The presence of restriction enzymes in bacterial cells has been predicted by either classical phage restriction-modification (R-M) tests, direct in vitro enzyme assays or more recently from bacterial genome sequence analysis. We have applied phage R-M test principles to the transformation of plasmid DNA and established a plasmid R-M test. To validate this test, six plasmids that contain BamHI f...

The regulation of cell motility is central to living systems. Consequently, cell migration assays are some of the most frequently used in vitro assays. This article provides a comprehensive, detailed review of in vitro cell migration assays both currently in use and possible with existing technology. Emphasis is given to two-dimensional migration assays using densely organized cells such as the...

Despite the advent of a considerable number of new antibiotics, treatment of intracellular pathogens still represents a major pharmaceutical challenge. The antibiotic concentration in those specialized niches are often subtherapeutic, for which high doses of antibiotics must often be used. This is not only costly but may also increase localized or systemic side effects. There is therefore an ur...

objective: piwil2, a member of ago/piwi gene family containing piwi and paz domains, has been shown to be ectopically expressed in different cancer cells, especially its remarkable expression in cancer stem cells (cscs), and is also known to be essential for germ line stem cell self-renewal in various organisms. the hypothesis that csc may hold the key to the central problem of clinical oncolog...

The objective of this study was to compare the in vitro release of indomethacin and hydrocortisone from self-emulsifying drug delivery systems (SEDDS) and aqueous or oily suspensions. SEDDS carriers were obtained by dissolving Cremophor EL, Tween 20 or Span 80 in Miglyol oil. The release experiment was performed over 6 h using a dialysis cellulose membrane and acceptor fluid imitating compositi...

Arginine is used in supra-physiological concentrations as an insulin secretagogue, in both in vitro and in vivo studies. To investigate the pharmacokinetics of arginine in humans, we have developed a rapid, automated assay of arginine in serum, based on our manual enzymatic method (Clin Chim Acta 1988, 176; 185-94). The limit of linearity of the automated assay was an arginine concentration of ...

In a previous study, we identified several structurally unrelated scaffolds of the Rho kinase inhibitor using pharmacophore information obtained from the results of a high-throughput screening and structural information from a homology model of Rho kinase. 1H-Indazole is one of the candidate scaffolds on which a new series of potent Rho kinase inhibitors could be developed. In this study, the d...

The sumoylation reaction is mechanistically similar to ubiquitination. It is ATP-dependent and in vitro can be completed in two steps using purified E1 (SAE1/SAE2), E2 (ubc9), and SUMO. Even without the inclusion of an E3 ligase, many substrates can be modified at the same lysines in vitro as in vivo. Here we describe a simplified in vitro sumoylation protocol using recombinant sumoylation subs...