Mucin competition was carried out by pretreating rAAV with bovine submandibular gland mucin (Sigma) for 30 min at 20°C. AAV vectors, in the presence or absence of mucin, were added to Cos-7 cells in equal volumes of DMEM for 1 h at 37°C. Cells were rinsed twice with DMEM and incubated at 37°C. Two thousand […]
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ΔOD595 for triplicate experiments is shown: ΔnanT (A) and ΔyjhB (B). ΔOD595 for triplicate experiments is shown: BW25113 (B), ΔyjhC (C), and complemented yjhC (D). 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
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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
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In some experiments the bacterial surface was desialylated with 13· In the earlier set of experiments that utilized neuraminidase, we noted that, following neuraminidase treatment, there were typically fewer cells in the dish, indicating that cells had lost cell-cell and/or cell-matrix adhesions. If you adored this article so you would like to get more info
The first step of AAV infection often requires the attachment of the capsid to carbohydrate moieties on the cell surface. Of the 11 well-characterized AAV serotypes, heparan sulfate proteoglycan and sialic acid have been suggested to be the attachment receptors for AAV type 2 and types 4 and 5, respectively. In this report, we identify
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Previous work using spectrophotometric assays reported that E. coli YjhC showed a weak interaction with Neu5Ac, with an apparent Km of 68.8 mm (20). Here, we used ESI-MS to assess the activity of YjhC against 2,7-anhydro-Neu5Ac or Neu5Ac. The glycan array binding data provide independent support of AAV1 interaction with α2,3 and α2,6 trisaccharides. Our
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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
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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
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To assay for oxidoreductase activity, the purified recombinant proteins were incubated in 100-μl reactions at 37 °C overnight with 1 mg/ml 2,7-anhydro-Neu5Ac or Neu5Ac in 20 mm sodium phosphate buffer, pH 7.5, in the presence 500 μm NADH. The structure was acquired from a crystal grown in the JCSG Plus screen (100 mm sodium citrate,
It was beforehand believed that influenza pandemics occurred at 10- to 14-yr intervals, however it has been over 30 years since H3N2 viruses appeared. Viruses have been first isolated in Hong Kong in July 1968. Widespread disease with increased excess mortality was observed in the United States during the winter of 1968-1969. Should you have
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