Preparation of benzoboroxole–modified gold surfaces for selective oligosaccharide and glycoprotein recognition

Recognition of oligosaccharides is associated with very limited specificity due to their strong solvation in water and the high degree of subtle structural variations between them. Here, oligosaccharide recognition sites are created on material surfaces with unmatched, binary on–off binding behaviour, sharply discriminating a target oligosaccharide over closely related carbohydrate structures. The basis for the superselective binding behaviour relies on the highly efficient generation of a pure, high order complex of the oligosaccharide target with synthetic carbohydrate receptor sites, in which the spatial arrangement of the multiple receptors in the complex is preserved upon material surface incorporation. The synthetic binding scaffolds can be easily tailored to recognise different oligosaccharides and glycoconjugates, opening up a realm of possibilities for their use in a wide field of applications, ranging from life sciences to diagnostics. In this thesis, a modular synthetic approach is to be created that can harness both the construction of high–yield, complex oligosaccharide–synthetic carbohydrate receptor assemblies and the precise generation of surface–confined templated binding sites. Therefore, recognition sites of unparalleled oligosaccharide discrimination are to be created. The synthesis and characterisation of various self–assembled monolayers (SAMs) and oligosaccharide–synthetic carbohydrate receptor complexes are to be described alongside molecular imprinting studies which will determine the binding of various saccharides. Surface plasmon resonance (SPR) is to be used and from these experiments, the selectivity and sensitivity of the binding can be calculated

Modulation of macrophage and epithelial cell immune defences by probiotic bacteria: immune stimulation versus suppression

Abstract Probiotic bacteria are live organisms, if consumed in adequate amounts might confer health benefits. These bacteria, such as Lactic acid bacteria (LAB), include a number of strains that have specific health promoting activi-ties, attributed to their immunomodulatory and anti-inflammatory properties. Gut mucosal macrophage subsets play a fundamental role in driving muco-sal immune responses. These include, tolerance, associated with an M2-, regulatory macrophage phenotype and inflammatory activation with an M1-like phenotype. The cross-link between mucosal tolerance and inflammatory cytokine suppression, and augmentation of IL-10 production in the gut relate to endotoxin tolerance. Endotoxin tolerance is a context; it could present an example for cell drive through a hypo-responsive state. An example is mu-cosal inflammatory pathologies, such as Crohn’s disease. When tolerance is broken, causing the destruction of gut mucosal tissue. This is where the macrophage phenotype, has been transformed from a regulatory M2- to an inflammatory M1-like phenotype. This is seen as a reaction to both, patho-genic and commensal bacteria. This investigation was aimed at assessing the activities of live probiotic bacteria; Lactobacillus salivarius strain MS13 and Lactobacillus plantarum strain C28 in the immunomodulation of macro-phage subsets in health, inflammation, and endotoxin tolerance. M1- and M2-like macrophages were generated in vitro from the THP-1 monocyte cell line by differentiation with PMA and Vitamin D3, respectively. Additionally, differentiated epithelial cells (Caco-2) were obtained by long term culturing for 21 days. The role of Lactobacillus strains C28 and MS13 to modulate epi-thelial barrier integrity and macrophage-epithelial cell inflammation was in-vestigated. TNFα, IL-1β, IL-18, IL-23, IL-12, IL-6, IL-8, and IL-10 were quanti-fied by ELISA and RT-PCR, whereas TLR-2, TLR-4, Tollip, SOCS3, STAT3 and TRAIL by RT-PCR. This study revealed that, first, live C28 and MS13 stimulated the proinflammatory cytokine by M2-like macrophages as well as the anti-inflammatory cytokine in a homeostatic status; whereas in an in-flammatory environment, C28 and MS13 differentially upregulated TNFα and IL-1β by M1 and M2-like macrophages induced by E.coli K12-LPS. Both strains downregulated K12-LPS induced IL-10 by M2-like macrophages. The response of stimulated M1 and M2 macrophages to C28 and MS13, was to differentially induce the gene expression of TLR-2, TLR-4, Tollip, NLRP3, SOCS-3, STAT3 and TRAIL. Second, the repeat-stimulation/tolerisation of M1 and M2 macrophages by live probiotic bacteria revealed, TNFα, IL-1β, IL-23, IL-18, IL-6 and IL-10 were upregulated in M1-like macrophages by C28, whereas MS13 upregulated TNFα, IL-1β, IL-18, and downregulated IL-12, IL-6, and IL-10. On the other hand, the tolerisation of M2-like macrophages by C28 and MS13 resulted in the downregulation of TNFα and IL-12p35 and upregulation of IL-1β, IL-18, IL-23, IL-12, IL-6, and IL-10. These findings were linked with the differential macrophage subset upregulation of TLR-4, NLRP3, STAT-3 and TRAIL gene expression. On the other hand, TLR-2, Tol-lip and SOCS-3 were downregulated in tolerised macrophage subsets by C28 and MS13. Furthermore, the role of lactobacilli strains C28 and MS13 in the modulation of endotoxin tolerance was to; upregulate TNF-α, IL-18, IL-23 and IL-10 by M1 and M2-like macrophages. This investigation also focused on the induction of the zona-occludin-1 (Zo-1), human β defensin-2 (hBD-2), and cytokine production IL-8 by Caco-2 cells. Trans epithelial electrical re-sistance (TEER) and RT-PCR measured the main cytokines studied pro-duced by Caco-2, were IL-8, also the epithelial barrier function. Live probiotic C28 and MS13 suppressed the production of IL-8 (in the presence or ab-sence TNFα and IL-1β). Moreover, in the co-culture of Caco-2 with macro-phage subsets, MS13 enhanced the expression of hBD-2 and ZO-1. These findings allow for the better understanding of live probiotic roles on macro-phage subsets functions and endotoxin tolerisation mechanisms, which may be beneficial for the development of in vivo models of probiotic bacteria and therapeutic targeting of inflammatory bowel disease.Iraqi Cultural attach

NOTIFICATION !!!

All the content of this special edition is retrieved from the conference proceedings published by the European Scientific Institute, ESI. http://eujournal.org/index.php/esj/pages/view/books The European Scientific Journal, ESJ, after approval from the publisher re publishes the papers in a Special edition

NOTIFICATION !!!

All the content of this special edition is retrieved from the conference proceedings published by the European Scientific Institute, ESI. http://eujournal.org/index.php/esj/pages/view/books The European Scientific Journal, ESJ, after approval from the publisher re publishes the papers in a Special edition