Summary Monoclonal antibodies were raised against human placental soluble E-cadherins and used in an immunoenzymometric assay to detect soluble E-cadherins in biological fluids. The E-cadherin assay was accurate enough to quantitate the concentration of soluble E-cadherin in the cell culture supernatants. Immunoreactive E-cadherins, identified as existing in the soluble form in normal serum, were shown to have apparent lower molecular mass (approximately 80 kDa) than intact molecules of E-cadherin. We found that the immunoreactive E-cadherin levels in the serum of the studied cancer patients were significantly elevated (mean ± s.d. 3.80 ± 2.36 tig ml', P<0.0001) when compared with the normal levels (1.99 ± 0.50 1tg ml-'). We also found that serum E-cadherin levels in the 22 patients with gastric cancer (3.51 ± 1.78 tLg ml-, P <0.02) or the 11 patients with hepatocellular cancer (5.55 ± 3.11 Lg ml-', P<0.001) were significantly higher than those in the 26 diabetic patients (2.33 ± 1.58 iLg ml-'). Of the 54 cancer patients, 53.7% exhibited an elevated amount of soluble E-cadherin in serum. Thus, it is evident that soluble E-cadherin in circulation can be used as a prospective tumour marker that accurately reflects the progressive regeneration of E-cadherin at tumour sites, potentially induced by tumour-associated proteolytic degradation.

Laminin-5 (LN-5), consisting of alpha3-, beta3-, and gamma2-chains, is a component of the cell adhesion complex containing hemidesmosomes and anchoring fibrils. This protein is a major constituent of the extracellular matrix and has recently proved to be an invasion marker for epithelial cells in many immunohistochemical surveys, indicating that it is frequently expressed in the invading edges of epithelial tumour cells. Additionally, intracellular accumulation of monomeric gamma2-chains has been widely observed in the invasive carcinoma cells, but its mechanism was not entirely understood. Epithelial carcinoma cells prefer to adhere onto the LN-5-rich basement membranes using the specific integrins as receptors. Induction of cell migration is an important function of LN-5 and the enhanced activity is observed in its truncated form after proteolytic shedding of the N-terminal fragments of gamma2-chains. This processing was demonstrated to be mediated mainly by several kinds of matrix metalloproteinases. The degraded fragments of gamma2-chains, released from invading carcinomas, can be immunodetected in biological fluids and potentially utilized in the clinical diagnosis of various epithelial cancers. Here, we summarize the previous clinical investigations of LN-5 in epithelial tumour progression, and also discuss what it can regulate in the cell physiological events.

P-selectin is an integral membrane glycoprotein stored in the secretory granules of platelets and endothelial cells. To determine whether soluble P-selectin may be present in the circulation of healthy humans, we used a sandwich immunoassay to assess citrated plasma from 50 subjects. P-selectin was present in concentrations ranging from 19 to 521 ng/ml (mean +/- SD = 121 +/- 84 ng/ml). The apparent molecular weight of P-selectin immunoisolated from platelet-poor plasma was similar to that of the detergent-soluble form isolated from platelet membrane. Plasma levels of P-selectin were unaffected by the following procedures: (1) drawing of blood in the presence of protease inhibitors; (2) stimulation of platelet-rich plasma with aggregating agents; (3) ultracentrifugation at 100,000 g for 120 min at 4 degrees C or filtration through a 0.22 micron membrane; or (4) preincubation of platelet-poor plasma with immobilized anti-platelet glycoprotein Ib monoclonal antibodies. It appeared that plasma P-selectin did not result from the in vitro activation of platelets, nor was it derived from platelet microparticles. We also found that plasma P-selectin levels were significantly elevated in patients with thrombotic thrombocytopenic purpura (12 patients, 332 +/- 184 ng/ml, P < 0.001) and haemolytic uraemic syndrome (17 patients, 297 +/- 191 ng/ml, P < 0.0001), as compared to the normal levels. Thus, these data should facilitate the study of the pathophysiological significance of circulating P-selectin.

von Willebrand factor (vWF) is a multimeric plasma protein with ABO (H) blood group sugar chains. We investigated a total of 330 plasmas from normal individuals having various ABO genotypes, with special reference to vWF antigen and its platelet glycoprotein-Ib-related biological activities, termed ristocetin cofactor (RCof) and botrocetin cofactor (BCof). RCof reflects the biological activity of higher vWF multimers, while BCof reflects that of vWF of multimers of all sizes. Plasmas from normal individuals carrying one O gene (genotypes AO and BO) had slightly, but proportionally lower levels of vWF antigen, RCof, and BCof than those carrying no O gene (genotypes AA, AB, and BB). Normal plasmas from individuals carrying two O genes (genotype OO) showed much lower values for these parameters than the other plasmas, as previously reported. However, multimeric analysis of plasma vWF antigen revealed no differences among the different genotypes.

Basic fibroblast growth factor (bFGF) may be involved in the development and repair of dentine and pulp because bFGF, its related peptides, and FGF receptors are expressed in dental mesenchymal cells. In this study, we examined the effects of bFGF on DNA synthesis, osteonectin/SPARC levels, alkaline phosphatase (ALPase) activity, their mRNA levels, and calcium levels in cultures of human pulp cells. Pulp cells were isolated from three healthy upper wisdom teeth of three patients and maintained separately. These cells produced SPARC, ALPase, and calcified nodules and there was a close correlation between the SPARC-synthetic activity of the cell lines and their levels of ALPase and calcification. The levels of SPARC, ALPase and calcium deposits in the three pulp cell cultures were 10-250 times those of human foreskin fibroblasts. Western blots showed that the pulp cells produced 38-kDa SPARC. Northern blots showed that the pulp cells expressed flg (FGF receptor type 1) transcripts throughout all culture stages, irrespective of the presence or absence of bFGF. The addition of bFGF to the pulp cultures suppressed the increases in ALPase activity, SPARC synthesis, and their mRNA levels, although it increased the incorporation of [3H]thymidine into DNA> 10-fold. The effects of bFGF on ALPase activity and SPARC synthesis were reversible. Furthermore, bFGF abolished the calcification of the extracellular matrix; the calcium content of bFGF-free cultures. These findings suggest that bFGF is a potent mitogen for human pulp cells and that it inhibits the expression of the odontoblast phenotype by the cells at least partly at pretranslational levels.