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Preliminary studies have demonstrated structural differences in the glycocalyx of pulmonary artery endothelial cells compared with pulmonary microvascular endothelial cells. In addition, if the interaction between sialic acid and the AAV6 capsid is more efficient than that of AAV1 for cell entry, this might account for the increased transduction of AAV6 compared to AAV1 in liver cells. To determine if N-linked sialic acid is necessary for efficient AAV1 and AAV6 transduction, we tested AAV1 and...
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The reaction was followed by acquiring 1D NMR experiments at 15-min intervals over 24 h. Gene functions were inferred from BLAST searches followed by gene linkage and cluster analysis. Deletion of yjhC resulted in loss of growth on 2,7-anhydro-Neu5Ac but not on Neu5Ac (Fig. 5C), which could be complemented in trans with yjhC (Fig. 5D), suggesting that the gene encodes an equivalent protein to RgNanOx. To test this hypothesis, the YjhC protein was recombinantly expressed and purified, and its ac...
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In addition, we determined by inhibitor (N-benzyl GalNAc)- and cell line-specific (Lec-1) studies that AAV1 and AAV6 require N-linked and not O-linked sialic acid. E. coli can transport and catabolize the common sialic acid, Neu5Ac, as a sole source of carbon and nitrogen but also related sialic acids, N-glycolylneuraminic acid (Neu5Gc) and 3-keto-3-deoxy-d-glycero-d-galactonononic acid (KDN), which are transported via the sialic acid transporter NanT and catabolized using the sialic acid aldol...
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4-Keto-DANA exists as an equilibrium between two ring-flipped forms; however, the form with the equatorial glycerol and N-acetyl substituents would be expected to predominate. Preferably, the substantial identity exists over a region of the sequences that is at least about 50 residus in length, more preferably over a region of at least about 100 residus, and most preferably the sequences are substantially identical over at least about 150 residus. The phrase "substantially identical," in the co...
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This was followed by the addition of neuraminidase solution or buffer control to each well and continued resistance measurement 4 (or more) h. AAV6 relies more on sialic acid or sialic acid-containing glycoproteins than AAV1 for cell entry and/or subsequent steps of infection. Neuraminidases are enzymes that cleave via hydrolysis α(2-3)-, α(2-6)-, and α(2-8)-linked terminal sialic acid residues bound to Gal, GlcNac, GalNAc, AcNeu, or GlyNeu residues of oligosaccharides, glycolipids, and glycopr...
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Dendritic cells (DCs) play an essential role in immunity against bacteria by phagocytosis and by eliciting adaptive immune responses. The phagocytosis of different pathogenic E. coli isolates was also enhanced by sialidase, which suggests that modifications on MDDC sialic acids may be considered in the development of MDDC-based antibacterial therapies. Although at first it was surprising that there was still lectin binding following neuraminidase treatment, especially by the neuraminidase from ...
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The resulting weight gain during the last 2 min of raising the venous pressure was divided by the concomitant increment of capillary pressure, and values were normalized to predicted lung weight. The amounts of activated Rac1 or Cdc42 (arbitrary units) are based on the optical density values obtained after subtracting negative control values. A "recombinant expression cassette" or simply an "expression cassette" is a nucleic acid construct, generated recombinantly or synthetically, with nucleic...
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