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1.
Molecules ; 21(5)2016 Apr 30.
Article in English | MEDLINE | ID: mdl-27144557

ABSTRACT

Bullfrog oil is a natural product extracted from the Rana catesbeiana Shaw adipose tissue and used in folk medicine for the treatment of several diseases. The aim of this study was to evaluate the extraction process of bullfrog oil, to develop a suitable topical nanoemulsion and to evaluate its efficacy against melanoma cells. The oil samples were obtained by hot and organic solvent extraction processes and were characterized by titration techniques and gas chromatography mass spectrometry (GC-MS). The required hydrophile-lipophile balance and the pseudo-ternary phase diagram (PTPD) were assessed to determine the emulsification ability of the bullfrog oil. The anti-tumoral activity of the samples was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for normal fibroblast (3T3) and melanoma (B16F10) cell lines. Both extraction methods produced yielded around 60% and the oil was mainly composed of unsaturated compounds (around 60%). The bullfrog oil nanoemulsion obtained from PTPD presented a droplet size of about 390 nm and polydispersity = 0.05 and a zeta potential of about -25 mV. Both the bullfrog oil itself and its topical nanoemulsion did not show cytotoxicity in 3T3 linage. However, these systems showed growth inhibition in B16F10 cells. Finally, the bullfrog oil presented itself as a candidate for the development of pharmaceutical products free from cytotoxicity and effective for antineoplastic therapy.


Subject(s)
Antineoplastic Agents/isolation & purification , Biological Products/therapeutic use , Melanoma, Experimental/drug therapy , Oils/therapeutic use , Rana catesbeiana , 3T3 Cells , Animals , Antineoplastic Agents/therapeutic use , Antineoplastic Agents/toxicity , Biomedical Research/trends , Cell Line, Tumor , Drug Screening Assays, Antitumor , Emulsions , HeLa Cells , Humans , Mice , Oils/chemistry , Oils/isolation & purification , Oils/toxicity
2.
Int Immunopharmacol ; 10(1): 34-42, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19804847

ABSTRACT

Caripia montagnei is a basidiomycete species which contains polysaccharides with immunomodulatory properties. An extract of this mushroom underwent removal of the fat content by organic solvent and subsequently proteolysis. The aqueous phase obtained after proteolysis was precipitated with methanol yielding a fraction containing carbohydrates (98.7+/-3.3%) and protein (1.3+/-0.25%). Chemical analysis, infrared spectroscopy and nuclear magnetic resonance (NMR) showed that the carbohydrate fraction contained (63.3+/-4.1) of beta-glucans and proteins (2.2+/-0.3%). These glucans (50mg/kg of body weight) significantly reduced the inflammatory infiltrate produced by thioglycolate-induced peritonitis by 75.5+/-5.2%, when compared to Wy-14643 (60.3+/-6.1%), PFOA (37.8+/-2.8%) and clofibrate (52.2+/-3.2%), p<0.001, which are of the peroxisome proliferator-activated receptor (PPAR-alpha). L-NAME, a nitric oxide synthase inhibitor, reduced the plantar edema in Wistar rats by 91.4+/-1.3% (p<0.001). A significant reduction in nitric oxide (NO) levels was observed in the exudates when the glucans was used in comparison to carrageenan. The C. montagnei glucans did not present signs of inducing cytotoxicity. A decrease in IL-1ra, IL-10 and IFN-gamma in the peritonitis model was observed. Thus, the results suggest that glucans from the C. montagnei mushroom is an effective immunomodulator and may have potential for anti-inflammatory properties.


Subject(s)
Anti-Inflammatory Agents/administration & dosage , Basidiomycota/immunology , Complex Mixtures/administration & dosage , Edema/immunology , Glucans/administration & dosage , Peritonitis/immunology , Animals , Anti-Inflammatory Agents/adverse effects , Anti-Inflammatory Agents/chemistry , Ascitic Fluid/chemistry , Ascitic Fluid/immunology , Ascitic Fluid/pathology , Caprylates/administration & dosage , Caprylates/pharmacology , Carrageenan/metabolism , Cell Movement/drug effects , Clofibrate/administration & dosage , Clofibrate/pharmacology , Complex Mixtures/adverse effects , Complex Mixtures/chemistry , Cytokines/biosynthesis , Cytokines/genetics , Edema/chemically induced , Edema/drug therapy , Fluorocarbons/administration & dosage , Fluorocarbons/pharmacology , Glucans/adverse effects , Glucans/chemistry , Leukocytes/drug effects , Leukocytes/immunology , Leukocytes/metabolism , Leukocytes/pathology , Magnetic Resonance Spectroscopy , Male , Mice , Nitric Oxide/analysis , Peritonitis/chemically induced , Peritonitis/drug therapy , Pyrimidines/administration & dosage , Pyrimidines/pharmacology , Rats , Rats, Wistar , Thioglycolates/metabolism
3.
Protein Pept Lett ; 12(7): 695-700, 2005 Oct.
Article in English | MEDLINE | ID: mdl-16522172

ABSTRACT

A beta-glucuronidase was purified from Pomacea sp. eggs by ammonium sulfate fractionation, DEAE-BioGel and Heparin-Sepharose chromatographies. This enzyme showed a Mr 180 kDa, with subunits of 90 kDa. The kinetic parameters were: pH 4.0, temperature 60 degrees C, Km 2.7 x 10(-6) and Vmax 15.3 microM/h, activator Mg+2, and inhibitor: lactone of D-saccharic acid. beta-glucuronidase is an exoglucuronidase involved in glycosaminoglycans metabolism with kinetics parameters similar to those found in mammals.


Subject(s)
Glucuronidase/isolation & purification , Glucuronidase/metabolism , Mollusca/embryology , Mollusca/enzymology , Animals , Embryonic Development , Enzyme Stability , Glucuronidase/chemistry , Hydrogen-Ion Concentration , Kinetics , Molecular Weight , Mollusca/classification , Temperature
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