Water-compatible molecularly imprinted polymers obtained via high-throughput synthesis and experimental design

A technique allowing high-throughput synthesis and evaluation of molecularly imprinted polymer sorbents at a reduced scale (mini-MIPs) was developed and used for the optimization of MIPs for use in pure aqueous environments. The technique incorporated a 4-port liquid-handling robot for the rapid dispensing of monomers, templates, solvents and initiator into the reaction vessels of a 96-well plate. A library of 80 polymers, each ca. 50 mg, could thus be prepared in 24 h. The MIP rebinding capacity and selectivity could be rapidly assessed in the batch mode by quantifying nonbound fractions in parallel using a UV monochromator plate reader. This allowed a complete evaluation of the binding characteristics of an 80 polymer library in approximately 1 week. With the objective of optimizing a polymer imprinted with the local anaesthetic Bupivacaine for use in pure aqueous systems, a polymer library was prepared by varying the original poly(MAA-co-EDMA) MIP composition. The variable factors were the added amount of the hydrophilic comonomer, 2-hydroxyethyl methacrylate (HEMA), the cross-linking ratio, and the porogen. This optimization resulted in polymers showing high imprinting factors (IF = K(MIP)/K(NIP)) in water as a result, mainly, of reduced binding to the nonimprinted polymer. Normal scale batches of these materials showed strong retention of the template and low nonspecific binding when assessed as chromatographic stationary phases using pure phosphate buffer, pH 7.4, as mobile phase, by equilibrium batch rebinding experiments and as sorbents for extractions of the analyte from blood plasma samples.     

Single nucleotide polymorphism analysis by allele-specific primer extension with real-time bioluminescence detection in a microfluidic device

A microfluidic approach for rapid bioluminescent real-time detection of single nucleotide polymorphism (SNP) is presented. The method is based on single-step primer extension using pyrosequencing chemistry to monitor nucleotide incorporations in real-time. The method takes advantage of the fact that the reaction kinetics differ between matched and mismatched primer-template configurations. We show here that monitoring the initial reaction in real time accurately scores SNPs by comparing the initial reaction kinetics between matched and mismatched configurations. Thus, no additional treatment is required to improve the sequence specificity of the extension, which has been the case for many allele-specific extension assays. The microfluidic approach was evaluated using four SNPs. Three of the SNPs included primer-template configurations that have been previously reported to be difficult to resolve by allele-specific primer extension. All SNPs investigated were successfully scored. Using the microfluidic device, the volume for the bioluminescent assay was reduced dramatically, thus offering a cost-effective and fast SNP analysis method.     

Structural and functional model of methane hydroxylase of membrane-bound methane monooxygenase from Methylococcus capsulatus (Bath)

Computer analysis of a wide range of primary sequences showed that -, -, and -peptides of membrane-bound methane hydroxylase contained 2, 7, and 6 transmembrane helices respectively. Conservative amino acid residues participating in complex formation were revealed. The - and -peptides are suggested to contain mononuclear copper ions with the ligand environment mainly consisting of His residues. The Cu sites are located in the hydrophilic region and are responsible for ESR signals. The active site of -peptide in which the activation of O₂ and oxidation of CH₄ occur is localized in the hydrophobic region close to the membrane surface. This site is formed by the amino acid residues of four transmembrane helices and one loop between them and is suggested to be a binuclear Cu—Fe or Fe—Fe center. The Cu site of -peptide transfers electrons to the active site of -peptide, and the Cu site of -peptide is either involved in this process or only stabilizes the protein structure.     

Remote sample characterization based on fluorescence monitoring

The possibilities and limitations of remote sample characterization using induced fluorescence are discussed. General equations for remote sensing of fluorescence are presented and discussed. The implications of different elements of a system for remote fluorescence sensing on different parameters of the equations are treated. Background light influence and signal processing are considered. Pulsed lasers, flashlamps and cw lamps as excitation sources are compared when matched to the proper detection scheme employing boxcar integration or lock-in detection. Model experiments have been performed to demonstrate different measurement strategies. Examples from remote oil-slick characterization are chosen as illustrations.     

Coherence effects in deep inelastic scattering

A superfield action for the relativistic massless superparticle as a spinning particle is presented in the new gauge. The symmetries in the relativistic superparticle theory, that is to say, the invariances under, the reparametrization and the local supersymmetry-transformation in the parameter space, are manifest in this formalism. It is clear how these two kinds of transformations descend from a unified origin to be called “super-reparametrization”, which is a restricted form of the general coordinate transformation in the superspace. The action is manifestly invariant under these transformation by its constructions The minimal coupling with electromagnetic field is also constructed in the superfield formalism, and a manifestly invariant superfield action for the interacting superparticle is presented in our gauge. The formalism is extended to the construction of an action for theN=2 superparticle.     

A new material for hydrogen storage; ScAl0.8Mg0.2

A novel aluminium rich alloy for hydrogen storage has been discovered, ScAl_0.8Mg_0.2, which has very promising properties regarding hydrogen storage capacity, kinetics and stability towards air oxidation in comparison to hydrogen absorption in state-of-the-art intermetallic compounds. The absorption of hydrogen was found to be very fast, even without adding any catalyst, and reversible. The discovered alloy crystallizes in a CsCl-type structure, but decomposes to ScH₂ and Al(Mg) during hydrogen absorption. Detailed analysis of the hydrogen absorption in ScAl_0.8Mg_0.2 has been performed using in situ synchrotron radiation powder X-ray diffraction, neutron powder diffraction and quantum mechanical calculations. The results from theory and experiments are in good agreement with each other.