Water potential of aqueous polyethylene glycol

Water potential (Ψω) values were determined for aqueous colloids of four molecular sizes of polyethylene glycol (PEG) using freezing-point depression and vapor-pressure deficit methods. A significant third-order interaction exists between the method used to determine Ψω, PEG molecular size, and concentration. At low PEG concentrations, freezing-point depression measurements result in higher (less negative) values for Ψω than do vapor-pressure deficit measurements. The reverse is true at high concentrations. PEG in water does not behave according to van't Hoff's law. Ψω is related to molality for a given PEG but not linearly. Moreover, Ψω varies with the molecular size of the PEG. It is suggested that the Ψω of PEG in water may be controlled primarily by the matric forces of ethylene oxide subunits of the PEG polymer. The term matricum is proposed for PEG in soil-plant-water relation studies.     

A new method for the determination of critical polyethylene glycol concentration for selective precipitation of DNA fragments

Separation strategies based on size-selective precipitation of DNA fragments with polyethylene glycol (PEG) have been used for achieving desired DNA interval in automated sample preparation for next-generation sequencing. By varying PEG concentration, DNA fragments of different sizes can be precipitated onto surfaces of carboxyl-coated paramagnetic particles selectively, and therefore, the desired DNA interval can be obtained. However, one of the crucial points in this approach is to determine the critical PEG concentration for DNA fragment of a certain size. The aim of this work was to develop a convenient and reliable method for accurately determining the critical PEG concentration. In our method, at a fixed concentration of sodium chloride (NaCl), recovered DNA samples obtained with different PEG concentrations were directly quantified, and their concentrations as a function of the PEG concentration were fitted by the logistic function. The critical PEG value was easily and accurately determined from the fitted logistic function. The repeatability and stability of the critical PEG value were assessed, showing an excellent reliability of the method. Based on this method, critical PEG values of different-size DNA fragments were determined at different NaCl concentrations. The effectiveness of the method was also demonstrated by selective precipitation of DNA fragments.     

Micropatterned thin film honeycomb materials from regiospecifically modified cellulose

Thin film honeycomb materials were prepared from regioselectively modified celluloses. The method uses water condensation at the surface of a cellulosic solution as an ordered template to form honeycomb structures. Pore size and distribution is controlled by several factors, one of which is the hydrophilicity of the cellulosic used. The amphiphilic nature of the celluloses was modified with varying lengths of ethylene glycol side chains using 2,6-thexyldimethylsilyl cellulose. It was found that the side chains do affect the honeycomb formation, with longer ethylene glycol chains leading to increased pore uniformity but having little influence on the pore size.     

Poroelastic bulk properties of the tectorial membrane measured with osmotic stress

The equilibrium stress-strain relation and the pore radius of the isolated tectorial membrane (TM) of the mouse were determined. Polyethylene glycol (PEG), with molecular mass (MM) in the range 20-511 kDa, added to the TM bathing solution was used to exert an osmotic pressure. Strain on the TM induced by isosmotic PEG solutions of different molecular masses was approximately the same for MM > or = 200 kDa. However, for MM < or = 100 kDa, the TM strain was appreciably smaller. We infer that for the smaller molecular mass, PEG entered the TM and exerted a smaller effective osmotic pressure. The pore radius of the TM was estimated as 22 nm. The equilibrium stress-strain relation of the TM was measured using PEG with a molecular mass of 511 kDa. This relation was nonlinear and was fit with a power function. In the radial cochlear direction, the transverse stiffness of the TM was 20% stiffer in the inner than in the outer region. TM segments from the basal region had a larger transverse stiffness on average compared to sections from the apical-middle region. These measurements provide a quantitative basis for a poroelastic model of the TM.     

Molecular weight determination of heparin and dermatan sulfate by size exclusion chromatography with a triple detector array

The determination of molecular weight (M) and molecular weight distribution (MD) of heparins by a novel approach, consisting of a high performance size exclusion chromatography (HP-SEC) combined with a triple detector array (TDA) is described. HP-SEC/TDA permits the evaluation of MD of polymeric samples through a combined and simultaneous action of three on-line detectors, right-angle laser light scattering (RALLS), refractometer (RI), and viscometer. The method does not require any chromatographic column calibration, thus overcoming also the difficulty to obtain adequate reference standards. It permits the size determination also of small molecules, even when scattering dissimmetry is not observable. Unfractionated heparins, eight fractions of a size fractionated heparin, and dermatan sulfates were analyzed by HP-SEC/TDA. The M values found for the heparin fractions were used to build up a calibration curve of a conventional HP-SEC system: the results obtained analyzing unfractionated heparin samples with both HP-SEC/TDA and HP-SEC were in excellent agreement, suggesting the possibility to use the TDA data to generate standard samples with known MD and intrinsic viscosity [eta]. Moreover, HP-SEC/TDA can successfully be employed also for the determination of the Mark-Houwink a and k parameters.     

DIATOM SILICA BIOMINERALIZATION: AT NANOSCALE LEVEL A CHEMICALLY UNIFORM PROCESS

Using a high-brilliance synchrotron X-ray source, combined small- and wide-angle X-ray scattering (SAXS and WAXS) was applied to study nanoscale characteristics, in particular pore size in the range of 3 to 65 nm, of a variety of unialgal cultures of centric and pennate diatoms, and of mixed diatom populations sampled in the field. Results of scattering analysis were compared with details of pore size, structure and orientation visible at the electron microscopic level. WAXS patterns did not reveal any crystalline phase or features of microcrystallinity (resolution 0.07 to 0.51 nm), which implies a totally amorphous character of the SiO₂ matrix of the frustule material. SAXS data (resolution 3 to 65 nm) provided information on geometry, size, and distribution of pores in the silica. Overall, two pore regions were recognized that were common to the silica of all samples: the smallest (d less than 10 nm) regularly spaced and shaped spherically, the larger (up to 65 nm) being cylinders or slits. Apparently, at a nanoscale level diatomaceous silica is quite homologous among species, in agreement with the chemical principles of silica polymerization under the conditions of pH and precursor concentrations inside the silicon deposition vesicle. The final frustule "macro"-morphology is of course species-specific, being determined genetically. Synthetically-derived MCM-type silicas have a similarly organized pore distribution in an amorphous silica matrix as we found in all diatom species studied. We therefore suggest that organic molecules of a kind used as structure-directing agents to produce these artificial silicas play a role in the nucleation of the silica polymerization reaction and the shaping of pore morphology inside the silicon deposition vesicle of diatoms. Structure-directing molecules now await isolation from the SDV, followed by identification and characterisation by molecular techniques.     

Modeling pore size distribution in cellulose rolled stationary phases

Rolled stationary phases are fabrics (i.e., nonparticulate phases) that rapidly separate proteins from salts on the basis of size exclusion. Pore size and pore size distributions in the stationary phase determine how different size molecules distribute between the stationary and mobile phases in liquid chromatography columns. The potential for size exclusion chromatography by fabrics is not initially obvious because their interlaced structures are atypical for size exclusion supports. A simple logistic model fits the pore size distribution of a rolled stationary phase when pore sizes were measured using PEG, Dextran, D2O, glucose, and NaCl probes. When the fabric is treated with cellulase enzymes, the water-accessible pores uniformly decrease and peak retention is lower. The logistic function model captures this result and enables comparison of pore size distribution curves between enzyme-treated and untreated fabrics in rolled stationary phase columns.     

Partitioning of Differently Sized Poly(ethylene glycol)s into OmpF Porin

To understand the physics of polymer equilibrium and dynamics in the confines of ion channel pores, we study partitioning of poly(ethylene glycol)s (PEGs) of different molecular weights into the bacterial porin, OmpF. Thermodynamic and kinetic parameters of partitioning are deduced from the effects of polymer addition on ion currents through single OmpF channels reconstituted into planar lipid bilayer membranes. The equilibrium partition coefficient is inferred from the average reduction of channel conductance in the presence of PEG; rates of polymer exchange between the pore and the bulk are estimated from PEG-induced conductance noise. Partition coefficient as a function of polymer weight is best fitted by a “compressed exponential” with the compression factor of 1.65. This finding demonstrates that PEG partitioning into the OmpF channel pore has sharper dependence on polymer molecular weight than predictions of hard-sphere, random-flight, or scaling models. A 1360-Da polymer separates regimes of partitioning and exclusion. Comparison of its characteristic size with the size of a 2200-Da polymer previously found to separate these regimes for the α-toxin shows good agreement with the x-ray structural data for these channels. The PEG-induced conductance noise is compatible with the polymer mobility reduced inside the OmpF pore by an order of magnitude relatively to its value in bulk solution.     

Pretreatment for enhanced hydrolysis of cellulosic biomass

This review will cover a number of physical and chemical pretreatment methods for cellulosic substrates which enhance their hydrolysis by cellulase or consumption by microorganisms. While the emphasis is on the literature of the last two years, some earlier work is cited which has influenced the work in the pretreatment field. In order to interpret the effects of a pretreatment method, emphasis in the past has been on crystallinity index (CI) and lignin content. Although these parameters happen often to correlate with the rate or extent of hydrolysis, it is suggested that a more basic parameter is the pore size distribution of the wet substrate and the associated surface area available to the cellulase that is the major factor in determining the effectiveness of a pretreatment method.     

The osmotic flow of an electrolyte through a charged porous membrane

A pore model in which the pore wall has a continuous distribution of electrical charge is used to investigate the osmotic flow through a charged permeable membrane separating electrolyte solutions of unequal concentrations. The pore is treated as a long, circular, cylindrical duct. The analysis is based on a continuum formulation in which a dilute electrolyte solution is described by the coupled Nernst-Planck/Poisson creeping flow equations. Account is taken of the significant size of the electrolyte ions (assumed to be rigid spheres) when compared with the diameter of the membrane pores. Analytical solutions for the ion concentrations, hydrostatic pressure and electrostatic potential in the electrolyte solutions are given and an intra-pore flow solution is derived. A mathematical expression for the osmotic reflection coefficient as a function of the solute ion: pore diameter ratio λ and the solute fluxes is obtained. Approximate solutions are quoted which relate the solute fluxes and the solution electrostatic potentials at the membrane surfaces to the bulk solution concentrations, the membrane pore charge and pore geometry. The osmotic reflection coefficient is thus determined as a function of these parameters.     

PEG blocking of single pores arising on phase transitions in unmodified lipid bilayers

Changes in ionic permeability of bilayer lipid membranes (BLM) from dipalmitoyl phosphatidylcholine at temperature of phase transition in 1 M LiCl solution in the presence of polyethyleneglycols (PEG) of various molecular masses are studied. The transition of ionic membrane channels from conducting to blocked nonconducting state using polymers makes it possible to calibrate lipid pores. It is shown that low-molecular weight glycerol and PEG with molecular weights of 300 and 600 decrease the amplitude of current fluctuations through the membrane, the decrease being proportional to the size of the polymer molecule incorporated. The addition of PEG with molecular masses of 1450, 2000, and 3350 decrease the current fluctuations to the basal noise level. The result is considered as a complete blockade of ion channel conductivity. In the presence of rather large polymers, such as PEG with molecular masses of 6000 and 20000, which are hardly incorporated in the pore, single current fluctuations occur again; however, their amplitudes are somewhat smaller than in the absence of PEG. It is assumed that a complete blockade of the conductivity of lipid ionic channels by PEG with molecular masses of 1450, 2000, and 3350 is due to dehydration of the pore gap and the conversion of the hydrophilic pore to a hydrophobic one.     

Size‐selective DNA separation: Recovery spectra help determine the sodium chloride (NaCl) and polyethylene glycol (PEG) concentrations required

In the presence of sodium chloride (NaCl), DNA fragments can be size‐selectively separated by varying the final concentration of polyethylene glycol (PEG). This separation strategy in combination with the use of paramagnetic particles provides a valuable platform for achieving the desired DNA size interval, which is important in automated library preparation for high‐throughput DNA sequencing. Here, we report the establishment of recovery spectra of DNA fragments that enable the determination of suitable NaCl and PEG concentrations for size‐selective separation. Firstly, at a given NaCl concentration, the recovery equation was obtained by fitting the DNA recovery ratios versus the PEG concentrations using the logistic function to determine the required parameters. Secondly, the slope function of the recovery equation was achieved by deducing its first derivative. Therefore, the recovery spectrum can be generated using the slope function based on those parameters. According to the recovery spectra of different length DNA fragments, suitable NaCl and PEG concentrations can be determined, respectively, by calculating their resolution values and recovery ratios. The strategy was effectively applied to the size‐selective separation of 532‐, 400‐, and 307‐bp fragments at the selected reagent concentrations with recoveries of 96.9, 64.7, and 85.9%, respectively. Our method enables good predictions of NaCl and PEG concentrations for size‐selective DNA separation.     

Effect of poly(ethylene glycol) on the Ca2+-induced fusion of didodecyl phosphate vesicles

This paper reports a study of the effect of the dehydrating agent poly(ethylene glycol) (PEG) on didodecyl phosphate (DDP) bilayers and on the fusion activity of DDP vesicles as a function of the molecular weight of PEG. PEG 8K in a concentration of 10 wt % does not induce fusion. However, Ca2+-induced fusion is promoted as reflected by a lowering of the Ca2+ threshold concentration. This effect can most likely be attributed to the dehydrating capacity of the polymer. Interestingly, low concentrations (0.1 wt %) of PEG 20 K induce a moderate fusion capacity. At higher concentrations (0.5 wt %) fusion is inhibited, irrespective of the presence of Ca2+. These molecular weight dependent effects can be rationalized by taking into account that the clouding temperature differs for PEGs of different molecular weights. In the case of PEG 20K a microscopic phase separation will occur at the bilayer-water interface because PEG-PEG interactions and presumably PEG-DDP interactions are favored over PEG-water interactions. As a consequence, the DDP vesicle surface becomes covered with PEG 20K, resulting in a steric stabilization of the vesicles. This will impede or prevent, depending on the polymer concentration, the vesicles from approaching each other sufficiently close for fusion to occur.     

Comparison of Hydrolysis Efficiency and Enzyme Adsorption of Three Different Cellulosic Materials in the Presence of Poly(ethylene Glycol)

The addition of non-ionic surfactants has recently been confirmed to positively affect the enzymatic hydrolysis of cellulosic materials. However, the functional mechanisms of these surfactants remain unclear. This work investigated the influence of poly(ethylene glycol) (PEG) on the enzymatic hydrolysis of three cellulosic materials, namely, acid steam-exploded corn straw, pure microcrystalline cellulose (Avicel PH101), and bagasse sulfite pulp (BSP). The results showed that PEG addition led to varied effects on the enzymatic hydrolysis of different cellulosic materials. Addition of PEG was most effective on the enzymatic hydrolysis of PH101 and weakly effective on the hydrolysis of BSP. We further investigated PEG concentrations and enzymatic activities in the supernatant during hydrolysis and found that the positive effects of PEG treatment might contribute to its influence on enzyme desorption from different substrates. We also found that the efficiency of PEG depended on its capacity to bind to different substrates. PEG exhibited stronger affinity to pure cellulose than to the two other lignocellulosic substrates. These findings are helpful in further revealing the mechanism of surfactants and improving the enzymatic hydrolysis process.     

PEG molecular weight and lateral diffusion of PEG-ylated lipids in magnetically aligned bicelles

Lateral diffusion coefficients of PEG-ylated lipids with three different molecular weight PEG groups (1000, 2000 and 5000) were measured in magnetically-aligned bicelles using the stimulated echo (STE) pulsed field gradient (PEG) (1)H nuclear magnetic resonance (NMR) method. At concentrations below the PEG "mushroom-to-brush" transition, all three PEG-ylated lipids exhibited unrestricted lateral diffusion, with lateral diffusion coefficients comparable to those of corresponding non-PEG-ylated lipids and independent of PEG molecular weight. At concentrations above this transition, lateral diffusion slowed progressively with increasing concentration of PEG-ylated lipid as a result of surface crowding. As well, the lateral diffusion coefficients exhibited a pronounced decrease with increasing PEG group molecular weight and a diffusion-time dependence indicative of obstructed diffusion. We conclude that, while lateral diffusion of PEG-ylated lipids within lipid bilayers is determined primarily by the hydrophobic anchoring group, when crowding at the lipid bilayer surface becomes significant, the size of the extra-membranous domain, in this case the PEG group, can influence lateral diffusion, leading to decreased diffusivity with increasing size and producing obstructed diffusion at high crowding. These findings imply that similar considerations will pertain to lateral diffusion of membrane proteins with large extra-membranous domains.     

Through pore diameter in the cell wall of Chara corallina.

Determination of pore size of the cell wall of Chara corallina has been made by using the polyethylene glycol (PEG) series as the hydrophilic probing molecules. In these experiments, the polydispersity of commercial preparation of PEGs was allowed for. The mass share (gamma(p)) of polyethylene glycol preparation fractions penetrating through the pores was determined using a cellular 'ghost', i.e. fragments of internodal cell walls filled with a 25% solution of non-penetrating PEG 6000 and tied up at the ends. In water, such a 'ghost' developed a hydrostatic pressure close to the cell turgor which persisted for several days. The determination of gamma(p), for polydisperse polyethylene glycols with different average molecular mass (M) was calculated from the degree of pressure restoration after water was replaced by a 5-10% polymer solution. Pressure was recorded using a dynamometer, which measures, in the quasi-isometric mode, the force necessary for the partial compression of the 'ghost' in its small fragment. By utilizing the data on the distribution of PEG 1000, 1450, 2000, and 3350 fractions over molecular mass (M), it was found that gamma(p), for these polyethylene glycols corresponded to the upper limit of ML=800-1100 D (hydrodynamic radius of molecules, r(h)=0.85-1.05 nm). Thus, the effective diameter of the pores in the cell wall of Chara did not exceed 2.1 nm.     

Molecular dynamics simulations of EMI-BF₄ in nanoporous carbon actuators

An artificial muscle composite material consisting of carbide derived carbon (CDC) and 1-ethyl-3-methylimidazolium tetrafluoroborate (EMI-BF₄) ionic liquid was modeled using molecular dynamics (MD) simulations, in order to determine the molecular structural rearrangements causing actuation. CDC was represented as separate curved graphene-like flakes with charges of +2, 0 or −2 on each flake, with 24–27 aromatic rings each. The charge distribution in the flakes was determined by PM6 semi-empirical optimization. The pore size distribution of CDC and the density of the material were comparable to experimental data. Molecular structure analysis revealed a preferential parallel orientation for the cations over the negatively charged CDC surfaces, while cationic rotations and reorientations could be observed for positively charged CDC. Changes in the pore occupancy for each ionic type were observed for pore sizes between 4 and 7 ?, which, together with the replacement of large cations with smaller anions, could explain the volume decrease in the anodes (and, vice versa, the volume increase in the cathodes) in this type of actuator.     

PEG molecular weight and lateral diffusion of PEG-ylated lipids in magnetically aligned bicelles

Lateral diffusion coefficients of PEG-ylated lipids with three different molecular weight PEG groups (1000, 2000 and 5000) were measured in magnetically-aligned bicelles using the stimulated echo (STE) pulsed field gradient (PEG) ¹H nuclear magnetic resonance (NMR) method. At concentrations below the PEG “mushroom-to-brush” transition, all three PEG-ylated lipids exhibited unrestricted lateral diffusion, with lateral diffusion coefficients comparable to those of corresponding non-PEG-ylated lipids and independent of PEG molecular weight. At concentrations above this transition, lateral diffusion slowed progressively with increasing concentration of PEG-ylated lipid as a result of surface crowding. As well, the lateral diffusion coefficients exhibited a pronounced decrease with increasing PEG group molecular weight and a diffusion-time dependence indicative of obstructed diffusion. We conclude that, while lateral diffusion of PEG-ylated lipids within lipid bilayers is determined primarily by the hydrophobic anchoring group, when crowding at the lipid bilayer surface becomes significant, the size of the extra-membranous domain, in this case the PEG group, can influence lateral diffusion, leading to decreased diffusivity with increasing size and producing obstructed diffusion at high crowding. These findings imply that similar considerations will pertain to lateral diffusion of membrane proteins with large extra-membranous domains.     

Size fractionation of double-stranded DNA by precipitation with polyethylene glycol

We show that DNA molecules of differing molecular mass are separable by selective precipitation with polyethylene glycol (PEG+.. Higher molecular mass DNA precipitates at lower PEG concentrations than lower molecular mass DNA. Double-stranded DNA can be fractionated at least in the range of 3 times 10-7 to 1 times 10-5 daltons. The effects on PEG concentration, sodium chloride concentration, DNA concentration, pH, divalent ions, precipitation time, and centrifugal force have been determined. These studies show PEG precipitation offers a size fractionation method for DNA which is convenient, of high capacity, and applicable over a wide range of conditions. However, resolution is not high and separation of two species approaches 100% only if they differ in molecular mass by at least a factor of two.     

Effect of pretreatments and fermentation on pore size in cellulosic materials

Surface area has been proposed as a major factor determining the extent of enzymatic hydrolysis of cellulose. We used cornstalk residue (CR) and Solka Floc BW-300 (SF) as substrates and NaOH (a cellulose swelling agent) and iron sodium tartrate (FeTNa, intercolates between cellulose microfibrils) as pretreatments to study the effect of surface area on extent of fermentation. Micropore sizes (8-130 A) were determined by a solute exclusion technique using glucose, cellobiose, and polyethylene glycols as molecular probes. The pore size distributions follow the logistic model function: I = a/[1+exp(b - cX)] where I is pore volume; X = log D; D is the molecular probe diameter; and a, b, and c are constants. The pore volumes of CR (1.9 mL/g) and SF (1.6 mL/g) are increased to 2.1 mL/g by pretreatment with NaOH. Pretreatment of SF with NaOH and cornstalk residue with FeTNa caused an upward shift in the pore size distribution. Fermentation of untreated CR by rumen microbes resulted in a 46% loss of dry matter while increasing the internal pore size and decreasing the pore volume to 0.9 mL/g. Fermentation of NaOH pretreated CR resulted in a 73% loss of dry matter with little change in pore size, total pore volume, or fiber composition. Fiber analysis indicated that selective utilization of hemicellulose over cellulose in both fermentations was small. The data show that: (1) removal of hemicellulose and lignin increases dry matter disappearance upon fermentation of the remaining material; (2) relative to the size of bacterial cellulases (40-160 A), the pretreatments have little effect on increasing accessibility of surface internal to the cellulose particles; and (3) the micropore changes caused by NaOH or FeTNa treatment do not explain the enchanced fermentation obtained for treated cornstalk residue. These observations infer that external or macropore surface properties may be a significant factor in determining the extent of utilization of the solid substrates by cellulolytic microorganisms.