Effect of laundry surfactants on surface charge and colloidal stability of silver nanoparticles.

The stability of silver nanoparticles (Ag NPs) potentially released from clothing during a laundry cycle and their interactions with laundry-relevant surfactants [anionic (LAS), cationic (DTAC), and nonionic (Berol)] have been investigated. Surface interactions between Ag NPs and surfactants influence their speciation and stability. In the absence of surfactants as well as in the presence of LAS, the negatively charged Ag NPs were stable in solution for more than 1 day. At low DTAC concentrations (≤1 mM), DTAC-Ag NP interactions resulted in charge neutralization and formation of agglomerates. The surface charge of the particles became positive at higher concentrations due to a bilayer type formation of DTAC that prevents from agglomeration due to repulsive electrostatic forces between the positively charged colloids. The adsorption of Berol was enhanced when above its critical micelle concentration (cmc). This resulted in a surface charge close to zero and subsequent agglomeration. Extended DLVO theory calculations were in compliance with observed findings. The stability of the Ag NPs was shown to depend on the charge and concentration of the adsorbed surfactants. Such knowledge is important as it may influence the subsequent transport of Ag NPs through different chemical transients and thus their potential bioavailability and toxicity.

TD-11 workshop report: characterization of monoclonal antibodies to S100 proteins.

Fourteen monoclonal antibodies with specificity against native or recombinant antigens within the S100 family were investigated with regard to immunoreactivity. The specificities of the antibodies were studied using ELISA tests, Western blotting epitope mapping using competitive assays, and QCM technology. The mimotopes of antibodies against S100A4 were determined by random peptide phage display libraries. Antibody specificity was also tested by IHC and pair combinations evaluated for construction of immunoradiometric assays for S100B. Out of the 14 antibodies included in this report eight demonstrated specificity to S100B, namely MAbs 4E3, 4D2, S23, S53, 6G1, S21, S36, and 8B10. This reactivity could be classified into four different epitope groups using competing studies. Several of these MAbs did display minor reactivity to other S100 proteins when they were presented in denatured form. Only one of the antibodies, MAb 3B10, displayed preferential reactivity to S100A1; however, it also showed partial cross-reactivity with S100A10 and S100A13. Three antibodies, MAbs 20.1, 22.3, and S195, were specific for recombinant S100A4 in solution. Western blot revealed that MAb 20.1 and 22.3 recognized linear epitopes of S100A4, while MAb S195 reacted with a conformational dependent epitope. Surprisingly, MAb 14B3 did not demonstrate any reactivity to the panel of antigens used in this study.

Layer-by-layer assemblies of chitosan and heparin: effect of solution ionic strength and pH.

The growth of polysaccharide multilayers consisting of positively charged chitosan (CH) and negatively charged heparin (HEP) was monitored in situ by employing a quartz crystal microbalance (QCM-D) and dual-polarization interferometry (DPI). The main focus was on how the physicochemical properties of the solution affect the growth and structure of the resulting multilayer film. These results showed that when increasing the ionic strength of the polysaccharide solutions at a fixed pH, both the "dry" (optical) (DPI) mass and wet (QCM) mass of the adsorbed multilayer film increased. The same effect was found when increasing the pH while keeping the ionic strength constant. Furthermore, the growth of multilayers showed an exponential-like behavior independent of the solution conditions that were used in this study. It was also established that chitosan was the predominant species present in the chitosan-heparin multilayer film. We discuss the viscoelastic properties of the adsorbed layers and their variation during the multilayer buildup. Interestingly and contrary to common interpretation of the QCM-D results, we found that under one particular solution condition (pH 4.2 and 30 mM NaCl) the increase in the dissipation of oscillation energy from the adsorbed layer was a consequence of layer stiffening rather than indicating a more hydrated and viscous film. On the basis of the widely used Voigt viscoelastic model for an adsorbed layer, we show that it is the film viscosity and shear that define the layer viscoelasticity (structure) of the film and not the absolute value of energy dissipation, which in fact can be very misleading.

Equal effect of vancomycin lock with or without heparin in treatment of central venous catheter related blood stream infections - an observational study in palliative home care.

BACKGROUND: Catheter related blood stream infections is a complication in patients with central venous catheter. The aim of this study was to compare vancomycin-heparin-lock and non-heparin, high-dose vancomycin-lock as treatment in patients admitted to palliative home care. With non-heparin, high-dose vancomycin-lock a high concentration of 5 mg/mL vancomycin is attained instead of only 500 ug/mL when dissolved in heparin. The non-heparin method also has the advantage of being easier and cheaper but might entail an increased risk of clotting. METHODS: Medical records from patients enrolled at a palliative home care unit in Stockholm between 2016 and 2018 were reviewed retrospectively. Three divisions used vancomycin-heparin-lock and the other 3 divisions used non-heparin, high-dose vancomycin-lock. Inclusion criteria were a central venous catheter related blood stream infection treated with one of the two methods for >7 days and a follow-up blood culture at the end of treatment. RESULTS: Twenty-five patients fulfilled the inclusion criteria, 12 treated with vancomycin-heparin-lock and 13 with non-heparin, high-dose vancomycin-lock. There was no significant difference in resolved infections between the two treatments, 6 of 12 for vancomycin-heparin-lock and 10 of 13 for non-heparin, high-dose vancomycin-lock (p = .23). In the non-heparin group one central venous catheter was removed due to clotting although the infection had resolved. Overall, removal of central venous catheter was similar in the two groups (6 of 12 and 4 of 13, p = .43). CONCLUSION: The current study does not support superiority of one treatment over the other. However, larger, randomized studies are needed, before firm conclusions can be drawn.

Polymer dynamics in layer-by-layer assemblies of chitosan and heparin.

The layer-by-layer deposition method has been used to build a multilayer thin film with two polysaccharides, chitosan CH (weak polycation) and heparin HEP (strong polyanion), on planar quartz surfaces. The film structure and dynamics in aqueous solution were studied with fluorescence resonance energy transfer (FRET) and total internal reflection fluorescence (TIRF). Particular emphasis was placed on the effect of deposition conditions, i.e., pH and salt concentration, on the out-of-plane (vertical) diffusion of fluorescence labeled chitosan in the chitosan/heparin (CH/HEP) film. FRET analysis showed that CH molecules diffused within the film with a diffusion coefficient that was not significantly sensitive to the deposition pH and solution ionic strength. A pH-sensitive label bound to CH embedded within the CH/HEP film was sensitive to the charge of the outermost polymer layer even when buried under 14 alternate layers of CH and HEP. A consideration of the results obtained with both fluorescence techniques showed that the structure of the CH/HEP thin film was highly interpenetrated without clear boundaries between each layer. These results are consistent with the hypothesis that the previously observed exponential-like film growth of CH and HEP in terms of layer thickness and deposited amount versus deposition cycle can be attributed to out-of-plane diffusion of CH molecules in the multilayer.

Soluble complexes in aqueous mixtures of low charge density comb polyelectrolyte and oppositely charged surfactant probed by scattering and NMR.

A low charge density polyelectrolyte with a high graft density of 45 units long poly(ethylene oxide) side-chains has been synthesized. In this comb polymer, denoted PEO(45)MEMA:METAC-2, 2 mol% of the repeating methacrylate units in the polymer backbone carry a permanent positive charge and the remaining 98 mol% a 45 unit long PEO side-chain. Here we describe the solution conformation of this polymer and its association with an anionic surfactant, sodium dodecylsulfate, SDS. It will be shown that the polymer can be viewed as a stiff rod with a cross-section radius of gyration of 29 A. The cross section of the rod contracts with increasing temperature due to decreased solvency of the PEO side-chains. The anionic surfactant associates to a significant degree with PEO(45)MEMA:METAC-2 to form soluble complexes at all stoichiometries. A cooperative association is observed as the free SDS concentration approaches 7 mM. At saturation the number of SDS molecules associated with the polymer amounts to 10 for each PEO side-chain. Two distinct populations of associated surfactants are observed, one is suggested to be molecularly distributed over the comb polymer and the other constitutes small micellar-like structures at the periphery of the aggregate. These conclusions are reached based on results from small-angle neutron scattering, static light scattering, NMR, and surface tension measurements.

Isolation of the promoters of Atlantic salmon MHCII genes.

The major histocompatibility complex class II (MHCII) has a central role in the immune response of vertebrates with its function of presenting antigenic peptides to the T-cell receptors. We have isolated the promoters and intron 1 of MHCIIalpha and MHCIIbeta genes of Atlantic salmon. To isolate these promoters, we constructed an Atlantic salmon ( Salmo salar) promoter finder kit (analogous to the commercially available "human promoter finder kit"). By nucleotide sequence alignment of known MHCII promoter regions, we identified the 3 conserved regulatory X, X2, and Y boxes in the salmon promoters. The W box was not found. In contrast, a salmon-specific putative W box was identified. Both of the isolated Atlantic salmon MHCIIalpha and beta promoters (included in patent applications by Genomar A/S, Oslo, Norway) were found to be functional since they both gave positive yellow fluorescence protein signal when inserted as promoters in the pEYFP-1 reporter plasmid and transfected into the salmon head kidney cell line (SHK-1).

Adsorption of IgG on/in a PAH/PSS multilayer film: Layer structure and cell response.

The binding of immunogloblulins (IgG) (mouse monoclonal recognizing IFNγ) on precoated polystyrene or silica surfaces by the layer-by-layer technique has been investigated with QCM-D and DPI. The aim of the work was to increase the sensitivity of the conventional enzyme-linked immunosorbent spot (ELISpot) assay. The polyelectrolytes used to build the multilayers were poly(allylamine hydrochloride) (PAH)/poly(sodium 4-styrenesulfonate) (PSS) alternately adsorbed from 150mM NaCl. The multilayer build up is linear and the internal structure of the PAH/PSS multilayer is compact and rigid as observed by low relative water content (20-25%) and high layer refractive index (n∼1.5) after the formation of five bilayers. Incorporation of IgG within the PAH/PSS multilayer did not give rise to overcharging and did not affect the linear build up. ELISpot test on PAH/PSS multilayer modified polystyrene wells showed that the cytokine response was significantly smaller than on the regular PVDF backed polystyrene wells. This may be due to the compact and rigid nature of the PAH/PSS multilayer, which does not allow formation of the kind of three dimensional support needed to achieve bioactive IgG binding to the surface. Immunological tests of the polyelectrolyte multilayers in the absence of IgG showed that PSS terminated PAH/PSS multilayer did not induce any cytokine response whereas PAH terminated did, which suggests that PSS totally covers the surface from the cells point of view.

Adsorption of lysozyme, beta-casein and their layer-by-layer formation on hydrophilic surfaces: Effect of ionic strength.

The adsorbed amount and layer structure of lysozyme, beta-casein and mixed layers of the two proteins were studied on hydrophilic silica and quartz surfaces using the following techniques: ellipsometry, quartz crystal microbalance with dissipation monitoring (QCM-D) and total internal reflection fluorescence (TIRF). Particular emphasis was put on the effect of solution ionic strength on the layer formation. Both lysozyme and beta-casein showed a higher affinity for the silica surface when adsorbed from a solution of low ionic strength even though beta-casein and silica are negatively charged at the pH used. No beta-casein remained adsorbed after rinsing with a 150mM buffer solution. The adsorbed amount of lysozyme on silica exceeded a monolayer coverage irrespective of the solution conditions and displayed a rigid structure. beta-Casein forms more than a single layer on pre-adsorbed lysozyme; an inner flat layer and an outer layer with an extended structure, which largely desorbs on rinsing. The build-up through sequential adsorption of lysozyme and beta-casein is favoured at intermediate and high ionic strength. The total adsorbed amount increased slightly with each deposition cycle and the mixed lysozyme/beta-casein layers contain higher amounts of protein compared to those of pure lysozyme or beta-casein. Sequential adsorption gives rise to a proteinaceous layer consisting of both lysozyme and beta-casein. The protein layers are probably highly interpenetrated with no clear separation between them.

Comparison of the adsorption kinetics and surface arrangement of "as received" and purified bovine submaxillary gland mucin (BSM) on hydrophilic surfaces.

The effect of bovine serum albumin (BSA) as impurity in a commercial bovine submaxillary gland mucin preparation (BSM; Sigma M3895) on the adsorption of BSM to hydrophilic surfaces (mica and silica) has been studied in terms of adsorption kinetics, amount and structure of the formed adlayer. The Surface Force Apparatus (SFA) was used to gain information about the extended and compressed structure of adsorbed "as received" BSM, purified BSM, BSA extracted from the "as received" BSM and mixtures of the latter purified proteins. The adsorbed amount was estimated using a combination of X-ray Photoelectron Spectroscopy (XPS), Enzyme-Linked Immuno Sorbent Assay (ELISA), Enzyme-Linked Lectin Assay (ELLA), Dual Polarization Interferometry (DPI) and Quartz Crystal Microbalance (QCM-D) measurements. Under the used conditions, purified BSM showed very low affinity for silica and only small amounts were found to adsorb on mica. Initially, the BSM molecules adopted an extended conformation on the mica surface with tails extending into the bulk phase. These tails were irreversibly compressed into a very thin (10A) layer upon applying a high load. "As received" BSM formed considerably thicker compressed layers (35A); however, the extended layer structure was qualitatively the same. When mixtures of purified BSM and BSA were coadsorbed on mica, a 9wt-% albumin content gave a comparable layer thickness as the "as received" BSM and from XPS data we draw the conclusion that the albumin content in the layer adsorbed from "as received" BSM was approximately 5wt-%. Adsorption from an equal amount of BSM and BSA revealed that even though the amount of BSM is scarce in the mixed layer, the few BSM molecules have a drastic effect on the adsorbed thickness and structure. Clearly, this study shows the importance of characterizing the mucin used since differences in purity give rise to different adsorption behaviours in terms of both adsorbed amount and layer structure.

Interactions between chitosan and SDS at a low-charged silica substrate compared to interactions in the bulk--the effect of ionic strength.

The effect of ionic strength on association between the cationic polysaccharide chitosan and the anionic surfactant sodium dodecyl sulfate, SDS, has been studied in bulk solution and at the solid/liquid interface. Bulk association was probed by turbidity, electrophoretic mobility, and surface tension measurements. The critical aggregation concentration, cac, and the saturation binding of surfactants were estimated from surface tension data. The number of associated SDS molecules per chitosan segment exceeded one at both salt concentrations. As a result, a net charge reversal of the polymer-surfactant complexes was observed, between 1.0 and 1.5 mM SDS, independent of ionic strength. Phase separation occurs in the SDS concentration region where low charge density complexes form, whereas at high surfactant concentrations (up to several multiples of cmc SDS) soluble aggregates are formed. Ellipsometry and QCM-D were employed to follow adsorption of chitosan onto low-charged silica substrates, and the interactions between SDS and preadsorbed chitosan layers. A thin (0.5 nm) and rigid chitosan layer was formed when adsorbed from a 0.1 mM NaNO3 solution, whereas thicker (2 nm) chitosan layers with higher dissipation/unit mass were formed from solutions at and above 30 mM NaNO3. The fraction of solvent in the chitosan layers was high independent of the layer thickness and rigidity and ionic strength. In 30 mM NaNO3 solution, addition of SDS induced a collapse at low concentrations, while at higher SDS concentrations the viscoelastic character of the layer was recovered. Maximum adsorbed mass (chitosan + SDS) was reached at 0.8 times the cmc of SDS, after which surfactant-induced polymer desorption occurred. In 0.1 mM NaNO3, the initial collapse was negligible and further addition of surfactant lead to the formation of a nonrigid, viscoelastic polymer layer until desorption began above a surfactant concentration of 0.4 times the cmc of SDS.

Stress-specific role of fission yeast Gcn5 histone acetyltransferase in programming a subset of stress response genes.

Gcn5 is a coactivator protein that contributes to gene activation by acetylating specific lysine residues within the N termini of histone proteins. Gcn5 has been intensively studied in the budding yeast, Saccharomyces cerevisiae, but the features of genes that determine whether they require Gcn5 during activation have not been conclusively clarified. To allow comparison with S. cerevisiae, we have studied the genome-wide role of Gcn5 in the distantly related fission yeast, Schizosaccharomyces pombe. We show that Gcn5 is specifically required for adaptation to KCl- and CaCl(2)-mediated stress in S. pombe. We have characterized the genome-wide gene expression responses to KCl stress and show that Gcn5 is involved in the regulation of a subset of stress response genes. Gcn5 is most clearly associated with KCl-induced genes, but there is no correlation between Gcn5 dependence and the extent of their induction. Instead, Gcn5-dependent KCl-induced genes are specifically enriched in four different DNA motifs. The Gcn5-dependent KCl-induced genes are also associated with biological process gene ontology terms such as carbohydrate metabolism, glycolysis, and nicotinamide metabolism that together constitute a subset of the ontology parameters associated with KCl-induced genes.

Squamous cell carcinoma antigen isoforms in serum from cervical cancer patients.

Squamous cell carcinoma antigen (SCCA) is a serological marker of squamous cell carcinomas (SCC). To study whether any of the SCCA isoforms would provide additional and more specific/sensitive clinical information than total SCCA, immunoassays specific for the different forms of SCCA (free SCCA2, total SCCA2, total SCCA1 and total SCCA) were developed. SCCA isoforms were determined before therapy and in follow-up samples from patients with cervical cancer and in serum samples from healthy females. Serum samples from patients with benign skin diseases (psoriasis and eczema) were also selected based on elevated SCCA levels. Rising levels of SCCA1 and SCCA2 were seen in patients with recurrence or progressive disease at the end of the study. The rise in SCCA2 was usually more prominent than that in SCCA1. The dominating serological form of SCCA was free SCCA2 both in healthy controls and in patients with cervical cancer. Both SCCA1 and SCCA2 were elevated in serum from cervical cancer patients and followed the clinical course of the disease during therapy monitoring. SCCA2 did not show improved tumor specificity as compared to SCCA1. A larger study is however necessary to make firm conclusions.

Mucin-chitosan complexes at the solid-liquid interface: multilayer formation and stability in surfactant solutions.

The adsorption of a biologically important glycoprotein, mucin, and mucin-chitosan complex layer formation on negatively charged surfaces, silica and mica, have been investigated employing ellipsometry, the interferometric surface apparatus, and atomic force microscopy techniques. Particular attention has been paid to the effect of an anionic surfactant sodium, dodecyl sulfate (SDS), with respect to the stability of the adsorption layers. It has been shown that mucin adsorbs on negatively charged surfaces to form highly hydrated layers. Such mucin layers readily associate with surfactants and are easily removed from the surfaces by rinsing with solutions of SDS at concentrations > or =0.2 cmc (1 cmc SDS in 30 mM NaCl is equal to 3.3 mM). The mucin adsorption layer is negatively charged, and we show how a positively charged polyelectrolyte, chitosan, associates with the preadsorbed mucin to form mucin-chitosan complexes that resist desorption by SDS even at SDS concentrations as high as 1 cmc. Thus, a method of mucin layer protection against removal by surfactants is offered. Further, we show how mucin-chitosan multilayers can be formed.