Hybrid phospholipid bilayer coatings for separations of cationic proteins in capillary zone electrophoresis

Protein separations in CZE suffer from nonspecific adsorption of analytes to the capillary surface. Semipermanent phospholipid bilayers have been used to minimize adsorption, but must be regenerated regularly to ensure reproducibility. We investigated the formation, characterization, and use of hybrid phospholipid bilayers (HPBs) as more stable biosurfactant capillary coatings for CZE protein separations. HPBs are formed by covalently modifying a support with a hydrophobic monolayer onto which a self‐assembled lipid monolayer is deposited. Monolayers prepared in capillaries using 3‐cyanopropyldimethylchlorosilane (CPDCS) or n‐octyldimethylchlorosilane (ODCS) yielded hydrophobic surfaces with lowered surface free energies of 6.0 ± 0.3 or 0.2 ± 0.1 mJ m^?2, respectively, compared to 17 ± 1 mJ m^?2 for bare silica capillaries. HPBs were formed by subsequently fusing vesicles comprised of 1,2‐dilauroyl‐sn‐glycero‐3‐phosphocholine or 1,2‐dioleoyl‐sn‐glycero‐3‐phosphocholine to CPDCS‐ or ODCS‐modified capillaries. The resultant HPB coatings shielded the capillary surface and yielded reduced electroosmotic mobility (1.3–1.9 × 10^?4 cm² V^?1s^?1) compared to CPDCS‐ and ODCS‐modified or bare capillaries (3.6 ± 0.2 × 10^?4 cm² V^?1s^?1, 4.8 ± 0.4 × 10^?4 cm² V^?1s^?1, and 6.0 ± 0.2 × 10^?4 cm² V^?1s^?1, respectively), with increased stability compared to phospholipid bilayer coatings. HPB‐coated capillaries yielded reproducible protein migration times (RSD ≤ 3.6%, n ≥ 6) with separation efficiencies as high as 200 000 plates/m.     

Algorithms for Symmetric Differential Systems

Over-determined systems of partial differential equations may be studied using differential—elimination algorithms, as a great deal of information about the solution set of the system may be obtained from the output. Unfortunately, many systems are effectively intractable by these methods due to the expression swell incurred in the intermediate stages of the calculations. This can happen when, for example, the input system depends on many variables and is invariant under a large rotation group, so that there is no natural choice of term ordering in the elimination and reduction processes. This paper describes how systems written in terms of the differential invariants of a Lie group action may be processed in a manner analogous to differential—elimination algorithms. The algorithm described terminates and yields, in a sense which we make precise, a complete set of representative invariant integrability conditions which may be calculated in a ``critical pair' completion procedure. Further, we discuss some of the profound differences between algebras of differential invariants and standard differential algebras. We use the new, regularized moving frame method of Fels and Olver [11], [12] to write a differential system in terms of the invariants of a symmetry group. The methods described have been implemented as a package in \MAPLE. The main example discussed is the analysis of the (2+1 )-d'Alembert—Hamilton system u_{xx}+u_{yy}- u_{zz}&=& f(u), u_x^2+u_y^2- u_z^2&=&1. (1) We demonstrate the classification of solutions due to Collins [7] for f\ne 0 using the new methods. October 13, 1999. Final version received: May 18, 2001.     

Hydrodynamically limited precision of gradient techniques in flow injection analysis

Based on the law of error propagation, a general expression is derived to study theoretically the hydrodynamically limited precision associated with each single element of fluid in a concentration profile in flow injection analysis. Convolution of the injection and residence time distribution functions is used to obtain response functions for a homogeneously stirred mixing chamber and for a straight capillary tube. The effects of stochastic variations on overall precision by sample introduction, pumping and timing are elucidated and compared with experimental findings. Resulting practical implications for the use of these two dispersing elements are outlined.     

Enhancing the enzymatic hydrolysis of cellulosic materials using simultaneous ball milling

One of the limiting factors restricting the effective and efficient bioconversion of softwood-derived lignocellulosic residues is the recalcitrance of the substrate following pretreatment. Consequently, the ensuing enzymatic process requires relatively high enzyme loadings to produce monomeric carbohydrates that are readily fermentable by ethanologenic microorganisms. In an attempt to circumvent the need for larger enzyme loadings, a simultaneous physical and enzymatic hydrolysis treatment was evaluated. A ball-mill reactor was used as the digestion vessel, and the extent and rate of hydrolysis were monitored. Concurrently, enzyme adsorption profiles and the rate of conversion during the course of hydrolysis were monitored. α-Cellulose, employed as a model substrate, and SO₂-impregnated steam-exploded Douglas-fir wood chips were assessed as the cellulosic substrates. The softwood-derived substrate was further posttreated with water and hot alkaline hydrogen peroxide to remove >90% of the original lignin. Experiments at different reaction conditions were evaluated, including substrate concentration, enzyme loading, reaction volumes, and number of ball beads employed during mechanical milling. It was apparent that the best conditions for the enzymatic hydrolysis of α-cellulose were attained using a higher number of beads, while the presence of air-liquid interface did not seem to affect the rate of saccharification. Similarly, when employing the lignocellulosic substrate, up to 100% hydrolysis could be achieved with a minimum enzyme loading (10 filter paper units/g of cellulose), at lower substrate concentrations and with a greater number of reaction beads during milling. It was apparent that the combined strategy of simultaneous ball milling and enzymatic hydrolysis could improve the rate of saccharification and/or reduce the enzyme loading required to attain total hydrolysis of the carbohydrate moieties.     

Comparison of 14C-amino acid mixture and [35S]methionine labeling of cellular proteins from mouse fibroblast C3H10T1/2 cells by two-dimensional gel electrophoresis.

Total cellular proteins from mouse C3H10T1/2 fibroblasts were compared by two-dimensional (2-D) gel electrophoresis after radiolabeling with [35S]methionine (35S-Met) or 14C-amino acids (14C-AA). 35S-Met labeling of protein was three to four times greater than 14C-AA incorporation over a 24 h period. Automated comparative analysis of replicate fluorographs after 6, 12, and 24 h of labeling showed considerable homology between radiolabeling methods. More than 88% percent of 35S-Met and 14C-AA-labeled proteins were common at each time point. However, the total number of 35S-Met-labeled proteins dropped from 6 to 24 h while the number of 14C-AA-labeled proteins increased. Additionally, twenty-one proteins were uniquely labeled by 14C-AA that were not detectable by 35S-Met over the labeling period. Densitometric analysis showed that several 35S-Met and 14C-AA-labeled proteins exhibited time-related differences in radiolabel incorporation while most proteins remained relatively constant. Protein patterns of silver-stained gels from 6 to 24 h were highly registered and showed few qualitative differences. Proteins detected in radiolabeled gels were generally, but not always, found in silver-stained gels. Thus, 35S-Met appears better suited for short-term radiolabeling of cellular protein while more comprehensive labeling of protein occurs with 14C-AA during prolonged incubation of cell cultures under present experimental conditions.     

High precision mean-field results for lattice gauge theories: with special attention paid to the gauge dependence of mean-field perturbation theory to finite order

We do mean-field perturbation theory for U(1) lattice gauge theory in the axial gauge, and evaluate corrections from fluctuations up to fourth order for the free energy and plaquette energy. Comparing with similar results previously obtained in the Feynman gauge we find, to those orders studied, a gauge dependence of the size of the first correction term neglected with one exception. This gauge dependence decreases rapidly as the order of the approximation is increased. To any finite order, results in axial gauge are better approximations than results in the Feynman gauge. We speculate why. Assuming it to be generally true, we evaluate the first correction beyond the one-loop mean-field approximation to the free energy of SU(2) gauge theory with Wilson action in the axial gauge. This correction brings the mean-field result very close to Monte Carlo results for β > 1.6. It also makes the mean-field result identical, within a narrow margin, to ressumed strong coupling results in the interval 1.6 < β < 2.4, thus showing the absence of a phase transition.For both groups studied, we find that the asymptotic series of mean-field perturbation theory give much better approximations than do ordinary weak coupling series.     

On finite element subspaces over quadrilateral and hexahedral meshes for incompressible viscous flow problems

Summary Optimal rates of convergence for the approximate solution of the stationary Stokes equations are obtained for finite element schemes which use piecewise constants to approximate the pressure and piecewise linear or piecewise bilinear or trilinear polynomials to approximate the velocity over fairly general quadrilateral or hexahedral meshes.This research was supported by NSF Grant MC80-16532