A series of novel amphiphilic chitosan derivatives, cholic acid modified N-(2-hydroxy)-propyl-3-trimethylammonium chitosan chloride (HTCC-CA) with different quaternization degrees and cholic acid substitutions were synthesized in this study. HTCC-CA is biocompatible and forms particles in aqueous solution. The binding with superoxide dismutase (SOD) at pH 6.8 destroys the original aggregates of HTCC-CA and produces smaller SOD/HTCC-CA complex nanoparticles via electrostatic and hydrophobic interactions. The SOD loading efficiency and loading capacity of HTCC-CA can reach to more than 90% and 45%, respectively. Confocal laser scanning microscopy observation and flow cytometry analysis reveal that SOD/HTCC-CA complex nanoparticles greatly enhance the cellular internalization of the loaded SOD. The SOD activities and malonaldehyde concentrations in the serum and organs of the rats, administrated intravenously with free SOD, free HTCC-CA, and SOD/HTCC-CA nanoparticles, were assayed to evaluate the antioxidant efficiency in vivo. The results demonstrate that free HTCC-CA is effective to scavenge superoxide radicals in the blood circulation and SOD/HTCC-CA nanoparticles have better antioxidant efficiency than free SOD as well as free HTCC-CA.
A general strategy for constructing high-effective upconversion nanocapsules based on triplet-triplet annihilation (TTA) was developed by loading both sensitizer and annihilator into BSA-dextran stabilized oil droplets. This strategy can maintain high translational mobility of the chromophores, avoid luminescence quenching of chromophore by aggregation, and decrease the O2-induced quenching of TTA-based upconversion emission. Pt(II)-tetraphenyl-tetrabenzoporphyrin (PtTPBP) and BODIPY dyes (BDP-G and BDP-Y with the maximal fluorescence emission at 528 and 546 nm, respectively) were chosen as sensitizer/annihilator couples to fabricate green and yellow upconversion luminescent emissive nanocapsules, named UCNC-G and UCNC-Y, respectively. In water under the atmospheric environment, interestingly, UCNC-G and UCNC-Y exhibit intense upconversion luminescence (UCL) emission (?ex = 635 nm) with the quantum efficiencies (?UCL) of 1.7% and 4.8%, respectively, whereas very weak UCL emission (?UCL < 0.1%) was observed for the corresponding previous reported SiO2-coating nanosystems because of aggregation-induced fluorescence quenching of annihilators. Furthermore, application of theses upconversion nanocapsules for high-contrast UCL bioimaging in vivo of living mice without removing the skin was demonstrated under 635-nm excitation with low power density of 12.5 mW cm(-2).
A preparative reversed-phase high performance liquid chromatography-electrospray ionization mass spectrometry (RP-HPLC/ESI-MS) method to obtain low cost casein peptides was developed. Casein was hydrolyzed by trypsin and the hydrolysate was analyzed by RP-HPLC/ESI-MS firstly. The gradient elution program in an analytical column was optimized to achieve a better separation. The effect of pH of the mobile phase on the separation was also investigated. Then, the optimal analytical conditions were applied in a preparative column directly. In each loading of the hydrolysate, several pure peptide fractions were collected automatically by the inductions of ultraviolet absorbance and mass spectrometry signals together. The process is simple and effective. The influence of the loading amount in a preparative column was investigated intensively. For the collection consisting of hydrophilic peptides that were poorly separated in the primary process, secondary separation was performed. By changing the gradient elution, several different pure peptides were obtained.
In this research, a simple, green and effective strategy was developed to produce long-term stable oil in water emulsion from soy protein and soy polysaccharide. Soy protein and soy polysaccharide formed dispersible complexes at pH around 3.25 aqueous solution through electrostatic and hydrophobic interactions. A high pressure homogenization produced the protein/polysaccharide complex emulsion having a droplet size about 250 nm. A heat treatment of the emulsion resulted in the protein denaturation, forming irreversible oil-water interfacial films composed of soy protein/soy polysaccharide complexes. The droplets of the emulsion were characterized by dynamic light scattering, ?-potential, transmission electron microscopy, polysaccharide digestion via pectinase, and confocal laser scanning microscopy observation via dual fluorescence probes. As a result of the polysaccharide being fixed on the droplet surface, the emulsions exhibited long-term stability in the media containing pH values of 2-8 and 0.2 mol/L NaCl. The stable soy protein/soy polysaccharide complex emulsion is a suitable food-grade delivery system in which lipophilic bioactive compounds can be encapsulated.
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