Sildenafil inhibits the ROS production by xanthine oxidase

Xanthine oxidase (XO) catalyzes the hydroxylation of a wide variety of substrates, including purines, pirimidines, pterins d aldehydes, to acids1. At relatively high oxygen pressure, it generates reactive oxygen species (ROS) as superoxides and hydroxyl radicals. The XO, detected in endothelial and epithelial cell outer surface, has been involved in ischemia/reperfusion injury1,2. Furthermore, XO-ROS production has been implicated in chronic hearth failure, inflammatory diseases, LDL oxidation, atherosclerosis, hypertension, cancer, aging1. Allopurinol, a hypoxanthine analogue developed as xanthine oxidase inhibitor 30 years ago, and oxypurinol, its oxidation product, have proved to be effe…

Proposed reaction mechanism of xanthine dehydrogenase in the oxidation of retinol to retinoic acid in mammary epithelial cells

Low levels of xanthine dehydrogenase and cellular retinol binding protein in tumor mammary epithelial cells are responsible of the retinoic acid deficit

Hypoxanthine and purine compounds in plasma from patients with multiple sclerosis and psoriasis

Development and validation of a fast and sensitive chromatographic assay for all-trans-retinol and tocopherols in human serum/plasma using liquid-liquid extraction.

A sensitive HPLC assay for all-trans-retinol, a-tocopherol, and g-tocopherols in human serum and plasma is reported. Sample preparation is performed in one step and involves precipitation of proteins and extraction of lipids with two volumes of an ethanol–chloroform mixture (3:1, v/v) without I.S. addition. After removal of the precipitated protein, 20 ml aliquots of the supernatant (equivalent to 6.7 ml of serum or plasma) were injected into the HPLC system and analyzed using fluorometric detection. RP-HPLC was performed using a C18 S3 ODS2 column with a methanol–water step gradient (97:3 to 100) at 1.0 ml/min. The quantification limit expressed as nanograms of analyte per milliliter of se…

Micellar electrokinetic capillary chromatography of polyamines and monoacetylpolyamines.

A selective procedure for qualitative and quantitative analysis of ten polyamines by micellar electrokinetic chromatography (MEKC) was developed. Benzoylated polyamines and acetylpolyamines in micellar phase of SDS (10 mM) were separated at 258C by 20 mM borate buffer pH 8.5, containing 8% ethanol, with an applied voltage of 25 kV (5 mA) and then detected at 198 nm. The experimental factors and operational parameters were optimized by performing analysis at different surfactant concentrations, pH, voltage and temperature with and without ethanol. The repeatibility of migration times and peak heights is a peculiarity of the method here described.

Polyamines and ripening of photoreceptor outer-segment in chicken embryo.

Polyamines and their related monoacetyl derivatives were studied in rod outer segment (ROS) and cone outer segment (COS) of photoreceptor cells from chick embryo retina during eye development (7th-18th days). Putrescine was found to be necessary, in the second phase of retinogenesis, to sustain both ROS and COS differentiation and, after acetylation, gamma-aminobutyric acid synthesis. On the Other hand, spermidine and even more spermine intervene in the third phase of development when photoreceptors mature. Moreover, the presence of Nl-acetylspermidine already at the 7th day indicates that in the outer segment of photoreceptor cells too, as in the whole retina, putrescine synthesis comes ab…

Xanthine dehydrogenase processes retinol to retinoic acid in human thyroid epitelial cells

The pattern of polyamine and GABA in chicken embryo retina photoreceptor membranes

Quantification and metabolic aspects of trans fatty acids

SCOPE To inquire with certainty of the analytical data on the effects of fat acids. INTRODUCTION The consumption of trans isomers of unsaturated fatty acids has been associated with untoward metabolic effects. Several clinical investigations demonstrated that trans fatty acids increase plasma LDL-cholesterol and lipoprotein (a) and reduce HDL-cholesterol concentrations. These alterations of plasma lipid profiles indicate an atherogenic effect of trans fatty acids. Moreover, in infant and children, trans fatty acids in blood plasma may impair human growth. METHODOLOGY We have put to point one methodology analytics in order to dose trans fatty acids in the blood. Such methodology analytics pr…

The tumor breast cells MCF-7 can synthesize retinoic acid startinf from retinaldehyde

The acetylation of spermidine catalyzed by P/CAF regulates its acetyltransferase activity versus histones

Transcriptional regulation in eukaryotes occurs within a chromatin setting and is strongly inhibited by nucleosomal barriers imposed by histone proteins. Among the well-known covalent modifications of chromatin, the reversible acetylation of specific lysine residues of histones, catalyzed by several acetyltransferase and deacetylases, has been linked to many biological processes including transcriptional regulation. Indeed, many transcriptional coactivators possess histone acetyltransferase (HAT) activity (1). Functional interactions between HAT and polyamines have been previously reported. In particular, it has been shown that polyamines facilitate oligomerization of nucleosomal arrays in …

Interactions between polyamines, acetylpolyamines and PCAF during histone acetylation.

Polyamines are involved in several gene regulatory functions, as the transcription process. In order to clarify their effect, in an initial “in vitro” biochemical study, we considered their influence on the nuclear PCAF acetyltransferase (PCAF-HAT). In a model system, using both commercial histones and laboratory histone preparations from mammary epithelial cells, we evidenced the inhibitory effect played by 40-180 uM spermidine (Spd) versus PCAF, assayed on 250 nM histone and 40 uM acetyl-CoA. At the same concentrations N8-acetylspermidine (N8-Ac.Spd), the acetylated form of spermidine located in the nucleus, exerts a significant activating effect on the PCAF-HAT. In addition, N8-Ac.Spd se…

Estrogen Inhibits Retinoic Acid Biosynthesis In Tumor Breast Epithelial Cell Lines

Protective Effect of Sildenafil against Estradiol-induced ROS production

Several reports suggest that xanthine dehydrogenase (XDH) and its oxidase form (XO) play an important role in various forms of ischemic and vascular injuries. Recently we have demonstrated that 17β-estradiol (E2) induces a significant decrease of the expression and activity of XDH and its conversion to XO in human mammary epithelial cells. E2 is known to induce upregulation of eNOS gene expression in aortic endothelial cells. In light of the ability of XO-derived O2•¯ to combine with •NO to yield ONOO¯, and considering that ONOO¯ converts XDH to XO, it is important to protect tissues against the XO increased activity and ROS increased production, that would in turn react with •NO to augment…

Low levels of both xanthine dehydrogenase and cellular retinol binding protein are responsible of the retinoic acid deficiency in malignant human mammary epithelial cells (abstract)