Rapid detection of CEA mRNA in peritoneal washes using One-Step Nucleic acid Amplification (OSNA) for gastric cancer patients.

Background: Carcinoembryonic antigen (CEA) mRNA expression in peritoneal washes from gastric cancer patients has been reported as an indicator for survival or peritoneal recurrence. The whole process of CEA mRNA detection is time- and labor-intensive. We report the potential of One-Step Nucleic acid Amplification (OSNA) as a rapid and simple system for CEA mRNA detection in peritoneal washes.

Structural and mechanistic roles of three consecutive Pro residues of porcine NADH-cytochrome b(5) reductase for the binding of beta-NADH.

Well-conserved three consecutive Pro residues (Pro247-249) in the NADH-binding subdomain of NADH-cytochrome b(5) reductase were proposed to form a basal part of the NADH-binding site. To investigate the structural and mechanistic roles of these residues, we expressed site-directed mutants for a soluble domain of the porcine enzyme where each of the residues was replaced with either Ala or Leu residue, respectively, using a heterologous expression system in Escherichia coli. Six mutants (P247A, P247L, P248A, P248L, P249A, and P249L) were produced as a fusion protein containing a 6xHis-tag sequence at the NH(2)-terminus and were purified to homogeneity with a stoichiometric amount of bound FAD.