3β-ol).

Article Snippet: 5-Cholesten-3β 7α-diol (7α-OH-cholesterol), 4-cholesten-7α-ol-3-one (7α-OH-4-cholesten-3-one), and 4-cholesten-3-one were purchased from Steraloids (Newport, RI).

Techniques: Expressing, Northern Blot

Journal:

Article Title: Identification of bile acid precursors as endogenous ligands for the nuclear xenobiotic pregnane X receptor

doi: 10.1073/pnas.0237082100

Figure Lengend Snippet: Identification of bile acid intermediates that activate PXR. (A) CV-1 cells were cotransfected with mouse or human PXR and the XREM-luciferase reporter (17), mouse FXR and the FXREIBABPx3-tk-luciferase reporter (34), mouse CAR and DR3rat CYP3A1-tk-luciferase, or mouse VDR and mSPPx3-tk-luc (19). Cells were exposed to various ligands, all provided at 10 μM, except 4-cholesten-3-one (lower bar of the pair; 33 μM), CDCA (100 μM), TCPOBOP (0.5 μM), and 1,25-dihydroxyvitamin D3 (100 nM) for ≈40 h before assaying for luciferase activity. Transfection efficiency was corrected by analysis of β-galactosidase activity because of the cotransfection of a constitutive lacZ expression reporter. See Fig. ​Fig.11 for the structures of bile acid intermediates. (B) Dose–response analysis of 5β-cholestan-3α,7α,12α-triol and 7α-OH-4-cholesten-3-one activation of mouse PXR and human PXR. Transfection experiments were performed as in A, except secreted placental alkaline phosphatase was used to correct for transfection activity and results are expressed as relative luciferase units (RLU).

Article Snippet: 5-Cholesten-3β 7α-diol (7α-OH-cholesterol), 4-cholesten-7α-ol-3-one (7α-OH-4-cholesten-3-one), and 4-cholesten-3-one were purchased from Steraloids (Newport, RI).

Techniques: Luciferase, Activity Assay, Transfection, Cotransfection, Expressing, Activation Assay

Journal:

Article Title: Identification of bile acid precursors as endogenous ligands for the nuclear xenobiotic pregnane X receptor

doi: 10.1073/pnas.0237082100

Figure Lengend Snippet: 5β-Cholestan-3α,7α,12α-triol and 7α-hydroxy-4-cholesten-3-one directly bind mouse PXR as determined by fluorescence polarization assay. A fluorescein-tagged SRC1 peptide (ILRKLLQE) was incubated with bacterially expressed, purified GST-mPXR (black bars) or GST (white bars) in the presence of vehicle (DMSO), PCN (10 μM), or bile acid intermediates at 10 μM. Ligand-induced recruitment of the fluorescein-tagged SRC1 peptide was monitored by an increase in millipolarization fluorescence units (mP). Data shown represent the mean ± SEM of six samples.

Article Snippet: 5-Cholesten-3β 7α-diol (7α-OH-cholesterol), 4-cholesten-7α-ol-3-one (7α-OH-4-cholesten-3-one), and 4-cholesten-3-one were purchased from Steraloids (Newport, RI).

Techniques: Fluorescence, Incubation, Purification