Yury A. Skorik

Evaluation of various chitin-glucan derivatives from Aspergillus niger as transition metal adsorbents.

A number of chelating resins were prepared by chemical derivatization of the chitin-glucan (CG) complex isolated from Aspergillus niger biomass, namely chitosan-glucan (CsG), O-carboxymethyl-chitin-glucan (CM-CG), O-(2-sulfoethyl)chitin-glucan (SE-CG), and N-(2-carboxyethyl)chitosan-glucan (CE-CsG). The chemical modification was confirmed by FT-IR and elemental analysis. Nanosecond electron beam irradiation was used to produce insoluble resins and to preserve the reactive functional groups. Batch experiments were carried out to evaluate the adsorption selectivity and capacity of the resins toward transition metal ions (Cu(2+), Ni(2+), Co(2+), Zn(2+)). The resins showed good adsorption capability with the following selectivity series: Co(2+)Zn(2+). The total metal adsorption capacities of CG, CsG, CM-CG, SE-CG, and CE-CsG resins at pH 6.5 (ammonium acetate buffer) were found to be 0.205, 0.382, 1.752, 0.319, and 0.350 mmol g(-1), respectively. Our results suggest that, depending on the type of chemical modification, the chitin-glucan complexes can be used either for selective Cu(2+) removal (CsG) or for total transition metal adsorption (CM-CG) from aqueous effluents.

Carboxyalkylation of chitosan in the gel state.

This study presents a new approach for direct carboxyalkylation of chitosan in the gel state by using aza-Michael addition and substitution reactions. Various reagents were applied including acrylic and crotonic acids, and α-, β-, γ-, δ-, and ɛ-halocarboxylic acids. The reaction of chitosan with γ- and δ-halocarboxylic acids showed no target product formation either in solution or in the gel state. In the case of acrylic, crotonic, α- and β-halocarboxylic acids, the reaction performed in the gel state (concentration of chitosan 20-40%) shows higher degree of substitution at lower reaction time and temperature than in diluted solutions (concentration of chitosan 0.5-2%). The results were discussed in terms of kinetics of the target and side reactions. (1)H and (13)C NMR confirmed that in all cases the carboxyalkylation of chitosan proceeds exclusively at the amino groups.

Bis[N-(2-hydroxyethyl)-β-alaninato]-copper(II)

The Cu(II) ion in the title complex, [Cu(C5H10NO3)2] or [Cu(He-ala)2] [He-ala = N-(2-hydroxyethyl)-beta-alaninate], resides at the inversion centre of a square bipyramid comprised of two facially arranged tridentate He-ala ligands. Each He-ala ligand binds to a Cu(II) ion by forming one six-membered beta-alaninate chelate ring in a twist conformation and one five-membered ethanolamine ring in an envelope conformation, with Cu-N = 2.017 (2) angstroms, Cu-O(COO) = 1.968 (1) angstroms and Cu-O(OH) = 2.473 (2) angstroms. The [Cu(He-ala)2] molecules are involved in a network of O-H...O and N-H...O hydrogen bonds, forming layers parallel to the (10-1) plane. The layers are connected into a three-dimensional structure by van der Waals interactions, so that the molecular centres form pseudo-face-centered close packing.

The interaction of amino acids, peptides, and proteins with DNA

Amino acids that carry charges on their side groups can bind to double stranded DNA (dsDNA) and change the strength of the double helix. Measurement of the DNA melting temperature (Tm) confirmed that acidic amino acids (Glu, Asp) weaken the H-bonds between DNA strands, whereas basic amino acids (Arg, Lys) strengthen the interaction between the strands. A rank correlation exists between the amino acid isoelectric points and the observed changes in Tm. A similar dependence of the hyperchromic effect on the isoelectric point of a protein (pepsin, insulin, cortexin, and protamine) was observed for DNA-protein complexes at room temperature. Short peptides (KE, AEDG, and KEDP) containing a mixture of acidic and basic amino acid residues also affect Tm and the stability of the double helix. A model for binding Glu and Lys to dsDNA was explored by a docking simulation. The model shows that Glu, in an untwisted shape, binds to dsDNA in its major groove and disrupts three H-bonds between the strands, thereby destabilizing the double helix. Lys, in an untwisted shape, binds to the external side of the dsDNA and forms two bonds with O atoms of neighboring phosphodiester groups, thereby strengthening the DNA helix.

Pervaporation multilayer membranes based on a polyelectrolyte complex of λ-carrageenan and chitosan

A polyelectrolyte complex (PEC) was prepared from chitosan (CS) and λ-carrageenan (λ-CAR) using a layer-by-layer deposition of polyion solutions on a plated nonporous support. This material was then used as a multilayer membrane for the pervaporation separation of aqueous ethanol solutions. The fabricated complex film (25-30μm thick) was a multilayer system (λ-CAR-PEC-CS) containing a polycation CS (MW 3.1×105, DDА 0.93), a polyanion λ-CAR (MW 3.5×105, extracted from the alga Chondrus armatus), and a PEC layer formed between the two polyion layers. X-ray diffraction indicated a significant structuring of the film in the region of the composite PEC-CS bilayer. The structural and morphological characteristics of the CS surface in the multilayer membrane, as revealed by atomic force microscopy, were close to the characteristics of the dense CS film. However, this structure changed following pervaporation (i.e., the distinct spherical structures on the surface disappeared). Similarly, the initially loose surface of λ-CAR in the composite changed to an ordered domain after pervaporation. The transport properties of the pervaporation membranes were tested by examining the separation of ethanol-water mixtures of different compositions. The flux increased with an increase in the weight percentage of water in the feed mixture, but the separation capacity of the membrane was unchanged. In a range of feed concentrations of 50-94wt%, the membrane mainly releases water with a corresponding concentration in the permeate of 99.9-99.8wt% and substantial fluxes of 0.003-1.130kgm-2h-1 at 40°C. The obtained results indicate significant prospects for the use of non-gelling type CARs for the formation of highly effective pervaporation membranes.

Influence of chitosan-chitin nanofiber composites on cytoskeleton structure and the proliferation of rat bone marrow stromal cells

Chitosan scaffolds have gained much attention in various tissue engineering applications, but the effect of their microstructure on cell-material spatial interactions remains unclear. Our objective was to evaluate the effect of chitosan-based matrices doping with chitin nano-whiskers (CNW) on adhesion, spreading, cytoskeleton structure, and proliferation of rat bone marrow stromal cells (BMSCs). The behavior of BMSCs during culture on chitosan-CNW films was determined by the molecular mass, hydrophobicity, porosity, crosslinking degree, protonation degree and molecular structure of the composite chitosan-CNW films. The shape, spreading area, cytoskeleton structure, and proliferation of BMSCs on chitosan matrices with a crystalline structure and high porosity were similar to that observed for BMSCs cultured on polystyrene tissue culture plates. The amorphous polymer structure and high swelling led to a decrease in the spreading area and cell proliferation. Thus, we can control the behavior of cells in culture (adhesion, spreading, and proliferation) by changing the physico-chemical properties of the chitosan-CNW films.

Polymorphic Modifications of Chitosan

This work describes the analysis of the crystal structures of chitosan, its main polymorphic modifications, and its characteristic mutual chain packing and hydrogen bond systems in the crystalline regions of conformers. The analysis takes into consideration the crystal structures of chitosan complexes (salts) with organic and inorganic acids and their structural transformations. Notably, the transformation of the hydrated form of chitosan into anhydrous is found to be irreversible and occurs either at high temperatures or through a less stable form of hydrated salts. The interaction with polyanions during the formation of multilayer films can be considered as a way to form the anhydrous crystalline form of chitosan.

N -[4-( N,N,N -trimethylammonium)benzyl]chitosan chloride: Synthesis, interaction with DNA and evaluation of transfection efficiency

А novel cationic chitosan derivative, N-[4-(N,N,N-trimethylammonium)benzyl]chitosan chloride (TMAB-CS), with different degrees of substitution (DS) was synthesized by a chemoselective interaction of 4-formyl-N,N,N-trimethylanilinium iodide with chitosan amino groups using a reductive amination method. Several factors (pH, reactant ratio, reaction time, and chitosan structure) were studied for their effects on the DS of the resulting TMAB-CS. The obtained derivatives were characterized by 1H NMR and FTIR spectroscopy. Turbidimetric titration showed enhanced solubility over a wide pH range even for low-substituted TMAB-CS. TMAB-CS provided strong DS-dependent binding of plasmid DNA. Dynamic light scattering measurements revealed the formation of stable polyplexes with hydrodynamic diameters of 200-300nm and ζ-potential of 20-30mV. TMAB-CS with relatively low DS (25%) demonstrated more pronounced transfection efficiency (up to 2000 cell/cm2) of plasmid DNA into the HEK293 cell line promoted by free TMAB-CS. The positive effects of lower DS can be related to a better polyplex dissociation within the cell. The cytotoxicity of TMAB-CS was comparable to that of the initial chitosan at concentrations up to 300ng/μL, even at high DS.

Comparative Study of Diethylaminoethyl-Chitosan and Methylglycol-Chitosan as Potential Non-Viral Vectors for Gene Therapy

In this paper, we compared the transfection efficiency and cytotoxicity of methylglycol-chitosan (MG-CS) and diethylaminoethyl-chitosan (DEAE-CSI and DEAE-CSII with degrees of substitution of 1.2 and 0.57, respectively) to that of Lipofectamine (used as a reference transfection vector). MG-CS contains quaternary amines to improve DNA binding, whereas the DEAE-CS exhibits pH buffering capability that would ostensibly enhance transfection efficiency by promoting endosomal escape. Gel retardation assays showed that both DEAE-CS and MG-CS bound to DNA at a polysaccharide:DNA mass ratio of 2:1. In Calu-3 cells, the DNA transfection activity was significantly better with MG-CS than with DEAE-CS, and the efficiency improved with increasing polysaccharide:DNA ratios. By contrast, the efficiency of DEAE-CSI and DEAE-CSII was independent of the polysaccharide:DNA ratio. Conversely, in the transfection-recalcitrant JAWSII cells, both Lipofectamine and MG-CS showed significantly lower DNA transfection activity than in Calu-3 cells, whereas the efficiency of DEAE-CSI and DEAE-CSII was similar in both cell lines. The toxicity of DEAE-CS increased with increasing concentrations of the polymer and its degree of substitution, whereas MG-CS demonstrated negligible cytotoxicity, even at the highest concentration studied. Overall, MG-CS proved to be a more efficient and less toxic transfection agent when compared to DEAE-CS.