Low Concentrations Of PEG Research Articles

Sustainable Valorisation and Efficient Downstream Processing of Giant Reed by High-Pressure Carbon Dioxide Pretreatment.

This work investigated the catalytic high-pressure CO2 pretreatment of giant reed. CO2 is a renewable resource; its use does not generate chemical wastes and it can be easily removed and recycled. The effect of the addition of low concentrations of FeCl3 (0.16 wt %) and PEG 400 (1.0 wt %) on the hemicellulose hydrolysis to xylose and xylo-oligosaccharides (XOS) is reported for the first time. Under the optimised pretreatment conditions, the xylan conversion of 82 mol % and xylose and XOS yields of 43 and 20 mol % were achieved, respectively. The solid residues obtained from different pretreatments were used as the substrate for the enzymatic hydrolysis to give glucose. The total glucose yield achieved under the optimised two-step process was 67.8 mol % with respect to the glucan units in the biomass. The results demonstrated that PEG-assisted FeCl3 -catalysed scCO2 pretreatment can produce xylose- or XOS-rich hydrolysates and improve the enzymatic hydrolysis of biomass.

Effects of osmotic stress on Chenopodium quinoa seed germination.

Quinoa is a pseudo-cereal native to the Andes known for its exceptional adaptation to extreme soil and climate conditions. The study of the response of Chenopodium quinoa to osmotic stress and the contribution to improving its yield aims to ensure production in arid and semi-arid regions as an alternative crop. The aim of this study is to test the germination capacity of 5 selected varieties; CQ-57, UDEC-2; ICBA-4, ICBA-5 and 115-R subjected to varying osmotic pressures using solutions of PEG-6000 and NaCl. Seeds are incubated at 25 °C for 10 days. Results showed that osmotic stress increases the lag phase but does not affect the germination rate at low concentrations of PEG and NaCl. While for high concentrations the germination rates decrease as compared to controls. Radicale and hypocotyl lengths decreased with increasing the degree of osmotic stress. The effects of osmotic stress are comparable in the 5 varieties. At high concentration of PEG or NaCl, ICBA-5 and CQ-57 exhibited the highest value of germination rate showing the capacity of these varieties to establish itself in regions suffering from salinity or water deficit.

Enhanced CO2 permeability in Matrimid® 5218 mixed matrix membranes for separating binary CO2/CH4 mixtures

Abstract Membrane gas separation is an emerging technology used for the separation of CO2. Matrimid® 5218, one of the current polymers used in membrane gas separation, has good selectivity, but poor CO2 permeability. We wondered if its CO2 permeability could be enhanced by the addition of a CO2-philic additive (PEG 200) and ZIF-8 nanoparticles. ZIF-8 particles were synthesized with a nanoparticle size of 33.83 ± 6.2 nm. These particles were characterized by SEM and XRD. Dense film-casting method was used to prepare novel ternary mixed matrix membranes with low PEG concentrations (4 wt%) at different filler loadings (10–40 wt%). In CO2/CH4 binary mixtures, binary and ternary MMMs membranes were tested at fixed feed composition (50:50) and different feed pressures (from 2 to 8 bar). The MMMs were also characterized using SEM, EDX, DSC, and TGA. Finally, the incorporation of 30 wt% of ZIF-8 nanoparticles leads to increase the CO2 permeability in binary (up to 31.47 Barrer) and ternary MMMs (up to 33.12 Barrer); pointing out that the addition of PEG and ZIF-8 enhanced the CO2 permeability in the neat Matrimid® membranes (7.16 Barrer). Our results suggest that such ternary MMMs have considerable potential for use in CO2 separation applications.

How Low Can You Go? Low Densities of Poly(ethylene glycol) Surfactants Attract Stealth Proteins.

It is now well-established that the surface chemistry and "stealth" surface functionalities such as poly(ethylene glycol) (PEG) chains of nanocarriers play an important role to decrease unspecific protein adsorption of opsonizing proteins, to increase the enrichment of specific stealth proteins, and to prolong the circulation times of the nanocarriers. At the same time, PEG chains are used to provide colloidal stability for the nanoparticles. However, it is not clear how the chain length and density influence the unspecific and specific protein adsorption keeping at the same time the stability of the nanoparticles in a biological environment. Therefore, this study aims at characterizing the protein adsorption patterns depending on PEG chain length and density to define limits for the amount of PEG needed for a stealth effect by selective protein adsorption as well as colloidal stability during cell experiments. PEG chains are introduced using the PEGylated Lutensol AT surfactants, which allow easy modification of the nanoparticle surface. These findings indicate that a specific enrichment of stealth proteins already occurs at low PEG concentrations; for the decrease of unspecific protein adsorption and finally the colloidal stability a full surface coverage is advised.

Molecular simulation and experimental studies of the miscibility of polylactic acid/polyethylene glycol blends

Computer simulation and experiments were performed to investigate the miscibility of PLA/PEG blends with different PEG concentrations. Flory-Huggins interaction (χ) parameter used to predict the miscibility for the blends was estimated by molecular dynamic simulation of fully atomistic model. The calculated χ parameter and radial distribution function suggest that the PLA and PEG blends are likely miscible at low PEG concentrations (10–30 wt%), but they become apparently immiscible at higher PEG content (>50 wt%). This result is consistent with density distribution of PLA and PEG beads calculated from dissipative particle dynamics simulation of coarse-grained model. To support the computational results, experiments based on differential scanning calorimetry (DSC) and rheometry were also performed. The DSC thermograms of 90:10, 80:20, and 70:30 (wt/wt) of PLA/PEG blends showed a single glass transition and PLA melting peak, indicating PLA/PEG is miscible over this composition. In rheometry, frequency (ω) dependence of storage moduli (G′) at low frequencies for 75:25 and 70:30 blends indicate that these samples are near the phase separation point.

Preparation and characterization of polyethylene glycol/poly(L-lactic acid) blends

Abstract The effect of incorporation of poly(ethylene glycol) (PEG) on thermomechanical and hydrophilicity properties of poly(L-lactic acid) (PLLA) was investigated. PEG/PLLA blends, containing 10, 20, 30 and 40 wt% PEG, were prepared by melt-extrusion in a co-rotating twin-screw extruder. By DSC analysis, it was observed that the Tg of PLLA phase in PEG/PLLA blends decreased accompanied by a significant decrease in Tcc and increase in their melting enthalpy. Therefore, the addition of PEG enhances the crystallization ability of PLLA phase due to its lubricating effect which increased mobility of PLLA chains. From TGA it was observed that low concentrations of PEG (10 & 20 wt%) increase the Tonset of thermal degradation, probably due to improved heat resistance of the crystalline phase. At higher PEG content, the Tonset decreases, as the lubricating effect becomes the controlling mechanism for the initiation of degradation process. Decrease in tensile strength and modulus was recorded especially in PLLA blends with PEG content higher than 20 wt%. The elongation at break decreases reaching a maximum at 20 wt% PEG and then dropped again. To investigate the effect of PEG on the wetting ability of PLLA, water contact angle measurements were performed. The results indicate that the introduction of PEG lowers the contact angle values in PEG/PLLA film surfaces, as compared to pure PLLA, suggesting improved hydrophilic properties.

Improved CO2 separation performance of Matrimid®5218 membrane by addition of low molecular weight polyethylene glycol

AbstractPolyethylene glycols have received worldwide attention as a highly permeable CO2‐philic polymer in carbon dioxide separation applications. In this study, we investigated the influence of a low molecular weight polyethylene glycol (PEG 200) on physicochemical, morphological, and gas separation properties of Matrimid®5218 as a novel polymer blend. Both symmetric and asymmetric Matrimid flat sheet membranes with 0–20 wt.% PEG were prepared via a dense film‐casting method. The miscibility of blends at low PEG concentrations (3–5 wt.%) was confirmed by differential scanning calorimetry (DSC) and polarized light microscopy (PLM). Moreover, the blends were partially miscible at higher PEG concentrations (10–20 wt.%). Matrimid/PEG molecular interactions were further studied by FTIR and XRD analysis. SEM images indicated an impressive influence of PEG on the symmetric structure of Matrimid membrane. An asymmetric structure with a dense thin skin layer and a highly porous sub‐layer was observed for the blends containing 10–20 wt.% PEG. The CO2 permeability and CO2/CH4 selectivity of the best‐yield CO2‐selective blend membrane (Matrimid/PEG (95:5)) incvreased about 25% (from 7.68 to 9.62 Barrer) and 15% (from 35 to 40) compared to pure Matrimid, respectively. Furthermore studying the plasticization pressures of the membranes revealed that the more PEG content in blend the less the plasticization pressure obtained. © 2015 Society of Chemical Industry and John Wiley & Sons, Ltd

Probing the structure of PEGylated-lipid assemblies by coarse-grained molecular dynamics

Poly(ethylene glycol) (PEG) and various zwitterionic species are now used routinely as components in soft biomaterials. The study of mixtures composed of PEGylated-lipids with zwitterionic lipids provides an excellent opportunity to understand molecular interactions in such systems. To this end, we examine the liposome/bicelle/micelle phase behavior of such mixtures in bulk solution and as monolayers at the air–water interface, utilizing a coarse-grained molecular-dynamics (CG-MD) approach rooted in experimental parameters that inform the nature of the short-range interactions. Specifically, we examine mixtures of PEGylated lipid and dimyristoylphosphatidylcholine (DMPC) lipid monolayers and find the standard force fields yield calculated surface tensions in agreement with experiment. Long-time CG-MD simulations (∼0.5 microsecond) of the self-assembly for different concentrations of PEG indicate the critical micellar size for the aggregate to form a bicelle from a spherical micellar nucleus. At lower PEG concentrations we find self-assembly of the PEGylated lipid into a liposome coexisting with a mixture of smaller bicelles. Examination of large individual bicelles (1000 lipids) at increasing concentrations of PEG indicates that the degree of PEG de-mixing towards the outer radius of the bicelle is correlated with the overall concentration of PEG in the bicelle. The CG-MD approach used herein should be a useful complement to experimental studies designed to probe solute (drug) interactions with such membrane systems.

Potentiation of proline accumulation in oilseed rape leaf discs exogenously supplied with combinations of PEG and cryoprotective agents is associated with overproduction of ABA

Physiological processes involved in the control of the magnitude of the stress-induced proline (Pro) response of higher plants are not fully understood. Here we are dealing with Pro accumulation by rape leaf discs (RLDs) treated in vitro, under light conditions, with dual unusual systems containing low concentrations of the non-permeant polymer PEG 6000 (PEG) added with readily permeant cryoprotective agents (CPAs). At osmotically active concentrations, dimethylsulfoxide (DMSO), glycerol, 1,3-propanediol, ethylene glycol and dimethylformamide (DMF) behaved as very poor inducers of the Pro response when provided alone. On the contrary when all those substances (apart DMF) were supplied at onset of treatments, in combination with PEG, Pro levels greatly exceeded what could be predicted through simple additivity of the effects of those substances provided individually. This suggested potentiation effects at the level of some component(s) of Pro metabolism involved in stress-induced Pro accumulation. We have also demonstrated that exogenous ABA could substitute for DMSO (but not PEG), this other binary system also inducing (through potentiation) high rate of Pro accumulation. The strinking similarity between the responses induced with PEG+DMSO and PEG+ABA suggested that DMSO induced increases in the endogenous amount of ABA which might be, at its turn, acting as exogenously supplied ABA. Additional amounts of ABA actually accumulated in leaf discs treated with the dual system PEG+DMSO. Other CPAs tested in this study, except dimethylformamide (DMF), might also be acting on the same way. Potentiating effects, associated with enhanced amount of ABA, were also found to result from combinations of PEG with a range of organic and inorganic substances which mimick to some extent the cytosolute composition of plant cells. Together, our results gain some insight into the physiological mechanisms involved in the control of stress-induced Pro accumulation and strongly suggest their relationship with stress-induced changes in ABA content.

Thermodynamic studies of the ionic liquid 1-hexyl-3-methylimidazolium chloride [C 6mim][Cl] in polyethylene glycol aqueous solutions

Vapour pressure osmometery (VPO) measurements at T = 308.15 K for {[C 6mim][Cl] + water}, {[C 6mim][Cl] + (0.005, 0.0155, and 0.0263) mol · kg −1 PEG2000 + water} and {[C 6mim][Cl] + (0.0017, 0.0052, and 0.0088) mol · kg −1 PEG6000 + water} systems and isopiestic measurements at T = 298.15 K for {[C 6mim][Cl] + PEG2000 + water} and {[C 6mim][Cl] + PEG6000 + water} systems have been carried out. The VPO measurements were carried out at very low concentrations of PEG and from which the values of the water activities, osmotic coefficients, vapour pressure and activity coefficients were obtained. The data obtained from the VPO method show that over the whole concentration range of the ionic liquid (IL), the activity coefficients of [C 6mim][Cl] in the presence of PEG2000 are increased. Although, at high IL concentrations, the values of the activity coefficient of [C 6mim][Cl] in the presence of PEG6000 are also increased, however for low concentrations of IL the values of the activity coefficient of [C 6mim][Cl] in pure water are larger than those in aqueous PEG6000 solutions. For a known IL concentration, the values of water activity coefficient for the binary {[C 6mim][Cl] + water} system are larger than those for the ternary {[C 6mim][Cl] + PEG + water} systems and decrease by increasing the concentration of PEG or decreasing the molar mass of PEG. The constant water activity lines of the all ternary systems obtained from the isopiestic method show positive deviation from the linear isopiestic relation (Zdanovskii–Stokes–Robinson rule) derived using the semi-ideal hydration model. The results have been interpreted in terms of the solute–water and solute–solute interactions.

Determining the optimal model for improving seed germination and seedling vigor of perennial grasses

در اصلاح و احیاء مراتع توجه به مرحله جوانه زنی و استقرارگراس های پایا که حساس به تنشهای محیطی می باشد ضروری به نظر می رسد. بدین منظور تکنیک پرایمینگ بذر جهت بهبود سرعت جوانه زنی و بنیه گیاهچه پنج گراس چند ساله فصل سرد مرتعی شامل Bromus confinis، Elymus elongatum، Agropyron pectiniform، Festuca ovinaو Festuca arundinaceae با استفاده از ماده اسمتیک پلی اتیلن گلایکول (PEG 8000) در غلظتهای 0، 5/0 ، 1، 5/1 ، 2 و 5/2 مگاپاسکال (MP) برای دوره­های نگهداری صفر، 2، 4، 6 و 8 روز (D) مورد ارزیابی قرار گرفت. این آزمایش به صورت فاکتوریل در قالب طرح کاملا تصادفی در 4 تکرار اجرا شد. نتایج تجزیه واریانس نشان داد که تفاوت بین تمامی عوامل مورد مطالعه و اثرات متقابل آنها برای سرعت جوانه زنی و اجزاء گیاهچه بسیار معنی داربود (P<0.01). تاثیر پرایمینگ بذر بر افزایش سرعت جوانه زنی نسبت به شاهد (بذر پرایم نشده) مثبت بود و به طور خاص برای گونه های B. confinis، pectiniforme .A و E. elongatum (2 روز نگهداری و 2 مگاپاسکال) به ترتیب 9/127، 8/77 و 5/66 درصد افزایش و برای گونه F. arundinaceae (6 روز و 5/1 مگاپاسکال) 5/96 % افزایش را سبب شد و کمترین تاثیر در گونه F. ovina ( 4 روز و 5/2 مگاپاسکال) با 15 % افزایش نسبت به شاهد مشاهده شد. روابط رگرسیونی نشان داد که با افزایش غلظت PEG وزن خشک ریشه­چه و ساقه­چه برای تمامی گونه­ها به جز F. arundinaceaeو F. ovinaکاهش یافت. همچنین بیشترین کاهش وزن خشک ریشه­چه و ساقه­چه را دو گونه دانه درشت E. elongatum و B. confinesبا افزایش دوره نگهداری بذر (بیشتر از دو روز) و کاهش غلظت PEG نشان دادند. به طورکلی نتایج این تحقیق نشان داد که افزایش غلظت PEG تا حدود 2 و 5/2 مگاپاسکال و کاهش مدت نگهداری بذر تا 2 روز می­تواند در موفقیت تکنیک پرایمینگ بذر گراس­ها نقش موثرتری ایفا کند و شاید گونه­های دانه ریز پاسخ بهتری به پرایمینگ بذر جهت بهبود جوانه زنی و استقرار نسبت به دانه درشت نشان دهند.

EFFECT OF PEG INDUCED OSMOTIC STRESS ON PROLINE, PROTEIN AND RELATIVE WATER CONTENT IN IN VITRO PLANTS OF MACROTYLOMA UNIFLORUM (LAM.) VERDC.

In vitro plantlets of Macrotyloma uniflorum were obtained on L 2 IBA+BAP under water stress conditions induced by supplementing the medium with PEG at various concentrations ranging from 5% to 30% (w/v). Soluble proteins, amino acids, proline, water potential and relative water content were estimated in both control and stress induced in vitro shoots. Induced water stress has decreased the relative water content and water potential. However, accumulation of proteins, amino acids and proline was high in in vitro shoots raised on the medium supplemented with PEG at 25%. The in vitro shoots adapted to low concentrations of PEG showed more growth when compared to the plants adapted to high concentration of PEG. Importance of stress induced in vitro plants of Macrotyloma uniflorum in gene transfer technology to raise stress tolerance plants is discussed.

Structural and Thermodynamic Aspects of Ionic Solvation in Concentrated Aqueous Poly(ethylene glycol)

Solvation of ions in concentrated aqueous poly(ethylene glycol) (PEG) has been studied from thermodynamic and structural viewpoints using ion-transfer voltammetry at the interface between aqueous and nitrobenzene phases and X-ray absorption fine structure (XAFS). Systematic changes in the ion-transfer potential from water to aqueous PEG have been confirmed for several ions relative to the corresponding potential of tetraethylammonium ion (Et4N+), which is almost independent of PEG concentration. The results obtained for alkali cations strongly suggest the involvement of their complexation with PEG even in relatively diluted PEG solutions. It has been implied that the solvation circumstances of Br- and ClO4- are drastically altered when the PEG concentration becomes higher than particular critical values (e.g., 30-50% PEG200), where free water molecules are diminished because of the hydration of PEG. XAFS measurements have also been performed for K+ and Br- to get direct evidence for these findings. Although the spectra at the K K-edge clearly indicate the presence of a PEG complex of K+ in relatively diluted PEG solutions ( approximately 33% PEG200), an obvious increase in its ion-transfer potential has been detected at lower PEG concentrations, indicating that complexes formed at the interface rather than in bulk solution are transferred into an organic phase. Br- is fully hydrated in 0-50% PEG solutions, whereas some water molecules are replaced by PEG when the PEG concentration increases. Increasing the PEG concentration causes decreases in the coordination number from 6 in water to 2-3 in neat PEG. Thus, the present approach not only has elucidated the structural and thermodynamic aspects of ionic solvation in aqueous PEG but also has provided the information of the hydration of PEG.

Investigation into the influence of polymeric stabilizing excipients on inter-particulate forces in pressurised metered dose inhalers

Colloid probe atomic force microscopy (AFM) was utilised to quantify the cohesive forces of salbutamol sulphate in a model non-pressurised fluorinated liquid (mHFA), in the presence of increasing concentrations of poly(ethylene glycol) (PEG; molecular weight (MW) 200, 400 and 600). In addition, samples of PEG 400 (0.05–0.5%, v/w), were analysed in the presence of 0.001% (w/w) of poly(vinyl pyrrolidone) (PVP). In the absence of any stabilizing agents, strong attractive forces were present between particles. Increasing the concentration of the different MW PEG solutions in the mHFA system (up to 0.5%, v/w), significantly decreased the force of interaction (ANOVA, p < 0.05). The decrease in cohesion was particularly evident at very low concentrations of PEG (0.05–0.1%, v/w). Further data analysis ( p < 0.05) suggested that the reduction in the force of cohesion was dependent on the concentration and molecular weight of PEG. The addition of low concentration of PVP to the PEG 400-mHFA system had the most significant influence on drug particle cohesion. In the presence of PVP, increasing addition of PEG 400 (0.05–0.5%, v/w) to the mHFA, resulted in no significant reduction in the force of cohesion ( p > 0.05). Clearly, an understanding of the conformation of polymer molecules at interfaces is of vital importance when controlling the stability/flocculation behaviour of sterically stabilized pMDI suspensions. In this context, the use of the colloid probe AFM technique has provided a quantitative insight into the interactions of these complex systems and may be an invaluable asset during the early phase of formulation product development.

Synthesis of Polyglycidol Hydrogel Films Crosslinked with Carboxyl-terminated Poly(ethylene glycol)

With the attempt to largely increase the water content while keeping mechanical properties, hyperbranched polyglycidol (PGLD) hydrogel films were prepared using the oligomeric crosslinking agent, carboxyl-terminated poly(ethylene glycol) (PEG) of different molecular weights. Both the swelling and mechanical (tensile) properties of PGLD hydrogels crosslinked with PEG were measured and compared with the previous glutaric acid crosslinking system. The resultant PGLD films showed higher gel fraction and greater swelling capacity even at low PEG concentrations compared to those crosslinked with glutaric acid, probably due to the combined effect of the hydrophilicity and long chain effect of the PEG crosslinker. The values of the Young’s modulus and tensile strength were also higher for crosslinked films from PEG than from glutaric acid at a given (lower) crosslinker concentration. In addition, both the swelling and tensile properties of the PGLD hydrogels were largely affected by the crosslinker concentration.

DNA Ψ-Condensation and Reentrant Decondensation: Effect of the PEG Degree of Polymerization

Psi-condensation of DNA fragments of about 4 kbp was induced by poly(ethylene glycol) (PEG), with degrees of polymerization ranging from 45 to 182, and univalent salt (NaCl). Using circular dichroism spectroscopy, we were able to accurately determine the critical amount of PEG needed to induce condensation, as a function of the NaCl concentration. A significant dependence on the PEG degree of polymerization was found. Phase boundaries determined for the multimolecular condensation were very similar to those observed previously for the monomolecular collapse, with two asymptotic regimes at low and high salt concentrations. We analyze our data using a theoretical model that properly takes into account both the polyelectrolyte nature of the DNA and the liquid crystallinity of the condensed phase. The model assumes that all PEG is excluded from the condensates and shows reentrant decondensation only at low salt. We also systematically study reentrant decondensation and find a very strong dependence on PEG molecular weight. At low PEG molecular weight, decondensation occurs at relatively low concentrations of PEG, and over a wide range of salt concentrations. This suggests that in the reentrant decondensation the flexible polymers used are not completely excluded from the condensed phase.

The effects of poly(ethylene glycol) on the solution structure of human serum albumin

Protein physical and chemical properties can be altered by polymer interaction. The presence of several high affinity binding sites on human serum albumin (HSA) makes it a possible target for many organic and polymer molecules. This study was designed to examine the interaction of HSA with poly(ethylene glycol) (PEG) in aqueous solution at physiological conditions. Fourier transform infrared, ultraviolet-visible, and CD spectroscopic methods were used to determine the polymer binding mode, the binding constant, and the effects of polymer complexation on protein secondary structure. The spectroscopic results showed that PEG is located along the polypeptide chains through H-bonding interactions with an overall affinity constant of K = 4.12 x 10(5) M(-1). The protein secondary structure showed no alterations at low PEG concentration (0.1 mM), whereas at high polymer content (1 mM), a reduction of alpha-helix from 59 (free HSA) to 53% and an increase of beta-turn from 11 (free HSA) to 22% occurred in the PEG-HSA complexes (infrared data). The CDSSTR program (CD data) also showed no major alterations of the protein secondary structure at low PEG concentrations (0.1 and 0.5 mM), while at high polymer content (1 mM), a major reduction of alpha-helix from 69 (free HSA) to 58% and an increase of beta-turn from 7 (free HSA) to 18% was observed.

Genotypic variation in the photosynthetic competence of Sorghum bicolor seedlings subjected to polyethylene glycol-mediated drought stress

Eleven varieties of Sorghum bicolor, subjected to PEG-mediated drought stress were compared for their photosynthetic performance. The varieties differed in their relative water content over a range of PEG concentrations (0-25%). CO2 assimilation, stomatal conductance and the quantum yield of PSII electron transport decreased with increasing PEG concentrations in all varieties. However the intercellular CO2 concentration showed a nonlinear PEG concentration-dependent change. At lower PEG concentrations there was a decrease in the levels of intercellular CO2 concentration in all varieties that could be attributed to stomatal closure. At higher PEG concentrations, some varieties showed an increase in the intercellular CO2 concentration, indicating an inhibition of photosynthetic activity due to non-stomatal effects, while others did not. It was seen that the varieties differed in the stress thresholds at which stomatal and metabolic limitations to photosynthesis occur. These differences in the photosynthetic adaptation of Sorghum varieties could be useful in identifying genotypes showing large differences in photosynthetic adaptation, which could be useful in mapping photosynthetic traits for drought stress tolerance.

Modulation of Graves' disease autoantibody stimulation of recombinant human thyrotrophin receptor

Background: The cAMP response of porcine thyrocytes to the immunoglobulin (Ig) fraction from Graves' sera is increased if these fractions are prepared with higher than usual concentrations of polyethylene glycol (PEG; 22.5% rather than 13.5%), leading to the suggestion that additional factors might exist in serum which influence the ability of autoantibodies to stimulate thyroid cells. Methods: We characterised the stimulatory activity of fractions prepared by differential PEG precipitation of Graves' sera, using heterologous eukaryotic cells expressing recombinant human thyrotrophin (TSH) receptor. Results: We found no evidence that material soluble in 13.5% PEG but precipitated by 22.5% PEG was stimulatory either on its own or in combination with the immunoglobulin-containing material precipitated by 13.5% PEG. Indeed, the stimulatory effect was reproduced by simply including low concentrations of PEG (1–4%) in the diluted sera from the majority of Graves' patients (12/18 tested) added to the target cells. Intriguingly, PEG had no effect on basal levels, with normal sera or stimulation by thyrotrophin. Conclusions: The increase in stimulation reported when using immunoglobulins prepared with higher concentrations of PEG is attributable to a direct effect of the extra PEG (which had not been removed from the preparations) during the stimulation of the cells. Because the effect is observed with recombinant heterologous cells, it must be caused by interaction between PEG and the autoantibodies and/or the receptor—it cannot involve any other thyroid-specific molecule, although an involvement of other molecules widely distributed on many cell types is possible.

Interaction between Poly(ethylene glycol) and C12E8 Investigated by Dynamic Light Scattering, Time-Resolved Fluorescence Quenching, and Calorimetry

Dynamic light scattering (DLS), time-resolved fluorescence quenching (TRFQ), and isothermal titration microcalorimetry have been used to show that, in dilute solution, low molecular weight poly(ethylene glycol) (PEG, M-w = 12 kDa) interacts with the nonionic surfactant octaethylene glycol n-dodecyl monoether, C12E8, to form a complex. Whereas the relaxation time distributions for the binary C12E8/water and PEG/water systems are unimodal, in the ternary mixtures they may be either uni- or bimodal depending on the relative concentrations of the components. At low concentrations of PEG or surfactant, the components of the relaxation time distribution are unresolvable, but the distribution becomes bimodal at higher concentrations of either polymer or surfactant. For the ternary system in excess surfactant, we ascribe, on the basis of the changes in apparent hydrodynamic radii and the scattered intensities, the fast mode to a single micelle, the surface of which is associated with the polymer and the slow mode to a similar complex but containing two or three micelles per PEG chain. Titration microcalorimetry results show that the interaction between C12E8, and PEG is exothermic and about 1 kJ mol(-1) at concentrations higher than the CMC Of C12E8. The aggregation number, obtained by TRFQ, is roughly constant when either the PEG or the C12E8 concentration is increased at a given concentration of the second component, owing to the increasing amount of surfactant micelles inside the complex. (Less)