Low Serum Paraoxonase; A Risk Factor for Coronary Artery Disease?

Objective: Serum Paraoxonase (PON) is a calcium dependent esterase which hydrolyses organophosphates. However, evidence exists for protective effect of PON against oxidative damage. PON has three genetic polymorphism (PON-1, PON-2, PON-3). The PON-1 has shown to provide resistance to development of atherosclerosis by protecting lipoprotein against oxidative modifications. Therefore, a study was performed to assess the relationship between Coronary Artery Disease (CAD) and PON-1 activity. Methods: A comparative cross-sectional study was performed with 33 angiogram positive patients and 48 healthy volunteers. Blood samples were collected after a 10 hours fast from controls and from patients who were awaiting angiogram test. Samples were analyzed for PON-1 activity using ELISA method. Data were analyzed using SPSS 15.0 statistical software. Results: The PON-1 concentration of angiogram positive cases and healthy volunteers (controls) were in the range 7.9 – 78.7 ug/ml and 14.9 – 395.2 ug/ml respectively. Angiogram positive cases showed significantly low (mean 36.7 ug/ml; P < 0.05) PON-1 activity when compared to controls (mean 111.5 ug/ml) suggesting that the low PON-1 activity has a potential to lead to an increased risk of CAD. Conclusion: The PON-1 activity may be a useful sensitive marker for assessment of CAD

Desaturase reactions complicate the use of norcarane as a mechanistic probe. Unraveling the mixture of twenty-plus products formed in enzyme-catalyzed oxidations of norcarane.

Norcarane, bicyclo[4.1.0]heptane, has been widely used as a mechanistic probe in studies of oxidations catalyzed by several iron-containing enzymes. We report here that, in addition to oxygenated products, norcarane is also oxidized by iron-containing enzymes in desaturase reactions that give 2-norcarene and 3-norcarene. Furthermore, secondary products from further oxidation reactions of the norcarenes are produced in yields that are comparable to those of the minor products from oxidation of the norcarane. We studied oxidations catalyzed by a representative spectrum of iron-containing enzymes including four cytochrome P450 enzymes, CYP2B1, CYPDelta2B4, CYPDelta2E1, and CYPDelta2E1 T303A, and three diiron enzymes, soluble methane monooxygenase (sMMO) from Methylococcus capsulatus (Bath), toluene monooxygenase (ToMO) from Pseudomonas stutzeri OX1, and phenol hydroxylase (PH) from Pseudomonas stutzeri OX1. 2-Norcarene and 3-norcarene and their oxidation products were found in all reaction mixtures, accounting for up to half of the oxidation products in some cases. In total, more than 20 oxidation products were identified from the enzyme-catalyzed reactions of norcarane. The putative radical-derived product from the oxidation of norcarane, 3-hydroxymethylcyclohexene (21), and the putative cation-derived product from the oxidation of norcarane, cyclohept-3-enol (22), coelute with other oxidation products on low-polarity GC columns. The yields of product 21 found in this study are smaller than those previously reported for the same or similar enzymes in studies where the products from norcarene oxidations were ignored, and therefore, the limiting values for lifetimes of radical intermediates produced in the enzyme-catalyzed oxidation reactions are shorter than those previously reported

Products from enzyme-catalyzed oxidations of norcarenes.

Recent studies revealed that norcarane (bicyclo[4.1.0]heptane) is oxidized to 2-norcarene (bicyclo[4.1.0]-hept-2-ene) and 3-norcarene (bicyclo[4.1.0]hept-3-ene) by iron-containing enzymes and that secondary oxidation products from the norcarenes complicate mechanistic probe studies employing norcarane as the substrate (Newcomb, M.; Chandrasena, R. E. P.; Lansakara-P., D. S. P.; Kim, H.-Y.; Lippard, S. J.; Beauvais, L. G.; Murray, L. J.; Izzo, V.; Hollenberg, P. F.; Coon, M. J. J. Org. Chem. 2007, 72, 1121-1127). In the present work, the product profiles from the oxidations of 2-norcarene and 3-norcarene by several enzymes were determined. Most of the products were identified by GC and GC-mass spectral comparison to authentic samples produced independently; in some cases, stereochemical assignments were made or confirmed by 2D NMR analysis of the products. The enzymes studied in this work were four cytochrome P450 enzymes, CYP2B1, CYPDelta2E1, CYPDelta2E1 T303A, and CYPDelta2B4, and three diiron-containing enzymes, soluble methane monooxygenase (sMMO) from Methylococcus capsulatus (Bath), toluene monooxygenase (ToMO) from Pseudomonas stutzeri OX1, and phenol hydroxylase (PH) from Pseudomonas stutzeri OX1. The oxidation products from the norcarenes identified in this work are 2-norcaranone, 3-norcaranone, syn- and anti-2-norcarene oxide, syn- and anti-3-norcarene oxide, syn- and anti-4-hydroxy-2-norcarene, syn- and anti-2-hydroxy-3-norcarene, 2-oxo-3-norcarene, 4-oxo-2-norcarene, and cyclohepta-3,5-dienol. Two additional, unidentified oxidation products were observed in low yields in the oxidations. In matched oxidations, 3-norcarene was a better substrate than 2-norcarene in terms of turnover by factors of 1.5-15 for the enzymes studied here. The oxidation products found in enzyme-catalyzed oxidations of the norcarenes are useful for understanding the complex product mixtures obtained in norcarane oxidations

Effects of ethephon pre-treatment on the response of Elymus repens to fluazifop-butyl

Growth inhibition of pot-grown Elymus repens (L.) Gould by fluazifop-butyl was occasionally enhanced by pre-treatment with ethephon. Studies with C-14-labelled fluazifop-butyl showed that ethephon increased the accumulation of C-14 in basal buds and proximal parts of the rhizome system. This potential modification in the distribution of fluazifop-butyl within E. repens rhizomes may have reduced regeneration and recovery of the E. repens, particularly from the basal buds, although there was no increase in the proportion of C-14 translocated into the rhizome. Also, ethephon not only activated the sinks in the rhizome but in the foliage of E. repens as well, and could initiate buds without sinks. The effects of ethephon on fluazifop-butyl phototoxicity were irregular in their occurrence and degree, and possible reasons for this are discussed