Journal: Pharmaceuticals

Article Title: Cuban Policosanol (Raydel ® ) Exerts Higher Antioxidant and Anti-Glycation Activities than Chinese Policosanol (BOC Sciences) in Reconstituted High-Density Lipoproteins: In Vivo Anti-Inflammatory Activities in Zebrafish and Its Embryos

doi: 10.3390/ph17040406

Figure Lengend Snippet: Electrophoresis of rHDL containing each policosanol (PCL) after synthesis with palmitoyloleoyl-phosphatidylcholine (POPC), free cholesterol (FC), and apolipoprotein A-I (apoA-I). The molar ratio of POPC:FC:apoA-I:PCL = 95:5:1:1. ( A ) Native electrophoresis of each rHDL under a non-denatured state on 0.6% agarose (15 μg of protein/lane) to compare electromobility depending on the three three-dimensional structures of apoA-I/HDL and its negative charge. The apoA-I in rHDL was also visualized via Coomassie brilliant blue staining (final 1.25%). The red and black arrowheads indicate the band position of rHDL-0 and rHDL-1, respectively. ( B ) Electrophoretic patterns of each rHDL in a denatured state on 14% SDS-PAGE (5 μg of protein/lane). The black arrowhead indicates phospholipid (PL) and policosanol (PCL) debris. Lane M, molecular weight marker (Bio-Rad 161-0374, precision plus protein standards). Lane A, lipid-free apoA-I—lane 0, rHDL alone; lane 1, rHDL containing PCL-1; lane 2, rHDL containing PCL-2. The gel was subjected to staining with Coomassie brilliant blue (0.125%) to facilitate visualization of apoA-I and phospholipids.

Article Snippet: Wheat germ policosanol (WPG) from Garuda International (Santa Cruz, CA, USA), which consisted of 2.1% tetracosanol (C24), 8.4% hexacosanol (C26), 67.9% octacosanol (C28), 12.6% triacontanol (C30), 5.9% dotriacontanol (C32), and 3.1% tetratriacontanol (C34), failed to lower the plasma cholesterol levels after four weeks of consumption with 20 mg of WPG in a chocolate pellet [ ].

Techniques: Electrophoresis, Staining, SDS Page, Molecular Weight, Marker

Journal: Pharmaceuticals

Article Title: Cuban Policosanol (Raydel ® ) Exerts Higher Antioxidant and Anti-Glycation Activities than Chinese Policosanol (BOC Sciences) in Reconstituted High-Density Lipoproteins: In Vivo Anti-Inflammatory Activities in Zebrafish and Its Embryos

doi: 10.3390/ph17040406

Figure Lengend Snippet: Transmission electron microscopy (TEM) images of each rHDL-incorporating policosanol, along with particle diameter measurements. A comparison of the particle morphology was conducted at 150 K magnification, revealing a consistent discoidal shape and a rouleaux pattern across all rHDL preparations. The inset graph illustrates the mean particle diameter ± SEM among various rHDL preparations. ns, no significant.

Article Snippet: Wheat germ policosanol (WPG) from Garuda International (Santa Cruz, CA, USA), which consisted of 2.1% tetracosanol (C24), 8.4% hexacosanol (C26), 67.9% octacosanol (C28), 12.6% triacontanol (C30), 5.9% dotriacontanol (C32), and 3.1% tetratriacontanol (C34), failed to lower the plasma cholesterol levels after four weeks of consumption with 20 mg of WPG in a chocolate pellet [ ].

Techniques: Transmission Assay, Electron Microscopy, Comparison

Journal: Pharmaceuticals

Article Title: Cuban Policosanol (Raydel ® ) Exerts Higher Antioxidant and Anti-Glycation Activities than Chinese Policosanol (BOC Sciences) in Reconstituted High-Density Lipoproteins: In Vivo Anti-Inflammatory Activities in Zebrafish and Its Embryos

doi: 10.3390/ph17040406

Figure Lengend Snippet: Comparative antioxidant potential of rHDL-containing policosanol to prevent cupric-ion-mediate oxidative damage of LDL. ( A ) Electrophoresis of LDL (10 μg of protein) treated with rHDL (0.5 μg of protein) comprising different origins of policosanols in 0.5% agarose gel using Tris-EDTA buffer (pH 8.0) at 50 V for 1 h. The separated bands of the apo-B fraction of LDL were stained using Coomassie brilliant blue (final 1.25%). The band positions of native LDL and oxidized LDL are indicated by a blue arrowhead and a red arrowhead, respectively. Lane N, native LDL; lane O, oxidized LDL (LDL + Cu 2+ ); lane 0, LDL + Cu 2+ + rHDL-0; lane 1, LDL + Cu 2+ + rHDL-1; lane 2, LDL + Cu 2+ + rHDL-2. ( B ) Quantification of thiobarbituric acid reactive substances (TBARS) in LDL challenged with cupric ion and subsequently treated with rHDL comprising policosanol. The values are represented as malondialdehyde (MDA, μM) in LDL using the MDA standard. The values in the bar graph represent the mean ± SD of three independent experiments. A pairwise statistical difference was established using a t -test by comparing the results with the ox-LDL value. *, p < 0.05 versus ox-LDL; **, p < 0.01 versus ox-LDL; ns, no significant.

Article Snippet: Wheat germ policosanol (WPG) from Garuda International (Santa Cruz, CA, USA), which consisted of 2.1% tetracosanol (C24), 8.4% hexacosanol (C26), 67.9% octacosanol (C28), 12.6% triacontanol (C30), 5.9% dotriacontanol (C32), and 3.1% tetratriacontanol (C34), failed to lower the plasma cholesterol levels after four weeks of consumption with 20 mg of WPG in a chocolate pellet [ ].

Techniques: Electrophoresis, Agarose Gel Electrophoresis, Staining

Journal: Pharmaceuticals

Article Title: Cuban Policosanol (Raydel ® ) Exerts Higher Antioxidant and Anti-Glycation Activities than Chinese Policosanol (BOC Sciences) in Reconstituted High-Density Lipoproteins: In Vivo Anti-Inflammatory Activities in Zebrafish and Its Embryos

doi: 10.3390/ph17040406

Figure Lengend Snippet: Anti-glycation activity of rHDL-containing policosanol in glycated HDL through fructose (Fruc, final 250 mM) treatment under 5% CO 2 at 37 °C. ( A ) Fluorescence spectroscopic analysis (Ex = 370 nm, Em = 440 nm) of HDL (2 mg/mL of protein), which was co-treated with fructose (final 250 mM) and each rHDL (2 mg/mL of apoA-I) containing policosanol (final 3 μg/mL) during 96 hr incubation. The data are expressed as the mean ± SD from three independent experiments with duplicate samples. Each rHDL treatment was compared with HDL + Fruc via a paired t -test. ( B ) Electrophoretic patterns of the HDL (5 μg/lane) after incubation with fructose and each rHDL after 96 hr incubation (15% SDS-PAGE). Yellow numbers indicate the band intensity in each lane. Lane 0, HDL alone at 0 hr incubation; lane 1, HDL alone at 96 hr incubation; lane 2, HDL + Fruc; lane 3, HDL + Fruc + rHDL-0; lane 4, HDL + Fruc + rHDL-1; lane 5, HDL + Fruc + rHDL-2.

Article Snippet: Wheat germ policosanol (WPG) from Garuda International (Santa Cruz, CA, USA), which consisted of 2.1% tetracosanol (C24), 8.4% hexacosanol (C26), 67.9% octacosanol (C28), 12.6% triacontanol (C30), 5.9% dotriacontanol (C32), and 3.1% tetratriacontanol (C34), failed to lower the plasma cholesterol levels after four weeks of consumption with 20 mg of WPG in a chocolate pellet [ ].

Techniques: Activity Assay, Fluorescence, Incubation, SDS Page

Journal: Pharmaceuticals

Article Title: Cuban Policosanol (Raydel ® ) Exerts Higher Antioxidant and Anti-Glycation Activities than Chinese Policosanol (BOC Sciences) in Reconstituted High-Density Lipoproteins: In Vivo Anti-Inflammatory Activities in Zebrafish and Its Embryos

doi: 10.3390/ph17040406

Figure Lengend Snippet: Evaluation of the comparative survivability and wound healing effects of recombinant high-density lipoprotein (rHDL) formulations containing Cuban (Raydel) or Chinese (BOC sciences) policosanol against carboxymethyllysine (CML)-impaired wounds in zebrafish. ( A ) Assessment of zebrafish survival within 48 h post-treatment. ( B ) The percentage of wound healing observed over the 48 h post-treatment period and calculated by comparing the stained wound area at different time points to the initial wound stained area at 0 h. The p -value documented the pairwise statistical variance retrieved from the ANOVA, employing the Dunnett’s test for post hoc analysis. ***, p < 0.001 versus CML + PBS; *, p < 0.05 versus CML + PBS; ns, non-significant. ( C ) Visual representation of the wounded area stained with 0.1% methylene blue. Wound healing was determined by a reduced methylene-blue-stained area (blue color). Red scale bar indicates 2 mm.

Article Snippet: Wheat germ policosanol (WPG) from Garuda International (Santa Cruz, CA, USA), which consisted of 2.1% tetracosanol (C24), 8.4% hexacosanol (C26), 67.9% octacosanol (C28), 12.6% triacontanol (C30), 5.9% dotriacontanol (C32), and 3.1% tetratriacontanol (C34), failed to lower the plasma cholesterol levels after four weeks of consumption with 20 mg of WPG in a chocolate pellet [ ].

Techniques: Recombinant, Staining

Journal: Pharmaceuticals

Article Title: Cuban Policosanol (Raydel ® ) Exerts Higher Antioxidant and Anti-Glycation Activities than Chinese Policosanol (BOC Sciences) in Reconstituted High-Density Lipoproteins: In Vivo Anti-Inflammatory Activities in Zebrafish and Its Embryos

doi: 10.3390/ph17040406

Figure Lengend Snippet: Comparative histological evaluation of different policosanol-embedded recombinant high-density lipoproteins (rHDLs) against carboxymethyl lysine (CML)-impaired cutaneous wounds in adult zebrafish. ( A ) Hematoxylin and eosin (H&E) staining. The black and red arrows symbolize densely and loosely arranged muscular tissue, respectively. The blue arrows symbolize the presence of granulation tissue. ( B ) Dihydroethidium (DHE) fluorescent staining for detecting reactive oxygen species (ROS). ( C ) Acridine orange (AO) staining to examine the extent of apoptosis. [Scale bar = 200 μm]. ( D ) Image J-based quantification of ROS and AO fluorescent intensity. Values in the bar graph represent mean ± SEM of three independent experiments. * and *** denote statistical significance at the p < 0.05 and p < 0.001 levels, respectively, for DHE fluorescent intensity. While ### signifies a p < 0.001 significance for AO fluorescent intensity compared to the CML-alone group, ns indicates a non-significant difference between the groups.

Article Snippet: Wheat germ policosanol (WPG) from Garuda International (Santa Cruz, CA, USA), which consisted of 2.1% tetracosanol (C24), 8.4% hexacosanol (C26), 67.9% octacosanol (C28), 12.6% triacontanol (C30), 5.9% dotriacontanol (C32), and 3.1% tetratriacontanol (C34), failed to lower the plasma cholesterol levels after four weeks of consumption with 20 mg of WPG in a chocolate pellet [ ].

Techniques: Recombinant, Staining

Journal: Pharmaceuticals

Article Title: Cuban Policosanol (Raydel ® ) Exerts Higher Antioxidant and Anti-Glycation Activities than Chinese Policosanol (BOC Sciences) in Reconstituted High-Density Lipoproteins: In Vivo Anti-Inflammatory Activities in Zebrafish and Its Embryos

doi: 10.3390/ph17040406

Figure Lengend Snippet: Comparative effect of different recombinant high-density lipoproteins (rHDLs) containing policosanol against carboxymethyllysine (CML)-induced interleukine-6 (IL-6) production in the hepatic tissue of adult zebrafish. ( A ) Immunohistochemistry (IHC) for IL-6 detection ( a1 – a5 ) at 400× magnification). The ( b1 – b5 ) images represent the IHC-stained area (brown color) interchanged with red color (at brown-color threshold values 20 (lower limit) and 120 (upper limit) using image J software) [Scale bar = 100 μm]. ( B ) Image-J-based quantification of IL-6-stained area (brown color interchanged to red color). The values in the bar graph represent mean ± SEM. *** denotes statistical significance at p < 0.001 compared to the CML + PBS injected group; ns signifies a non-significant different between the groups.

Article Snippet: Wheat germ policosanol (WPG) from Garuda International (Santa Cruz, CA, USA), which consisted of 2.1% tetracosanol (C24), 8.4% hexacosanol (C26), 67.9% octacosanol (C28), 12.6% triacontanol (C30), 5.9% dotriacontanol (C32), and 3.1% tetratriacontanol (C34), failed to lower the plasma cholesterol levels after four weeks of consumption with 20 mg of WPG in a chocolate pellet [ ].

Techniques: Recombinant, Immunohistochemistry, Staining, Software, Injection

Journal: Pharmaceuticals

Article Title: Cuban Policosanol (Raydel ® ) Exerts Higher Antioxidant and Anti-Glycation Activities than Chinese Policosanol (BOC Sciences) in Reconstituted High-Density Lipoproteins: In Vivo Anti-Inflammatory Activities in Zebrafish and Its Embryos

doi: 10.3390/ph17040406

Figure Lengend Snippet: The comparative effect of different policosanol-embedded recombinant high-density lipoproteins (rHDLs) against carboxymethyl lysine (CML)-impaired kidney damage of adult zebrafish. ( A ) Hematoxylin and eosin (H&E) staining. PT and DT symbolize the proximal and distal tubules, respectively. The red arrows point to the lumen cell debris. ( B ) Dihydroethidium (DHE) fluorescent staining for detecting reactive oxygen species (ROS). ( C ) Acridine orange (AO) staining to examine the extent of apoptosis. [Scale bar = 100 μm at 400× magnification]. ( D ) Image-J-based quantification of DHE and AO fluorescent intensity. Values in the bar graph represent mean ± SEM. * and *** denote statistical significance at the p < 0.05 and p < 0.001 levels, respectively, for AO fluorescent intensity compared to the CML + PBS treated group. While ### signifies a p < 0.001 significance for DHE fluorescent intensity compared to the CML + PBS-treated group, ns indicates a non-significant difference between the groups.

Article Snippet: Wheat germ policosanol (WPG) from Garuda International (Santa Cruz, CA, USA), which consisted of 2.1% tetracosanol (C24), 8.4% hexacosanol (C26), 67.9% octacosanol (C28), 12.6% triacontanol (C30), 5.9% dotriacontanol (C32), and 3.1% tetratriacontanol (C34), failed to lower the plasma cholesterol levels after four weeks of consumption with 20 mg of WPG in a chocolate pellet [ ].

Techniques: Recombinant, Staining