Supramolecular Composite Materials from Cellulose, Chitosan, and Cyclodextrin: Facile Preparation and Their Selective Inclusion Complex Formation with Endocrine Disruptors

We have successfully developed a simple one-step method of preparing high-performance supramolecular polysaccharide composites from cellulose (CEL), chitosan (CS), and (2,3,6-tri-O-acetyl)-α-, β-, and γ-cyclodextrin (α-, β-, and γ-TCD). In this method, [BMIm+Cl–], an ionic liquid (IL), was used as a solvent to dissolve and prepare the composites. Because a majority (\u3e88%) of the IL used was recovered for reuse, the method is recyclable. XRD, FT-IR, NIR, and SEM were used to monitor the dissolution process and to confirm that the polysaccharides were regenerated without any chemical modifications. It was found that unique properties of each component including superior mechanical properties (from CEL), excellent adsorption for pollutants and toxins (from CS), and size/structure selectivity through inclusion complex formation (from TCDs) remain intact in the composites. Specifically, the results from kinetics and adsorption isotherms show that whereas CS-based composites can effectively adsorb the endocrine disruptors (polychlrophenols, bisphenol A), their adsorption is independent of the size and structure of the analytes. Conversely, the adsorption by γ-TCD-based composites exhibits a strong dependence on the size and structure of the analytes. For example, whereas all three TCD-based composites (i.e., α-, β-, and γ-TCD) can effectively adsorb 2-, 3-, and 4-chlorophenol, only the γ-TCD-based composite can adsorb analytes with bulky groups including 3,4-dichloro- and 2,4,5-trichlorophenol. Furthermore, the equilibrium sorption capacities for the analytes with bulky groups by the γ-TCD-based composite are much higher than those by CS-based composites. Together, these results indicate that the γ-TCD-based composite with its relatively larger cavity size can readily form inclusion complexes with analytes with bulky groups, and through inclusion complex formation, it can strongly adsorb many more analytes and has a size/structure selectivity compared to that of CS-based composites that can adsorb the analyte only by surface adsorption

Recyclable Synthesis, Characterization, and Antimicrobial Activity of Chitosan-based Polysaccharide Composite Materials

We have successfully developed a simple and totally recyclable method to synthesize novel, biocompatible, and biodegradable composite materials from cellulose (CEL) and chitosan (CS). In this method, [BMIm+Cl−], an ionic liquid (IL), was used as a green solvent to dissolve and synthesize the [CEL+CS] composites. Since, the IL can be removed from the composites by washing them with water, and recovered by distilling the washed solution, the method is totally recyclable. Spectroscopic and imaging techniques including XRD, FTIR, NIR, and SEM were used to monitor the dissolution, to characterize and to confirm that CEL and CS were successfully regenerated. More importantly, we have successfully demonstrated that [CEL+CS] composite is particularly suited for many applications including antimicrobial property. This is because the composites have combined advantages of their components, namely superior chemical and mechanical stability (from CEL) and bactericide (from CS). Results of tensile strength measurements clearly indicate that adding CEL into CS substantially increase its tensile strength. Up to 5× increase in tensile strength can be achieved by adding 80% of CEL into CS. Results of in vitro antibacterial assays confirm that CS retains its antibacterial property in the composite. More importantly, the composites reported here can inhibit growth of wider range of bacteria than other CS-based materials prepared by conventional methods; that is over 24 h period, the composites substantially inhibited growth of bacteria such as MRSA, VRE, S. aureus, E. coli. These are bacteria that are often found to have the highest morbidity and mortality associated with wound infections. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013

Polysaccharide Ecocomposite Materials: Materials: Synthesis, Characterization and Application for Removal of Pollutants and Bacteria

A novel, simple and totally recyclable method has been developed for the synthesis of nontoxic, biocompatible and biodegradable composite materials from cellulose and chitosan. In this method, [BMIm+Cl-], an ionic liquid (IL), was used as a solvent to dissolve and synthesize the [CEL+CS] composite materials. Since the IL can be removed from the materials by washing them with water, and recovered from the washed solution, the method is totally recyclable. XRD, FTIR, NIR and SEM were used to characterize the materials and to confirm that CEL and CS were successfully regenerated by the method without any chemical transformation. More importantly, we have successfully demonstrated that [CEL+CS] material can serve as an effective adsorbent for removal of various endocrine disruptors including polychlorophenols and bisphenol A. This is because the composites have combined advantages of their components, namely superior chemical stability and mechanical stability (from CEL) and excellent adsorption capability for pollutants (from CS)

Chitosan-cellulose Composite Materials: Preparation, Characterization and Application for Removal of Microcystin

We developed a simple and one-step method to prepare biocompatible composites from cellulose (CEL) and chitosan (CS). [BMIm+Cl−], an ionic liquid (IL), was used as a green solvent to dissolve and prepare the [CEL + CS] composites. Since majority (\u3e88%) of IL used was recovered for reuse by distilling the aqueous washings of [CEL + CS], the method is recyclable. XRD, FTIR, NIR, 13C CP-MAS-NMR and SEM were used to monitor the dissolution and to characterize the composites. The composite was found to have combined advantages of their components: superior mechanical strength (from CEL) and excellent adsorption capability for microcystin-LR, a deadly toxin produced by cyanobacteria (from CS). Specifically, the mechanical strength of the composites increased with CEL loading; e.g., up to 5× increase in tensile strength was achieved by adding 80% of CEL into CS. Kinetic results of adsorption confirm that unique properties of CS remain intact in the composite, i.e., it is not only a very good adsorbent for microcystin but also is better than all other available adsorbents. For example, it can adsorb 4× times more microcystin than the best reported adsorbent. Importantly, the microcystin adsorbed can be quantitatively desorbed to enable the composite to be reused with similar adsorption efficiency

Interferon-γ induces immunoproteasomes and the presentation of MHC I-associated peptides on human salivary gland cells.

A prominent histopathological feature of Sjögren's syndrome, an autoimmune disease, is the presence of lymphocytic infiltrates in the salivary and lachrymal glands. Such infiltrates are comprised of activated lymphocytes and macrophages, and known to produce multiple cytokines including interferon-gamma (IFN-γ). In this study, we have demonstrated that IFN-γ strongly induces the expression of immunoproteasome beta subunits (β1i, β2i and β5i) and immunoproteasome activity but conversely inhibits the expression of proteasome beta subunits (β1, β2 and β5) in human salivary gland (HSG) cells. Mass spectrometric analysis has revealed potential MHC I-associated peptides on the HSG cells, including a tryptic peptide derived from salivary amylase, due to IFN-γ stimulation. These results suggest that IFN-γ induces immunoproteasomes in HSG cells, leading to enhanced presentation of MHC I-associated peptides on cell surface. These peptide-presenting salivary gland cells may be recognized and targeted by auto-reactive T lymphocytes. We have also found that lactacystin, a proteasome inhibitor, inhibits the expression of β1 subunit in HSG cells and blocks the IFN-γ-induced expression of β1i and immunoproteasome activity. However, the expression of β2i and β5i in HSG cells is not affected by lactacystin. These results may add new insight into the mechanism regarding how lactacystin blocks the action of proteasomes or immunoproteasomes

Enantiomeric Selective Adsorption of Amino Acid by Polysaccharide Composite Materials

A composite containing cellulose (CEL) and chitosan (CS) synthesized by a simple and recyclable method by using butylmethylimmidazolium chloride, an ionic liquid, was found to exhibit remarkable enantiomeric selectivity toward the adsorption of amino acids. The highest adsorption capacity and enantiomeric selectivity are exhibited by 100% CS. A racemic amino acid can be enantiomerically resolved by 100% CS in about 96–120 h. Interestingly, adsorption by 50:50 CEL/CS is more similar to that by 100% CS than to 100% CEL. Specifically, whereas 100% CEL has the lowest adsorption capacity and enantiomeric selectivity, 50:50 CEL/CS has sufficient enantiomeric selectivity to enable it to be used for chiral resolution. This is significant because in spite of its high enantiomeric selectivity 100% CS cannot practically be used because it has relatively poor mechanical properties and undergoes extensive swelling. Adding 50% CEL to CS substantially improves the mechanical properties and reduces its swelling while it retains sufficient enantiomeric selectivity to enable it to be used for routine chiral separations. The kinetic results indicate that the enantiomerically selective adsorption is due not to the initial surface adsorption but rather to the subsequent stage in which the adsorbate molecules diffuse into the pores within the particles of the composites and consequently are adsorbed by the interior of each particle. The strong intermolecular and intramolecular hydrogen bond network in CEL enables it to adopt a very dense structure that makes it difficult for adsorbate molecules to diffuse into its interior, thereby leading to low enantiomeric selectivity. Compared to hydroxy groups, amino groups cannot form strong hydrogen bonds. The hydrogen bond network in CS is not as extensive as in CEL, and its inner structure is relatively less dense than that of CEL. Adsorbate molecules can, therefore, diffuse from the outer surface to its inner structure relatively more easily than in CEL, thereby leading to higher enantiomeric selectivity for 100% CS

Discriminating Pulmonary Hypertension Caused by Monocrotaline Toxicity from Chronic Hypoxia by Near-Infrared Spectroscopy and Multivariate Methods of Analysis

A new method has been developed for the determination of tissue pathology caused by chronic hypoxia and monocrotaline toxicity. The method is based on the use of near-infrared (NIR) spectrophotometry to measure spectra of lung tissue from normal chronic hypoxia (CH) and monocrotaline (MCT) models of pulmonary hypertension (PH), followed by analysis using multivariate methods, that is, principal component analysis (PCA) and partial least squares (PLS). Synergistic use of NIR with the PCA/PLS method makes it possible, for the first time, not only to divide different lung tissue samples into their respective groups (normal, CH, and MCT) but also to gain insight into mechanisms of PH caused by CH and MCT toxicity. Specifically, MCT metabolites and other hypertensive conditions are known to produce subtle and minor chemical changes in the compositions of tissue (e.g., proteins, carbohydrates, lipids). Although these changes were detected by the NIR technique, they were too small to be discerned through visual inspection of the spectra. However, they can be accurately classified and properly assigned by the PCA/PLS method. The fact that different tissue types can be accurately divided into their corresponding groups by the NIR and PCA/PLS methods suggests that chemical alterations and mechanisms of pulmonary vascular remodeling and PH induced by MCT are different from those induced by CH

Axolotls' and Mices' Oral-Maxillofacial Trephining Wounds Heal Differently

The Ambystoma maxicanum (axolotl) regenerates strikingly from wounds and amputations. Comparing its healing ability to non-regenerative species such as the mouse should help narrow in on mechanisms to improve human wound healing. Here, the tongue and intermandibular soft tissues of both mice (C57BL/6NCrl) and axolotls were wounded with a 2-2.5 mm punch biopsy. The study aimed to compare the differences between these 2 species following surgical resection with regard to the macroscopic and histological characteristics. These include wound closure times, epithelial wound sealing and thickness as well as acute immune marker myeloperoxidase (MPO) response over 30 days. Post surgery, mice visually showed greater haemorrhage; their wounds immediately collapsed while it took 14 days for the axolotls mandibular void to close. The epithelium sealed the axolotls' wound margins within 24 h with a maximal mean thickness of 0.42 +/- 0.13-fold normalized to unwounded skin. In mice, the epithelium separately sealed the ventral and dorsal sides, respectively at 7 and 7-30 days with mean maximal epithelial thicknesses reaching 13 +/- 5.6 and 3.0 +/- 0.63-fold. Mean MPO-positive cell values peaked in axolotls at 14 +/- 1.5-fold between hours 6-12; while in mice, it peaked at 8.7 +/- 0.9-fold between hours 24-96. We conclude that axolotls form smaller blood clots, have a faster and thinner epithelial cell migrating front, and a shorter MPO-positive cell response in comparison to mice. These observations may help refine future oral and facial wound-healing research and treatment.Peer reviewe

Evaluation of pre-analytical factors affecting plasma DNA analysis.

Pre-analytical factors can significantly affect circulating cell-free DNA (cfDNA) analysis. However, there are few robust methods to rapidly assess sample quality and the impact of pre-analytical processing. To address this gap and to evaluate effects of DNA extraction methods and blood collection tubes on cfDNA yield and fragment size, we developed a multiplexed droplet digital PCR (ddPCR) assay with 5 short and 4 long amplicons targeting single copy genomic loci. Using this assay, we compared 7 cfDNA extraction kits and found cfDNA yield and fragment size vary significantly. We also compared 3 blood collection protocols using plasma samples from 23 healthy volunteers (EDTA tubes processed within 1 hour and Cell-free DNA Blood Collection Tubes processed within 24 and 72 hours) and found no significant differences in cfDNA yield, fragment size and background noise between these protocols. In 219 clinical samples, cfDNA fragments were shorter in plasma samples processed immediately after venipuncture compared to archived samples, suggesting contribution of background DNA by lysed peripheral blood cells. In summary, we have described a multiplexed ddPCR assay to assess quality of cfDNA samples prior to downstream molecular analyses and we have evaluated potential sources of pre-analytical variation in cfDNA studies