Selectively Charged and Zwitterionic Analogues of the Smallest Immunogenic Structure of Streptococcus Pneumoniae Type 14.

Zwitterionic polysaccharides (ZPs) have been shown in recent years to display peculiar immunological properties, thus attracting the interest of the carbohydrate research community. To fully elucidate the mechanisms underlying these properties and exploit the potential of this kind of structures, in depth studies are still required. In this context, the preparation of two cationic, an anionic, as well as two zwitterionic tetrasaccharide analogues of the smallest immunogenic structure of Streptococcus pneumoniae type 14 (SP14) capsular polysaccharide are presented. By exploiting a block strategy, the negative charge has been installed on the non-reducing end of the lactose unit of the tetrasaccharide and the positive charge either on the non-reducing end of the lactosamine moiety or on an external linker. These structures have then been tested by competitive ELISA, showing that the structural variations we made do not modify the affinity of the neutral compound to binding to a specific antibody. However, lower efficacies than the natural SP14 compound were observed. The results obtained, although promising, point to the need to further elongate the polysaccharide structure, which is likely too short to cover the entire epitopes.

Potential of Biocellulose Carrier Impregnated with Essential Oils to Fight Against Biofilms Formed on Hydroxyapatite.

In this research, bacterial cellulose (BC), one of the most promising biopolymers of the recent years, was saturated with thyme, eucalyptus and clove essential oils (EOs) and applied against staphylococcal and pseudomonal biofilms formed on hydroxyapatite (HA). BC dressings were thoroughly analyzed with regard to their physical properties. Moreover, the exact composition and ability of particular EO molecules to adhere to HA was assessed. Additionally, cytotoxicity of oil-containing, cellulose-based dressings towards osteoblasts and fibroblasts as well as their impact on reactive oxygen species (ROS) production by macrophages was assessed. The results revealed the high ability of BC dressings to absorb and subsequently release EOs from within their microstructure; the highest number of compounds able to adhere to HA was found in the thyme EO. The eucalyptus EO displayed low, while thyme and clove EOs displayed high cytotoxicity towards fibroblast and osteoblast cell lines. The clove EO displayed the highest eradication ability toward staphylococcal, while the thyme EO against pseudomonal biofilm. Taken together, the results obtained indicate the suitability of EO-saturated BC dressings to eradicate pseudomonal and staphylococcal biofilm on HA surface and moreover, to not trigger reactive oxygen species production by immune system effector cells. However, due to cytotoxic effects of thyme and clove EOs towards cell lines in vitro, the eucalyptus EO-saturated BC dressing is of highest potential to be further applied.

Carbohydrate Specificity and Isotypes of Monoclonal and Polyclonal Antibodies to Conjugated Tetrasaccharide, a Synthetic Analogue of Repeating Unit of Capsular Polysaccharide of Streptococcus Pneumoniae Serotype 14.

We studied carbohydrate specificity and isotypes of antibodies to BSA-conjugated tetrasaccharide, a repeating unit of the capsular polysaccharide of Streptococcus pneumoniae serotype 14, in mouse polyclonal sera and hybridoma-synthesized products. Natural IgM antibodies to the tetrasaccharide containing epitopes similar to surface carbohydrate structures of mammalian and human cells in low titers were determined in native mouse serum by ELISA using biotinylated tetrasaccharide and synthetic capsular polysaccharide as the solid-phase antigens. Polyclonal sera to the conjugated tetrasaccharide contained IgM and all subclasses of IgG antibodies, which were detected in a higher titer when the biotinylated tetrasaccharide was used as a solid phase antigen compared to synthetic capsular polysaccharide. Monoclonal antibodies to S. pneumoniae serotype 14 tetrasaccharide were identified in an equivalent titer using either biotinylated tetrasaccharide or synthetic capsular polysaccharide. Monoclonal antibodies obtained in vitro belonged to IgM isotype and cross-reacted with secondary full-size IgG antibodies. In the serum of mice inoculated with hybridoma, IgM and IgG2a antibodies recognizing the tetrasaccharide epitope in the structure of synthetic capsular polysaccharide were simultaneously determined.

Modification of Bacterial Cellulose with Quaternary Ammonium Compounds Based on Fatty Acids and Amino Acids and the Effect on Antimicrobial Activity.

In the present work, bacterial cellulose (BC) membranes have been modified with bioactive compounds based on long chain dimer of C18 linoleic acid, referred to as the dilinoleic acid (DLA) and tyrosine (Tyr), a natural amino acid capable of forming noncovalent cation-π interactions with positively charged ethylene diamine (EDA). This new compound, [EDA][DLA-Tyr], has been synthesized by simple coupling reaction, and its chemical structure was characterized by 1H NMR and Fourier transform infrared spectroscopy. The antimicrobial activity of a new compound against S. aureus and S. epidermidis, two cocci associated with skin and wound infections, was assessed. The [EDA][DLA-Tyr] impregnated BC exhibited strong and long-term antimicrobial activity against both staphylococcal species. The results showed a 57-66% and 56-60% reduction in S. aureus and S. epidermidis viability, respectively, depending on [EDA][DLA-Tyr] concentration used. Importantly, [EDA][DLA-Tyr] molecules were released gradually from the BC pellicle, while a reference antibiotic, erythromycine (ER), did not show any antibacterial activity against S. aureus and S. epidermidis after 48 h of soaking in deionized water. Thus, a combination of [EDA][DLA-Tyr] and BC could be a promising new class of wound dressing displaying both biocompatibility and antimicrobial activity.

Affinity binding of proteins to the modified bacterial cellulose nanofibers.

The potential of the modified bacterial cellulose (BC) nanofibers was determined bearing metal ion coordination interactions to enhance the protein adsorption and binding capacity. Thus, a household synthesized metal chelating monomer, namely N-methacryloyl-l-histidine methylester (MAH), and a commercial metal chelating monomer, namely 4-vinylimidazole (VIm), were used to complex with metal ions Cu(II) and Ni(II) respectively for the synthesis of the modified BC nanofibers. The modified nanofibers were characterized by FT-IR, SEM and EDX measurements. The protein adsorption tests were carried out using hemoglobin as a model protein and it was determined that the maximum adsorption capacity of hemoglobin onto the modified BC nanofibers was found as 47.40mg/g. The novel strategy for the preparation of metal chelated nanofibers was developed.

A multipurpose natural and renewable polymer in medical applications: Bacterial cellulose.

Bacterial cellulose (BC) produced by some bacteria, among them Gluconacetobacter xylinum, which secrets an abundant 3D networks fibrils, represents an interesting emerging biocompatible nanomaterial. Since its discovery BC has shown tremendous potential in a wide range of biomedical applications, such as artificial skin, artificial blood vessels and microvessels, wound dressing, among others. BC can be easily manipulated to improve its properties and/or functionalities resulting in several BC based nanocomposites. As example BC/collagen, BC/gelatin, BC/Fibroin, BC/Chitosan, etc. Thus, the aim of this review is to discuss about the applicability in biomedicine by demonstrating a variety of forms of this biopolymer highlighting in detail some qualities of bacterial cellulose. Therefore, various biomedical applications ranging from implants and scaffolds, carriers for drug delivery, wound-dressing materials, etc. that were reported until date will be presented.

Acetylation of bacterial cellulose catalyzed by citric acid: Use of reaction conditions for tailoring the esterification extent.

Bacterial cellulose (BC) nanoribbons were partially acetylated by a simple direct solvent-free route catalyzed by citric acid. The assay of reaction conditions within chosen intervals (i.e. esterification time (0.5-7h), catalyst content (0.08-1.01mmol/mmol AGU), and temperature (90-140°C)), illustrated the flexibility of the methodology proposed, with reaction variables which can be conveniently manipulated to acetylate BC to the required degree of substitution (DS) within the 0.20-0.73 interval. Within this DS interval, characterization results indicated a surface-only process in which acetylated bacterial cellulose with tunable DS, preserved fibrous structure and increased hydrophobicity could be easily obtained. The feasibility of reusing the catalyst/excess acylant in view of potential scale-up was also illustrated.

Effects of reaction conditions on cellulose structures synthesized in vitro by bacterial cellulose synthases.

Cellulose was synthesized by cellulose synthases extracted from the Komagataeibacter xylinus (formerly known as Gluconacetobacter xylinus). The effects of temperature and centrifugation of the reaction solution on the synthesis products were investigated. Cellulose with number-average degree of polymerization (DPn) roughly in the range 60-80 and cellulose II crystal structure was produced under all conditions. The amount of cellulose varied with temperature and centrifugation, and the centrifugation at 2000 × g also slightly reduced the DPn. Cellulose production was maximal around the temperature 35 °C and without centrifugation. At higher temperatures and during centrifugation at 2000 × g the proteins started to denature, causing differences also in the morphology of the cellulosic aggregates, as seen with electron microscopy. These observations serve as a basis for discussions about the factors affecting the structure formation and chain length of in vitro synthesized cellulose.

Bacterial cellulose-poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) composites for optoelectronic applications.

Electrically conducting bacterial cellulose (BC) membranes were prepared by ex situ incorporation of poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) ( PEDOT: PSS) into BC pellicles. The BC pellicles were immersed into an aqueous solution of PEDOT: PSS for 6, 12, 18, or 24h, and the resultant composites were vacuum dried at ambient temperature. The structural features of the composites were determined using X-ray photoelectron spectroscopy (XPS), field-emission scanning electron microscopy (FE-SEM), Fourier-transform infrared (FTIR) spectroscopy, and X-ray diffraction (XRD). XPS confirmed synthesis of the composites, and SEM showed uniform incorporation of PEDOT: PSS into the BC matrix. The FTIR spectra of the composites exhibited characteristic bands for both BC and PEDOT: PSS, and XRD analysis showed a slight decrease in crystallinity during composite preparation. The electrical conductivity of the composites was 12.17S/cm for incorporation of 31.24 wt% PEDOT: PSS into the BC matrix. These highly conducting BC-PEDOT:PSS composites are expected to find potential applications in optoelectronic devices such as biosensors, organic light-emitting diodes, and solar cells.

Chemical modification, characterization and bioactivity of a released exopolysaccharide (r-EPS1) from Lactobacillus plantarum 70810.

In the present study, a released exopolysaccharide (r-EPS1) from L. plantarum 70810 was modified by acetylation, phosphorylation and carboxymethylation. Scanning electron micrograph (SEM) and thermogravimetric analysis (TGA) showed the r-EPS1 derivatives had different surface morphology and thermal behavior. Compared with r-EPS1, the derivatives exhibited stronger antioxidant and antitumor activities. The study provided experimental evidences that chemical modification could be an effective way to improve the bioactivity of exopolysaccharide from L. plantarum 70810. It is noted that these derivatives could be explored as novel potential antioxidant and antitumor agents.

Synthesis and immunochemical evaluation of a non-methylated disaccharide analogue of the anthrax tetrasaccharide.

Anthrax tetrasaccharide is an oligosaccharide expressed at the outermost surface of the Bacillus anthracis spores, featuring three rhamnoses and a rare sugar called anthrose. This motif has now been identified as a plausible component of future human vaccines against anthrax. We report herein the synthesis of a 2-O-demethylated-ß-D-anthropyranosyl-(1→3)-α-L-rhamnopyranose disaccharide analogue of this tetrasaccharide from a cyclic sulfate intermediate. This disaccharide conjugated to BSA induces an anti-native tetrasaccharide IgG antibody response when administered in BALB/c mice. Moreover, induced sera bound to native B. anthracis endospores. These results suggest that the disaccharide analogue, easily amenable for a synthetic scale-up, could be used in a glycoconjugate antigen formulation.