Oxidized lipoproteins activate NF-kappaB binding activity and apoptosis in PC12 cells

For the detailed analysis of energy metabolism, a HPLC method is described allowing the single-run separation and quantification of most metabolites from glycolysis and the Krebs cycle including the high energy phosphates. With a detection limit in the picomolar range this method is even applicable when only small sample sizes of tissue are obtained.     

Design and synthesis of potent, selective inhibitors of endothelin-converting enzyme

Endothelin-1 is the most potent peptidic vasoconstrictor discovered to date. The final step of posttranslational processing of this peptide is the conversion of its precursor by endothelin-converting enzyme-1 (ECE-1), a metalloprotease which displays high amino acid sequence identity with neutral endopeptidase 24.11 (NEP) especially at the catalytic center. A series of potent and selective arylacetylene-containing ECE-1 inhibitors have been prepared. (S, S)-3-Cyclohexyl-2-[[5-(2, 4-difluorophenyl)-2-[(phosphonomethyl)amino]pent-4-ynoyl]amino] propio nic acid (47), an arylacetylene amino phosphonate dipeptide, was found to inhibit ECE-1 and NEP with IC50 values of 14 nM and 2 microM, respectively. Similarly, (S)-[[1-[(2-biphenyl-4-ylethyl)carbamoyl]-4-(2-fluorophenyl)but-3- yny l]amino]methyl]phosphonic acid (56), an arylacetylene amino phosphonate amide, had IC50's of 33 nM and 6.5 microM for ECE-1 and NEP, respectively. Slight modification of the aryl moiety was found to have dramatic effects on ECE-1/NEP selectivity. The 2-fluoro dipeptide analogue, (S, S)-2-[[5-(2-fluorophenyl)-2-[(phosphonomethyl)amino]pent-4-ynoyl]+ ++amin o]-4-methylpentanoic acid (40), showed a 72-fold selectivity for ECE-1 over NEP, while the 3-fluoro dipeptide analogue, (S, S)-2-[[5-(3-fluorophenyl)-2-[(phosphonomethyl)amino]pent-4-ynoyl]+ ++amin o]-4-methylpentanoic acid (22), was equipotent for ECE-1 and NEP. Several of these inhibitors were shown to be potent in blocking ET-1 production in vivo as demonstrated by the big ET-1-induced pressor response in rats. These potent inhibitors are the most selective for ECE-1 reported to date and are envisaged to have a variety of therapeutic applications.     

Study and characterization of different developmental stages of Leishmania major

The complete developmental cycle of Leishmania major in axenic culture was achieved by simply changing the temperature whether sudden from 22 degrees C to 37 degrees C or stepwise 22 degrees C, 29 degrees C and then 37 degrees C. The morphology by light microscopy, GPI isoenzyme pattern and PCR amplification of minicircle of kinetoplast DNA of the different stages were studied. The amastigotes obtained from the foot pads of mice were compared to those obtained from axenic culture. The GPI isoenzyme pattern and the PCR amplification products showed distinct differences between the promastigotes and the amastigotes. The amastigotes of the two sources also showed differences after temperature changes.