July 2024

Neuraminidases and other reagents were obtained from Sigma-Aldrich. To determine whether sialic acids play a role in endothelial barrier function, cells were treated with neuraminidases to hydrolyze sialic acid moieties. Two components of glycocalyx structure that may play a role in determining function in glycoproteins are 1) the terminal sugar residue(s) on the oligosaccharide chain(s) […]

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

To confirm correct sequences in plasmids constructed, the plasmids can be analyzed by standard techniques such as by restriction endonuclease digestion, and/or sequencing according to known methods. Yeast Integrating plasmids (e.g., YIp5) and Yeast Replicating plasmids (the YRp series plasmids) and pGPD-2. Media can be rich media, e.g., Luria broth or terrific broth, or synthetic

HepG2 (A) and Pro-5 (B) cells were transduced with a constant amount of AAV1-luc or AAV6-luc vectors in the presence of a 200-fold-excess amount of competing AAV1, AAV6, or AAV2 encapsidated λ phage DNA-containing vector at 37°C for 1 h. However, more and more studies suggest that AAV2-based vectors are rate limiting in certain tissues

Background was washed out with deionized water and gel was scanned with the default instrument settings for analysis (Lexmark CX727 scanner). Inhibition of glycolipid synthesis was carried out as described previously (36) with some modifications. Resialylation was carried out with 50 mU/ml sialyltransferase in α-MEM for 2 h at 37°C. In the control groups, Lec-2

We stained cell lines with lectins that have been used to recognize the following three different epitopes: (i) WGA recognizes all sialic acids, (ii) MAA recognizes α2,3 sialic acid, and (iii) SNA recognizes α2,6 sialic acid. In this report, we demonstrate that AAV1 and AAV6 use both α2,3 and α2,6 N-linked sialic acids for binding

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