Confirmation of oxidative stress and fatty acid disturbances in two further Papillon-Lefèvre syndrome families with identification of a new mutation.

BACKGROUND: We have previously reported oxidative and fatty acids disturbances in one Papillon-Lefèvre syndrome (PLS) family. This Mendelian condition characterized by palmar plantar keratosis and severe aggressive periodontitis, is caused by mutations in the cathepsin C (CTSC) gene. In this study, we have analysed two further unrelated PLS families to confirm this association. METHODS: Mutations were identified by direct sequencing of CTSC. Biochemical analyses were performed in probands and their relatives in order to determine plasma levels of vitamin E, CoQ10 , lipid hydroperoxides (HP) and fatty acid patterns. RESULTS: Pathogenic CTSC mutations were identified in both families including a new mutation (c504C>G). Both probands showed low levels of vitamin E and CoQ10 , and high levels of lipid HP, and also very low levels of docohexaenoic acid. CONCLUSIONS: The previously reported oxidative and fatty acids disturbances were confirmed as a feature of this condition in two further families. There are low levels of antioxidant markers and high levels of oxidative markers, in addition of low levels of some anti-inflammatory fatty acids in persons suffering PLS and some of their relatives.

Periodontitis is associated with altered plasma fatty acids and cardiovascular risk markers.

BACKGROUND AND AIMS: In periodontitis it has been found that some perturbation exists in lipid biomarkers, such as increased serum total cholesterol and low-density lipoprotein cholesterol. Nevertheless, the relationship between fatty acids and periodontitis has been demonstrated only in a few studies and remains controversial. The aim of this investigation was to explore the effects of periodontitis on a cluster of traditional and novel cardiovascular risk factors such as plasma-lipids profile, types of plasma fatty acids, adhesion molecules and systemic inflammatory markers. METHODS AND RESULTS: At a university dental school, 56 patients all over 35 years old were enrolled and invited to participate in the study. Total plasma fatty acids, saturated, n-6 polyunsaturated and monounsaturated fatty acids, peroxidability index, soluble VCAM, TNF-alpha, cholesterol, triacylglycerols, and VLDL-c were significantly higher in the periodontitis group compared to the non-periodontitis group. CONCLUSIONS: This close association found between plasma triacylglycerols, LDL-c, saturated fatty acids, polyunsaturated fatty acids, total amount of fatty acids and coenzyme Q(10) with some periodontal data such as periodontal probing depth, recession of the gingival margin and clinical attachment level (Pearson correlation between 0.3 and 0.6), leads to the conclusion that there is an inter-relationship between periodontitis, plasma fatty acids profile and the increase in metabolic risk factors for cardiovascular diseases.

Olive oil and health: summary of the II international conference on olive oil and health consensus report, Jaén and Córdoba (Spain) 2008.

Olive oil (OO) is the most representative food of the traditional Mediterranean Diet (MedDiet). Increasing evidence suggests that monounsaturated fatty acids (MUFA) as a nutrient, OO as a food, and the MedDiet as a food pattern are associated with a decreased risk of cardiovascular disease, obesity, metabolic syndrome, type 2 diabetes and hypertension. A MedDiet rich in OO and OO per se has been shown to improve cardiovascular risk factors, such as lipid profiles, blood pressure, postprandial hyperlipidemia, endothelial dysfunction, oxidative stress, and antithrombotic profiles. Some of these beneficial effects can be attributed to the OO minor components. Therefore, the definition of the MedDiet should include OO. Phenolic compounds in OO have shown antioxidant and anti-inflammatory properties, prevent lipoperoxidation, induce favorable changes of lipid profile, improve endothelial function, and disclose antithrombotic properties. Observational studies from Mediterranean cohorts have suggested that dietary MUFA may be protective against age-related cognitive decline and Alzheimer's disease. Recent studies consistently support the concept that the OO-rich MedDiet is compatible with healthier aging and increased longevity. In countries where the population adheres to the MedDiet, such as Spain, Greece and Italy, and OO is the principal source of fat, rates of cancer incidence are lower than in northern European countries. Experimental and human cellular studies have provided new evidence on the potential protective effect of OO on cancer. Furthermore, results of case-control and cohort studies suggest that MUFA intake including OO is associated with a reduction in cancer risk (mainly breast, colorectal and prostate cancers).

Breeding strawberry (Fragaria X ananassa Duch) to increase fruit nutritional quality.

In the last few years the nutritional value of fruit has been widely studied and requested by the consumers, especially for protection against cardiovascular events, cancer and others as well as for the general health benefits it can provide. These benefits can be ascribed mainly to the Total Antioxidant Capacity (TAC) of fruit which is determined by a complex combination of bioactive compounds, generally related to specific characteristics of the genotype and of its interaction with the environment. Strawberry breeding programs are currently used to acquire new varieties improved for specific agronomic (Yield and Size), qualitative (Firmness, Sugars Content and Acidity) and sensorial (Colour and Aroma) characteristics, all combined to increased disease resistance and plant adaptability. Nowadays, besides to all these parameters is necessary to look for the specific bioactive components well known for their effect on human health. This aspect is now highly requested by the consumer. By testing different commercial strawberry varieties, a wide difference among their production and fruit quality, mostly related to their adaptability to the local evaluation conditions, is generally detected. Regarding their nutritional values, only few varieties nowadays differ for their higher phenol content and total antioxidant capacity. By taking into account this knowledge different breeding programs specifically addressed to increase fruit nutritional value, have been started. A first approach is based on the use of inter-specific crosses within wild and cultivated genotypes, so to increase variability for these characters, while the second by considering the intra-specific crosses within cultivars already known for their agronomic behavior and good quality performance. Herewith, we report on the major aspects related to breeding program aimed to increase strawberry fruit nutritional quality and what we can expect for the improvement of consumer acceptability of this fruit.

Oxidative stress markers in oral lichen planus.

Oral lichen planus (OLP) is a relatively common disorder whose cause is still unknown. Oral cancer is preceded in most cases by pre malignant lesions-leukoplasia, submucous fibrosis and lichen planus. Free radicals and reactive oxygen species play important roles in both pathogenesis of lichen planus and carcinogenesis. Thus monitoring systemic and saliva compounds important for the antioxidant defence (oxidative balance) could be important for the clinician's treatment strategy. Thorough medical management and early active treatment are necessary to improve symptoms and might also be a relevant prevention strategy from squamous cell carcinoma risk, although data to fully support this statement still need investigation. The principal aim of this study was to determine the systemic uric acid, GGT, and albumin levels as well as the levels of uric acid and albumin in 20 patients diagnosed with lichen planus and 20 controls. Extensive medline search failed to reveal any study of this type. Our results showed a significant decrease of saliva (p < 0.005) uric acid and an increase in serum gamma glutamyl transpherase (GGT) (p < 0.01) as well as in the total antioxidant capacity of saliva in patient group with respect to the control one. The preliminary conclusion of our study is that uric acid, the most important salivary antioxidant and GGT could be considered in the future as useful markers of oxidative stress for elaboration of treatment strategy and monitoring.

Nifedipine and cyclosporin affect fibroblast calcium and gingiva.

It has been stated that cyclosporin and nifedipine produce gingival overgrowth. However, the specific pathogenic mechanism remains uncertain. We used an experimental rat model to test the hypothesis that changes in collagen metabolism and numbers of gingival blood vessels are not mediated by intracellular calcium concentration (ratiometric Fura-2 AM measurement) in gingival fibroblasts. In the cyclosporin group, both width (364.2 +/- 67.5 mum) and microvessel density (number of vessels/mm(2), stained with anti-CD34 antibody) (41.6 +/- 5.1) of gingiva were statistically different when compared with those in the control group (width = 184.3 +/- 35.2 mum, microvessel density = 19.6 +/- 2.4). The nifedipine group showed the highest content of collagen (proportion of total stroma occupied by collagen, stained with Picro-Mallory) (nifedipine group = 66.3 +/- 9.4, cyclosporin group = 55.2 +/- 7.9, control group = 30.1 +/- 10.2). Freshly cultured fibroblasts from the cyclosporin group exhibited higher ratiometric values of fluorescence than did both the control and nifedipine groups (p = 0.03). Our results support the hypothesis that changes in gingival collagen metabolism are not mediated by calcium intracellular oscillations.

The relationship between insulin resistance and periodontitis is not affected by Mediterranean diet in a Spanish population.

OBJETIVE: To examine the insulin resistance measured by surrogate indices in subjects with and without periodontitis and to find out any correlation among dietary intake with insulin resistance. DESIGN: Fifty-five patients were recruited to participate in this cross-sectional study. Insulin resistance measured by the homoeostasis model assessment (HOMA-IR) and the quantitative insulin sensitivity check index moreover glycaemia, creatinine, uric acid, high density lipoproteins, low density lipoproteins, very low density lipoproteins and triglycerides among others. True periodontal disease was elucidated through the examination of probing pocket depth, clinical attachment level, recession of the gingival margin and gingival bleeding. The statistical analyses used were the student's T-test for independent variables, Kolmogorov-Smirnov if variations were homogeneous; if not, the Mann-Whitney U Test was applied instead. Correlations between variables were assessed using Pearson's correlation coefficients. True periodontal disease was confirmed through the greater values of probing pocket depth, clinical attachment level, gingival margin and gingival bleeding in the periodontitis group in comparison with non-periodontitis group. RESULTS: Insulin resistance was evidenced by the greater values of HOMA-IR as well as by the lower quantitative insulin sensitivity check index values in the periodontitis group. Fasting insulin, glucose, uric acid, creatinine, low density lipoproteins, triglycerides and very low density lipoprotein levels were significant higher in periodontitis group. Pearson's correlations did not show any association among diet data and insulin resistance parameters in periodontitis patients. CONCLUSION: A putative systemic relationship between insulin resistance and periodontitis exists but it does not seem conceivable any effect of diet over such relationship.

In vitro antioxidant activities of antioxidant-enriched toothpastes.

Several forms of periodontal diseases (PD) are often associated with modified phagocytosing leukocytes and contemporary free radical production. Host antioxidant defenses could benefit from toothpastes used as adjuncts to counteract plaque-associated bacteria. The aim of the present study was to determine possible antioxidant activity (AA) of 12 differently antioxidant-enriched toothpastes, regardless of their efficacy as antimicrobial agents. Toothpastes were enriched alternatively with sodium ascorbyl phosphate, alpha-tocopherol acetate, pycnogenol, allantoin and methyl salycilate or a mixture of these. AA was tested in a cell-free system with a ABTS-decolorization assay improved by means of a flow injection analysis device. Comet assay, using NCTC 2544 keratinocytes, was performed to test if it was possible to identify any protection against in vitro DNA fragmentation provoked by a challenge with H(2)O(2) in cultures pre-incubated with toothpaste extracts. Only toothpastes containing sodium ascorbyl phosphate displayed clear AA with I(50) values ranging between 50 and 80 mg of toothpaste/ml water. COMET analysis of cells challenged with H(2)O(2) in presence of toothpaste extracts revealed a limited protection exerted by sodium ascorbyl phosphate. The results described herein indicate that toothpastes containing sodium ascorbyl phosphate possess AA. All the data were obtained in systems in vitro and the demonstration of in vivo AA is desirable. These findings could be useful in the treatment and maintenance of some forms of PD and should be considered when arranging new toothpaste formulations.

Interaction between reactive oxygen species and coenzyme Q10 in an aprotic medium: a cyclic voltammetry study.

The involvement of coenzyme Q (CoQ) as an antioxidant agent in several oxidative processes both in vitro and in vivo is nowadays pointed out by several biochemical and clinical studies, but the chemical mechanisms of this action are not yet unequivocally established. Electrochemistry provides very useful techniques for the analysis of the kinetics and thermodynamics, and mechanisms of chemical phenomena involving electron transfers, e.g. in the case of radical reactions. In the present study we used cyclic voltammetry to investigate the interactions between oxygen radicals and ubiquinone in aprotic medium, a condition similar to that existing in the biological membranes. The results obtained showed that ubiquinone is more easily reduced than oxygen, ruling out the possibility of an electron transfer from semiquinone to oxygen to produce superoxide radicals. On the contrary, it was demonstrated that fully reduced quinone is able to scavenge the superoxide radical, by reduction to peroxide ion, lowering actually the oxidative potential in the medium.

Metabolic and antioxidant markers in the plasma of sportsmen from a Mediterranean town performing non-agonistic activity.

It has been widely indicated that several pathological conditions depend upon concomitant risk factors rather than a unique one and that also the putative protective factors do not act alone. For these reasons it could be useful to consider subjects that present sufficiently homogeneous lifestyles (i.e. nutrition and physical activity). We carried out an investigation in a free-living community in order to clarify the possible correlations and differences among plasma metabolic and antioxidant markers in non-agonistic athletes. When subjects were divided in two main groups according to age (35-44 and 45-54 years) without considering the activity they performed, Duncan's analysis of variance revealed that they showed similar characteristics and only triglyceride levels were different. A clear negative correlation was found between vitamin E and VO2max in both age groups, a negative correlation was also found between CoQ10 and VO2max in the younger subjects and finally CoQ10 and vitamin E were also positively correlated in this first group. It appears, therefore, that people with a higher aerobic capacity have lower circulating levels of antioxidants.

Exogenous CoQ10 supplementation prevents plasma ubiquinone reduction induced by HMG-CoA reductase inhibitors.

The biosynthetic pathway of the CoQ polyisoprenoid side chain, starting from acetyl-CoA and proceeding through mevalonate and isopentenylpyrophosphate, is the same as that of cholesterol. We performed this study to evaluate whether vastatins (hypocholesterolemic drugs that inhibit HMG-CoA reductase) modify blood levels of ubiquinone. Thirty-four unrelated outpatients with hypercholesterolemia (IIa phenotype) were treated with 20 mg of simvastatin for a 6-month period (group S) or with 20 mg of simvastatin plus 100 mg CoQ10 (group US). The following parameters were evaluated at time 0, 45, 90, 135 and 180 days: total plasma cholesterol (TC), HDL-cholesterol, LDL-cholesterol (LDL-C), triglycerides (TG), apo A1, apo B and CoQ10 in plasma and platelets. In the S group, there was a marked decrease in TC and LDL-C (from 290.3 mg/dl to 228.7 mg/dl for TC and from 228.7 mg/dl to 167.6 mg/dl for LDL-C) and in plasma CoQ10 levels from 1.08 mg/dl to 0.80 mg/dl. In contrast, in the US group we observed a significant increase of CoQ10 in plasma (from 1.20 to 1.48 mg/dl) while the hypocholesterolemic effect was similar to that observed in the S group. Platelet CoQ10 also decreased in the S group (from 104 to 90 ng/mg) and increased in the US group (from 95 to 145 ng/mg). This study demonstrates that simvastatin lowers both LDL-C and apo B plasma levels together with the plasma and platelet levels of CoQ10, and that CoQ10 therapy prevents both plasma and platelet CoQ10 decrease, without affecting the cholesterol lowering effect of simvastatin.

Coenzyme Q content depends upon oxidative stress and dietary fat unsaturation.

The presence of Coenzyme Q (CoQ) in food, its role in cellular bioenergetics and antioxidant protection and the key role played by dietary fatty acids on membrane structure support the interest for a wide research concerning the relationship between dietary fats, CoQ content and biochemical behaviour. Several models of peroxidative stress 'in vivo' have been extensively investigated in our laboratory, with particular regards to the influence of dietary fat upon mitochondrial CoQ levels. First studies showed that the unsaturation degree of dietary fat leads to different CoQ9 and CoQ10 mitochondrial contents. The highest levels were found using polyunsaturated fat. A significant CoQ9 decrease after adriamycin peroxidative induction was found when dietary fat was polyunsaturated; on the contrary, a light increase was found in the case of monounsaturated fat. Another example of oxidative stress is that produced by food frying. The results obtained were in some cases similar to those of the previous experimental design: in fact monounsaturated dietary fats increased CoQ mitochondrial contents, whereas the polyunsaturated ones decreased CoQ levels. Finally, the combined effect of physical exercise and dietary fats on tissue and plasma CoQ levels has been studied. CoQ levels did not change during aerobic performances when dietary fat was monounsaturated whereas light increases were detected in the case of polyunsaturated fats. On the contrary, in anaerobic conditions, CoQ levels clearly increased with monounsaturated fats and no alterations were found in the case of polyunsaturated ones.

Metabolic implications of coenzyme Q10 in red blood cells and plasma lipoproteins.

Plasma coenzyme Q10 (CoQ10) is currently assayed in our laboratory for its well-known diagnostic meaning; in fact plasma CoQ10 levels are inversely related to metabolic demand. Definite levels of CoQ10 are also found in white and red blood cell components, as well as in platelets. Plasma and erythrocyte CoQ10 has a well assessed antioxidant role, which was demonstrated through a series of experiments. Erythrocytes previously enriched with exogenous CoQ10 were found more resistant to a hemolysis induced by a free radical initiator. Several enzymatic activities of erythrocyte ghosts were also protected by different side chain CoQ homologues, both when reduced and, although at a lesser extent, in the oxidized state. CoQ was not effective in preventing metal-catalyzed oxidation of erythrocyte membrane enzymes, and this effect is likely to be due to lack of interaction of CoQ with the metal target. Moreover CoQ was able to protect isolated enzymes and erythrocyte membrane bound enzymes from the inactivating effect of free radicals generated by water sonolysis or radiolysis. As far as plasma lipoproteins are concerned it is well known that LDL isolated from healthy volunteers supplemented with CoQ10 are more resistant to peroxidation induced by an azoinitiator. We started to systematically investigate CoQ10 and vitamin E levels in isolated human LDL and HDL. Both CoQ10 and vitamin E concentrations, referred to protein, were found higher in LDL than in HDL. Susceptibility to exogenously applied peroxidation did not correlate with the endogeneous content of the two antioxidants, possibly on the basis of different lipid content of these lipoproteins.

Peroxidative extent and coenzyme Q levels in the rat: influence of physical training and dietary fats.

Sport practice is widely recognized as capable of producing peroxidative damages, even of severe intensity. Dietary manipulations can also modify membrane susceptibility to peroxidation. In previous experiments we found that, while dietary virgin olive oil successfully protects mitochondrial and microsomal membranes from endogenous, xenobiotics-induced peroxidation, dietary polyunsaturated oils lead to increased peroxidative levels. In the latter conditions, cell machinery tries to counteract the structural and functional changes which have occurred, by modulating enzyme activities and concentrations, by increasing biosynthesis of coenzyme Q and by mobilizing cholesterol. In the present study we hypothesized that combining these two aspects could give useful information on the membrane response to peroxidation phenomena that daily occur throughout the lifespan. Rats fed different dietary oils as only fat source underwent a carefully designed training program and were killed at different times following acute or chronic exercise. Results show that peroxidation related to chronic training and to an acute bout of exercise sum up with peroxidative effects induced by dietary factors. The above mentioned phenomena occurred simultaneously with increased tissue levels of coenzyme Q, possibly triggered within a physiological reactive antioxidant strategy.

Lipid peroxidation and antioxidants in newborns.

The aim of this investigation was to elucidate a possible correlation between lipid peroxidation, antioxidant concentrations and erythrocyte membrane fluidity in plasma from newborns. Ten healthy newborns were recruited. Venous blood samples were collected at birth, and thereafter at 3 and 72 h postnatal age. The following parameters were assessed: hydroperoxides, Coenzyme Q10 and alpha-tocopherol both in plasma and in erythrocyte membranes, and fluorescence polarization (as a tool for assessing membrane fluidity). Hydroperoxides were shown to be high in erythrocyte membranes at birth and significantly decreased at 3 and 72 h after birth. In the erythrocyte membranes, coenzyme Q10 content showed an opposite behaviour with respect to the plasma compartment. Membrane fluidity appeared unchanged even in the presence of the above mentioned modifications.

Dietary restriction affects antioxidant levels in rat liver mitochondria during ageing.

Six experimental groups of young (7-month-old) and aged (24-32-month-old) rats, underwent different dietary manipulations (i.e. dietary restriction and/or a vitamin E-depleted diet), and their liver mitochondria were assayed for several antioxidants and peroxidation markers. Glutathione levels were affected both by age and dietary treatment. Coenzyme Q9 and C0Q10 showed the highest levels in the oldest rats where ageing, as well as other oxidative stresses, could induce ubiquinone biosynthesis.

Coenzyme Q homologs and vitamin E in synaptic and non-synaptic occipital cerebral cortex mitochondria in the ageing rat.

The coenzyme Q8 (CoQ8) and alpha-tocopherol contents of different mitochondrial fractions were investigated from occipital cerebral cortices of different ages. The highest CoQ8 and vitamin E concentrations were found in non-synaptic free mitochondria (FM) fractions. In several cases heavy mitochondria (HM) fractions displayed the lowest values. Occipital cerebral cortex mitochondria contained higher CoQ9 and lower CoQ10 amounts than those typical for other brain regions.

Kinetic alterations of cytochrome-c oxidase in cystic fibrosis.

We compared the kinetics of cytochrome-c oxidase (cytochrome-c:oxygen oxidoreductase, EC 1.9.3.1) in fibroblasts derived from normal and cystic fibrosis individuals. The Km of the enzyme for reduced cytochrome c was significantly increased in CF cells; the change, however, was observed only at temperatures above 25 degrees C. The Vmax values were comparable in both types of individuals.

Changes in mitochondrial and microsomal rat liver coenzyme Q9 and Q10 content induced by dietary fat and endogenous lipid peroxidation.

The influence of different kinds of dietary fat (8%) and of endogenous lipid peroxidation with regard to coenzyme Q9 (CoQ9) and coenzyme Q10 (CoQ10) concentrations in mitochondria and microsomes from rat liver has been investigated by means of an HPLC technique. Although the different diet fats used did not produce any effect on microsomes, it was possible to show that each experimental diet differently influenced the mitochondrial levels of CoQ9 and CoQ10. The highest mitochondrial CoQ content was found in case of a diet supplemented with corn oil. An endogenous oxidative stress induced by adriamycin was able to produce a sharp decrease in mitochondrial CoQ9 levels in the rats to which corn oil was administered. The results suggest that dietary fat ought to be considered when studies concerning CoQ mitochondrial levels are carried out.

Measurement of the lateral diffusion coefficients of ubiquinones in lipid vesicles by fluorescence quenching of 12-(9-anthroyl)stearate.

The lateral diffusion coefficients of some ubiquinone homologues have been measured in phospholipid vesicles exploiting the fluorescence quenching of the probe 12-(9-anthroyl)stearate by the quinones. Diffusion coefficients higher than 10(-6) cm2 X s-1 have been found at 25 degrees C, compatible with the localization of the ubiquinones in the low-viscosity midplane region of the bilayer.