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1. (Article ID: 5494)
 
Sengyee S, Yoon SH, West TE, Ernst RK, Chantratita N
Lipopolysaccharides from Different Burkholderia Species with Different Lipid A Structures Induce Toll-Like Receptor 4 Activation and React with Melioidosis Patient Sera
Infection and Immunity 87(17) (2019) e00692-19
 

Lipopolysaccharides (LPSs) of Gram-negative bacteria comprise lipid A, core, and O-polysaccharide (OPS) components. Studies have demonstrated that LPSs isolated from the pathogenic species Burkholderia pseudomallei and Burkholderia mallei and from less-pathogenic species, such as Burkholderia thailandensis, are potent immune stimulators. The LPS structure of B. pseudomallei, the causative agent of melioidosis, is highly conserved in isolates from Thailand; however, the LPSs isolated from other, related species have not been characterized to enable understanding of their immune recognition and antigenicities. Here, we describe the structural and immunological characteristics of the LPSs isolated from eight Burkholderia species and compare those for B. pseudomallei to those for the other seven species. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), gas chromatography (GC), SDS-PAGE, Toll-like receptor 4 (TLR4) stimulation, and immunoblot analysis were performed on these Burkholderia species. MALDI-TOF profiles demonstrated that Burkholderia lipid A contains predominantly penta-acylated species modified with 4-amino-4-deoxy-arabinose residues at both terminal phosphate groups. The lipid A could be differentiated based on mass differences at m/z 1,511, 1,642, 1,773, and 1,926 and on fatty acid composition. LPSs of all species induced TLR4-dependent NF-?B responses; however, while SDS-PAGE analysis showed similar LPS ladder patterns for B. pseudomallei, B. thailandensis, and B. mallei, these patterns differed from those of other Burkholderia species. Interestingly, immunoblot analysis demonstrated that melioidosis patient sera cross-reacted with OPSs of other Burkholderia species. These findings can be used to better understand the characteristics of LPS in Burkholderia species, and they have implications for serological diagnostics based on the detection of antibodies to OPS.

Lipopolysaccharide, Burkholderia, antibody, O-polysaccharide, serology, melioidosis, innate immunity, LPS structure, TLR4, Diagnostics, bacterial infection

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2. (Article ID: 7265)
 
Nie SP, Cui SW, Phillips AO, Xie MY, Phillips GO, Al-Assaf S, Zhang XL
Elucidation of the structure of a bioactive hydrophilic polysaccharide from Cordyceps sinensis by methylation analysis and NMR spectroscopy
Carbohydrate Polymers 84(3) (2011) 894-899
 

The structure of a bioactive hydrophilic polysaccharide fraction from Cordyceps sinensis (CBHP) was studied using methylation analysis and 2D NMR spectroscopy. Monosaccharide composition analysis revealed that it consists mainly of glucose (95.19%) with trace amounts of mannose (0.91%) and galactose (0.61%). The results of methylation analysis indicated that α-1,4 linked Glcp is the main linkage type (65.7%), followed by t-Glcp (20.7%), 1,2,3,6-Glcp (4.1%), 1,2,4,6-Glcp (3.0%), 1,3,6-Glcp (2.0%), 1,4,6-Glcp (1.6%), and 1,2-Manp (1.9%) and 1,3-Galp (1.0%). Based on 1D and 2D NMR analysis, a preliminary structure is proposed: The backbone is composed of Glcp joined by 1→4 linkages and 1→3 linkages; the branching points are located at O-2 or O-6 of Glcp with α- terminal-d-Glcp as side chain. The trace amounts of 1,2-Manp and 1,3-Galp linkages are probably located randomly in the side chains. A schematic structure is proposed as following: structure.

NMR, structure, methylation analysis, Cordyceps sinensis, hydrophilic polysaccharide

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