To determine the role of cathepsin L in Echinoderms, starfish (Asterina pectinifera) cathepsin L (ApCtL) was cloned. The results of RT-PCR analysis indicated that the expression of ApCtL in all of the tissues. The pro-mature enzyme of ApCtL, proApCtL, was expressed in Escherichia coli, and cathepsin activity was detected by cleaving of synthetic fluorogenic peptide substrates and gelatin zymogr...

The permeability characteristics of biomaterials are critical parameters for a variety of implants. To analyse the permeability of membranes made from crosslinked ultrathin gelatin membranes and the transmigration of cells across the membranes, we combined three technical approaches: (1) a two-chamber-based permeability assay, (2) cell culturing with cytochemical analysis and (3) biochemical en...

References 1. Tanumihardjo SA, Cheng JC, Permaesih D, Muherdiyantiningsih, Rustan E, Muhilal, et al. Refinement of the modified-relative-dose-response test as a method for assessing vitamin A status in a field setting: experience with Indonesian children. Am J Clin Nutr 1996;64: 966–71. 2. de Pee S, Yuniar Y, West CE, Muhilal. Evaluation of biochemical indicators of vitamin A status in breast-f...

We have developed a two-dimensional (2D-) gel system of zymography and reverse zymography for the detection and characterization of proteases and protease inhibitors. Isoelectric focusing (IEF) agarose gels with pH gradients were employed for separation in the first-dimension and sodium dodecyl sulfate (SDS)-polyacrylamide gel copolymerized with gelatin used for the second dimension. Proteases ...

BACKGROUND The matrix-degrading proteinases are believed to play an important role in the invasion and metastasis of hepatocellular carcinoma (HCC), but no one has ever seen the in situ matrix-degrading activity in HCCs. PURPOSE To demonstrate the cellular localization of actual gelatinolytic activity and to investigate the invasive potential of human HCC. EXPERIMENTAL DESIGN HCC cases (30)...

We report the effect of nickel chloride on fibroblast-mediated collagen gel contraction, fibroblast migration and pattern of matrix metalloproteinase (MMP) activities expressed by human embrzonary lung fibroblasts (HELFs). After the treatment with nickel chloride, gelatin zymography, casein zymography and also western blot assay revealed a significant decrease of MMP-2 and MMP-1 active forms. M...

PURPOSE To conduct zymographic analysis to study the matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) in vitreous samples of patients undergoing pars plana vitrectomy as part of the treatment of vitreoretinal disease. METHODS Forty-two vitreous samples were collected at the time of pars plana vitrectomy. Diagnoses included severe (exudative) age-related macu...

The uses of shark collagen as a matrix for cell culture and as a substrate for zymography were investigated. Fibroblasts were cultured on a gel matrix of shark type I collagen at 30 degrees C. The collagen gel had contracted by 4 days of incubation. Individual fibroblasts were visible against the transparent background of the contracted collagen as long, lean star-shaped cells. The matrix metal...

An aqueous extract of human placenta exhibits strong gelatinase/collagenase activity in zymography. 2-D gel electrophoresis of the extract with gelatin zymography in the second dimension displayed a single spot, identified as ubiquitin-like component upon MALDI/TOF MS/MS analysis. Immunoblot indicated presence of ubiquitin and absence of collagenase in the extract. Collagenase activity of the u...