Hypnea musciformis-mediated Ag/AgCl-NPs inhibit pathogenic bacteria, HCT-116 and MCF-7 cells' growth in vitro and Ehrlich ascites carcinoma cells in vivo in mice.

In the present study, Ag/AgCl-NPs were biosynthesised using Hypnea musciformis seaweed extract; NPs synthesis was confirmed by a change of colour and observation of a razor-sharp peak at 424 nm by UV-visible spectroscopy. Synthesised nanoparticles were characterised by transmission electron microscopy, energy-dispersive X-ray spectroscopy, X-ray powder diffraction and Fourier transform infrared spectroscopy. Bacterial cell growth inhibition proves that the Ag/AgCl-NPs have strong antibacterial activity and cell morphological alteration was observed in treated bacterial cells using propidium iodide (PI). Ag/AgCl-NPs inhibited Ehrlich ascites carcinoma (EAC) cells, colorectal cancer (HCT-116) and breast cancer (MCF-7) cell line in vitro with the IC50 values of 40.45, 24.08 and 36.95 µg/ml, respectively. Initiation of apoptosis in HCT-116 and MCF-7 cells was confirmed using PI, FITC-annexin V and Hoechst 33342 dye. No reaction oxygen species generation was observed in both treated and untreated cell lines. A significant increase of ATG-5 gene expression indicates the possibility of autophagy cell death besides apoptosis in MCF-7 cells. The initiation of apoptosis in EAC cells was confirmed by observing caspase-3 protein expression. Ag/AgCl-NPs inhibited 22.83% and 51% of the EAC cell growth in vivo in mice when administered 1.5 and 3.0 mg/kg/day (i.p.), respectively, for 5 consequent days.

Moringa oleifera seed lectin inhibits Ehrlich ascites carcinoma cell growth by inducing apoptosis through the regulation of Bak and NF-κB gene expression.

A Moringa oleifera seed lectin (MOSL) was purified by using chitin column with the molecular mass of 17±1kDa. The lectin agglutinated mouse, cow and human erythrocytes and the hemagglutination activity was inhibited by methyl-α-d-mannopyranoside, methyl-ß-d-galactopyranoside, lactose and glucose. The lectin exhibited 100% hemagglutination activity at the pH range from 8.0 to 9.0 and temperature range from 30 to 60°C. Additionally, the lectin gradually lost its activity in the presence of urea but the activity abolish completely when treated with EDTA. MOSL showed mild toxicity against brine shrimp nauplii with a LC50 value of 131.0µg/ml. Antiproliferative activity was studied against Ehrlich ascites carcinoma (EAC) cells and 71.08% cell growth inhibition was observed in vitro at 200µg/ml. The lectin was injected (i.p.) into EAC mice at the doses of 2.0 and 4.0mg/kg/day for five consecutive days and 25.38% and 55% of cell growth inhibition was observed, respectively. MOSL caused the cell cycle arrest at G2/M phase as determined by FACS flow cytometry. The cell growth inhibition was due to the induction of apoptosis in the EAC cells which was confirmed by cell morphological study, caspase-3 inhibitor and activation of Bak and suppression of Bcl-2 and NF-κB genes expression.

Purification and characterization of a novel chitinase from Trichosanthes dioica seed with antifungal activity.

Chitinases are a group of enzymes that show differences in their molecular structure, substrate specificity, and catalytic mechanism and widely found in organisms like bacteria, yeasts, fungi, arthropods actinomycetes, plants and humans. A novel chitinase enzyme (designated as TDSC) was purified from Trichosanthes dioica seed with a molecular mass of 39±1 kDa in the presence and absence of ß-mercaptoethanol. The enzyme was a glycoprotein in nature containing 8% neutral sugar. The N-terminal sequence was determined to be EINGGGA which did not match with other proteins. Amino acid analysis performed by LC-MS revealed that the protein was rich in leucine. The enzyme was stable at a wide range of pH (5.0-11.0) and temperature (30-90 °C). Chitinase activity was little bit inhibited in the presence of chelating agent EDTA (ethylenediaminetetraaceticacid), urea and Ca(2+). A strong fluorescence quenching effect was found when dithiothreitol and sodium dodecyl sulfate were added to the enzyme. TDSC showed antifungal activity against Aspergillus niger and Trichoderma sp. as tested by MTT assay and disc diffusion method.

Characterization of lecithin isolated from anchovy (Engraulis japonica) residues deoiled by supercritical carbon dioxide and organic solvent extraction.

UNLABELLED: Lecithin was isolated and characterized from anchovy (Engraulis japonica) deoiled residues using supercritical carbon dioxide (SC-CO(2)) at a semibatch flow extraction process and an organic solvent (hexane) extraction. SC-CO(2) extraction was carried out to extract oil from anchovy at different temperatures (35 to 45 °C) and pressures (15 to 25 MPa). Extraction yield of oil was influenced by physical properties of SC-CO(2) with temperature and pressure changes. The major phospholipids of anchovy lecithin were quantitatively analyzed by high-performance liquid chromatography. Phosphatidylcholine (PC) (68%± 1.00%) and phosphatidylethanolamine (PE) (29%± 0.50%) were the main phospholipids. Thin layer chromatography was performed to purify the individual phospholipids. The fatty acid compositions of lecithin, PC, and PE were analyzed by gas chromatography. A significant amount of eicosapentaenoic acid and docosahexaenoic acid were present in both phospholipids of PC and PE. Emulsions of lecithin in water were prepared through the use of a homogenizer. Oxidative stability of anchovy lecithin was high in spite of its high concentration of long-chain polyunsaturated fatty acids. PRACTICAL APPLICATION: Lecithin can be totally metabolized by humans, so is well tolerated by humans and nontoxic when ingested. Lecithin from anchovy contain higher amounts of ω-3 fatty acids especially EPA and DHA, it may have positive outcome to use in food and pharmaceutical industries.