2.3 Syndecan-1 assay:
Plasma concentrations of the glycocalyx component syndecan-1 (SD-1), a marker of glycocalyx shedding activated by acute inflammation (Mulivor et al. , 2004; Nieuwdorp et al. , 2006) , were determined by ELISA (Abcam biotechnology, UK). Assay was performed according to the manufacturer’s instructions. Briefly, standards and samples were prepared and were pipetted into a 96- well plate coated with primary antibody for SD-1. The plate was incubated at room temperature for 60 min on a mechanical shaker followed by a wash step and addition of the enzyme streptavidin-HRP. Finally, a color-forming substrate for that enzyme was added in each well and incubated again for 15min in the dark, then absorbance was assessed using a spectrophotometer (Bio-Rad, USA) at 450nm wavelength.