Hydrophilic monolayer formation of adsorbed cationic starch and cationic hydroxyethyl cellulose derivatives on polyester surfaces

Cationic starch, cationic cellulose derivatives, and hydrophobically modified cationic cellulose were physically adsorbed from aqueous solution onto oppositely charged hydrophobic polyester (poly(ethylene terephthalate)) fabric and nonwoven, and this resulted in hydrophilic surface properties. Surface coverage of the polysaccharides occurred primarily by strong electrostatic interactions, and the surface characteristics were evaluated by measuring the time required for a water droplet to be absorbed into the polyester material as well as by electron spectroscopy for chemical analysis (ESCA). From a comparison of the adsorption characteristics we assess the polysaccharide-dependent and substrate-dependent adsorption behavior and discuss the similarities and differences in the hydrophilic properties and wettability observed. In particular, the temperature of the cationic polysaccharide solutions in which the substrate was immersed, the configuration of the polymer in solution, and the presence of hydrophobic substituents on the cationic moiety have a considerable effect on the polysaccharide affinity and its adsorption on the surface, irrespective of the substrate type (fabric or nonwoven). We also evaluate the relative contribution of the polyelectrolyte molecular weight, concentration in solution, and degree of charge density along the polymer chain which determine the range of interactions and alter surface hydroplilicity dependent on the type of substrate.     

The relationship between surface tension and the industrial performance of water-soluble polymers prepared from acid hydrolysis lignin, a saccharification by-product from woody materials

In this study, water-soluble anionic and cationic polymers were prepared from sulfuric acid lignin (SAL), an acid hydrolysis lignin, and the relationship between the surface tension of these polymers and industrial performance was examined. The SAL was phenolized (P-SAL) to enhance its solubility and reactivity. Sulfonation and the Mannich reaction with aminocarboxylic acids produced water-soluble anionic polymers and high-dispersibility gypsum paste. The dispersing efficiency increased as the surface tension decreased, suggesting that the fluidity of the gypsum paste increased with the polymer adsorption on the gypsum particle surface. Water-soluble cationic polymers were prepared using the Mannich reaction with dimethylamine. The cationic polymers showed high sizing efficiency under neutral papermaking conditions; the sizing efficiency increased with the surface tension. This suggests that the polymer with high hydrophilicity spread in the water and readily adhered to the pulp surface and the rosin, showing good retention.     

Effect of anticoagulation with citrate versus heparin on the adsorption of coagulation factors to blood purification resins with different charge

In liver failure, hydrophobic toxins accumulate in the blood circulation. To support hepatic function, extracorporeal blood purification systems have been developed, in which both cationic and neutral adsorbents are used to remove albumin-bound metabolites from blood. An issue of these systems is the additional removal of coagulation factors containing negatively charged γ-carboxyglutamate (Gla) domains, which, in physiological conditions, are shielded by calcium ions. We hypothesized that complexation of calcium ions by citrate leads to exposure of negative Gla domains, resulting in their binding to the positively charged adsorbents. The data presented here confirm that the binding of coagulation factors containing Gla domains to positively charged polymers is enhanced in the presence of citrate as compared to heparin. This effect increased with increasing charge density of the polymer and has important implications for the clinical application of positively charged polymers.     

The Effect of PS Content on the Ability of Natural Membranes to Fuse with Positively Charged Liposomes and Lipoplexes

Supramolecular aggregates containing cationic lipids have been widely used as transfection mediators due to their ability to interact with negatively charged DNA molecules and biological membranes. First steps of the process leading to transfection are partly electrostatic, partly hydrophobic interactions of liposomes/lipoplexes with cell and/or endosomal membrane. Negatively charged compounds of biological membranes, namely glycolipids, glycoproteins and phosphatidylserine (PS), are responsible for such events as adsorption, hemifusion, fusion, poration and destabilization of natural membranes upon contact with cationic liposomes/lipoplexes. The present communication describes the dependence of interaction of cationic liposomes with natural and artificial membranes on the negative charge of the target membrane, charges which in most cases were generated by charging the PS content or its exposure. The model for the target membranes were liposomes of variable content of PS or PG (phosphatidylglycerol) and erythrocyte membranes in which the PS and other anionic compound content/exposure was modified in several ways. Membranes of increased anionic phospholipid content displayed increased fusion with DOTAP (1,2-dioleoyl-3-trimethylammoniumpropane) liposomes, while erythrocyte membranes partly depleted of glycocalix, its sialic acid, in particular, showed a decreased fusion ability. The role of the anionic component is also supported by the fact that erythrocyte membrane inside-out vesicles fused easily with cationic liposomes. The data obtained on erythrocyte ghosts of normal and disrupted asymmetry, in particular, those obtained in the presence of Ca²⁺, indicate the role of lipid flip-flop movement catalyzed by scramblase. The ATP-depletion of erythrocytes also induced an increased sensitivity to hemoglobin leakage upon interactions with DOTAP liposomes. Calcein leakage from anionic liposomes incubated with DOTAP liposomes was also dependent on surface charge of the target membranes. In all experiments with the asymmetric membranes the fusion level markedly increased with an increase of temperature, which supports the role of membrane lipid mobility. The decrease in positive charge by binding of plasmid DNA and the increase in ionic strength decreased the ability of DOTAP liposomes/lipoplexes to fuse with erythrocyte ghosts. Lower pH promotes fusion between erythrocyte ghosts and DOTAP liposomes and lipoplexes. The obtained results indicate that electrostatic interactions together with increased mobility of membrane lipids and susceptibility to form structures of negative curvature play a major role in the fusion of DOTAP liposomes with natural and artificial membranes.     

Surfactant-mediated amyloidogenesis behavior of stem bromelain; a biophysical insight

Neurodegenerative disorders are mainly associated with amyloid fibril formation of different proteins. Stem bromelain (SB), a cysteine protease, is known to exist as a molten globule state at pH 10.0. It passes through the identical surrounding (pH 10.0) in the gut epithelium of intestine upon oral administration. Protein–surfactant complexes are widely employed as drug carriers, so the nature of surfactant toward protein is of great interest. The present work describes the effect of cationic surfactants (CTAB & DTAB) and their hydrophobic behavior toward amyloidogenesis behavior of SB at pH 10.0. Multiple approaches including light scattering, far UV-CD, turbidity measurements, and dye binding assay (ThT, Congo red and ANS) were performed to measure the aggregation propensity of SB. Further, we monitored the hydrodynamic radii of aggregates formed using dynamic light scattering technique. Structure of fibrils was also visualized through fluorescence microscopy as well as TEM. At pH 10.0, low concentration of CTAB (0–200 μM) induced amyloid formation in SB as evident from a prominent increase in turbidity and light scattering, gain in β-sheet content, and enhanced ThT fluorescence intensity. However, further increase in CTAB concentration suppressed the fibrillation phenomenon. In contrast, DTAB did not induce fibril formation at any concentration used (0–500 μM) due to lower hydrophobicity. Net negative charge developed on protein at high pH (10.0) might have facilitated amyloid formation at low concentration of cationic surfactant (CTAB) due to electrostatic and hydrophobic interactions.     

The synthesis and screening of 1,4,5,8-naphthalenetetracarboxylic diimide-peptide conjugates with antibacterial activity

We have employed an initial combinatorial approach followed by systematic lead optimization to investigate a series of novel molecules that exhibit antimicrobial activity against Gram-negative and Gram-positive bacteria. The new molecules contain various sequences of amino acids, generally L-lysine and glycine, attached to the 1,4,5,8-naphthalenetetracarboxylic diimide aromatic unit. Systematic structure-activity studies found that increasing positive charge enhanced activity and molecules containing one naphthalenetetracarboxylic diimide unit as well as at least seven lysine residues were optimum for antimicrobial activity. The naphthalenetetracarboxylic diimide derivatives were found to be inactive against mammalian cell lines, making them excellent antimicrobial candidates. Our results indicate that combining positive charge with aromatic and/or hydrophobic elements may be an interesting new approach to antimicrobial agents and adds an important new dimension to the field of cationic peptides.     

Medium Effects on Minimum Inhibitory Concentrations of Nylon-3 Polymers against E. coli

Minimum inhibitory concentrations (MICs) against E. coli were measured for three nylon-3 polymers using Luria-Bertani broth (LB), brain-heart infusion broth (BHI), and a chemically defined complete medium (EZRDM). The polymers differ in the ratio of hydrophobic to cationic subunits. The cationic homopolymer is inert against E. coli in BHI and LB, but becomes highly potent in EZRDM. A mixed hydrophobic/cationic polymer with a hydrophobic t-butylbenzoyl group at its N-terminus is effective in BHI, but becomes more effective in EZRDM. Supplementation of EZRDM with the tryptic digest of casein (often found in LB) recapitulates the LB and BHI behavior. Additional evidence suggests that polyanionic peptides present in LB and BHI may form electrostatic complexes with cationic polymers, decreasing activity by diminishing binding to the anionic lipopolysaccharide layer of E. coli. In contrast, two natural antimicrobial peptides show no medium effects. Thus, the use of a chemically defined medium helps to reveal factors that influence antimicrobial potency of cationic polymers and functional differences between these polymers and evolved antimicrobial peptides.     

Synthesis of 4% Glu-containing Val1 and Ile1-polypentapeptides: Model protein systems for demonstrating mechanochemical coupling

The synthesis of 4% Glu-polypentapeptide (PPP) (i.e., 4 Glu residues per 100 amino acid residues) and 4% Glu-Ile¹-PPP, in which Val¹ is substituted by a more hydrophobic Ile residue, is carried out by copolymerizing thep-nitrophenyl-active esters of GE(OMe)GVP and GE(OMe)GIP with their parent pentamers GVGVP and GVGIP in 1:4 ratios, respectively. After removal of the methyl ester on the side chain of Glu, these polymers exhibited a remarkablepH dependence of the temperature for their inverse temperature transitions, which are followed as turbidity development at 300 nm. On -irradiation crosslinking, the elastomeric bands obtained exhibited apH-mediated contraction and relaxation. Thus, for the first time, mechanochemical coupling is demonstrated in a synthetic polypeptide system. That the basic mechanism involves the hydrophobic effect (chemical modulation of an inverse temperature transition) and not ion-ion electrostatic repulsion is also discussed.     

Charged polymers modulate retrovirus transduction via membrane charge neutralization and virus aggregation

The specific mechanisms of charged polymer modulation of retrovirus transduction were analyzed by characterizing their effects on virus transport and adsorption. From a standard colloidal perspective two mechanisms, charge shielding and virus aggregation, can potentially account for the experimentally observed changes in adsorption behavior and biophysical parameters due to charged polymers. Experimental testing revealed that both mechanisms could be at work depending on the characteristics of the cationic polymer. All cationic polymers enhanced adsorption and transduction via charge shielding; however, only polymers greater than 15 kDa in size were capable of enhancing these processes via the virus aggregation mechanism, explaining the higher efficiency enhancement of the high molecular weight molecules. The role of anionic polymers was also characterized and they were found to inhibit transduction via sequestration of cationic polymers, thereby preventing charge shielding and virus aggregation. Taken together, these findings suggest the basis for a revised physical model of virus transport that incorporates electrostatic interactions through both virus-cell repulsive and attractive interactions, as well as the aggregation state of the virus.     

Synthesis and characterization of methylated poly(L-histidine) to control the stability of its siRNA polyion complexes for RNAi

Poly(L-histidine) (PLH) with dimethylimidazole groups has been synthesized as a pH-sensitive polypeptide to control the stability of its small interfering RNA (siRNA) polyion complexes for RNA interference (RNAi). The resulting methylated PLH (PLH-Me) was water-soluble despite deprotonation of the imidazole groups at physiological pH, as determined by acid-base titration and solution turbidity measurement. Agarose gel retardation assay proved that the quaternary dimethylimidazole groups worked as cationic groups to retain siRNA. The stability of the PLH-Me/siRNA complexes has depended on the content of hydrophobic groups, that is, τ/π-methylimidazole groups as well as deprotonated imidazole groups. PLH-Me exhibited no significant cytotoxicity despite the existence of cationic dimethylimidazole groups. By use of PLH-Me as a pH-sensitive siRNA carrier, the PLH-Me/siRNA complexes mediated efficient siRNA delivery attributed to the dimethylimidazole groups, and the gene silencing depended on the content balance among dimethyl, τ/π-methyl, and unmodified imidazole groups. These results suggest that PLH-Me controls the stability of siRNA polyion complexes by enhancing noncytotoxic siRNA delivery by optimizing the content balance of dimethyl, τ/π-methyl, and unmodified imidazole groups.     

A comparison of the effects of cationic, anionic, and neutral amphipathic agents on the contractile behaviour of frog skeletal muscle. I. Twitches and potential threshold for contraction

Cationic, anionic, and neutral amphipathic agents displayed striking differences as well as similarities in their effects on the contractile function of frog skeletal muscle. Slowed repolarization during the action potential appeared to account for twitch potentiation by low concentrations of alkyl trimethylammonium and by small n-alkanols (propanol, butanol). Small n-alkanols also caused a decrease in the potential threshold for K contractures and slower relaxation of submaximum K contractures as well as enhancement of chloride withdrawal and caffeine contractures, but these effects were not observed with larger alkanols. For the ionic amphipathic agents, the direction of the changes in the relation between Ko and K-contracture tension could be accounted for on the basis of the expected changes in surface charge, but the effects of these two types of agents on the rate of relaxation of submaximum K contractures were disproportionate and with the cationic series were opposite in direction to those produced by inorganic divalent cations. The reductions in the amplitude of chloride-withdrawal contractures by cationic as well as anionic amphipaths indicated that both types of agents can impair excitation-contraction coupling. Similar depressant effects on caffeine contractures demonstrate that these responses also can be influenced by events restricted to the external lamina of the sarcolemma. It is concluded that opposite effects can be produced by similar perturbations in different regions of the sarcolemma and that electrostatic as well as hydrophobic interactions can make an important contribution to the effects of amphipathic agents on twitches and contractures in skeletal muscle.     

Intermolecular interactions and morphology of aqueous polymer/surfactant mixtures containing cationic chitosan and nonionic sorbitan esters

In this study, the impact of surfactant molecular composition (saturated sorbitan monolaurate or unsaturated sorbitan monooleate) on polymer/surfactant assemblies was examined. Specifically, the associations between the cationic chitosan and the uncharged surfactants were monitored by surface tension, turbidity, and conductivity measurements. Bright field, confocal laser scanning, and transmission electron microscopy revealed that nanometer-sized chitosan/surfactant aggregates comprised of a chitosan-rich shell and a chitosan-poor core agglomerate at high surfactant concentrations to yield micrometer-scaled supramolecular structures with highly ordered internal structure. The size and architecture of these chitosan/surfactant assemblies were dependent on the structure and concentration of the surfactant employed. The association mechanism among chitosan, surfactant, and the chitosan/surfactant aggregates was discussed in terms of the semirigid polyelectrolyte character of chitosan and the hydrophobic character of sorbitan esters. This study provides important insight into the structural and physical parameters of surfactant that govern the formation of multicompartment polymer/surfactant assemblies.     

Interaction of horse heart and thermus thermophilus type c cytochromes with phospholipid vesicles and hydrophobic surfaces

The binding of horse heart cytochrome c (cyt-c) and Thermus thermophilus cytochrome c(552) (cyt-c(552)) to dioleoyl phosphatidylglycerol (DOPG) vesicles was investigated using Fourier transform infrared (FTIR) spectroscopy and turbidity measurements. FTIR spectra revealed that the tertiary structures of both cytochromes became more open when bound to DOPG vesicles, but this was more pronounced for cyt-c. Their secondary structures were unchanged. Turbidity measurements showed important differences in their behavior bound to the negatively charged DOPG vesicles. Both cytochromes caused the liposomes to aggregate and flocculate, but the ways they did so differed. For cyt-c, more than a monolayer was adsorbed onto the liposome surface prior to aggregation due to charge neutralization, whereas cyt c(552) caused aggregation at a protein/lipid ratio well below that required for charge neutralization. Therefore, although cyt-c may cause liposomes to aggregate by electrostatic interaction, cyt-c(552) does not act in this way. FTIR-attenuated total reflection spectroscopy (FTIR-ATR) revealed that cyt-c lost much of its secondary structure when bound to the hydrophobic surface of octadecyltrichlorosilane, whereas cyt-c(552) folds its domains into a beta-structure. This hydrophobic effect may be the key to the difference between the behaviors of the two cytochromes when bound to DOPG vesicles.     

Algal Flocculation with Synthetic Organic Polyelectrolytes

The feasibility of removing algae from water and wastewater by chemical flocculation techniques was investigated. Mixed cultures of algae were obtained from both continuous- and batch-fed laboratory reactors. Representative cationic, anionic, and nonionic synthetic organic polyelectrolytes were used as flocculants. Under the experimental conditions, chemically induced algal flocculation occurred with the addition of cationic polyelectrolyte, but not with anionic or nonionic polymers, although attachment of all polyelectrolyte species to the algal surface is shown. The mechanism of chemically induced algal flocculation is interpreted in terms of bridging phenomena between the discrete algal cells and the linearly extended polymer chains, forming a three-dimensional matrix that is capable of subsiding under quiescent conditions. The degree of flocculation is shown to be a direct function of the extent of polymer coverage of the active sites on the algal surface, although to induce flocculation by this method requires that the algal surface charge must concurrently be reduced to a level at which the extended polymers can bridge the minimal distance of separation imposed by electrostatic repulsion. The influence of pH, algal concentration, and algal growth phase on the requisite cationic flocculant dose is also reported.     

Ammonium Compounds with Localized and Delocalized Charge as Reversible Inhibitors of Cholinesterases of Different Origin

Five derivatives of benzimidazole, compounds with delocalized charge in cationic group, are studied and turned out to be reversible inhibitors of hydrolysis of acetylthiocholine under action of acetylcholinesterase from human erythrocytes, butyrylcholinesterase from horse blood serum, and cholinesterases from brain of the brown frog Rana temporaria and from optical ganglion of the Pacific squid Todarodes pacificus. It was only for acetylcholinesterase from erythrocyte as well as (with propyonylthiocholine as substrate) from squid that sensitivity to the studied benzimidazole derivatives correlated with degree of localization of the charge in the cationic group; this confirms the current concepts of functioning of the enzyme active center. A comparative study of 9 ammonium inhibitors with localized cation in their molecules, including the complete sterical analogue of the benzimidazole derivatives, benzimidazolinium iodide, has revealed both quantitative and qualitative differences.     

华南热带人工林土壤有机碳含量及其稳定性特征

研究华南热带多雨地区小良人工森林生态系统定位站不同人工林的碳固持特征,测定了林地表土有机碳含量、有机碳随土层深度的变化以及有机碳在不同粒级团聚体上的分布。结果表明：无植被覆盖的光裸地储存了极低水平的有机碳,在光裸地上种植桉树林后,如果地表枯落物长期人为取走,则土壤有机碳积累大大低于未受干扰桉林。当桉林进一步改造为阔叶混交林后,30-40a林龄混交林土壤有机碳含量均高于16.5gkg^-1,但仍只有邻近天然林土壤有机碳水平的76%,因而,这些人工混交林土壤具有进一步积累碳的潜力。光裸地在1m剖面上的含碳量均极低,并无显著的深度变化,约2-3gkg^-1,未保护的桉林含量水平在4-5gkg^-1,同样表土与心土层没有明显差别,通过地表输入的凋落物对土壤有机碳的影响主要集中在表层0.5m范围内。以豹皮樟Litsea rotundifolia,阴香Cinnamomum burmanni,潺槁木姜子Litsea glutinosa,陈氏钓樟Lindera chuni等为优势种的混交林,深层土壤含碳量显著高于其它混交林,并与村边林相似。土壤有机碳储量与土壤团聚体发育有一定的相关性,非水稳性团聚体以最小粒径（〈0.25mm）的含碳浓度最高,其它粒级的有机碳浓度相似,未成熟人工林,小团聚体的碳浓度较高,成熟林则在不同粒级中较均匀地分配有机碳,但实际碳储量于最大粒级的非水稳性团聚体中最多。成熟森林土壤水稳性团聚体含碳浓度大大高于非水稳性团聚体,水稳性团聚体在0.25-0.5mm及0.5-1.0mm两个粒级上有最大的碳积累量。结果表明,建立特定类型的混交林是增加林地碳汇功能的一个有效途径,非水稳性的大团聚体与水稳性的小团聚体的有效发育可能是成熟森林高固碳量的重要机理。     

Cationic polyelectrolyte hydrogel fosters fibroblast spreading, proliferation, and extracellular matrix production: Implications for tissue engineering

Fibrous encapsulation is known to occur to many prosthetic implants and is thought to be due to the cells not adhering adequately to the surface. For developing new materials able to enhance cellular adhesion by mimicking extracellular matrix components, polyelectrolyte polymers, characterized by tunable surface charges, have been proposed. Here we demonstrate that panoply of cell functions over a two-dimensional substratum is influenced by surface charge. We have at first generated structurally related polyelectrolyte substrata varying in their positive surface charge amount and subsequently evaluated a variety of behaviors of human primary fibroblasts seeded on these polymers. The proportion of adherent, spreading, and proliferating cells was increased significantly on cationic hydrophilic surfaces when compared with the neutral base surface. The extent of cell spreading correlated with cytoskeleton organization as assessed using immunofluorescence techniques. In the key experiment, the presence of cationic charges on cell adhesion-resistant neutral surface increased the synthesis of collagen I and III, the release of their metabolites, and the expression of their mRNA by fibroblasts. Interestingly, the scarce collagen deposits on neutral polymer consisted, for the most part, of collagen I while collagen III was present only in traces probably due to the secretion of metalloproteinase-2 by non-adherent fibroblasts. Taken together, these results show that polyelectrolyte films may promote the attachment of fibroblast cells as well as their normal secretory phenotype. Both effects could be potentially useful in integrating soft connective tissue to the implant, decreasing the chance of its fibrous encapsulation.     

Surface and aggregate properties of an amphiphilic derivative of carboxymethylchitosan

A new kind of amphiphilic derivative of carboxymethylchitosan, a group of (2-hydroxyl-3-butoxyl)propylcarboxymethylchitosans (HBP-CMCHS), has been synthesized, and the surface and aggregate properties have been studied by means of surface tension, surface pressure and fluorescence measurements. HBP-CMCHS can adsorb on the surface to decrease the surface tension of the solution. The adsorption film was quite stable, which can make the relative compressed pressure increase dramatically with the decrease of the surface area. In solution, hydrophobic aggregations were identified by the decrease in the ratio of the fluorescence emission intensity of the first and third pyrene vibronic peaks ( I(1)/ I(3)). Results showed that the aggregation began to form at a concentration similar to that of the polymer transfer to the air-water interface. Aggregate formation of the polymers is a gradually compact process with hydrophobic associations. Increase of DS and addition of NaCl to the HBP-CMCHS solution can make the surface tension decrease, make the aggregation occur at lower concentration, and make the aggregation more hydrophobic.     

Gene-transferring efficiencies of novel diamino cationic lipids with varied hydrocarbon chains

Utilizing three biocompatible components, a series of novel cationic lipids has been chemically synthesized and tested for their gene-transferring capabilities in 293 transformed kidney cells and B16BL6 mouse melanoma cells. The synthesized cationic lipids consisting of a core of lysine and aspartic acid with hydrocarbon chains of varied length were assigned the acronyms DLKD (O,O'-dilauryl N-lysylaspartate), DMKD (O,O'-dimyristyl N-lysylaspartate), DPKD (O,O'-dipalmityl N-lysylaspartate), and DSKD (O,O'-distearyl N-lysylaspartate). The gene-transferring capabilities of these cationic lipids were found to be dependent on the hydrocarbon chain length. Under similar experimental conditions, the order of gene transfection efficiency was DMKD > DLKD > DPKD > DSKD. Addition of cholesterol or dioleoyl phosphatidylethanolamine (DOPE) as a colipid did not change this order. Colipid addition affected the transfection efficiency positively or negatively depending on the length of the cationic lipid acyl chain. On the whole, the length of the hydrophobic carbon chain was a major factor governing the gene-transferring capabilities of this series of cationic lipids. The observed differences in transfection efficiency may be due to differing binding affinities to DNA molecules as well as differences in the surface charge potential of the liposome-DNA complexes (lipoplexes) in the aqueous environment.