In the present study we determined the effect of the lectins concanavalin A (Con A), phytohemagglutinin (PHA) and pokeweed (PWM), frequently used in cell proliferation assays, on the proliferation of goat peripheral blood mononuclear cells (PBMCs). The use of a standardized Percoll gradient proved to be highly effective for the separation/enrichment of these cells, without damaging the cell mem...

Polymorphonuclear eosinophilic leucocytes (PME) participate in wound healing processes, the inflammatory response, bronchial asthma, allergies and defence against invading parasites. We have examined the effects of thiopental, methohexital, propofol, etomidate and ketamine on PME chemotaxis in vitro. PME were isolated from venous blood samples of 10 healthy volunteers using multi-stage Percoll ...

To identify previously unknown peroxisomal proteins, we established an optimized method for isolating highly purified peroxisomes from etiolated soybean cotyledons using Percoll density gradient centrifugation followed by iodixanol density gradient centrifugation. Proteins in highly purified peroxisomes were separated by two-dimensional PAGE. We performed peptide mass fingerprinting of proteins...

Functional plasma membranes from the filamentous fungus Penicillium chrysogenum have been isolated with the objective of studying transport processes. The isolation procedure consists of three steps, namely homogenization of cells with a Braun MSK homogenizer, followed by Percoll gradient centrifugation and floatation of membranes in a three-step Nycodenz gradient. This method can be applied to...

Proline accumulation is a well-known response to water deficits in leaves. The primary cause of accumulation is proline synthesis. Delta(1)-Pyrroline-5-carboxylate reductase (PCR) catalyzes the final reaction of proline synthesis. To determine the subcellular location of PCR, protoplasts were made from leaves of Pisum sativum L., lysed, and fractionated by differential and Percoll density gradi...

Hepatic stellate cells (HSC) and endothelial cells of the liver sinusoids synthesize and degrade hyaluronan, respectively. The roles of these cell types in the biosynthesis and degradation of hyaluronan were studied during regeneration following partial hepatectomy. Pure cultures of HSC and liver endothelial cells (LEC) were obtained from regenerating liver at different stages using a Nycodenz ...

The phenomenon of increased spontaneous incorporation of 3H-thymidine (3H-TdR) into peripheral blood mononuclear cells in rheumatoid arthritis (RA) has been investigated. The activity was found to be short lived and affected less than 1% of cells. Using a Percoll density gradient we identified two populations of active cells. RA patients with active synovitis and increased 3H-TdR incorporation ...