Low Molecular Weight Shuttle Molecules Enhance Polychloramide Antimicrobial Activity.

Investigated were the influences of succinimide (SI), 5,5-dimethylhydantoin (DMH), and 3-(hydroxymethyl)-5,5-dimethylhydantoin (HDMH) on the biocidal activity of chlorinated, water-soluble polyamide prepared by the reaction of isopropylamine with poly(styrene-alt-maleic anhydride). The resulting polymer was a negatively charged, water-soluble polymer bearing a carboxylic acid and an isopropylamide moiety on nearly every repeat unit. Subsequent treatment with NaOCl chlorinated the polymers to up to 4.4% Cl while inflicting some polymer chain scission. SI, DMH, or HDMH increased the biocidal activity of polychloramides toward planktonic Escherichia coli and Staphylococcus aureus. Independent solution studies confirmed that oxidative chlorine spontaneously transferred from aqueous polychloramides to small molecules. We concluded that SI, DMH, and HDMH acted as shuttles that extracted oxidative Cl from the polymer chloramides and transported oxidative Cl more efficiently to microbial surfaces. Succinimide was the most effective shuttle. These results warn that some low molecular weight soluble molecules in antimicrobial testing solutions may exaggerate the effectiveness of the polymer or immobilized antimicrobial agents.

Modeling the Impact of Polychloramide Solution Properties on Bacterial Disinfection Kinetics.

Anionic water-soluble polychloramide biocides are of interest because, compared to conventional cationic antimicrobial polymers, anionic biocides are less likely to be sequestered or deactivated by contact with non-microbial soil. Although electrostatics can prevent anionic polymers from adsorbing on microbes, water-soluble polychloramides appear to transfer oxidative chlorine during transient contacts between polymer chains and microbe surfaces. The Chick-Watson model of disinfection kinetics has been modified to account for the contributions of polychloramide molecular weight (MW) and the polychloramide configuration in solution estimated from the overlap concentration, C*, below which dilute polymer chains exist as discrete objects in solution. The key assumption in the modeling was that the transfer rate of oxidative chlorine from polychloramide chains to microbe surfaces impacts the disinfection kinetics. Because both C* and MW are measurable, the polymer-modified Chick-Watson (PCW) model has one less unknown parameter than the two-parameter Chick-Watson equation. The PCW model predicts that lower MW polymers are more effective biocides compared with high MW counterparts. Additionally, polymers with more compressed configurations in solution are more effective biocides. Experimental evidence supports these conclusions. Based on the estimated time scale of bacteria/polymer collisions compared with disinfection kinetics, arguments are made that bacteria surfaces must be contracted many times by polychloramide chains to achieve sufficient Cl transfer to deactivate bacteria.

High Yield Poly(ethylene-alt-maleic acid) Grafting to Wood Pulp while Minimizing Fiber/Fiber Wet Adhesion.

Heating bleached kraft pulps treated with poly(ethylene-alt-maleic acid) (PEMAc) can lead to high yields of carboxylated polymer grafted to fibers. However, in many cases, the cured, dry pulp cannot be effectively repulped (redispersed in water) because the wet strength is too high. Impregnation with PEMAc solutions at pH 4 followed by high temperature (120-180 °C), catalyst-free curing for short times can give fixation yields >85% while maintaining repulpability. The combination of high fixation yields with low wet strength is possible because the extent of curing required for high grafting yields is less than the curing requirement for high wet strength. Two challenges in moving this technology to practicable applications are (1) identifying the optimum laboratory pulp curing conditions and (2) translating laboratory curing conditions to industrial processes. A modeling tool was developed to meet these challenges. The model is based on the observation that for curing conditions giving high fixation yields the wet tensile indices of grafted pulp sheets showed a power-law dependence on the ßΓ product where ß is the conversion of the succinic acid moieties in PEMAc to the corresponding succinic anhydride groups in the curing step and Γ is the amount of polymer applied to the pulp. For two PEMAc molecular weights and two pulp types, the power-law slopes were 0.6; however, the pre-exponential terms depended upon the specific polymer and pulp type combination. We propose that the relationships between the wet tensile index and ßΓ, from polymer-treated, laboratory pulp handsheets, can be used to predict if proposed curing conditions for larger-scale processes will produce a repulpable product.

Challenges to Achieving Strong but Fully Degradable Adhesive Joints between Wet Cellulose Surfaces.

The dramatic loss of strength upon exposure to water is one of the challenges preventing the widespread substitution of plastic packaging with paper and paperboard. Although treatment with conventional wet strength resins and other adhesive polymers can strengthen wet paper, it is at the expense of green credentials, including easy recycling or rapid composting. The goal of this work was to demonstrate the adhesive requirements for strong, wet cellulose-cellulose joints that can be recycled easily because the joint strength can be destroyed by the presence of a weak reducing agent. Cellulose membrane surfaces were first treated with a bound layer of carboxymethyl cellulose, modified to have covalently tethered hydrazide groups. Joints were fabricated by laminating two hydrazide-modified membranes with a polymeric adhesive bearing aldehyde functionality. Aldehydes spontaneously condense with hydrazide to give hydrazone bonds. When the adhesive was oxidized dextran, the wet laminates had an intermediate strength. Upon exposure to a reducing agent, the joint strength was reduced to nearly zero because every hydrazone moiety connecting two cellulose surfaces included a cleavable disulfide linkage. By contrast, glyoxalated cationic polyacrylamide gave very strong wet joints. However, the wet adhesion was dominated by polyelectrolyte complexation, and the presence of hydrazone linkages had little influence on the wet adhesion. We conclude that robust joint degradability will require cleavable linkages in the adhesive backbone while minimizing polyelectrolyte complexation.

Deposited Nanoparticles Can Promote Air Clogging of Piezoelectric Inkjet Printhead Nozzles.

Piezoelectric inkjet printing is susceptible to printhead clogging when printing with inks that contain dispersed particles. This paper investigates the mechanisms by which 28-530 nm nanoparticle dispersions induce printhead clogging without forming large aggregates or thick deposited layers on printhead surfaces. Printing experiments were combined with nanoparticle deposition studies and with experiments where inks were pumped through printheads at a constant flow rate with a syringe pump. Submonolayer coverages of hydrophobic cationic polystyrene nanoparticles adhering to printhead surfaces promote rapid clogging by trapped air that enters from the nozzle opening. We propose that the deposited particles distort the shape of the ink/air meniscus, possibly causing air entrainment, and promote air bubble adhesion to the interior printhead surfaces. The printer's purge-blot cleaning procedure removes air clogs, but the clogs quickly reform when printing is resumed because the adsorbed nanoparticles are not removed by the cleaning procedure. Nondepositing anionic hydrophobic nanoparticles cause much less clogging, possibly because of filtration of trace large aggregates. Colloidal stability is a necessary but not sufficient criterion for ink dispersions; the ink particles must not adsorb onto the printhead surfaces. Thus, alternate surface chemistries for the printhead and ink particle surfaces may be required to print hydrophobic ink materials.

Relating Redox Properties of Polyvinylamine-g-TEMPO/Laccase Hydrogel Complexes to Cellulose Oxidation.

The structure and electrochemical properties of adsorbed complexes based on mixtures of polyvinylamine-g-TEMPO (PVAm-T) and laccase were related to the ability of the adsorbed complexes to oxidize cellulose. PVAm-T10 with 10% of the amines bearing TEMPO moieties (i.e., DS = 10%), adsorbed onto gold sulfonate EQCM-D sensor surfaces giving a hydrogel film that was 7 nm thick, 89% water, and encasing laccase (200 mM) and TEMPO moieties (33 mM). For DS values >10%, all of the TEMPOs in the hydrogel film were redox-active in that they could be oxidized by the electrode. With hydrogel layers made with lower-DS PVAm-Ts, only about half of the TEMPOs were redox-active; 10% DS appears to be a percolation threshold for complete TEMPO-to-TEMPO electron transport. In parallel experiments with hydrogel complexes adsorbed onto regenerated cellulose films, the aldehyde concentrations increased monotonically with the density of redox-active TEMPO moieties in the adsorbed hydrogel. The maximum density of aldehydes was 0.24 µmol/m2, about 10 times less than the theoretical concentration of primary hydroxyl groups exposed on crystalline cellulose surfaces. Previous work showed that PVAm-T/laccase complexes are effective adhesives between wet cellulose surfaces when the DS is >10%. This work supports the explanation that TEMPO-to-TEMPO electron transport is required for the generation of aldehydes necessary for wet adhesion to PVAm.

A Colloidal Stability Assay Suitable for High-Throughput Screening.

A library of 32 polystyrene copolymer latexes, with diameters ranging between 53 and 387 nm, was used to develop and demonstrate a high-throughput assay using a 96-well microplate platform to measure critical coagulation concentrations, a measure of colloidal stability. The most robust assay involved an automated centrifugation-decantation step to remove latex aggregates before absorbance measurements, eliminating aggregate interference with optical measurements made through the base of the multiwell plates. For smaller nanoparticles (diameter <150 nm), the centrifugation-decantation step was not required as the interference was less than with larger particles. Parallel measurements with a ChemiDoc MP plate scanner gave indications of aggregation; however, the results were less sensitive than the absorbance measurements.

Stable Aqueous Foams from Cellulose Nanocrystals and Methyl Cellulose.

The addition of cellulose nanocrystals (CNC) greatly enhanced the properties of methylcellulose (MC) stabilized aqueous foams. CNC addition decreased air bubble size, initial foam densities and drainage rates. Mixtures of 2 wt % CNC + 0.5 wt % MC gave the lowest density foams. This composition sits near the onset of nematic phase formation and also near the overlap concentration of methylcellulose. More than 94% of the added CNC particles remained in the foam phase, not leaving with the draining water. We propose that the nanoscale CNC particles bind to the larger MC coils both in solution and with MC at the air/water interface, forming weak gels that stabilize air bubbles. Wet CNC-MC foams were sufficiently robust to withstand high temperature (70 °C for 6 h) polymerization of water-soluble monomers giving macroporous CNC composite hydrogels based on acrylamide (AM), 2-hydroxyethyl methacrylate (HEMA), or polyethylene glycol diacrylate (PEGDA). At high temperatures, the MC was present as a fibrillar gel phase reinforced by CNC particles, explaining the very high foam stability. Finally, our CNC-MC foams are based on commercially available forms of CNC and MC, already approved for many applications. This is a "shovel-ready" technology.

Printed paper sensors for serum lactate dehydrogenase using pullulan-based inks to immobilize reagents.

In this study, a paper-based point-of-care (POC) colorimetric biosensor was developed for the detection of lactate dehydrogenase in serum using a nonporous, oxygen impermeable reversibly gelling polysaccharide material based on pullulan. The pullulan could be printed onto paper surfaces along with all required assay reagents, providing a means for high-stability immobilization of all reagents on paper. Serum containing lactate dehydrogenase (LDH) was directly spotted on to the pullulan-coated bioactive paper and provided quantitative colorimetric data that was comparable to that obtained with a conventional plate-reader method. The paper strip was found to be highly stable and could be stored at 4 °C for at least 10 weeks with no loss in performance, as compared to a complete loss in performance within 1 day when the reagents were printed without the stabilizing polysaccharide. The ease of fabrication coupled with the high stability of the printed reagents provides a facile platform for easily manufactured POC sensors.

Emulsion/Surface Interactions from Quiescent Quartz Crystal Microbalance Measurements with an Inverted Sensor.

Interactions of three oil-in-water emulsion types with polystyrene-coated quartz crystal microbalance (QCM) sensor surfaces were probed with the QCM cell in both the conventional orientation (i.e., polystyrene surface on the bottom, "looking up") and the inverted orientation (polystyrene on top interior surface of sensor chamber, "looking down"). With the conventionally oriented QCM sensors, the adsorption of soluble and/or dispersed species quickly gave steady-state frequency and dissipation outputs. By contrast, the inverted sensors gave changing responses at long times because of the gravity driven buildup of a viscous consolidation layer next to but not necessarily bound to the sensor surface. Three emulsion types (a simple hexadecane/phosphatidylcholine emulsion, 2% homogenized milk, and a diluted commercial ophthalmic emulsion) displayed a wide range of behaviors. We propose that quiescent QCM measurement made with an inverted sample chamber is a new approach to probing emulsion behaviors near solid surfaces.

Polyvinylamine: a tool for engineering interfaces.

With the highest content of primary amine functional groups of any polymer, polyvinylamine (PVAm) is a potent tool for the modification of macroscopic and nanoparticle surfaces. Based on the free radical polymerization and subsequent hydrolysis of N-vinylformamide, PVAm is prepared as linear polymers (0.8 kDa to >1 MDa), microgels, macrogels, and copolymers. The amine groups serve as reaction sites for grafting PVAm to surfaces and for the preparation of derivatives. Coupling low-molecular-weight molecules and oligomers gives PVAm-X, where X includes hydrophobes, carbohydrate oligomers, proteins, TEMPO (2,2,6,6-tetramethyl-1-piperidinyloxy), phenylboronic acids, and fluorocarbons. This contribution highlights the use of PVAm and PVAm-X to modify solid surface properties. Where possible, the PVAm properties and applications as an interfacial agent are compared to those of linear polyethylenimine, polyallylamine, and chitosan.

Aminated thermoresponsive microgels prepared from the Hofmann rearrangement of amides without side reactions.

Thermoresponsive microgels bearing primary amine groups were prepared by the Hofmann rearrangement of methacrylamide groups present in cross-linked NIPMAM (N-isopropylmethacrylamide) microgels. Most thermoresponsive microgels are based on NIPAM (N-isopropylacrylamide). By substituting NIPMAM for NIPAM, and methacrylamide for acrylamide, side reactions and the generation of carboxyl groups were prevented during the Hofmann reaction. The Hofmann rearrangement is sufficiently slow under our conditions (1 h for a 51% conversion) to permit fine control of the primary amine contents in the microgels. When starting with PNIPMAM microgels containing both methacrylamide and acrylic acid residues, we prepared a series of amphoteric microgels spanning a range of amine contents, all from a common parent microgel. Therefore, every microgel in the series had the same microstructure, cross-link density, and molecular weight.

Tuning cellulose nanocrystal gelation with polysaccharides and surfactants.

Gelation of cellulose nanocrystal (CNC) dispersions was measured as a function of the presence of four nonionic polysaccharides. Addition of hydroxyethyl cellulose (HEC), hydroxypropyl guar (HPG), or locust bean gum (LBG) to CNC dispersions induced the gelation of dilute CNC dispersions, whereas dextran (DEX) did not. These behaviors correlated with adsorption tendencies; HEC, HPG, and LBG adsorbed onto CNC-coated quartz crystal microbalance sensors, whereas DEX did not adsorb. We propose that the adsorbing polysaccharides greatly increased the effective volume fraction of dilute CNC dispersions, driving more of the nanocrystals into anisotropic domains. SDS and Triton X-100 addition disrupted HEC-CNC gels whereas CTAB did not. Surface plasmon resonance measurements with CNC-coated sensors showed that SDS and Triton X-100 partially removed adsorbed HEC, whereas CTAB did not. These behaviors illustrate the complexities associated with including CNC dispersions in formulated products: low CNC contents can induce spectacular changes in rheology; however, surfactants and soluble polymers may promote gel formation or induce CNC coagulation.

Weak gelation of hydrophobic guar by albumin in simulated human tear solutions.

This initial study shows that hydrophobic modification of guar polymers used in eye drops forms weak gels with human serum albumin (HSA), suggesting that modified guar may offer advantages for treatment of dry eye diseases that lead to elevated HSA concentrations in tears. Specifically, hydroxypropyl guar samples were oxidized and derivatized with linear alkyl amines to give a series of modified guar polymers (MGuar) bearing hydroxypropyl, N-alkylamide, and carboxyl moieties. MGuar interactions with lysozyme and HSA were measured by binding and rheological methods as functions of the alkyl chain length and the extent of hydrophobic modification. HSA binds MGuar, giving weak gels, whereas lysozyme shows little tendency to bind MGuar or to interfere with HSA binding. Six mole percent substitution of decyl hydrophobes gave the strongest gels in the presence of HSA.

An inkjet-printed bioactive paper sensor that reports ATP through odour generation.

We describe an inkjet printed paper-based sensor that reports ATP by the enzyme catalysed hydrolysis of S-methyl-L-cysteine generating an odour (methyl mercaptan) that is easily detectable by the human nose.

Turning tryptophanase into odor-generating biosensors.

An odor-based sensor system that exploits the metabolic enzyme tryptophanase (TPase) as the key component is reported. This enzyme is able to convert an odorless substrate like S-methyl-L-cysteine or L-tryptophan into the odorous products methyl mercaptan or indole. To make a biosensor, TPase was biotinylated so that it could be coupled with a molecular recognition element, such as an antibody, to develop an ELISA-like assay. This method was used for the detection of an antibody present in nM concentrations by the human nose. TPase can also be combined with the enzyme pyridoxal kinase (PKase) for use in a coupled assay to detect adenosine 5'-triphosphate (ATP). When ATP is present in the low µM concentration range, the coupled enzymatic system generates an odor that is easily detectable by the human nose. Biotinylated TPase can be combined with various biotin-labeled molecular recognition elements, thereby enabling a broad range of applications for this odor-based reporting system.

Pullulan encapsulation of labile biomolecules to give stable bioassay tablets.

A simple and inexpensive method is reported for the long-term stabilization of enzymes and other unstable reagents in premeasured quantities in water-soluble tablets (cast, not compressed) made with pullulan, a nonionic polysaccharide that forms an oxygen impermeable solid upon drying. The pullulan tablets dissolve in aqueous solutions in seconds, thereby facilitating the easy execution of bioassays at remote sites with no need for special reagent handling and liquid pipetting. This approach is modular in nature, thus allowing the creation of individual tablets for enzymes and their substrates. Proof-of-principle demonstrations include a Taq polymerase tablet for DNA amplification through PCR and a pesticide assay kit consisting of separate tablets for acetylcholinesterase and its chromogenic substrate, indoxyl acetate, both of which are highly unstable. The encapsulated reagents remain stable at room temperature for months, thus enabling the room-temperature shipping and storage of bioassay components.

Targeted disinfection of E. coli via bioconjugation to photoreactive TiO2.

The selective control of pathogenic bacteria is an ongoing challenge. A strategy is proposed that combines targeted binding of the bacterium, using antibodies, with their photoactivated oxidative destruction. Photoactive colloidal TiO2 was first derivatized with E. coli antibodies (EA-TiO2). When mixtures of the organisms E. coli and Pseudomonas putida ( P. putida ) were exposed to modified EA-TiO2, the particles preferentially selected E. coli for surface binding. Two consequences arose from surface bioconjugation: bacteria were found to flocculate upon mixing at appropriate ratios of EA-TiO2/ E. coli , and EA-TiO2-bound E. coli underwent cell death after exposure to UV light. In the former case, flocculation of the bacteria was optimal at ~50 EA-TiO2 particles per E. coli . Selective flocculation provides an alternative strategy for pathogen removal. With respect to UV disinfection, as few as 26 EA-TiO2 particles per E. coli gave a 10 000-fold decrease in viable bacteria. Thus, it is possible to selectively target and kill one type of bacteria in a mixture of pathogens. The results give support to the proposal that photocatalytic TiO2 most effectively delivers an oxidizing agent when the titania is bound to the bacterial surface.

N-chlorinated poly(N-isopropylacrylamide) microgels.

The treatment of poly(N-isopropylacrylamide) (PNIPAM) microgels with aqueous bleach (NaClO) at pH 10.5 resulted in the partial conversion of the amide hydrogen to the corresponding chloramide. N-Chlorinated microgels poly(NIPAM-co-NIPAMCl) are more hydrophobic than the parent PNIPAM microgels. Thus, the volume phase transition temperature decreases with increasing chlorination. During chlorination, the microgels coagulate once they undergo a volume phase transition. The chlorination reaction stops once the microgels dehydrate and coagulate, presumably as a result of the decreased diffusion rate of the ClO(-) anion into the microgels. The microgels are reversibly dechlorinated by glutathione (GSH), first giving PNIPAM shell + poly(NIPAM-co-NIPAMCl) core microgels. Because GSH is an important redox actor in biological cells, this work suggests that chlorinated microgels may be employed to deliver active chlorine to targeted cells.

Facile phenylboronate modification of silica by a silaneboronate.

Macroscopic and colloidal silica surfaces were readily modified with alkoxysilaneboronate, IV, yielding silica surfaces with covalently bonded phenylboronic acid groups. XPS and neutron activation confirmed the presence of boron. The ability of these surfaces to specifically interact with polyols was demonstrated with polyol-coated latex and ARS, a dye that specifically couples to boronic acid groups immobilized on colloidal or macroscopic silica. This is a new, direct approach for introduction of phenylboronic acid groups onto silica surfaces.