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Brandon C. Durney Beth A. Bachert Hillary S. Sloane Slawomir Lukomski James P. Landers Lisa A. Holland 《Analytica chimica acta》2015
Phospholipid additives are a cost-effective medium to separate deoxyribonucleic acid (DNA) fragments and possess a thermally-responsive viscosity. This provides a mechanism to easily create and replace a highly viscous nanogel in a narrow bore capillary with only a 10 °C change in temperature. Preparations composed of dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) self-assemble, forming structures such as nanodisks and wormlike micelles. Factors that influence the morphology of a particular DMPC–DHPC preparation include the concentration of lipid in solution, the temperature, and the ratio of DMPC and DHPC. It has previously been established that an aqueous solution containing 10% phospholipid with a ratio of [DMPC]/[DHPC] = 2.5 separates DNA fragments with nearly single base resolution for DNA fragments up to 500 base pairs in length, but beyond this size the resolution decreases dramatically. A new DMPC–DHPC medium is developed to effectively separate and size DNA fragments up to 1500 base pairs by decreasing the total lipid concentration to 2.5%. A 2.5% phospholipid nanogel generates a resolution of 1% of the DNA fragment size up to 1500 base pairs. This increase in the upper size limit is accomplished using commercially available phospholipids at an even lower material cost than is achieved with the 10% preparation. The separation additive is used to evaluate size markers ranging between 200 and 1500 base pairs in order to distinguish invasive strains of Streptococcus pyogenes and Aspergillus species by harnessing differences in gene sequences of collagen-like proteins in these organisms. For the first time, a reversible stacking gel is integrated in a capillary sieving separation by utilizing the thermally-responsive viscosity of these self-assembled phospholipid preparations. A discontinuous matrix is created that is composed of a cartridge of highly viscous phospholipid assimilated into a separation matrix of low viscosity. DNA sample stacking is facilitated with longer injection times without sacrificing separation efficiency. 相似文献
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Nguyen CN Isaacson JI Shimmyo KB Chen A Stratt RM 《The Journal of chemical physics》2012,136(18):184504
It has been suggested that the most-efficient pathway taken by a slowly diffusing many-body system is its geodesic path through the parts of the potential energy landscape lying below a prescribed value of the potential energy. From this perspective, slow diffusion occurs just because these optimal paths become particularly long and convoluted. We test this idea here by applying it to diffusion in two kinds of well-studied low-dimensional percolation problems: the 2d overlapping Lorentz model, and square and simple-cubic bond-dilute lattices. Although the most efficient path should be at its most dominant with the high-dimensional landscapes associated with many-body problems, it is useful to examine simpler, low-dimensional, constant-potential-energy problems such as these ones, both because the simpler models lend themselves to more accurate geodesic-path-finding approaches, and because they offer a significant contrast to many of the models used in the traditional energy-landscape literature. Neither the continuum nor the lattice percolation examples are adequately described by our geodesic-path formalism in the weakly disordered (relatively-fast-diffusion) limit, but in both cases the formalism successfully predicts the existence of the percolation transition and (to a certain extent) the slow diffusion characteristic of near-percolation behavior. The numerical results for these models are not nearly accurate enough near their transitions to describe critical exponents, but the models do showcase the qualitative validity of the geodesic perspective in that they allow us to see explicitly how tortuous and sparse the optimal pathways become as the diffusion constants begin to vanish. 相似文献
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Melissa D. Carter M. Wade Calcutt Beth A. Malow Kristie L. Rose David L. Hachey 《Journal of mass spectrometry : JMS》2012,47(3):277-285
Melatonin (MEL) and its chemical precursor N‐acetylserotonin (NAS) are believed to be potential biomarkers for sleep‐related disorders. Measurement of these compounds, however, has proven to be difficult due to their low circulating levels, especially that of NAS. Few methods offer the sensitivity, specificity and dynamic range needed to monitor MEL and its precursors and metabolites in small blood samples, such as those obtained from pediatric patients. In support of our ongoing study to determine the safety, tolerability and PK dosing strategies for MEL in treating insomnia in children with autism spectrum disorder, two highly sensitive LC‐MS/MS assays were developed for the quantitation of MEL and precursor NAS at pg/mL levels in small volumes of human plasma. A validated electrospray ionization (ESI) method was used to quantitate high levels of MEL in PK studies, and a validated nanospray (nESI) method was developed for quantitation of MEL and NAS at endogenous levels. In both assays, plasma samples were processed by centrifugal membrane dialysis after addition of stable isotopic internal standards, and the components were separated by either conventional LC using a Waters SymmetryShield RP18 column (2.1 × 100 mm, 3.5 µm) or on a polyimide‐coated, fused‐silica capillary self‐packed with 17 cm AquaC18 (3 µm, 125 Å). Quantitation was done using the SRM transitions m/z 233 → 174 and m/z 219 → 160 for MEL and NAS, respectively. The analytical response ratio versus concentration curves were linear for MEL (nanoflow LC: 11.7–1165 pg/mL, LC: 1165–116500 pg/mL) and for NAS (nanoflow LC: 11.0–1095 pg/mL). Copyright © 2012 John Wiley & Sons, Ltd. 相似文献
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Conditional stability constants for the Cu-sensitive fluorescent dye Phen Green™ SK (PGSK) were determined for complexes containing both Cu(I) and Cu(II). Experimental conditions were optimized to minimize oxidation of Cu(I) to Cu(II). A binding constant of 1011.38 for PGSK and Cu(I) was determined using chloride as the competing ligand and an iterative procedure involving equilibrium calculations to fit experimental data. The new constant was tested by evaluating PGSK fluorescence in the presence of the strong Cu(I)-ligand neocuproine and thiourea. There was good agreement between experimental data and changes in fluorescence predicted by calculations using the new constant and published constants for the competing ligands. Using EDTA to buffer the free ion concentration of Cu2+, the conditional stability constant of PGSK with Cu(II) was measured to be 109.8. Both of these new constants are significantly higher than previously published values. Finally PGSK was used to examine the kinetics of Cu(I) dissociation from the biologically important ligand glutathione. 相似文献
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Joe D. Oxman Dwight W. Jacobs Matthew C. Trom Vishal Sipani Beth Ficek Alec B. Scranton 《Journal of polymer science. Part A, Polymer chemistry》2005,43(9):1747-1756
Free‐radical/cationic hybrid photopolymerizations of acrylates and epoxides were initiated using a three‐component initiator system comprised of camphorquinone as the photosensitizer, an amine as the electron donor, and a diaryliodonium salt. Thermodynamic considerations revealed that the oxidation potential of the electron donor must be less than 1.34 V relative to SCE for electron transfer with the photoexcited camphorquinone to take place. This electron transfer leads to the production of the active centers for the hybrid polymerization (two radicals and a cation). Further investigation revealed that only a subset of electron donors that meet the oxidation potential requirement resulted in polymerization of the epoxide monomer; therefore, a second requirement for the electron donor (pKb higher than 8) was established. Experiments performed using a combination of electron donors revealed that the onset of the hybrid system's cationic polymerization can be advanced or delayed by controlling the concentration and composition of the electron donor(s). These studies demonstrate that a single three‐component initiator system can be used to initiate and chemically control the sequential curing properties of a free‐radical/cationic hybrid photopolymerization and is a viable alternative to separate photoinitiators for each type of polymerization. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 1747–1756, 2005 相似文献
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Navzer Sachinvala D. V. Parikh Paul Sawhney SeChin Chang Jerzey Mirzawa William Jarrett Beth Joiner 《先进技术聚合物》2007,18(8):620-628
In this paper we discuss the preparation and comparative evaluation of silver (I) [Ag(I)] nonwoven and woven antimicrobial barrier fabrics generated from commercial calcium‐sodium alginates and laboratory prepared sodium carboxymethyl (CM) cotton nonwovens and CM‐cotton printcloth for potential use as wound dressings. Degrees of CM substitution (DS) in cotton nonwoven and printcloth samples by titrimetry were 0.38 and 0.10, respectively. Coordination of Ag(I) with carboxylates on fabrics was effected by ion exchange and nitrates were removed by washing to mitigate nitrate ion toxicity issues. Durability of silver coordinated fabrics was tested by soaking them in deionized water with slight agitation at 50°C. Ag(I) alginates and nonwoven Ag(I)‐CM‐cottons lost structural integrity in water. Ag‐CM‐cotton printcloth samples retained structural integrity even after four soak‐and‐dry cycles, were smooth to the touch when dry, and were smoother when moistened. They could be easily peeled from wound surfaces without inducing trauma. Solid‐state carbon‐13 (13C) nuclear magnetic resonance (NMR) spectrometry was used to observe changes in carbonyl resonances in Ag(I) alginates and Ag(I)‐CM‐printcloth, and the chemical shift positions of carbonyl resonances of uncoordinated and Ag(I) coordinated fabrics did not change. Inductively coupled plasma‐mass spectrometry (ICP‐MS) was used following fabric digestion to determine the total Ag(I) ion content in fabrics. Ag(I) alginates were found to hold about 10–50 mg Ag(I) per gram fabric; and Ag(I) cotton woven and nonwoven fabrics held about 5–10 mg Ag(I) ions per gram fabric. Kinetic release of Ag(I) after soaking once in physiological saline was studied with ICP‐MS to estimate the availability of Ag(I) upon a single exchange with Na(I) ions on wound surfaces. Alginates released between ~13 and 28% of coordinated Ag(I), and CM‐cotton nonwovens and CM‐cotton printcloth released ~14 and 3% of coordinated Ag(I) ions, respectively. Finally, Ag(I) alginates and Ag(I)‐CM‐cotton printcloth samples were evaluated against Gram‐positive Staphylococcus aureus and Gram‐negative Klebsiella pneumoniae. Ag(I) alginates suppressed 99.95% of bacterial growth in vitro. Even after four soak‐and‐dry cycles in deionized water Ag(I)‐CM‐cotton printcloth suppressed 99.99% of bacterial growth in vitro. Published in 2007 by John Wiley & Sons, Ltd. 相似文献
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