Control of Enzymatic Degradation of Microbial Polyesters by Plasma Modification

The rate of enzymatic degradation of surface‐modified microbial polyesters, poly[(R)‐3‐hydroxybutyrate] and poly[(R)‐3‐hydroxybutyrate‐co‐3‐hydroxyvalerate], was studied. The plasma treatments were carried out in a CF₃H or O₂ environment. It was found that the CF₃H plasma‐treated polyesters exhibited significant retardation of enzymatic erosion because of the surface fluorocarbon groups induced by CF₃H plasma. These surface fluorocarbon groups act as retardants on enzymatic degradation due to increased hydrophobicity and of the inactivity of enzymes. However, the increased surface hydrophilicity of polyesters induced by O₂ plasma results in no significant acceleration of the enzymatic erosion, which may be due to the thin modified layer.

Weight loss profiles of P(3HB) film exposed to CF₃H plasma as a function of plasma exposure time.     

Preliminary investigation of the application of on-line membrane extraction of trifluoroacetic acid as an aid to improvement of negative ion electrospray mass spectrometry data

We have recently investigated the biodegradation of a number of acidic aromatic compounds that give excellent chromatography using trifluoroacetic acid (TFA) based HPLC methods. Unfortunately HPLC methods using TFA are not usually compatible with detection by negative ion mass spectrometry as TFA suppresses ionisation of the analyte during the electrospray process. We present a preliminary investigation of the use of an anion-exchange micro-membrane suppressor to remove TFA on-line post column with the aim of improvement of mass spectral data using an aromatic acid as an example, Thus LC-MS using a TFA based HPLC method with negative ion mass spectral detection is shown to be possible with good sensitivity.     

Highly efficient Heck reactions of aryl bromides with n-butyl acrylate mediated by a palladium/phosphine-imidazolium salt system

[reaction: see text] A new phosphine-imidazolium salt, L.HBr (1, L = (1-ethylenediphenylphosphino-3-(mesityl))imidazol-2-ylidene), has been prepared. A combination of 0.5 mol % of Pd(dba)(2) and 0.5 mol % of L.HBr in the presence of 2 equiv of Cs(2)CO(3) as base has proven to be highly efficient in the Heck coupling reactions of aryl bromides (from electron-deficient to electron-rich aryl bromides) with n-butyl acrylate.     

Determination of triphenyltin hydroxide derivatives by capillary GC and tin-selective FPD

A new method for the determination of triphenyltin hydroxide using capillary column gas chromatography with a tin-selective flame photometric detector has been developed. Triphenyltin hydroxide and its potential metabolites are converted to methyl derivatives and separated on glass capillary columns coated with OV-101. Derivatization of triphenyltin hydroxide, triphenyltin chloride, diphenyltin dichloride, phenyltin trichloride, and bis-triphenyltin oxide is nearly quantitative with a minimum of redistribution products. The selectivity of the flame photometric detector is cearly demonstrated by the comparison of chromatographic profiles obtained from using both the flame photometric and flame ionization detectors. The use of this chromatographic system in the analysis of triphenyltin hydroxide in a fortified water sample demonstrates the potential use of this system in organotin residue chemistry.     

Immobilization of antibody fragment for immunosensor application based on surface plasmon resonance

Biosurface fabrication using the Fab′ fragment of immunoglobulin (IgG) was carried out by self-assembly (SA) technique. The pepsin-digested monoclonal antibody (Mab) against bovine insulin containing the F(ab′)₂ fragment and residual proteins was separated using affinity chromatography and dialysis. To prevent the nonspecific binding of F(ab′)₂ onto gold (Au) substrate, the native disulfide bridge was reduced using dithiothreitol (DTT) to convert F(ab′)₂ into Fab′, which made the immobilization to be carried out via the native thiol (–SH) group. The fabricated biosurface using SA technique showed the formation of stable thin film through AFM topography. Through the concentration change of DTT and Fab′, the absorption characteristics against the Au surface were investigated using surface plasmon resonance (SPR) with the flow cell. The amount of immobilized antibody fragment and the antigen binding capacity were regulated with respect to the reduction state and concentration of F(ab′)₂. Based on the biosurface of the fabricated Fab′, the insulin-detection was carried out by the measurement of SPR. The proposed antibody surface could successfully detect the bovine insulin at the concentration from 100 ng/mL to 10 μg/mL.     

Heavy hydrides: H2Te ultraviolet photochemistry

The room-temperature ultraviolet absorption spectrum of H2Te has been recorded. Unlike other group-6 hydrides, it displays a long-wavelength tail that extends to 400 nm. Dissociation dynamics have been examined at photolysis wavelengths of 266 nm (which lies in the main absorption feature) and 355 nm (which lies in the long-wavelength tail) by using high-n Rydberg time-of-flight spectroscopy to obtain center-of-mass translational energy distributions for the channels that yield H atoms. Photodissociation at 355 nm yields TeH(2Pi1/2) selectively relative to the TeH(2Pi3/2) ground state. This is attributed to the role of the 3A' state, which has a shallow well at large R(H-TeH) and correlates to H+TeH(2Pi1/2). Note that the 2Pi1/2 state is analogous to the 2P1/2 spin-orbit excited state of atomic iodine, which is isoelectronic with TeH. The 3A' state is crossed at large R only by 2A", with which it does not interact. The character of 3A' at large R is influenced by a strong spin-orbit interaction in the TeH product. Namely, 2Pi1/2 has a higher degree of spherical symmetry than does 2Pi3/2 (recall that I(2P1/2) is spherically symmetric), and consequently 2Pi1/2 is not inclined to form either strongly bonding or antibonding orbitals with the H atom. The 3A'<--X transition dipole moment dominates in the long-wavelength region and increases with R. Structure observed in the absorption spectrum in the 380-400 nm region is attributed to vibrations on 3A'. The main absorption feature that is peaked at approximately 240 nm might arise from several excited surfaces. On the basis of the high degree of laboratory system spatial anisotropy of the fragments from 266 nm photolysis, as well as high-level theoretical studies, the main contribution is believed to be due to the 4A" surface. The 4A"<--X transition dipole moment dominates in the Franck-Condon region, and its polarization is in accord with the experimental observations. An extensive secondary photolysis (i.e., of nascent TeH) is observed at 266 and 355 nm, and the corresponding spectral features are assigned. Analyses of the c.m. translational energy distributions yield bond dissociation energies D0. For H2Te and TeH, these are 65.0+/-0.1 and 63.8+/-0.4 kcalmol, respectively, in good agreement with predictions that use high-level relativistic theory.     

A microfabricated capillary electrophoresis chip with multiple buried optical fibers and microfocusing lens for multiwavelength detection

We present a new microfluidic device utilizing multiwavelength detection for high-throughput capillary electrophoresis (CE). In general, different fluorescent dyes are only excited by light sources with appropriate wavelengths. When excited by an appropriate light source, a fluorescent dye emits specific fluorescence signals of a longer wavelength. This study designs and fabricates plastic micro-CE chips capable of performing multiple-wavelength fluorescence detection by means of multimode optic fiber pairs embedded downstream of the separation channel. For detection purposes, the fluorescence signals are enhanced by positioning microfocusing lens structures at the outlets of the excitation fibers and the inlets of the detection fibers, respectively. The proposed device is capable of detecting multiple samples labeled with different kinds of fluorescent dyes in the same channel in a single run. The experimental results demonstrate that various proteins, including bovine serum albumin and beta-casein, can be successfully injected and detected by coupling two light sources of different wavelengths to the two excitation optic fibers. Furthermore, the proposed device also provides the ability to measure the speed of the samples traveling in the microchannel. The developed multiwavelength micro-CE chip could have significant potential for the analysis of DNA and protein samples.     

A surface plasmon resonance biosensor for detecting Pseudomonas aeruginosa cells with self-assembled chitosan-alginate multilayers

A self-assembled multilayer (SAMu) including the alginate layer was prepared for detecting Pseudomonas aeruginosa cells in a solution and its potential was evaluated with a BIAcore system. After layer-by-layer formation, the refractive units (RU) values monitored with the biosensor increased by the interaction between the layers. The responses by the binding of P. aeruginosa cells to the alginate-immobilized SAMu were visualized immediately upon injection of the cell suspension. The RU values after injection of the cells were measured with approximately 1152, 656 and 173 for 1 × 10⁹, 1 × 10⁸ and 1 × 10⁷ CFU/ml. This result suggests that the alginate-immobilized SAMu will have useful application for detecting P. aeruginosa cells in a biosensor analysis.     

Synthesis of the apoptosis inducing agent apoptolidin. Assembly of the C(16)-C(28) fragment

[reaction--see text] A stereoselective synthesis of the C(16)-C(28) fragment of the apoptosis inducing agent apoptolidin is described. Key steps include two propionate aldol reactions and a stereoselective Mukaiyama aldol addition of enolsilane 19 to beta-methoxy aldehyde 4.     

Electrophoretic characterization of a novel cysteine protease produced by Vibrio harveyi

Electrophoretic characterization of a novel cysteine protease produced by pathogenic luminous Vibrio harveyi, originally isolated from diseased tiger prawn Penaeus monodon in Taiwan, is demonstrated in the present study using native polyacrylamide gel electrophoresis (native PAGE), sodium dodecyl sulfate-PAGE (SDS-PAGE), crossed immunoelectrophoresis (CIE) and isoelectric focusing (IEF) gels. The protease has a pI of 6.4 and exhibits a fast-migrating feature in native-PAGE and CIE gels indicating that it is a negatively charged protease. The protease electrophoresed as a 22 kDa protein band in native- and SDS-PAGE (in SDS - buffer with or without the presence of 2-mercaptoethanol) while it electrophoresed as a 38 kDa protein band in SDS-PAGE when the samples were boiled for 10 min prior to electrophoresis. The results reveal that the enzyme is an SDS-resistant monomeric protease and its high negative charge is not influenced by SDS (detergent) without boiling the sample. The present results are useful in determining proteins of similar nature to this unique cysteine protease.