Glycocalyx in vivo measurement.

The endothelial glycocalyx (EG) lining the endoluminal surface of the capillaries has been proposed as a key component of the microcirculation and a major player in microvascular pathology. Recent advances in the understanding of its physiological role and clinical significance have been made upon the development of methods allowing EG assessment in clinical medicine. Laboratory methods can assess the amount of EG damage by measuring levels of its degradation products (e.g. syndecan-1, heparan sulphate and hyaluronan sulphate), mostly in the plasma, however, their physiological turnover disqualifies them from being the reliable index of EG damage. At the bedside, in vivo video microscopy tools technologies (e.g. Side-stream Dark Field imaging technology) allow indirect assessment of EG thickness in sublingual microcirculation by measuring the penetration extent (called Perfused Boundary Region) of flowing red blood cells into the EG.

Novel High-throughput Approach for Purification of Infectious Virions.

Viruses are extensively studied as pathogens and exploited as molecular tools and therapeutic agents. Existing methods to purify viruses such as gradient ultracentrifugation or chromatography have limitations, for example demand for technical expertise or specialized equipment, high time consumption, and restricted capacity. Our laboratory explores mutations in oncolytic reovirus that could improve oncolytic activity, and makes routine use of numerous virus variants, genome reassortants, and reverse engineered mutants. Our research pace was limited by the lack of high-throughput virus purification methods that efficiently remove confounding cellular contaminants such as cytokines and proteases. To overcome this shortcoming, we evaluated a commercially available resin (Capto Core 700) that captures molecules smaller than 700 kDa. Capto. Core 700 chromatography produced virion purity and infectivity indistinguishable from CsCl density gradient ultracentrifugation as determined by electron microscopy, gel electrophoresis analysis and plaque titration. Capto Core 700 resin was then effectively adapted to a rapid in-slurry pull-out approach for high-throughput purification of reovirus and adenovirus. The in-slurry purification approach offered substantially increased virus purity over crude cell lysates, media, or high-spin preparations and would be especially useful for high-throughput virus screening applications where density gradient ultracentrifugation is not feasible.