Nonviral vectors are highly desirable
for the development of efficient
gene delivery systems. In this study, we report the monomolecular
condensation of plasmid DNA and efficient cell transfection by imidazolium
gemini surfactants ([C12-4-C12im]Br2), which could be a potential nonviral vector for efficient gene
therapy. Homogeneous DNA/[C12-4-C12im]Br2 nanoparticles are formed with a diameter of approximately
100 nm and investigated by using atomic force microscopy. DNA condensates
evolve from supercoiled DNA molecules, to individual toroids, to close-packed
particles, and eventually to multimolecular aggregates with the increase
of [C12-4-C12im]Br2 concentrations.
Highly efficient gene transfection in vitro is demonstrated in human
embryonic kidney 293 (HEK293) and HeLa cells, which could be attributed
to the effective DNA condensation into uniform nanoparticles induced
by [C12-4-C12im]Br2. In addition,
the low cytotoxicity of [C12-4-C12im]Br2 at transfection concentration region verified by cell viability
assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide,
MTT assay) also supports [C12-4-C12im]Br2 as an effective gene vector. The high gene transfection efficiency
by [C12-4-C12im]Br2 as well as its
low cytotoxicity could shed light on the rational molecular design
of nonviral vectors for gene delivery systems.
Microbial Fuel Cells (MFCs) are robust devices capable of taping biological energy, converting pollutants into electricity through renewable biomass. The fabrication of nanostructured electrodes with good bio- and electrochemical activity, play a profound role in promoting power generation of MFCs. Au nanoparticles (AuNPs)-modified Boron-Doped Diamond (BDD) electrodes are fabricated by layer-by-layer (LBL) self-assembly technique and used for the direct electrochemistry of Shewanella loihica PV-4 in an electrochemical cell. Experimental results show that the peak current densities generated on the Au/PAH multilayer-modified BDD electrodes increased from 1.25 to 2.93 microA/cm(-2) as the layer increased from 0 to 6. Different cell morphologies of S. loihica PV-4 were also observed on the electrodes and the highest density of cells was attached on the (Au/PAH)6/BDD electrode with well-formed three-dimensional nanostructure. The electrochemistry of S. loihica PV-4 was enhanced on the (Au/PAH)4/BDD electrode due to the appropriate amount of AuNPsand thickness of PAH layer.
BackgroundThe substrate, serving as carbon and energy source, is one of the major factors affecting the performance of microbial fuel cells (MFCs). We utilized BIOLOG system to rapidly screen substrates for electricigens, and further evaluated influence of these substrates on electricity generation of Shewanella loihica PV-4 in MFCs.ResultsThree of most favorable substrates (lactate acid, formic acid and cyclodextrin) with OD590/750 of 0.952, 0.880 and 0.849 as well as three of most unfavorable substrates (galactose, arabinose and glucose) with OD590/750 of 0.248, 0.137 and 0.119 were selected by BIOLOG system under aerobic conditions. The chronoamperometry results showed that MFCs fed with these substrates exhibited different current behaviors. Cyclic voltammograms results showed that arabinose, galactose and glucose promoted electron transfer from outer membrane c-Cyts of cells to the electrode surface. Lactic acid, formic acid and cyclodextrin produced lower quantity of electric charge of 10.13 C, 9.83 C and 10.10 C, the corresponding OD600 value was 0.180, 0.286 and 0.152 in BES; while galactose, arabinose and glucose generated higher quantity of electric charge of 12.34 C, 13.42 C and 17.45 C, and increased OD600 values were 0.338, 0.558 and 0.409 in BES. SEMs results showed that plenty of plump and stretched cells as well as appendages were observed when lactic acid, formic acid, and cyclodextrin were utilized as substrates, while sparse cells in short shape were obtained when galactose, arabinose and glucose were used as substrates.ConclusionsThese results suggest that substrate not only has important role in electrochemical performances of MFCs but also in biological properties of electricigens. Lactic acid, formic acid, and cyclodextrin beneficial for cell growth under aerobic conditions are unfavourable for planktonic cell growth and current generation under anaerobic conditions, while consumptions of galactose, arabinose and glucose adverse to cell growth under aerobic conditions are favourable for planktonic cell growth and current generation under anaerobic conditions due to the increase of cell numbers with more outer membrane c-Cyts transferring electrons between the electrode surface and cells.
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