Guanidine improves DEAE anion exchange-based analytical separation of plasmid DNA

Elution of strong and weak anion exchangers with sodium chloride gradients is commonly employed for analysis of sample mixtures containing different isomers of plasmid DNA. Gradient elution of a weak anion exchanger (diethylaminoethyl) in the presence of guanidine hydrochloride (Gdn) roughly doubles resolution between open-circular (oc) and supercoiled (sc) isomers. It also improves resolution among sc, linear, and multimeric/aggregated forms. Sharper elution peaks with less tailing increase sensitivity about 30%. However, elution with an exclusively Gdn gradient to 900 mM causes more than 10% loss of plasmid. Elution with a sodium chloride gradient while maintaining Gdn at a level concentration of 300 mM achieves close to 100% recovery of sc plasmid while maintaining the separation improvements achieved by exclusively Gdn elution. Corresponding improvements in separation performance are not observed on a strong (quaternary amine) anion exchanger. Other chaotropic salts do not produce a favorable result on either exchanger, nor does the inclusion of surfactants or EDTA. Selectivity of the diethylaminoethyl-Gdn method is orthogonal to electrophoresis, but with better quantification than agarose electrophoresis, better quantitative accuracy than CE, and resolution approaching CE.     

Influence of antimicrobial finishes on the biodeterioration of cotton and cotton/polyester fabrics: Leaching versus bio-barrier formation

The influence of antimicrobial activity of two contemporary finishes, specifically a dispersion of colloidal silver (Ag) and 3-(trimethoxysilyl)-propyldimethyloctadecyl ammonium chloride (Si-QAC), on the degree of biodeterioration of 100% cotton (CO) fabric and fabric composed of a mixture of cotton and polyester (CO/PET) was studied. Ag was chosen for the leaching agent, while Si-QAC was used as the bio-barrier-forming agent. The biodeterioration of samples finished with different concentrations of Ag and Si-QAC was analysed from a standard soil burial test after 3, 6 and 12 days of exposure to soil microflora. SEM micrographs revealed intensive biodeterioration of the unfinished cellulose fibres, while the highly biologically resistant polyester fibres remained undamaged. A controlled release of Ag successfully inhibited biodeterioration of the cellulose fibres in the CO and CO/PET fabrics when its concentration reached a lethal, biocidal concentration. Contrary to the effects of Ag, the bio-barrier formation of Si-QAC on CO and CO/PET fabrics was insufficient to protect the cellulose fibres during longer periods of soil burial, irrespective of its concentration. Intensive chemical changes to the cellulose were clearly seen from the FT-IR spectra of all of the samples. The resistance of the polyester component to biodeterioration did not provide any significant protection for the cotton component in CO/PET fabric.     

Bridging like-charged macroions through long divalent rodlike ions

Like-charged macroions in aqueous electrolyte solution can attract each other because of the presence of inter- and/or intramolecular correlations. Poisson-Boltzmann theory is able to predict attractive interactions if the spatially extended structure (which reflects the presence of intramolecular correlations) of the mobile ions in the electrolyte is accounted for. We demonstrate this for the case of divalent, mobile ions where each ion consists of two individual charges separated by a fixed distance. Variational theory applied to this symmetric 2:2 electrolyte of rodlike ions leads to an integro-differential equation, valid for arbitrary rod length. Numerical solutions reveal the existence of a critical rod length above which electrostatic attraction starts to emerge. This electrostatic attraction is distinct from nonelectrostatic depletion forces. Analysis of the orientational distribution functions suggests a bridging mechanism of the rodlike ions to hold the two macroions together. For sufficiently large rod length, we also observe "overcharging", that is, an over-compensation of the macroion charges by the diffuse layer of mobile rodlike ions. Our results emphasize the importance of the often rodlike internal structure that condensing agents such as polyamines, peptides, or polymer segments exhibit. The results were compared with Monte Carlo simulations.     

Modeling the influence of adsorbed DNA on the lateral pressure and tilt transition of a zwitterionic lipid monolayer

Certain lipid monolayers at the air-water interface undergo a second-order transition from a tilted to an untilted liquid-crystalline state of their lipid hydrocarbon chains at sufficiently large lateral pressure. Recent experimental observations demonstrate that in the presence of divalent cations DNA adsorbs onto a zwitterionic lipid monolayer and decreases the tilt transition pressure. Lowering of the tilt transition pressure indicates that the DNA condenses the lipid monolayer laterally. To rationalize this finding we analyze a theoretical model that combines a phenomenological Landau approach with an extension of the Poisson-Boltzmann model to zwitterionic lipids. Based on numerical calculations of the mean-field electrostatic free energy of a zwitterionic lipid monolayer-DNA complex in the presence of divalent cations, we analyze the thermodynamic equilibrium of DNA adsorption. We find that adsorbed DNA induces a 10% reduction of the electrostatic contribution to the lateral pressure exerted by the monolayer. This result implies a small but notable decrease in the tilt transition pressure. Additional mechanisms due to ion-ion correlations and headgroup reorientations are likely to further enhance this decrease.