Uric acid inhibition of dipeptidyl peptidase IV in vitro is dependent on the intracellular formation of triuret.

Uric acid affects endothelial and adipose cell function and has been linked to diseases such as hypertension, metabolic syndrome, and cardiovascular disease. Interestingly uric acid has been shown to increase endothelial progenitor cell (EPC) mobilization, a potential mechanism to repair endothelial injury. Since EPC mobilization is dependent on activity of the enzyme CD26/dipeptidyl peptidase (DPP)IV, we examined the effect uric acid will have on CD26/DPPIV activity.

Evaluation of stability and sensitivity of cell fluorescent labels when used for cell migration.

The directed migration of mammalian cells is a foundation of development and growth. A variety of processes such as tissue development, wound healing, pathogen recognition/destruction as well as cancer metastasis are the result of regulated or dysregulated cell migration. While the ability to measure a cell's propensity to migrate has clinical relevance in several settings, no universal protocol has been established to measure cell migration.

Induction of nitric oxide by erythropoietin is mediated by the {beta} common receptor and requires interaction with VEGF receptor 2.

Vascular endothelial growth factor (VEGF) and erythropoietin (EPO) have profound effects on the endothelium and endothelial progenitor cells (EPCs), which originate from the bone marrow and differentiate into endothelial cells. Both EPO and VEGF have demonstrated an ability to increase the number and performance properties of EPCs. EPC behavior is highly dependent on nitric oxide (NO), and both VEGF and EPO can stimulate intracellular NO.

Lambda chops: creation of site-directed mutants in insertable fragments utilizing Gateway technology.

We describe a method to produce site-directed mutations anywhere within cDNA by assembling mutagenized PCR fragments in proper orientation using lambda integration in an extension of Gateway technology to yield a full-length mutated gene. This process exploits the directionality of lambda insertion sequences ensuring integration and directionality of PCR product into a cloning vector. The process requires only two sequential integration steps to yield a mutagenized expression vector.

Nitric oxide cytoskeletal-induced alterations reverse the endothelial progenitor cell migratory defect associated with diabetes.

Stromal-derived factor-1 (SDF-1) is a critical chemokine for endothelial progenitor cell (EPC) recruitment to areas of ischemia, allowing these cells to participate in compensatory angiogenesis. The SDF-1 receptor, CXCR4, is expressed in developing blood vessels as well as on CD34+ EPCs. We describe that picomolar and nanomolar concentrations of SDF-1 differentially influence neovascularization, inducing CD34+ cell migration and EPC tube formation.

The 1.4-MDa apoptosome is a critical intermediate in apoptosome maturation.

Previously, we demonstrated that both 150 mM KCl and alkaline pH inhibit cytochrome c-mediated activation of procaspase-3 in a unique manner. To determine the mechanism of inhibition, we analyzed the effect of KCl and alkaline pH on the formation of apoptosomes (a large complex consisting of cytochrome c, Apaf-1, and procaspase-9/caspase-9) in vitro. Our results suggest that an initial approximately 700-kDa apoptosome matures through a 1.

Marker for real-time analysis of caspase activity in intact cells.

Apoptosis, or programmed cell death, is an important regulator of growth, development, defense, and homeostasis in multicellular organisms. A family of cysteine proteases known as caspases is central to many apoptotic pathways, and thus detection of their activity offers an effective means to assess apoptosis. However, currently available methods only allow the evaluation of in vivo caspase activity at a given time point or over a few hours.