Addressing of LnCaP Cell Using Magnetic Particles Assisted Impedimetric Microelectrode.

In this study, we provide a facile, effective technique for a simple isolation and enrichment of low metastatic prostate tumor cell LNCaP using biocompatible, magnetic particles asissted impedimetric sensing system. Hydrophobic cell membrane anchors (BAM) were generated onto magnetic particles which diameters vary from 50 nm to 5 µm and were used to capture LNCaP cells from the suspension. Finally, magnetic particle-LNCaP complex were addressed onto the surface of the interdigitated microelectrode (IDM). Cell viability was monitored by our laboratory developed-technique Electrical Cell Substrate Impedance Sensing (ECIS). The results reavealed that 50 nm-magnetic particles showed best performance in terms of cell separation and cell viability. This technique provides a simple and efficient method for the direct addressing of LNCaP cell on the surface and enhances better understanding of cell behavior for cancer management in the near future.

Fabrication of dual dye-doped silica nanotube as a fluorescent ratiometric pH sensor.

This study described a novel fabrication of fluorescence co-encapsulating silica nanotubes (F@SNT) and the further application of the as-synthesized nanostructure as a ratiometric pH sensor in buffer solution. Silica nanotubes (SNTs) embedded anodic alumina oxide (AAO) template was fabricated by sol-gel technique, tetramethyl rhodamine (TMR-the reference dye) was incorporated directly onto silica layer via hydrophobic interaction. Subsequently, fluorescent isothiocyanate (FITC-pH sensitive dye) was encapsulated inside poly-dimethylsiloxane (PDMS) matrix and the FITC-PDMS nanocomposite was doped into the hollow structure of SNT using nano-molding lithography. On removing AAO, free-standing SNTs were obtained and were subsequently applied as a ratiometric pH sensor in phosphate buffer solution. The dual dye-doped SNTs showed excellent fluorescence and a good pH sensing performance from pH 5.2-8.0. The results were distinguishable by the emission spectra and by fluorescent visualization. High photostability, sensitivity, biocompatibility with adjustable sizes make dual dye doped-SNT a promising nanostructure for bioapplications.

Purification and characterization of novel fibrinolytic proteases as potential antithrombotic agents from earthworm Perionyx excavatus.

Six protease fractions, namely FI, FII, FIII-1, FIII-2, FIII-3 and FIV, were isolated from Perionyx excavatus earthworm biomass by acetone precipitation, followed by serial chromatography using anion exchange, hydrophobic interaction and size exclusion chromatography. All fractions exhibited strong hydrolytic activity towards casein. The activity of six fractions towards fibrin, determined by fibrin plate assay, ranged from 44 to 831 plasmin unit.mg-1 and ranked as FIII-3 > FIII-2 > FI > FIII-1 > FIV > FII. Casein degradation was optimal at pH 7 and 11, and at 45-60°C. All fractions were considerably stable at high temperature and wide pH range. They were completely inhibited by phenylmethylsulfonyl fluoride (PMSF). The molecular weights (MW) and isoelectric points (pI) determined by 2D-electrophoresis were 27.5-34.5 kDa, and 4.3-5.2, respectively. Tandem mass spectrometry (MS) analysis was used to deduce the amino acid sequences of some peptides from FIII-1 and FIII-2. The sequences shared 16.9% and 13.2% similarity, respectively, with the fibrinolytic enzymes from two related earthworm species, Lumbricus rubellus and Eisenia fetida. The P. excavatus proteases were classified as serine proteases. They could perform rapid hydrolysis on both coagulated fibrous fibrin and soluble fibrinogen monomers without the presence of activators such as tPA or urokinase.