Activation of the epidermal growth factor receptor (EGFR) by a novel metalloprotease pathway

Neutrophil Elastase (NE) is a pro-inflammatory protease present at higher than normal levels in the lung during inflammatory disease. NE regulates IL-8 production from airway epithelial cells and can activate both EGFR and TLR4. TACE/ADAM17 has been reported to trans-activate EGFR in response to NE. Here, using 16HBE14o-human bronchial epithelial cells we demonstrate a new mechanism by which NE regulates both of these events. A high molecular weight soluble metalloprotease activity detectable only in supernatants from NE-treated cells by gelatin and casein zymography was confirmed to be meprin alpha by Western immunoblotting. In vitro studies demonstrated the ability of NE to activate meprin alpha, which in turn could release soluble TGFa and induce IL-8 production from 16HBE14o- cells. These effects were abrogated by actinonin, a specific meprin inhibitor. NE-induced IL-8 expression was also inhibited by meprin alpha siRNA. Immunoprecipitation studies detected EGFR/TLR4 complexes in NE-stimulated cells overexpressing these receptors. Confocal studies confirmed colocalization of EGFR and TLR4 in 16HBE14o- cells stimulated with meprin alpha. NF?B was also activated via MyD88 in these cells by meprin alpha. In bronchoalveolar lavage fluid from NE knock-out mice infected intra-tracheally with Pseudomonas aeruginosa meprin alpha was significantly decreased compared with control mice, and was significantly increased and correlated with NE activity, in bronchoalveolar lavage fluid from individuals with cystic fibrosis but not healthy controls. The data describe a previously unidentified lung metalloprotease meprin alpha, and its role in NE-induced EGFR and TLR4 activation and IL-8 production.

The influence of reusing ‘Formtex’ controlled permeability

The effects of the reuse of ‘Formtex’ Controlled Permeability Formwork (CPF) liner on strength and durability properties of concrete were investigated at two different water-cement ratios and the results are reported in this paper. Test blocks were cast using the CPF on one side and impermeable formwork (IF) on the opposite side of the mould so that direct comparisons could be made between the two. The strength was assessed using the Limpet pulloff tester and both the air permeability and the water absorption (sorptivity) were measured using the Autoclam Permeability System. Both these instruments measured the ‘covercrete’ properties. In addition, cores cut from the test specimens were subjected to an accelerated carbonation test and a chloride exposure test. The results showed that the ‘Formtex’ CPF increases the surface strength and the durability of concrete compared to the IF. There was an almost complete elimination of blowholes. The permeability of concrete decreased and its resistance to the ingress of both carbon dioxide and chlorides increased when CPF was used. The beneficial effects of the Formtex CPF were most evident in concrete of higher water-cement ratio. With the reuse of the Formtex liner twice, that is a total of three uses, the performance of the CPF to improve the properties of concrete remained almost the same. In this research the CPF liner was cleaned thoroughly between each use, which must be adhered to for site applications for reproducing the beneficial effects observed in the laboratory.

MMP expression and abnormal lung permeability are important determinants of outcome in IPF

Matrix metalloproteinases (MMPs) degrade all of the extracellular matrix components of the intersititium and may play a role in abnormal alveolar permeability, which is a feature of idiopathic pulmonary fibrosis (IPF). The aims of the present study were to evaluate MMP protein levels in patients with IPF and determine any relationship to treatment and markers of permeability.