Paola Astolfi

Antioxidant and antimicrobial capacity of several monofloral Cuban honeys and their correlation with color, polyphenol content and other chemical compounds.

Several monofloral Cuban honeys were analyzed to determine their total phenolic, flavonoid, ascorbic acid, amino acid, protein and carotenoid contents as well as their radical-scavenging activity and antimicrobial capacities. The total phenolic, flavonoid and carotenoid contents varied considerably, and the highest values were obtained for Linen vine (Govania polygama (Jack) Urb) honey, which is classified as an amber honey. The highest amino acid content was found in Morning glory (Ipomoea triloba L.) while Liven vine had the highest protein content. Similarly Linen vine honey had the highest antioxidant activity while the lowest was found in Christmas vine (Turbina corymbosa (L.) Raf). Ascorbic acid was absent. Hydroxyl radical formation was studied by EPR and spin trapping, and it was found in all honeys tested. The antimicrobial activity was screened using two Gram-positive and Gram-negative bacteria. S. aureus was the most sensitive microorganism while Pseudomonas aeruginosa presented higher minimum active dilution values. Bacillus subtilis and Escherichia coli were both moderately sensitive to honey antimicrobial activity. A correlation between radical-scavenging activity and total phenolic content was found. Correlation existed also between color vs phenolics content, vs flavonoid content or between phenolic vs flavonoid.

Sunscreens Cause Coral Bleaching by Promoting Viral Infections

Background Coral bleaching (i.e., the release of coral symbiotic zooxanthellae) has negative impacts on biodiversity and functioning of reef ecosystems and their production of goods and services. This increasing world-wide phenomenon is associated with temperature anomalies, high irradiance, pollution, and bacterial diseases. Recently, it has been demonstrated that personal care products, including sunscreens, have an impact on aquatic organisms similar to that of other contaminants. Objectives Our goal was to evaluate the potential impact of sunscreen ingredients on hard corals and their symbiotic algae. Methods In situ and laboratory experiments were conducted in several tropical regions (the Atlantic, Indian, and Pacific Oceans, and the Red Sea) by supplementing coral branches with aliquots of sunscreens and common ultraviolet filters contained in sunscreen formula. Zooxanthellae were checked for viral infection by epifluorescence and transmission electron microscopy analyses. Results Sunscreens cause the rapid and complete bleaching of hard corals, even at extremely low concentrations. The effect of sunscreens is due to organic ultraviolet filters, which are able to induce the lytic viral cycle in symbiotic zooxanthellae with latent infections. Conclusions We conclude that sunscreens, by promoting viral infection, potentially play an important role in coral bleaching in areas prone to high levels of recreational use by humans.

New mediators for the enzyme laccase: mechanistic features and selectivity in the oxidation of non-phenolic substrates

New mediators of laccase have been comparatively evaluated and ranked towards the benchmark aerobic oxidation of p-MeO-benzyl alcohol. The mechanism of oxidation of this non-phenolic substrate by each mediator, which is initially oxidised by laccase to the Medox form, has been assessed among three alternatives. The latter make the phenoloxidise laccase competent for the indirect oxidation of non-phenolic (and thus ‘unnatural’) substrates. Experimental characterisation of the mediators, by means of spectrophotometric, electrochemical and thermochemical survey, is reported. Clear-cut evidence for the formation of a benzyl radical intermediate in the oxidation of a particular benzyl alcohol with laccase and a N–OH mediator is attained by means of a trapping experiment. The selectivity of the laccase-catalysed oxidation of two competing lignin and polysaccharide model compounds has been assessed by using the highly proficient 4-MeO-HPI mediator, and found very high in favour of the former model. This evidence is in keeping with the operation of a radical hydrogen-abstraction process that efficiently cleaves the benzylic rather than the aliphatic C–H bond of the two models. Significant is the finding that catechol, i.e., a model of recurring phenolic structures in lignin, once oxidised to aryloxyl radical by laccase is capable to mediate a radical oxidation of non-phenolic compounds. This supports a fully-fledged role of laccase as a delignifying enzyme in nature by way of no other mediators than the very phenolic groups of lignin. Finally, an evaluation of the dissociation energy of the NO–H bond of HBT, which is not accessible experimentally, is provided by the use of a thermochemical cycle and theoretical calculations.

Phenolics from monofloral honeys protect human erythrocyte membranes against oxidative damage.

The aim of the present work was to analyze the phenolic extracts from two monofloral Cuban honeys for their in vitro total antioxidant capacity, phenolic compounds content and free radical scavenging activity. The phenolic extracts, rich in lipophilic compounds, were tested further for their ability to inhibit AAPH-induced oxidative damage (hemolysis, lipid peroxidation and cytosolic depletion of reduced glutathione and decrease of superoxide dismutase activity) in erythrocytes. Results indicate an important total antioxidant capacity measured by TEAC and ORAC assays, as well as a relevant radical scavenging activity performed by EPR. Moreover, 13 phenolic compounds were identified using HPLC-LC/MS with quercetin as the most abundant flavonoid. The results also show that both extracts were able to inhibit erythrocytes oxidative damage, and that this may likely be due to their incorporation into cell membranes and their ability to cross it and reach the cytosol. In fact, flavonoid uptake by erythrocytes was further confirmed by testing quercetin, which efficiently incorporated into erythrocytes. Overall, this study indicates that honey contains relevant antioxidant compounds responsible, at least in part, for its biological activity and that uptake of its flavonoids may provide defense and promote cell functions in erythrocytes.

Radical-scavenging Activity, Protective Effect Against Lipid Peroxidation and Mineral Contents of Monofloral Cuban Honeys

Several monofloral Cuban honeys were analyzed to determine their free radical-scavenging activity and from this the total antioxidant content was estimated. The protective effect against lipid peroxidation in an in vitro model of rat liver homogenates was evaluated and, lastly, the mineral content of the honeys, which can be related to the maintenance of intracellular oxidative balance, was determined. The scavenging capacities against hydroxyl and superoxide radicals were determined using the spin-trapping technique and the hypoxanthine/xanthine oxidase assay, respectively. Lipid peroxidation was evaluated through the production of TBARS and hydroperoxides. All honeys tested showed potential antioxidant activity with Linen vine displaying the highest scavenging capacity towards the DPPH, hydroxyl and superoxide radicals, while the least efficient was Christmas vine honey. Honeys also inhibited, in a concentration-dependent mode, lipid peroxidation in rat liver homogenates, with Linen vine resulting the best while the least effective was Christmas vine honey. The ability to scavenge free radicals and protect against lipid peroxidation may contribute to the ability of certain Cuban honeys to help in preventing/reducing some inflammatory diseases in which oxidative stress is involved. A total of eight minerals were identified and quantified as follows: cadmium, chromium, copper, nickel, iron, manganese, lead, and zinc. Minerals found in higher concentrations were iron, zinc and manganese.

Synthesis, structural and spectroscopic characterization and biomimetic properties of new copper, manganese, zinc complexes: identification of possible superoxide-dismutase mimics bearing hydroxyl radical generating/scavenging abilities.

A series of Cu(II), Zn(II) and Mn(II) coordination compounds has been synthesized by reaction of the corresponding metal salts and pyrazolyl-based ligands, i.e. the neutral 1-(2-(4-((2,2,2-tri(1H-pyrazol-1-yl)ethoxy)methyl)benzyloxy)-1,1-di(1H-pyrazol-1-yl)ethyl)-1H-pyrazole {p-C(6)H(4)[CH(2)OCH(2)C(pz)(3)](2), (L(1)), and the anionic hydridotris(3-phenyl-5-methylpyrazolyl)borate (L(2))(-), bis(pyrazolyl)acetate (L(3)) and bis(3,5-dimethylpyrazolyl)acetate (L(4))(-): the species [L(1)(CuCl(2))(2)] (1), [L(1)(Cu(OAc)(2))(2)] (2), [L(1)(Zn(OAc)(2))(2)] (3), [(CuCl(L(2))(Hpz(Ph,Me))] (4), [Mn(L(3))(2)].2H(2)O, (5), [ZnCl(L(3))(imH)].MeOH [CuCl(L(4))(imH)].2H(2)O (7) have been obtained (Hpz(Ph,Me)=3-phenyl-5-methylpyrazole, imH=imidazole). Complexes 1 and 4 have been structurally characterized, also using less conventional powder diffraction methods. The superoxide scavenging activity has been characterized by indirect assays including EPR analysis. All complexes exhibit superoxide scavenging activity with IC(50) in the microM range and they protect against the oxidative action of peroxynitrite in different ways. 1, 4 and 7 exert both an anti- and pro-oxidant effect depending on their concentration as evaluated by EPR and fluorescence methods. The pro-oxidative effects are absent in Zn(II) and Mn(II) complexes.

Strawberry consumption improves aging-associated impairments, mitochondrial biogenesis and functionality through the AMP-activated protein kinase signaling cascade

Dietary polyphenols have been recently proposed as activators of the AMP-activated protein kinase (AMPK) signaling pathway and this fact might explain the relationship between the consumption of polyphenol-rich foods and the slowdown of the progression of aging. In the present work, the effects of strawberry consumption were evaluated on biomarkers of oxidative damage and on aging-associated reductions in mitochondrial function and biogenesis for 8weeks in old rats. Strawberry supplementation increased antioxidant enzyme activities, mitochondrial biomass and functionality, and decreased intracellular ROS levels and biomarkers of protein, lipid and DNA damage (P<0.05). Furthermore, a significant (P<0.05) increase in the expression of the AMPK cascade genes, involved in mitochondrial biogenesis and antioxidant defences, was also detected after strawberry intake. These in vivo results were then verified in vitro on HepG2 cells, confirming the involvement of AMPK in the beneficial effects exerted by strawberry against aging progression.

Nitroxides and a nitroxide-based UV filter have the potential to photoprotect UVA-irradiated human skin fibroblasts against oxidative damage.

BACKGROUND Antioxidants are now being incorporated into sunscreens as additional topical measure for delaying the aging process and reducing photo-damage to skin induced by excessive UVA exposure. UVA radiation reaching the skin leads to the generation of ROS (reactive oxygen species) implicated in DNA damage and activation of matrix metalloproteinase-1 (MMP-1) responsible for collagen damage and photo-aging. Nitroxides are a class of compounds endowed with versatile antioxidant activity and recently, nitroxide-based UV filters in which a nitroxide moiety has been attached to the most popular UV filter present in sunscreens have been developed. OBJECTIVE This study explores the potential photo-protective effects of these compounds on ROS production and induction of MMP-1 in cultured human dermal fibroblasts exposed to UVA. For comparison, vitamin E was also tested. METHODS The effects were assessed by measuring intracellular ROS production using a ROS-index probe and MMP-1 mRNA expression levels using quantitative real-time PCR (qPCR). RESULTS Exposure of fibroblasts to 18J/cm(2) UVA lead to a two-fold increase in ROS production which was reduced to non-irradiated control levels in the presence of 50μM nitroxide compounds and vitamin E. Under the same conditions, a ten-fold increase in MMP-1 mRNA expression levels was observed 24h post-UVA treatment which was significantly reduced by all nitroxide compounds but not vitamin E. CONCLUSION The results of this study support the potential use of nitroxide compounds, including novel nitroxide-based UV filters, as a useful and alternative strategy for improving the efficacy of topical formulations against photo-aging and possibly photo-carcinogenesis.

Synthesis and application of a novel sunscreen-antioxidant.

Background information on the inefficacy of sunscreens to provide free radical protection in skin, despite their usefulness in preventing sunburn/erythema, prompted us to synthesize a compound which would display in the same molecule both UV-absorbing and antioxidant capacities. For this purpose, the UVB absorber, 2-ethylhexyl-4-methoxycinnamate (OMC) was combined with the piperidine nitroxide TEMPOL, which has antioxidant properties. The spectral properties of the new nitroxide-based sunscreen (MC–NO) as well as its efficacy to prevent photo-oxidative damage to lipids induced by UVA, natural sunlight and 4-tert-butyl-4-methoxydibenzoylmethane (BMDBM), a photo-unstable sunscreen which generates free radicals upon UV radiation, was studied. The results obtained demonstrate that MC–NO: (a) absorbs in the UVB region even after UVA irradiation; (b) acts as free radical scavenger as demonstrated by EPR experiments; (c) strongly reduces both UVA-, sunlight- and BMDBM-induced lipid peroxidation in liposomes, measured as reduced TBARS levels; and (d) has comparable antioxidant activity to that of commonly used vitamin E and BHT in skin care formulations. These results suggest that the use of the novel sunscreen-antioxidant or of other nitroxide-based sunscreens in formulations aimed at reducing photoinduced skin damage may be envisaged.

On the assessment of photostability of sunscreens exposed to UVA irradiation: from glass plates to pig/human skin, which is best?

Photostability of suncare products is a great area of interest since several sunscreens on the market are photounstable, and this is primarily a problem concerning the UVA region (320-400 nm). Here we report a comparative study on the photostability assessment of two commercial sunscreens with same SPF, spread onto glass plates or onto full thickness pig ear skin or human/pig SCE membranes, and exposed to 183 kJ/m(2) UVA. Absorbance spectra and lipid peroxidation (measured by TBARS production) were determined. The results indicate: (a) sunscreen performance consequent to UVA exposure is independent of whether it is spread onto a non-biological and chemically inert substrate such as glass, or on biological substrates such as skin/SCE membranes; (b) despite the same SPF, sunscreen performance and photostability can be very different; (c) the data on human SCE membranes are similar to those on pig SCE membranes, indicating the suitability of the latter as a model for human skin. However, since the results obtained using skin membranes, akin to the more realistic conditions of use in vivo, do not substantially differ from those obtained on glass plates, the method proposed here using the latter may be applied for rapid, inexpensive, efficacy screening of photostability of sunscreens. Photostability testing should be a mandatory requirement for safer sunscreen protection products, since the results clearly show that some are still far from perfect.