Biosorption of nickel, chromium and zinc by MerP-expressing recombinant Escherichia coli

Escherichia coli hosts able to over-express metal-binding proteins (MerP) originating from Gram-positive (Bacillus cereus RC607) and Gram-negative (Pseudomonas sp. K-62) bacterial strains were used to adsorb Ni(2+), Zn(2+) and Cr(3+) in aqueous solutions. The initial adsorption rate and adsorption capacity were determined to evaluate the performance of the biosorbents. With the expression of MerP protein, the metal adsorption capacity of the recombinant strains for Ni(2+), Zn(2+) and Cr(3+) significantly improved. The cells carrying Gram-positive merP gene (GB) adsorbed Zn(2+) and Cr(3+) at a capacity of 22.3 and 0.98 mmol/g biomass, which is 121% and 72% higher, respectively, over that of the MerP-free host cells. Adsorption capacity of the cells carrying Gram-negative merP gene (GP) also increased 144% and 126% for Zn(2+) and Cr(3+), respectively. Both recombinant strains also exhibited 24% and 5% enhancement in adsorption of Ni(2+) for GB and GP, respectively. The initial adsorption rate of the recombinant biosorbents was also higher than that of the MerP-free host, suggesting an increased metal-binding affinity with MerP expression. Severe cell damage on GB biosorbent was observed after Cr(3+) adsorption, probably due to the metal toxicity effect on the cells.     

Biodegradable quantum dot nanocomposites enable live cell labeling and imaging of cytoplasmic targets

Semiconductor quantum dots (QDs) offer great promise as the new generation of fluorescent probes to image and study biological processes. Despite their superior optical properties, QDs for live cell monitoring and tracking of cytoplasmic processes remain limited due to inefficient delivery methods available, altered state or function of cells during the delivery process and the requirement of surface-functionalized QDs for specific labeling of subcellular structures. Here, we present a noninvasive method to image subcellular structures in live cells using bioconjugated QD nanocomposites. By incorporating antibody-coated QDs within biodegradable polymeric nanospheres, we have designed a bioresponsive delivery system that undergoes endolysosomal to cytosolic translocation via pH-dependent reversal of nanocomposite surface charge polarity. Upon entering the cytosol, the polymer nanospheres undergo hydrolysis thus releasing the QD bioconjugates. This approach facilitates multiplexed labeling of subcellular structures inside live cells without the requirement of cell fixation or membrane permeabilization. As compared to conventional intracellular delivery techniques, this approach allows the high throughput cytoplasmic delivery of QDs with minimal toxicity to the cell. More importantly, this development demonstrates an important rational strategy for the design of a multifunctional nanosystem for biological applications.     

Nanoparticle-mediated cellular response is size-dependent

Nanostructures of different sizes, shapes and material properties have many applications in biomedical imaging, clinical diagnostics and therapeutics. In spite of what has been achieved so far, a complete understanding of how cells interact with nanostructures of well-defined sizes, at the molecular level, remains poorly understood. Here we show that gold and silver nanoparticles coated with antibodies can regulate the process of membrane receptor internalization. The binding and activation of membrane receptors and subsequent protein expression strongly depend on nanoparticle size. Although all nanoparticles within the 2-100 nm size range were found to alter signalling processes essential for basic cell functions (including cell death), 40- and 50-nm nanoparticles demonstrated the greatest effect. These results show that nanoparticles should no longer be viewed as simple carriers for biomedical applications, but can also play an active role in mediating biological effects. The findings presented here may assist in the design of nanoscale delivery and therapeutic systems and provide insights into nanotoxicity.     

Localized surface plasmon coupled fluorescence fiber-optic biosensor with gold nanoparticles

A novel fiber-optic biosensor based on a localized surface plasmon coupled fluorescence (LSPCF) system is proposed and developed. This biosensor consists of a biomolecular complex in a sandwich format of . It is immobilized on the surface of an optical fiber where a complex forms the fluorescence probe and is produced by mixing Cy5-labeled antibody and protein A conjugated gold nanoparticles (Au-PA). The LSPCF is excited by localized surface plasmon on the GNP surface where the evanescent field is applied near the core surface of the optical fiber. At the same time, the fluorescence signal is detected by a photomultiplier tube located beside the unclad optical fiber with high collection efficiency. Experimentally, this novel LSPCF biosensor is able to detect mouse immunoglobulin G (IgG) at a minimum concentration of 1 pg/mL (7 fM) during the biomolecular interaction of the IgG with anti-mouse IgG. The analysis is expanded by a discussion of the amplification of the LSPCF intensity by GNP coupling, and overall, this LSPCF biosensor is confirmed experimentally as a biosensor with very high sensitivity.     

Comprehensive three-dimensional gas chromatography with parallel factor analysis

Development of a comprehensive, three-dimensional gas chromatograph (GC3) instrument is described. The instrument utilizes two six-port diaphragm valves as the interfaces between three, in-series capillary columns housed in a standard Agilent 6890 gas chromatograph fitted with a high data acquisition rate flame ionization detector. The modulation periods for sampling column one by column two and column two by column three are set so that a minimum of three slices (more commonly four or five) are acquired by the subsequent dimension resulting in both comprehensive and quantitative data. A 26-component test mixture and quantitative standards are analyzed using the GC3 instrument. A useful methodology for three-dimensional (3D) data analysis is evaluated, based on the chemometric technique parallel factor analysis (PARAFAC). Since the GC3 instrument produces trilinear data, we are able to use this powerful chemometric technique, which is better known for the analysis of two-dimensional (2D) separations with multichannel detection (e.g., GC x GC-TOFMS) or multiple samples (or replicates) of 2D data. Using PARAFAC, we mathematically separate (deconvolute) the 3D data "volume" for overlapped analytes (i.e., ellipsoids), provided there is sufficient chromatographic resolution in each of the three separation dimensions. Additionally, PARAFAC is applied to quantify analyte standards. For the quantitative analysis, it is demonstrated that PARAFAC may provide a 10-fold improvement in the signal-to-noise ratio relative to a traditional integration method applied to the raw, baseline-corrected data. The GC3 instrument obtains a 3D peak capacity of 3500 at a chromatographic resolution of one in each separation dimension. Furthermore, PARAFAC deconvolution provides a considerable enhancement in the effective 3D peak capacity.     

Orthogonal polarization simultaneous readout for volume holograms with hybrid angle and polarization multiplexing in LiNbO(3)

The orthogonal polarization simultaneous readout technique in a hybrid-multiplexed memory using angular multiplexing and polarization multiplexing is presented. Twenty holograms were hybrid multiplexed in a single LiNbO(3) crystal with ten angular positions for angular multiplexing. In each angular position of the holographic memory, two images with orthogonal polarization are multiplexed in the same spatial location inside the LiNbO(3) via polarization multiplexing. These two orthogonally polarized images can be reconstructed simultaneously with a linear polarization reading beam, but they can be separated with a polarization beam splitter, and accordingly each can be viewed independently. The exposure schedule for holographic storage using the proposed hybrid-multiplexing technique is derived.     

Recovery of nitrotoluenes from wastewater by solvent extraction enhanced with salting-out effect

Toluene extraction enhanced by salting-out effect was employed to recover dinitrotoluene isomers and 2,4,6-trinitrotoluene (2,4,6-TNT) from wastewater of toluene nitration processes (e.g. dinitration or trinitration). The batchwise experiments were conducted to elucidate the influence of various operating variables on the extracting performance, including concentrations and species of inorganic salts, such as NaCl, KCl, Na(2)SO(4), K(2)SO(4) and MgSO4, acidity of wastewater, volume ratios of solvent versus wastewater and extraction stages in existence of inorganic salts. It was found that recovery of total organic compounds (TOC) was significantly elevated with increasing concentrations of salts, whose promoting effects were in the following order: NaCl>Na(2)SO(4)>K(2)SO(4)>MgSO4>KCl on the weight basis of wastewater. Besides, high volume ratio of toluene/wastewater (ca. 2.0) was more suitable for recovery of TOC from wastewater with or without addition of NaCl, of which extractable priority was as follows: 2,6-DNT>2,4-DNT>2,4,6-TNT. It is remarkable that TOC in wastewater would be almost completely recovered by sequential four stages toluene extraction, promoted continuously by salting-out effect.     

Spectral self-interference microscopy for low-signal nanoscale axial imaging

A theoretical and numerical analysis of spectral self-interference microscopy (SSM) is presented with the goal of expanding the realm of SSM applications. In particular, this work is intended to enable SSM imaging in low-signal applications such as single-molecule studies. A comprehensive electromagnetic model for SSM is presented, allowing arbitrary forms of the excitation field, detection optics, and tensor sample response. An evanescently excited SSM system, analogous to total internal reflection microscopy, is proposed and investigated through Monte Carlo simulations. Nanometer-scale axial localization for single-emitter objects is demonstrated, even in low-signal environments. The capabilities of SSM in imaging more general objects are also considered--specifically, imaging arbitrary fluorophore distributions and two-emitter objects. A data-processing method is presented that makes SSM robust to noise and uncertainties in the detected spectral envelope.     

Elucidating the mechanism of cellular uptake and removal of protein-coated gold nanoparticles of different sizes and shapes

We investigated the mechanism by which transferrin-coated gold nanoparticles (Au NP) of different sizes and shapes entered mammalian cells. We determined that transferrin-coated Au NP entered the cells via clathrin-mediated endocytosis pathway. The NPs exocytosed out of the cells in a linear relationship to size. This was different than the relationship between uptake and size. Furthermore, we developed a mathematical equation to predict the relationship of size versus exocytosis for different cell lines. These studies will provide guidelines for developing NPs for imaging and drug delivery applications, which will require "controlling" NP accumulation rate. These studies will also have implications in determining nanotoxicity.     

Optothermal property and decomposition characteristics of PtO(x) ultrathin film sandwiched between ZnS-SiO2 for super-resolution near-field recording

We have investigated the optothermal property and decomposition characteristics of PtO(x) ultrathin film protected by ZnS-SiO2 layers and effects of the constituent phases of PtO(x) on super-resolution capability and read stability of the super-RENS disk. All the ZnS-SiO2/PtO(x)/ZnS-SiO2 multilayers exhibited a steep reflectivity drop at the temperature range between 265 and 350 degrees C, corresponding to the decomposition of PtO(x). The decomposition temperature of the 4-nm-thick PtO(x) ultrathin film protected by ZnS-SiO2 layers was much lower than those obtained in thick PtO(x) films without protection. The activation energy for thermal decomposition was approximately 1.3 eV. Both the decomposition temperature and activation energy for thermal decomposition were unaffected by the constituent phases of PtO(x). Carrier to noise ratios (CNR) of over 40 dB for mark size of 150 nm were achieved in all super-resolution near-field structure (super-RENS) disks, while the super-resolution readout was limited to 2.5 x 10(3) approximately 4.5 x 10(4) cycles. The effect of constituent phases of PtO(x) on the super-resolution capability of super-RENS disk with a PtO(x) mask layer was minimal. However, as the constituent phases of PtO(x) mask layer transformed from a mixture of Pt and PtO, to pure PtO, and then to a mixture of PtO and PtO2, the readout stability of super-RENS disk increased dramatically since less heat was absorbed by the PtO(x) mask layer composed of PtO and PtO2 during the readout process, prohibiting the diffusion of materials inside the bubble to the GeSbTe phase change layer.