Regulation of L-selectin-dependent hydrodynamic shear thresholding by leukocyte deformability and shear dependent bond number.

Background: During inflammation leukocyte attachment to the blood vessel wall is augmented by capture of near-wall flowing leukocytes by previously adherent leukocytes. Adhesive interactions between flowing and adherent leukocytes are mediated by L-selectin and P-selectin Glycoprotein Ligand-1 (PSGL-1) co-expressed on the leukocyte surface and ultimately regulated by hydrodynamic shear thresholding.

Objective: We hypothesized that leukocyte deformability is a significant contributory factor in shear thresholding and secondary capture.

The Escherichia coli clamp loader can actively pry open the β-sliding clamp.

Clamp loaders load ring-shaped sliding clamps onto DNA. Once loaded onto DNA, sliding clamps bind to DNA polymerases to increase the processivity of DNA synthesis. To load clamps onto DNA, an open clamp loader-clamp complex must form.

A slow ATP-induced conformational change limits the rate of DNA binding but not the rate of beta clamp binding by the escherichia coli gamma complex clamp loader.

In Escherichia coli, the gamma complex clamp loader loads the beta-sliding clamp onto DNA. The beta clamp tethers DNA polymerase III to DNA and enhances the efficiency of replication by increasing the processivity of DNA synthesis. In the presence of ATP, gamma complex binds beta and DNA to form a ternary complex.

Temporal correlation of DNA binding, ATP hydrolysis, and clamp release in the clamp loading reaction catalyzed by the Escherichia coli gamma complex.

Clamp loaders are multisubunit complexes that use the energy derived from ATP binding and hydrolysis to assemble ring-shaped sliding clamps onto DNA. Sliding clamps in turn tether DNA polymerases to the templates being copied to increase the processivity of DNA synthesis. Here, the rate of clamp release during the clamp loading reaction was measured directly for the first time using a FRET-based assay in which the E.

L-selectin shear thresholding modulates leukocyte secondary capture.

Transient homotypic adhesions between flowing leukocytes and those previously adherent on the vessel wall has been proposed to amplify the accumulation of leukocytes at sites of inflammation. While adhesion of leukocytes to the vessel wall (primary capture) is mediated primarily by P-selectin on the endothelium and P-selectin Glycoprotein Ligand-1 (PSGL-1) on the leukocyte, the homotypic interactions leading to downstream leukocyte adhesion (secondary capture) are mediated primarily by reciprocal interactions between PSGL-1 and L-selectin on apposing leukocytes. One consequence of leukocyte secondary capture events are the formation of strings of adherent leukocytes as each recently captured leukocyte in turn captures another one flowing over its surface.

Enhancement of L-selectin, but not P-selectin, bond formation frequency by convective flow.

L-selectin-mediated leukocyte rolling has been proposed to require a high rate of bond formation compared to that of P-selectin to compensate for its much higher off-rate. To test this hypothesis, a microbead system was utilized to measure relative L-selectin and P-selectin bond formation rates on their common ligand P-selectin glycoprotein ligand-1 (PSGL-1) under shear flow. Using video microscopy, we tracked selectin-coated microbeads to detect the formation frequency of adhesive tether bonds.

A model for clinical estimation of perioperative hemorrhage.

The purpose of this study was to assess the accuracy of estimated blood loss (EBL) as a reliable predictor of actual blood loss during orthopedic procedures. Between 1999 and 2002, 198 orthopedic cases were reviewed. A retrospective review compiled preoperative and postoperative demographic and laboratory data from the surgical patients.