Endothelial and Smooth Muscle Cells from Abdominal Aortic Aneurysm Have Increased Oxidative Stress and Telomere Attrition

Background: Abdominal aortic aneurysm (AAA) is a complex multi-factorial disease with life-threatening complications. AAA is typically asymptomatic and its rupture is associated with high mortality rate. Both environmental and genetic risk factors are involved in AAA pathogenesis. Aim of this study was to investigate telomere length (TL) and oxidative DNA damage in paired blood lymphocytes, aortic endothelial cells (EC), vascular smooth muscle cells (VSMC), and epidermal cells from patients with AAA in comparison with matched controls. Methods: TL was assessed using a modification of quantitative (Q)-FISH in combination with immunofluorescence for CD31 or α-smooth muscle actin to detect EC and VSMC, respectively. Oxidative DNA damage was investigated by immunofluorescence staining for 7, 8-dihydro-8-oxo-2′-deoxyguanosine (8-oxo-dG). Results and Conclusions: Telomeres were found to be significantly shortened in EC, VSMC, keratinocytes and blood lymphocytes from AAA patients compared to matched controls. 8-oxo-dG immunoreactivity, indicative of oxidative DNA damage, was detected at higher levels in all of the above cell types from AAA patients compared to matched controls. Increased DNA double strand breaks were detected in AAA patients vs controls by nuclear staining for γ-H2AX histone. There was statistically significant inverse correlation between TL and accumulation of oxidative DNA damage in blood lymphocytes from AAA patients. This study shows for the first time that EC and VSMC from AAA have shortened telomeres and oxidative DNA damage. Similar findings were obtained with circulating lymphocytes and keratinocytes, indicating the systemic nature of the disease. Potential translational implications of these findings are discussed. © 2012 Cafueri et al

Iron Behaving Badly: Inappropriate Iron Chelation as a Major Contributor to the Aetiology of Vascular and Other Progressive Inflammatory and Degenerative Diseases

The production of peroxide and superoxide is an inevitable consequence of aerobic metabolism, and while these particular "reactive oxygen species" (ROSs) can exhibit a number of biological effects, they are not of themselves excessively reactive and thus they are not especially damaging at physiological concentrations. However, their reactions with poorly liganded iron species can lead to the catalytic production of the very reactive and dangerous hydroxyl radical, which is exceptionally damaging, and a major cause of chronic inflammation. We review the considerable and wide-ranging evidence for the involvement of this combination of (su)peroxide and poorly liganded iron in a large number of physiological and indeed pathological processes and inflammatory disorders, especially those involving the progressive degradation of cellular and organismal performance. These diseases share a great many similarities and thus might be considered to have a common cause (i.e. iron-catalysed free radical and especially hydroxyl radical generation). The studies reviewed include those focused on a series of cardiovascular, metabolic and neurological diseases, where iron can be found at the sites of plaques and lesions, as well as studies showing the significance of iron to aging and longevity. The effective chelation of iron by natural or synthetic ligands is thus of major physiological (and potentially therapeutic) importance. As systems properties, we need to recognise that physiological observables have multiple molecular causes, and studying them in isolation leads to inconsistent patterns of apparent causality when it is the simultaneous combination of multiple factors that is responsible. This explains, for instance, the decidedly mixed effects of antioxidants that have been observed, etc...Comment: 159 pages, including 9 Figs and 2184 reference

Evaluation of Functional Stability and Batch-to-Batch Reproducibility of a Castanea sativa Leaf Extract with Antioxidant Activity

A growing body of evidence suggests that free radicals are generated by UV irradiation being responsible for skin injury. In this regard, the topical use of formulations composed of plant extracts with antioxidant activity could represent a useful strategy for the prevention of photoaging and oxidative-stress-mediated diseases. The aim of this study was to assess the reproducibility of the extraction method and the functional stability of a Castanea sativa leaf extract in view of its application as topical antioxidant. Measurements of 1,1-diphenyl-2-picryl hydrazyl (DPPH) scavenging activity, total phenols (measured by the Folin Ciocalteu assay) and phenolic composition (high-performance liquid chromatography unit coupled to a UV detector) were carried out on three different batches. The influence of pH and temperature on the extract’s DPPH scavenging activity was assessed in aqueous and glyceric solutions (0.025% w/v) over a 3-month period. Minor differences were found between the three extract batches for all the evaluated parameters, and therefore the reproducibility of the extraction method can be inferred. pH presented a great influence in the extract functional stability. Major antioxidant activity decrease was found at pH 7.1, while lower changes were observed at pH 5. Glyceric solutions were stable throughout the test period. At 40°C and pH 5, a marked decrease of activity was observed. Again, glyceric solutions were the most stable, even at 40°C. Proper selection of pH and solvent is mandatory to ensure the stability of the studied extract after being incorporated in semisolid forms. In view of these results, glycerine is proposed as the best vehicle for topical formulations incorporating C. sativa leaf extract, which should have a pH around 5