The expression of proteinase-activated receptor (PAR)(2) in human hepatocellular carcinoma (HCC) was established by reverse transcription-polymerase chain reaction, confocal immunofluorescence and electron microscopy in permanent cell lines, primary HCC cell cultures and HCC tumor tissue. Stimulation of HCC cells with trypsin and the PAR(2)-selective activating peptide, 2-furoyl-LIGRLO-NH(2), increased cell invasion across Matrigel. Both effects were blocked by a PAR(2)-selective pepducin antagonist peptide (pal-PAR(2)) and by PAR(2) silencing with specific small interfering RNA (siRNA). PAR(2)-initiated HCC cell invasion was also blocked by inhibiting the hepatocyte growth factor receptor (Met receptor tyrosine kinase) with the receptor-targeted kinase inhibitors, SU 11274 and PHA 665752, or by downregulation of Met with specific siRNA. The involvement of Met in PAR(2)-mediated HCC invasive signaling was further supported by the finding that treatment of HCC cells with trypsin or the PAR(2)-selective agonist peptide, 2-furoyl-LIGRLO-NH(2), stimulated Met activation-phosphorylation. In addition, Met-dependent stimulation of p42/p44 mitogen-activated protein Kinases was found to be critical for the PAR(2)-Met receptor tyrosine kinase-invasive signaling axis in HCC cells. Our study establishes an important link between the PAR(2) and Met receptor tyrosine kinase signaling in promoting HCC cell invasion.

The aim of this research was to assess the treatment efficiencies for gully pot liquor of experimental vertical-flow constructed wetland filters containing Phragmites australis (Cav.) Trin. ex Steud. (common reed) and filter media of different adsorption capacities. Six out of 12 filters received inflow water spiked with metals. For 2 years, hydrated nickel and copper nitrate were added to sieved gully pot liquor to simulate contaminated primary treated storm runoff. For those six constructed wetland filters receiving heavy metals, an obvious breakthrough of dissolved nickel was recorded after road salting during the first winter. However, a breakthrough of nickel was not observed, since the inflow pH was raised to eight after the first year of operation. High pH facilitated the formation of particulate metal compounds such as nickel hydroxide. During the second year, reduction efficiencies of heavy metal, 5-days at 20°C N-Allylthiourea biochemical oxygen demand (BOD) and suspended solids (SS) improved considerably. Concentrations of BOD were frequently Ͻ20 mg/L. However, concentrations for SS were frequently Ͼ30 mg/L. These are the two international thresholds for secondary wastewater treatment. The BOD removal increased over time due to biomass maturation, and the increase of pH. An analysis of the findings with case-based reasoning can be found in the corresponding follow-up paper (Part B).

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The aim of this research was to assess the treatment efficiencies for gully pot liquor of experimental vertical-flow constructed wetland filters containing Phragmites australis (Cav.) Trin. ex Steud. (common reed) and filter media of different adsorption capacities. Six out of 12 filters received inflow water spiked with metals. For 2 years, hydrated nickel and copper nitrate were added to sieved gully pot liquor to simulate contaminated primary treated storm runoff. The findings were analyzed and discussed in a previous paper (Part A). Case-based reasoning (CBR) methods were applied to predict 5 days at 20°C N-Allylthiourea biochemical oxygen demand (BOD) and suspended solids (SS), and to demonstrate an alternative method of analyzing water quality performance indicators. The CBR method was successful in predicting if outflow concentrations were either above or below the thresholds set for water-quality variables. Relatively small case bases of approximately 60 entries are sufficient to yield relatively high predictions of compliance of at least 90% for BOD. Biochemical oxygen demand and SS are expensive to estimate, and can be cost-effectively controlled by applying CBR with the input variables turbidity and conductivity.