Reporter genes produce a protein product in transfected cells that can be easily measured in intact or lysed cells and they have been extensively used in numerous basic and applied research applications.Over the past 10 years,reporter gene assays have been widely accepted and used for analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like compounds in various types of matrices,such as biological,environmental,food and feed samples,given that high-resolution instrumental analysis techniques are impractical for large-scale screening analysis.The most sensitive cell-based reporter gene bioassay systems developed are the mechanism-based CALUX(Chemically Activated Luciferase Expression) and CAFLUX(Chemically Activated Fluorescent Expression) bioassays,which utilize recombinant cell lines containing stably transfected dioxin(AhR) responsive firefly luciferase or enhanced green fluorescent protein(EGFP) reporter genes,respectively.While the current CALUX and CAFLUX bioassays are very sensitive,increasing their lower limit of sensitivity,magnitude of response and dynamic range for chemical detection would significantly increase their utility,particularly for those samples that contain low levels of dioxin-like HAHs(i.e.,serum) .In this study,we report that the addition of modulators of cell signaling pathways or modification of cell culture conditions results in significant improvement in the magnitude and overall responsiveness of the existing CALUX and CAFLUX cell bioassays.
BASTON David S.KHAN ElaineSORRENTINO ClaudioDENISON Michael S.
The reactions of 4-pyrimidin-5-ylbenzoic acid (HL) with Co(NO3)2·6H2O and Ni(NO3)2·6H2O gave rise to two isomorphous mononuclear coordination complexes, namely, [CoL2(H2O)4] (1) and [NiL2(H2O)4] (2). Both compounds were characterized by IR spectroscopy and single-crystal X-ray diffraction. X-ray crystallographic analyses reveal that they crystallize in monoclinic, space group P21/n. In the crystal structures of these two complexes, only one nitrogen atom of L coordinates to the metal center, and neither carboxylate group nor another nitrogen atom of L links to Co(II) or Ni(II). 2D Layers with significant hydrogen bonds can be observed, and they are further linked by L to form a 3D supramolecular network. Nitrogen atoms are not involved in forming hydrogen bonds, but carboxylic groups of the ligands play an important role in the configuration of this hydrogen-bond layer.
