Metal phosphonates applied to biotechnologies: a novel approach to oligonucleotide microarrays

A new process for preparing oligonucleotide arrays is described that uses surface grafting chemistry which is fundamentally different from the electrostatic adsorption and organic covalent binding methods normally employed. Solid supports are modified with a mixed organic/inorganic zirconium phosphonate monolayer film providing a stable, well-defined interface. Oligonucleotide probes terminated with phosphate are spotted directly on to the zirconated surface forming a covalent linkage. Specific binding of terminal phosphate groups with minimal binding of the internal phosphate diesters has been demonstrated. The mixed organic/inorganic thin films have also been extended for use arraying DNA duplex probes, and therefore represent a viable general approach to DNA-based bioarrays. Ideas for interfacing mixed organic/inorganic interfaces to other bioapplications are also discussed.     

Reversed-phase liquid chromatography and argentation chromatography of the minor capsaicinoids

An investigation of the liquid chromatography of the minor capsaicinoids in a commercial capsaicinoid mixture is reported. Twelve stationary phases including C₈, C₁₈, C₃₀, phenyl, and cation-exchange chemistries were examined in combination with isocratic aqueous methanol and aqueous acetonitrile mobile phases. A phenyl stationary phase and aqueous acetonitrile mobile phase baseline-resolved 7 of 11 capsaicinoids, and selected ion chromatograms (LC–ESI-MS) demonstrated this was the most effective reversed-phase separation. Argentation chromatography with an alkyl or phenyl column and aqueous silver nitrate–methanol mobile phase revealed the presence of the 6-ene-8-methyl and 6-ene-9-methyl homocapsaicin isomers and the absence of 7-ene-9-methyl homocapsaicin. A mixed phenyl–cation-exchange stationary phase (charged with silver ion) enabled unique and useful separations of the capsaicinoids.     

A chemical inhibitor reveals the role of Raf kinase inhibitor protein in cell migration

Raf kinase inhibitor protein (RKIP) is a modulator of cell signaling that functions as an endogenous inhibitor of multiple kinases. We demonstrate here a positive role for RKIP in the regulation of cell locomotion. We discovered that RKIP is the relevant cellular target of locostatin, a cell migration inhibitor. Locostatin abrogates RKIP's ability to bind and inhibit Raf-1 kinase, and it acts by disrupting a protein-protein interaction, an uncommon mode of action for a small molecule. Small interfering RNA-mediated silencing of RKIP expression also reduces cell migration rate. Overexpression of RKIP converts epithelial cells to a highly migratory fibroblast-like phenotype, with dramatic reduction in the sensitivity of cells to locostatin. RKIP is therefore the compound's valid target and a key regulator of cell motility.     

Theoretical description of pressure shifts and quenching of excited atomic states in liquid helium

The pressure dependence of the linewidth and shift for the 2 ³S→2 ³P and 3 ³S→2 ³P transitions observed in electron-bombarded liquid helium is calculated by applying static line-broadening theory within the framework of a theoretical model. The calculated pressure shift, width and asymmetry are in good agreement with recent measurements over the pressure range of 1 to 25 atm. A new mechanism is proposed for the strong pressure quenching observed for several atomic and molecular emission lines.     

Neomycin binding to Watson-Hoogsteen (W-H) DNA triplex groove: a model

Neomycin is the most effective aminoglycoside (groove binder) in stabilizing a DNA triple helix. It stabilizes TAT, as well as mixed base DNA triplexes, better than known DNA minor groove binders (which usually destabilize the triplex) and polyamines. Neomycin selectively stabilizes the triplex (in the presence of salt), without any effect on the DNA duplex. (1) Triplex stabilization by neomycin is salt dependent (increased KCl and MgCl(2) concentrations decrease neomycin's effectiveness, at a fixed drug concentration). (2) Triplex stabilization by neomycin is pH dependent (increased pH decreases neomycin's effectiveness, at a fixed drug concentration). (3) CD binding studies indicate approximately 5-7 base triplets/drug apparent binding site, depending upon the structure/sequence of the triplex. (4) Neomycin shows nonintercalative groove binding to the DNA triplex, as evident from viscometric studies. (5) Neomycin shows a preference for stabilization of TAT triplets but can also accommodate CGC(+) triplets. (6) Isothermal titration calorimetry (ITC) studies reveal an association constant of approximately 2 x 10(5) M(-)(1) between neomycin and an intramolecular triplex and a higher K(a) for polydA.2polydT. (7) Binding/modeling studies show a marked preference for neomycin binding to the larger W-H groove. Ring I/II amino groups and ring IV amines are proposed to be involved in the recognition process. (8) The novel selectivity of neomycin is suggested to be a function of its charge and shape complementarity to the triplex W-H groove, making neomycin the first molecule that selectively recognizes a triplex groove over a duplex groove.     

Use of CF3,Br/Al,discharges for reactive ion etching of III-V semiconductors

The reactive ion etching of GaAs, InP, InGaAs, and InAlAs in CF₃Br/Ar discharges was investigated as a function of both plasma power density (0.56-1.3 W - cm^–2) and total pressure (10-40 mTorr) The etch rate of GaAs in 19CF₃Br:1Ar discharges at 10 m Torr increases linearly with power density, from 600 Å min^–1 at 0.56 W · cm^–2, to 1550 Å · min at 1.3 W · cm^–2. The in-based materials show linear increases in etch rates only for power densities above – 1.0 W · cm^–2. These etch rates are comparable to those obtained with CCI₂F₂:O₂ mixtures under the same conditions. Smooth surface morphologies and vertical sidewalls are obtained over a wide range of plasma parameters. Reductions in the near-surface carrier concentration in n-type GaAs are evident for etching with power densities of >0.8 W cm^–2, due to the introduction of deep level trapping centers. At 1.3 W· cm^–2, the Schottky barrier height of TiPtAu contacts on GaAs is reduced from 0.74 to 0.53 eV as a result of this damage, and the photoluminescent intensity from the material is degraded. Alter RIE, we detect the presence of both F and Br on the surface of all of the semiconductors. This contamination is worse than with CCl₂F₂-based mixtures. High-power etching with CF₃Br/Ar together with Al-containing electrodes can lead to the presence of a substantial layer of aluminum oxide on the samples if the moisture content in the reactor is appreciable.     

An Interlaboratory Comparison of a Standardised EDTA Extraction Procedure for the Analysis of Available Trace Elements in Two Quality Control Soils

Abstract

A standardised EDTA extraction procedure was tested collaboratively by six laboratories using two in-house reference soils identified as soil A and soil B. The extracts were analysed for Zn, Cu, Pb and Mn by Inductively Coupled Plasma Spectrometry. Concentrations of extractable elements in soil A were generally much lower than those found in soil B. All laboratories produced some extreme outlying results, most of these were produced in soil B. Results for Mn were the most variable, with a range of 63.4–100.3 μg g^?1 in soil A and 226.4–415.3 μg g^?1 in soil B. In both soils, one laboratory reported high values for Zn and Mn and, one laboratory, for soil B, produced values for all four elements which were consistently low.

If outlying results are ignored, the results from most laboratories were in reasonable agreement for all elements except Mn.     

Computational analysis of relative stabilities of polyazine N-oxides

There is considerable interest in polyazine N-oxides as potential frameworks for energetic compounds with relatively high enthalpies of formation and crystal densities. The N⁺→O^? linkages, if appropriately located, may diminish the destabilization associated with nitrogen catenation. We have computationally characterized 40 N-oxides of the isomeric diazines, triazines, and tetrazines in terms of their geometries, relative energies, and (for a representative selection) electrostatic potentials. The presence of N⁺→O^? linkages does partially counteract the destabilizing effects of nitrogen catenation, although the isomers with complete catenation remain the least stable. The stabilizing influence of N⁺→O^? groups, and the accompanying changes in bond lengths, can be understood in terms of resonance charge delocalization to the polyazine rings. The N(O)–N(O) bonds between nitrogens that both bear oxygens tend to be relatively weak. The electrostatic potentials above the polyazine rings become increasingly positive as there are more nitrogens and oxygens; eventually they are positive above all of the carbons and nitrogens and possibly even the oxygens, with negative regions only on the peripheries of the molecules. However, the nitrogens that bear oxygens always have more positive potentials than those that do not.