Automated glycan assembly of a S. pneumoniae serotype 3 CPS antigen

Vaccines against S. pneumoniae, one of the most prevalent bacterial infections causing severe disease, rely on isolated capsular polysaccharide (CPS) that are conjugated to proteins. Such isolates contain a heterogeneous oligosaccharide mixture of different chain lengths and frame shifts. Access to defined synthetic S. pneumoniae CPS structures is desirable. Known syntheses of S. pneumoniae serotype 3 CPS rely on a time-consuming and low-yielding late-stage oxidation step, or use disaccharide building blocks which limits variability. Herein, we report the first iterative automated glycan assembly (AGA) of a conjugation-ready S. pneumoniae serotype 3 CPS trisaccharide. This oligosaccharide was assembled using a novel glucuronic acid building block to circumvent the need for a late-stage oxidation. The introduction of a washing step with the activator prior to each glycosylation cycle greatly increased the yields by neutralizing any residual base from deprotection steps in the synthetic cycle. This process improvement is applicable to AGA of many other oligosaccharides

The Alignment of Liquid Crystals on the Film Surfaces of Soluble Aromatic Polyimides Bearing t-Butylphenyl and Trimethylsilylphenyl Side Groups

With the study goal of firstly elucidating the anisotropic interactions between oriented polymer chain segments and liquid crystal (LC) molecules, and secondly of determining the contributions of the chemical components of the polymer segments to the film surface topography, LC alignment, pretilt, and anchoring energy, we synthesized three dianhydrides, 1,4-bis(4'-t-butylphenyl)pyromellitic dianhydride (BBPD), 1,4-bis(4'-trimethylsilylphenyl)pyromellitic dianhydride (BTPD), and 2,2'-bis(4 ''-tert-butylphenyl)-4,4',5,5'-biphenyltetracarboxylic dianhydride (BBBPAn), and a series of their organosoluble polyimides, BBPD-ODA, BBPD-NMA, BBPD-MDA, BTPD-FDA, and BBBPAn-FDA, which contain the diamines 4,4'-oxydianiline (ODA), 4,4'-methylenediamine (MDA), and 4,4'-(hexafluoroisopropylidene)dianiline (FDA). All the polyimides were determined to be positive birefringent polymers, regardless of the chemical components. Although all the rubbed polyimide films exhibited microgrooves which were created by rubbing process, the film surface topography varied depending on the polyimides. In all the rubbed films, the polymer chains were unidirectionally oriented along the rubbing direction. However, the degree of in-plane birefringence in the rubbed film varied depending on the polyimides. The rubbing-aligned polymer chains in the polyimide films effectively induced the alignment of nematic LCs along their orientation directors by anisotropic interactions between the preferentially oriented polymer chain segments and the LCs. The azimuthal and polar anchoring energies of the LCs ranged from 0.45x10(-4) - 1.37x10(-4) J/m(2) and from 0.86x10(-5) - 4.26x10(-5) J/m(2), respectively, depending on the polyimides. The pretilt angles of the LCs were in the range 0.10-0.62 degrees. In summary, the soluble aromatic polyimides, reported here are promising LC alignment layer candidates for the production of advanced LC display devices.X116sciescopuskc

Extract of polygala tenuifolia alleviates stress-exacerbated atopy-like skin dermatitis through the modulation of protein kinase a and p38 mitogen-activated protein kinase signaling pathway

© 2017 by the authors; licensee MDPI, Basel, Switzerland. Atopic dermatitis (AD) and stress create a vicious cycle: stress exacerbates atopic symptoms, and atopic disease elicits stress and anxiety. Targeting multiple pathways including stress and allergic inflammation is, therefore, important for treating AD. In this study, we investigated the remedial value of Polygala tenuifoliaWilld. (PTW) for treating immobilization (IMO) stress-exacerbated atopy-like skin dermatitis and its underlying mechanism. Trimellitic anhydride (TMA) was applied to dorsal skin for sensitization and subsequently both ears for eliciting T-cell-dependent contact hypersensitivity in mice, which underwent 2 h-IMO stress and PTW administration for the latter 6 and 9 days in the ear exposure period of TMA, respectively. To elicit in vitro degranulation of human mast cell line-1 (HMC-1), 10 μM substance P (SP) and 200 nM corticotrophin-releasing factor (CRF) were sequentially added with 48 h-interval. PTW extract (500 μg/mL) was added 30 min before CRF treatment. IMO stress exacerbated TMA-induced scratching behavior by 252%, and increased their blood corticosterone levels by two-fold. Treatment with 250 mg/kg PTW significantly restored IMO stress-exacerbated scratching behavior and other indicators such as skin inflammation and water content, lymph node weights, and serum histamine and immunoglobulin E (lgE) levels. Furthermore, it also reversed TMA-stimulated expression of tumor necrosis factor (TNF)-α and interleukin (IL)-4 mRNAs in ear tissues. PTW significantly inhibited SP/CRF-stimulated degranulation of HMC-1 cells, subsequent tryptase secretion, and protein kinase A (PKA) activity. PTW also selectively inhibited p38 mitogen-activated protein kinase (MAPK) phosphorylation in SP/CRF-treated HMC-1 cells. PTW significantly inhibited HMC-1 cell degranulation and alleviated IMO stress-exacerbated atopic dermatitis symptoms by modulating the PKA/p38 MAPK signaling pathway

Automated glycan assembly of plant cell wall oligosaccharides

Synthetic cell wall oligosaccharides are promising molecular tools for investigating the structure and function of plant cell walls. Their well-defined structure and high purity prevents misinterpretations of experimental data, and the possibility to introduce chemical handles provides means for easier localization and detection. Automated glycan assembly as emerged has a powerful new method for the efficient preparation of oligosaccharide libraries. We recently made use of this technology to prepare a collection of plant cell wall glycans for cell wall research. In this chapter, detailed experimental procedures for the automated synthesis of oligosaccharides that are ready for use in biological assays are described

REASSESSMENT OF THE UNBOUND CONCENTRATIONS OF UNCONJUGATED BILIRUBIN IN RELATION TO NEUROTOXICITY IN VITRO.

3nonenoneOSTROW JD; PASCOLO L; TIRIBELLI C.Ostrow, Jd; Pascolo, L; Tiribelli, Claudi