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Di Tomo P.,University of Chieti Pescara | Canali R.,National Research Institute for Food and Nutrition | Ciavardelli D.,University of Chieti Pescara | Ciavardelli D.,Kore University of Enna | And 7 more authors.
Molecular Nutrition and Food Research | Year: 2012

Scope: Cardiovascular disease (CVD) is associated with vascular oxidative imbalance and inflammation. Increased reactive oxygen species (ROS) generation is associated with a functional inactivation of nitric oxide (NO) due to the reaction with O - 2, leading to peroxynitrite (ONOO -) formation and subsequent reduction in the beneficial effect of vascular NO bioavailability. Carotenoids'-rich diets have been associated with decreased risk of CVD, but the underlying mechanism is still unknown. Methods and results: In human umbilical vein endothelial cells (HUVECs), both β-carotene (BC) or lycopene (Lyc) significantly affected tumor necrosis factor-α (TNF-α)-induced inflammation, being associated with a significant decrease in the generation of ROS (spectrofluorometry) and nitrotyrosine (an index of ONOO - formation, cytofluorimetry), an increased NO/cGMP (cyclic guanosine monophosphate) levels (EIA), and a down-regulation of NF-κB-dependent adhesion molecule expression (Western blot and EMSA) and monocyte-HUVEC interaction (adhesion assay). Our results indicate that BC or Lyc treatment reduce the inflammatory response in TNF-α-treated HUVECs. This is due to the redox balance protection and to the maintenance of NO bioavailability. Conclusion: Our observations provide background for a novel mechanism for carotenoids' anti-inflammatory activity in the vasculature and may contribute to a better understanding of the protective effects of carotenoid-rich diets against CVD risk. © 2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.


Comitato R.,National Research Institute for Food and Nutrition | Leoni G.,National Research Institute for Food and Nutrition | Canali R.,National Research Institute for Food and Nutrition | Ambra R.,National Research Institute for Food and Nutrition | And 2 more authors.
Molecular Nutrition and Food Research | Year: 2010

The term Vitamin E is utilized to describe eight molecules, subdivided into two groups, tocopherols and tocotrienols (TTs). It has been shown that specific TTs affect the growth of sevERαl lines of tumour cells, and that this activity is not shared by tocopherols. In agreement with these observations, a TTs-rich fraction from palm oil (PTRF) was reported to inhibit prolifERαtion and induce apoptosis in sevERαl cancer cells. However, the molecular mechanism involved in TTs activity is still unclear. We have recently proposed that TTs pro-apoptotic activity involves estrogen receptor beta (ERβ) signalling. In this study, we report that, in MCF-7 breast cancer cell, expressing both ERα and ERβ, PTRF treatment increases ERβ nuclear translocation, as demonstrated by immunofluorescence experiments and significantly inhibits ERα expression (458.91-fold of change) and complete disappearing of the protein from the nucleus. Moreover, PTRF treatment induces ER-dependent genes expression (macrophage inhibitory cytokine-1, early growth response-1 and Cathepsin D) which is inhibited by the ER inhibitor, ICI 182.780, and induces DNA fragmentation. Finally, cDNA-array experiments suggest that the activation of specific pathways in cells treated with γ-TT with respect to γ-TT. Our data suggest a novel potential molecular mechanism for TTs activity. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Speciale A.,Messina University | Canali R.,National Research Institute for Food and Nutrition | Chirafisi J.,Messina University | Saija A.,Messina University | And 2 more authors.
Journal of Agricultural and Food Chemistry | Year: 2010

Oxidative stress and inflammation are considered to play a pivotal role In vascular endothelial dysfunction by triggering activation of transcription factors, such as NF-κB, functionally dependent on cellular redox status. The anthocyanin cyanidin-3-O-glucoside (C3G), as well as other phytochemicals recognized as potent antioxidants and free radical scavengers, may act as modulators of gene regulation and signal transduction pathways. This study demonstrates that C3G is able to protect human endothelial cells against alterations induced by TNF-α, including the activation of NF-κB, increased gene expression of adhesion molecules, leukocyte adhesion to endothelium, and Intracellular accumulation of H2O2 and lipid peroxidation byproducts. These observations contribute to provide a conceptual background for the understanding of the mechanisms underlying the role of C3G, as well as other dietary plant polyphenols, In the prevention of diseases associated with inflammation and oxidative stress, Including atherosclerosis. © 2010 American Chemical Society.


Speciale A.,Messina University | Anwar S.,Messina University | Canali R.,National Research Institute for Food and Nutrition | Chirafisi J.,Messina University | And 3 more authors.
Molecular Nutrition and Food Research | Year: 2013

Scope: Many dietary phytochemicals have been shown able to prevent a large spectrum of diseases, including cardiovascular disorders, with a mechanism commonly ascribed to an antioxidant effect. However, these in vivo beneficial effects are unlikely to be explained on the base of this mechanism. The discovery of specific genes regulated by the antioxidant responsive element (ARE) affected by antioxidants/electrophiles, led to the hypothesis that some phytochemicals may act as modulators of signal transduction pathways. The aim of the study was to investigate if in vitro pharmacological activation of Nrf2 pathway by cyanidin-3-O-glucoside (C3G) may be involved in its antiatherogenic effects. Methods and results: Herein, we investigated the in vitro effects of C3G on cell signaling pathways in human umbilical vein endothelial cells (HUVECs) challenged with tumor necrosis factor-α (TNF-α). Pretreatment with C3G prevented oxidative stress, improved antioxidant systems, and activated Nrf2/ARE pathway, at baseline and after TNF-α treatment. Furthermore, we demonstrated the involvement of specific mitogen-activated protein kinases (MAPKs) (ERK1/2) in C3G induction of Nrf2/ARE pathway. Finally, the inactivation of ERK1/2 activity by the inhibitor PD98059 abolished the increase of Nrf2 nuclear accumulation induced by C3G, and also increased NF-κB p65 nuclear translocation in TNF-α challenged cells. Conclusion: Our data confirm the hypothesis that natural Nrf2 and HO-1 inducers, such as C3G and other dietary phytochemicals, might be a potential therapeutic strategy to protect vascular system against various stressors preventing several pathological conditions. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Cimino F.,Messina University | Speciale A.,Messina University | Anwar S.,Messina University | Canali R.,National Research Institute for Food and Nutrition | And 4 more authors.
Genes and Nutrition | Year: 2013

The detrimental effects of high oxygen supplementation have been widely reported. Conversely, few is known about the effects of exposure to mild hyperoxic conditions, an interesting issue since the use of oxygen-enriched mixture is now increasingly used in clinical practice and especially for professional and recreational reasons. Our study investigated if in vitro exposure of human umbilical vein endothelial cells (HUVECs) to moderate hyperoxia (O2 32 %) induces cellular alterations, measured as changes in cell signaling pathways. Furthermore, by means of an ex vivo experimental model where human volunteers were used as bioreactors, we studied whether anthocyanin metabolites are able to protect HUVECs against mild hyperoxia-induced damage. We observed that the cytotoxic effect of mild hyperoxia came along with a significant decrease in nuclear accumulation of the transcription factor Nrf2, as well as in the expression of Nrf2-regulated antioxidant and cytoprotective genes. Furthermore, under normoxic conditions, anthocyanin metabolites appeared able to activate the Nrf2 pathway, through the involvement of specific kinases (ERK1/2); this adaptive effect may explain the protective effect observed in mild hyperoxia-exposed HUVECs following anthocyanin pretreatment. This study confirms that dietary anthocyanins and/or their metabolites can protect endothelial cells against mild hyperoxia-induced alterations acting as cell signaling modulators. © 2012 Springer-Verlag Berlin Heidelberg.


De Nicolo S.,CNR Institute of Neuroscience | Tarani L.,University of Rome La Sapienza | Ceccanti M.,University of Rome La Sapienza | Maldini M.,National Research Institute for Food and Nutrition | And 4 more authors.
Nutrition | Year: 2013

Objective: Polyphenols are chemicals derived from plants known to possess antioxidant and anti-inflammatory properties. High intake of fruit and vegetables is believed to be beneficial to human health. Various studies have suggested that dietary polyphenols may protect against cancer and cardiometabolic and neurodegenerative diseases. Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are neurotrophins that play key roles in brain cell development, growth, and survival. The aim of this study was to investigate whether or not administration of olive (Olea europaea L.) polyphenols could have an effect on NGF and BDNF content and the expression of their receptors, TrkA and TrkB, respectively, in the mouse brain. Methods: NGF and BDNF were measured by enzyme-linked immunosorbent assay. TrkA and TrkB were measured by Western blotting. Results: We found NGF and BDNF elevation in the hippocampus and olfactory bulbs and a decrease in the frontal cortex and striatum. These data were associated with potentiated expression of TrkA and TrkB in the hippocampus and olfactory bulbs but no differences between groups in the striatum and frontal cortex. Polyphenols did not affect some behavioral mouse parameters associated with stressing situations. Conclusions: Altogether, this study shows that olive polyphenols in the mouse may increase the levels of NGF and BDNF in crucial areas of the limbic system and olfactory bulbs, which play a key role in learning and memory processes and in the proliferation and migration of endogenous progenitor cells present in the rodent brain. © 2013 Elsevier Inc.


Sette S.,National Research Institute for Food and Nutrition | Le Donne C.,National Research Institute for Food and Nutrition | Piccinelli R.,National Research Institute for Food and Nutrition | Arcella D.,National Research Institute for Food and Nutrition | And 2 more authors.
Nutrition, Metabolism and Cardiovascular Diseases | Year: 2011

Background and aims: Italian National Food Consumption Survey, INRAN-SCAI 2005-06, is the third national food consumption survey performed in Italy. This study describes energy and nutrient intakes in Italy. Methods and results: A national cross-sectional food consumption survey was conducted using consecutive 3-day food records between October 2005 and December 2006. A sample of 3323 males and females aged 0.1-97.7 years living in private households was investigated. Individual food records were converted into energy and nutrient intakes with the use of recently updated national food composition databases. For each subject, intakes of energy and of 27 nutrients were calculated, including six minerals (i.e., iron, calcium, phosphorus, magnesium, potassium and zinc) and 10 vitamins (i.e., thiamine, riboflavin, vitamin C, vitamin B 6, retinol, β-carotene, vitamin A as retinol equivalents (REs), vitamin E, vitamin D and vitamin B 12). On average, 36% of calories appeared to derive from fat (11% from saturated fatty acids) and 45% from available carbohydrates (15% from soluble carbohydrates). Conclusions: The results of the INRAN-SCAI 2005-06 survey in terms of nutrient intakes provide an important piece of information for nutrition surveillance of the population and may also be used to identify priorities for further research. © 2010 Elsevier B.V..


Natella F.,National Research Institute for Food and Nutrition | Belelli F.,National Research Institute for Food and Nutrition | Ramberti A.,National Research Institute for Food and Nutrition | Scaccini C.,National Research Institute for Food and Nutrition
Journal of Food Biochemistry | Year: 2010

The effect of microwave, boiling and pressure cooking on total antioxidant capacity and total phenolics content of seven vegetables were studied. Total phenolics in raw and cooked vegetables were determined by the Folin-Ciocalteu method, whereas the total antioxidant capacity of vegetables extracts was evaluated using the Crocin method. After boiling, four out of seven vegetables (cauliflower, peas, spinach and Swiss chard) showed a significant decrease in their total phenolic content (P < 0.05). No decrease or a smaller decrease was observed for these four vegetables (P < 0.05) after pressure cooking and/or microwaving than after boiling. The total antioxidant capacity of potato and Swiss chard was not significantly affected by cooking procedures, whereas it decreased for spinach and peas, and it increased for tomato and carrots (P < 0.05). In addition, there was a statistically significant correlation between total phenolic content and total antioxidant capacity in cooked and uncooked vegetables, but the strength of the correlation increased when separating carotenoid-poor from carotenoid-rich vegetables. © 2010, The Author(s). Journal compilation © 2010, Wiley Periodicals, Inc.


Canali R.,National Research Institute for Food and Nutrition | Comitato R.,National Research Institute for Food and Nutrition | Ambra R.,National Research Institute for Food and Nutrition | Virgili F.,National Research Institute for Food and Nutrition
British Journal of Nutrition | Year: 2010

We have studied the effect of human serum, collected after red wine consumption (RWS), on TNF-α-dependent activation of transcription factors (NF-B, activator protein-1 (AP-1) and cAMP response element-binding proteins) and on the expression of selected genes involved in cell adhesion or fibrinolysis processes in human primary endothelial cells (human umbilical vein endothelial cells (HUVEC)). Our data indicate that RWS containing RW metabolites, isolated after 40min from an acute consume of wine (5ml/kg body weight), induces nuclear translocation of NF-κB and AP-1 in the absence of any further stimulus. On the other hand, TNF-α treatment in the presence of RWS is associated with a delay in transcription factor activation and to a negative modulation on the expression of specific genes. Moreover, RWS stimulates c-jun binding to the tissue-type plasminogen activator cAMP responsive element consensus site modulating the expression of the specific gene downstream. These results confirm that RW metabolites affect the activity of different transcription factors playing an important preconditioning role in the modulation of the inflammatory pathway in endothelial cells. This is the first report on the effects of a complex food matrix, on the molecular mechanisms associated with inflammatory response in HUVEC cultured in condition that reproduces the physiological environment occurring in vivo. © 2009 The Authors.


Maldini M.,National Research Institute for Food and Nutrition | Baima S.,National Research Institute for Food and Nutrition | Morelli G.,National Research Institute for Food and Nutrition | Scaccini C.,National Research Institute for Food and Nutrition | Natella F.,National Research Institute for Food and Nutrition
Journal of Mass Spectrometry | Year: 2012

Glucosinolates are an important class of secondary plant metabolites, possessing health-promoting properties. Young broccoli plants are a very good source of glucosinolates with concentrations several times greater than in mature plants. The aim of our study was to develop a liquid chromatography-mass spectrometry and liquid chromatography/tandem mass spectrometry qualitative and quantitative method for the measure of glucosinolates in broccoli sprouts. The described method provides high sensitivity and specificity, allowing a rapid and simultaneous determination of 14 glucosinolates. The proposed method has been validated for eight glucosinolates: glucobrassicin, glucoraphanin, glucoiberin, glucoerucin, progoitrin, gluconapin, sinigrin and glucocheirolin. The linear range was 1-150 μg ml-1, the intra-day and inter-day precision values are within 6% and 8% at the lower limit of quantification, while the overall recovery of the eight glucosinolates was 99 ± 9%. This validated method was used successfully for analysis of glucosinolates content of broccoli sprouts grown in different conditions. Copyright © 2012 John Wiley & Sons, Ltd.

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