Journal: Journal of Lipid Research

Article Title: Peroxisome proliferator-activated receptor α controls cellular cholesterol trafficking in macrophages

doi: 10.1194/jlr.m500326-jlr200

Figure Lengend Snippet: Fig. 2. NPC1 and NPC2 gene expression is regu- lated by peroxisome proliferator-activated receptor (PPAR) agonists in human macrophages in a dose- dependent manner. Quantitative PCR analysis of NPC1 (A) and NPC2 (B) was performed on RNA iso- lated from primary human macrophages treated or not with Wy14643 (50 M), GW7647 (600 nmol/l), fenofibric acid (50 mol/l), ciprofibrate (50 M), or bezafibrate (50 M) for 24 h. Induction of NPC1 (C) and NPC2 (D) expression in human macrophages treated for 24 h with increasing concentrations of Wy14643 (10, 25, and 50 mol/l). NPC1 and NPC2 mRNA levels were normalized to cyclophilin mRNA and are expressed relative to the levels in untreated cells set as 1. Results are expressed as mean SD. Sta- tistically significant differences between treatments are indicated (ANOVA followed by Student’s t-test; *P 0.05, **P 0.01).

Article Snippet: Total cellular RNA was extracted from 10-day-old primary human macrophages treated or not with OxLDL or AcLDL (50 g/ ml) or the PPAR ligands Wy14643 (10, 25, and 50 mol/l), fenofibric acid (50 mol/l), ciprofibrate (50 mol/l), bezafibrate (50 mol/l), and GW7647 (600 nmol/l) for 24 h using Trizol (Life Technologies, France).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Expressing

Journal: Journal of Lipid Research

Article Title: Peroxisome proliferator-activated receptor α controls cellular cholesterol trafficking in macrophages

doi: 10.1194/jlr.m500326-jlr200

Figure Lengend Snippet: Fig. 3. PPAR activation induces NPC1 and NPC2 protein expression. Cell proteins were isolated from dif- ferentiated human macrophages treated or not with fenofibric acid (50 mol/l) for 24 h, and NPC1 (A) and NPC2 (B) protein levels were measured by Western blot analysis, as described in Materials and Methods. Spe- cific signals were quantified by densitometry. NPC1 and NPC2 protein levels in fenofibric acid-treated macro- phages were normalized to those in untreated control cells set as 1. Results are expressed as mean SD of results obtained from three independent cell preparations. Statistically significant differences between treat- ments are indicated (Student’s t-test; *P 0.05).

Article Snippet: Total cellular RNA was extracted from 10-day-old primary human macrophages treated or not with OxLDL or AcLDL (50 g/ ml) or the PPAR ligands Wy14643 (10, 25, and 50 mol/l), fenofibric acid (50 mol/l), ciprofibrate (50 mol/l), bezafibrate (50 mol/l), and GW7647 (600 nmol/l) for 24 h using Trizol (Life Technologies, France).

Techniques: Activation Assay, Expressing, Isolation, Western Blot, Control

Journal: Journal of Lipid Research

Article Title: Peroxisome proliferator-activated receptor α controls cellular cholesterol trafficking in macrophages

doi: 10.1194/jlr.m500326-jlr200

Figure Lengend Snippet: Fig. 5. PPAR agonists increase outer-layer plasma membrane cholesterol content. Human macrophages were pretreated for 24 h and thereafter every 24 h with Wy14643 (50 mol/l), GW7647 (600 nmol/l), fenofibric acid (50 mol/l), ciprofibrate (50 mol/l), and bezafibrate (50 mol/l) and cholesterol-loaded by incubation with [3H]cholesterol-AcLDL (50 g/ml) for 48 h. Cells were then fixed with paraformaldehyde before incubation for 1 h at 37C in PBS with or without cholesterol oxidase from Pseudomonas fluores- cens (1 U/ml). Cellular lipids were then extracted by hexane-iso- propanol and separated by TLC. Spots corresponding to free cho- lesterol and 4-cholestenone were scraped, and radioactivity was measured by scintillation counting. Results are expressed relative to untreated cells set as 1. Statistically significant differences between treatments are indicated (Student’s t-test; **P 0.01, ***P 0.001). Error bars indicate mean and SD.

Article Snippet: Total cellular RNA was extracted from 10-day-old primary human macrophages treated or not with OxLDL or AcLDL (50 g/ ml) or the PPAR ligands Wy14643 (10, 25, and 50 mol/l), fenofibric acid (50 mol/l), ciprofibrate (50 mol/l), bezafibrate (50 mol/l), and GW7647 (600 nmol/l) for 24 h using Trizol (Life Technologies, France).

Techniques: Clinical Proteomics, Membrane, Incubation, Radioactivity

Journal: Pharmaceutics

Article Title: Intrinsic Dissolution Rate Profiling of Poorly Water-Soluble Compounds in Biorelevant Dissolution Media

doi: 10.3390/pharmaceutics12060493

Figure Lengend Snippet: Physicochemical properties of active pharmaceutical ingredients (APIs).

Article Snippet: All compounds were purchased from Sigma Aldrich (St. Louis, MO, USA) with the exceptions of carvedilol (H. Lundbeck A/S, Valby, Denmark), felodipine (AstraZeneca, Mölndal, Sweden), fenofibric acid (APIChem, Hangzhou, China) and danazol (Toronto Research Chemicals, Inc., Toronto, Canada).

Techniques:

Journal: Pharmaceutics

Article Title: Intrinsic Dissolution Rate Profiling of Poorly Water-Soluble Compounds in Biorelevant Dissolution Media

doi: 10.3390/pharmaceutics12060493

Figure Lengend Snippet: API apparent solubility, experimentally determined intrinsic dissolution rates (IDR) from disc (D) or suspensions (S) of the compounds in fasted (FaSSIF-V1 and FaSSIF-V2) and fed (FeSSIF-V1 and FeSSIF-V2) state simulated intestinal fluids.

Article Snippet: All compounds were purchased from Sigma Aldrich (St. Louis, MO, USA) with the exceptions of carvedilol (H. Lundbeck A/S, Valby, Denmark), felodipine (AstraZeneca, Mölndal, Sweden), fenofibric acid (APIChem, Hangzhou, China) and danazol (Toronto Research Chemicals, Inc., Toronto, Canada).

Techniques: Solubility, Dissolution

Journal: Pharmaceutics

Article Title: Intrinsic Dissolution Rate Profiling of Poorly Water-Soluble Compounds in Biorelevant Dissolution Media

doi: 10.3390/pharmaceutics12060493

Figure Lengend Snippet: Dissolution profiles of the acidic compounds in fasted state simulated small intestinal fluid (FaSSIF) (circles, blue lines) and fed state simulated small intestinal fluid (FeSSIF) (squares, red lines). Dissolution of fenofibric acid, indomethacin, and naproxen were measured from discs, while bezafibrate, mefenamic acid, and tolfenamic acid were suspension assays. The dissolution profiles from the discs were acquired over several hours due to the small disc surface area, while suspension measurements were completed within 20 min.

Article Snippet: All compounds were purchased from Sigma Aldrich (St. Louis, MO, USA) with the exceptions of carvedilol (H. Lundbeck A/S, Valby, Denmark), felodipine (AstraZeneca, Mölndal, Sweden), fenofibric acid (APIChem, Hangzhou, China) and danazol (Toronto Research Chemicals, Inc., Toronto, Canada).

Techniques: Dissolution, Suspension