Paolino Ninfali

Antioxidant capacity of vegetables, spices and dressings relevant to nutrition

Vegetables are the most important sources of phenolics in the Mediterranean diet. Phenolics, especially flavonoids, are suggested as being essential bioactive compounds providing health benefits. In this study, twenty-seven vegetables, fifteen aromatic herbs and some spices consumed in Central Italy (the Marches region) were studied to reveal total phenolic, flavonoid and flavanol content as well as their antioxidant capacity measured by the oxygen radical absorbance capacity (ORAC) method. A comparison in terms of antioxidant capacity was made between different salads, as well as between salads to which aromatic herbs had been added. Lemon balm and marjoram at a concentration of 1.5 % w/w increased by 150 % and 200 % respectively the antioxidant capacity of a salad portion. A 200 g portion of a salad enriched with marjoram corresponded to an intake of 200 (SD 10) mg phenolics and 4000 (SD 300) ORAC units (micromol Trolox equivalents). Olive oils and wine or apple vinegars were the salad dressings that provided the highest increase in antioxidant capacity. Among the spices tested, cumin and fresh ginger made the most significant contribution to the antioxidant capacity. The results are useful in surveying the antioxidant parameters of vegetables, herbs and spices produced and consumed in our geographical area as well as in quantifying the daily intake of phenolics and ORAC units. The results can be used in public health campaigns to stimulate the consumption of vegetables able to provide significant health protection in order to prevent chronic diseases.

Nutritional and functional potential of Beta vulgaris cicla and rubra.

Swiss chard (Beta vulgaris cicla, BVc) and beetroot (Beta vulgaris rubra, BVr) are vegetables of the Chenopodiaceae family, widely consumed in traditional western cooking. These vegetables represent a highly renewable and cheap source of nutrients. They can be cultivated in soils with scarce organic material and little light and water. BVc and BVr have a long history of use in folk medicine. Modern pharmacology shows that BVc extracts possess antihypertensive and hypoglycaemic activity as well as excellent antioxidant activity. BVc contains apigenin flavonoids, namely vitexin, vitexin-2-O-rhamnoside and vitexin-2-O-xyloside, which show antiproliferative activity on cancer cell lines. BVr contains secondary metabolites, called betalains, which are used as natural dyes in food industry and show anticancer activity. In this light, BVc and BVr can be considered functional foods. Moreover, the promising results of their phytochemicals in health protection suggest the opportunity to take advantage of the large availability of this crop for purification of chemopreventive molecules to be used in functional foods and nutraceutical products.

Comparison of bioactive phytochemical content and release of isothiocyanates in selected brassica sprouts

The consumption of brassica sprouts as raw vegetables provides a fair amount of glucosinolates (GLs) and active plant myrosinase, which enables the breakdown of GLs into health-promoting isothiocyanates (ITCs). This study reports the determination of the main constituents related to human health found in edible sprouts of two Brassica oleracea varieties, broccoli and Tuscan black kale, and two Raphanus sativus varieties, Daikon and Sango. Radish sprouts exhibited the highest ability to produce ITCs, with Daikon showing the greatest level of conversion of GLs into bioactive ITCs (96.5%), followed by Sango (90.0%). Tuscan black kale gave a value of 68.5%, whereas broccoli displayed the lowest with 18.7%. ITCs were not the exclusive GL breakdown products in the two B. oleracea varieties, since nitriles were also produced, thus accounting for the lower conversion observed. Measuring the release of plant ITCs is a valuable tool in predicting the potential level of exposure to these bioactive compounds after the consumption of raw brassica sprouts.

Posttranslational Regulation of Glucose-6-phosphate Dehydrogenase Activity in Tongue Epithelium

Expression of glucose-6-phosphate dehydrogenase (G6PD) activity is high in tongue epithelium, but its exact function is still unknown. It may be related either to the high proliferation rate of this tissue or to protection against oxidative stress. To elucidate its exact role, we localized quantitatively G6PD activity, protein and mRNA using image analysis in tongue epithelium of rat and rabbit, two species with different diets. Distribution patterns of G6PD activity were largely similar in rat and rabbit but the activities were twofold lower in rabbit. Activity was two to three times higher in upper cell layers of epithelium than in basal cell layers, whereas basal layers, where proliferation takes place, contained twice as much G6PD protein and 40% more mRNA than upper layers. Our findings show that G6PD is synthetized mainly in basal cell layers of tongue epithelium and that it is posttranslationally activated when cells move to upper layers. Therefore, we conclude that the major function of G6PD activity in tongue epithelium is the formation of NADPH for protection against oxidative stress and that diet affects enzyme expression in this tissue.

Fruit and Vegetable Antioxidants in Health

Publisher Summary In the history of human nutrition, one of the most widespread alimentary regimens linked to health protection is represented by the Mediterranean diet (MD). MD eating patterns consist of the wide use of whole grains, fruits, vegetables, nuts, fish, and olive oil. People obtain a wide range of antioxidants from the intake of a large variety of fresh fruits and vegetables. Investigations have shown that the risk of cancer and other chronic diseases is inversely related to the consumption of vegetables and fruits. Results are maximally oriented to attribute the highest protective role to the antioxidant compounds contained in fruits and vegetables. Processed fruits and vegetables show a wide range of phytochemical loss. The technology in the food industry should be used to reduce the loss of antioxidants and micronutrients to the minimum level by means of mild processes and the monitoring of each step of the transformation with due control assays. Functional foods, containing fruit and vegetable juices or extracts, are an important part of the healthy lifestyle, which includes a balanced diet and physical activity. To deliver their potential public health benefits, functional foods need to be quality controlled through the collaborative efforts of food-control organizations and the food industry, in order to market only those functional foods that are clearly supported with scientific evidence of nutritional value. The emerging field of nutrigenomics, or “personalized nutrition,” provides individual dietary recommendations and may one day have a greater ability to reduce the risk of disease.

Phytochemicals as innovative therapeutic tools against cancer stem cells

The theory that several carcinogenetic processes are initiated and sustained by cancer stem cells (CSCs) has been validated, and specific methods to identify the CSCs in the entire population of cancer cells have also proven to be effective. This review aims to provide an overview of recently acquired scientific knowledge regarding phytochemicals and herbal extracts, which have been shown to be able to target and kill CSCs. Many genes and proteins that sustain the CSCs’ self-renewal capacity and drug resistance have been described and applications of phytochemicals able to interfere with these signaling systems have been shown to be operatively efficient both in vitro and in vivo. Identification of specific surface antigens, mammosphere formation assays, serial colony-forming unit assays, xenograft transplantation and label-retention assays coupled with Aldehyde dehydrogenase 1 (ALDH1) activity evaluation are the most frequently used techniques for measuring phytochemical efficiency in killing CSCs. Moreover, it has been demonstrated that EGCG, curcumin, piperine, sulforaphane, β-carotene, genistein and the whole extract of some plants are able to kill CSCs. Most of these phytochemicals act by interfering with the canonical Wnt (β-catenin/T cell factor-lymphoid enhancer factor (TCF-LEF)) pathway implicated in the pathogenesis of several cancers. Therefore, the use of phytochemicals may be a true therapeutic strategy for eradicating cancer through the elimination of CSCs.

Vitexin-2-O-xyloside, raphasatin and (-)-epigallocatechin-3-gallate synergistically affect cell growth and apoptosis of colon cancer cells.

Cytotoxic effects of the combination of the food components vitexin-2-O-xyloside (X), raphasatin (4-methylsulphanyl-3-butenyl isothiocyanates; G) and (-)-epigallocatechin-3-gallate (E) were investigated in colon (LoVo and CaCo-2) and breast (MDA-MB-231 and MCF-7) cancer cells. Breast cancer cells were more resistant than colon cells to X, G and E inhibition. On the contrary, marked synergistic effects among X, G and E on cell growth were found in both colon cancer cells. Further analysis revealed a G0/G1 arrest of the phase cell progression and apoptosis, linked to modulation of Bax, Bcl2, caspase-9 and poly(ADP-ribose) polymerase as well as Reactive Oxygen Species (ROS) generation in both colon cancer cells, whereas apoptosis and ROS were not significantly detected in normal human lymphocytes. We conclude that the X, G and E mixture might act by mitochondrial pathway activation of apoptosis, possibly elicited by ROS and the mixture may be effective in the chemoprevention of colon cancer.

Total extract of Beta Vulgaris var. Cicla seeds versus its purified phenolic components: antioxidant activities and antiproliferative effects against colon cancer cells

INTRODUCTION Beta vulgaris var. cicla (BV) leaves contain chemopreventive compounds that have been investigated for new drug discovery. These compounds belong to the family of the apigenin-glycosides. Since the leaves are seasonal products containing high percentages of water, they are easily degradable during storage in fresh conditions. To be stored they require a drying process, consuming time and a large amount of energy. The extraction of apigenin-glycosides may also be conveniently performed from BV seeds, which represent a stable and year-long available biomass. OBJECTIVES The present report was undertaken to find a strategy of purification of bioactive flavonoids from BV seeds and test their ability to inhibit proliferation both on human colon cancer (RKO) cells and normal human fibroblasts (HF). MATERIALS AND METHODS The ethyl-acetate extract of BV seeds was fractionated on a Sephadex LH 20 column. A fraction of this extract, labeled as P4, exploited a marked antiproliferative activity on RKO cells. The components of P4 were purified on an RP₁₈ column chromatography and identified by HPLC-ESI-MS as 2,4,5-trihydroxybenzaldehyde, 2,5-dihydroxybenzaldehyde, vanillic acid, xylosylvitexin, glucopyranosyl-glucopyrasyl-rhamnetin and glucopyranosyl-xylosyl-rhamnetin. All of them were tested for cytostatic and cytotoxic activity on RKO and HF cells. RESULTS Xylosylvitexin exhibited the strongest antiproliferative activity on RKO cells, together with an enhancement of the apoptosis, an increase of cells in the G₁ phase and a reduction of cells in the S phase; on the contrary, the proliferation of HF was significantly stimulated. CONCLUSION Xylosylvitexin is the main and more efficient chemopreventive compound in BV seeds, but the natural cocktail of molecules, represented by P4 fraction, showed a better compromise between the antiproliferative activity on RKO cells and the enhancement of HF proliferation.

Vanadate affects glucose metabolism of human erythrocytes

Vanadate causes a rapid breakdown of 2,3-bisphosphoglycerate in intact erythrocytes. This metabolite is nearly stoichiometrically transformed into pyruvate, which changes the cell redox state and enhances the glycolytic flux. The results show that the vanadate effect on 2,3-bisphosphoglycerate, also evident in hemolysates, is attributable to the stimulation of a phosphatase activity of the phosphoglycerate mutase. In agreement with others (J. Carreras, F. Climent, R. Bartrons, and G. Pons (1982) Biochim. Biophys. Acta 705, 238-242), vanadate is thought to destabilize the phosphoryl form of this enzyme which shows competitive inhibition between the ion and 2,3-bisphosphoglycerate in the mutase reaction. A competitive inhibition between vanadate and glucose 1,6-bisphosphate is also found for phosphoglucomutase, without evidence for phosphatase activity toward the bisphosphate cofactor.

Chemical and cellular antioxidant activity of phytochemicals purified from olive mill waste waters.

The isolation and identification of a phytocomplex from olive mill waste waters (OMWW) was achieved. The isolated phytocomplex is made up of the following three phenolic compounds: hydroxytyrosol (3,4-DHPEA), tyrosol (p-HPEA) and the dialdehydic form of decarboxymethyl elenolic acid, linked with (3,4-dihydroxyphenyl)ethanol (3,4-DHPEA-EDA). The purification of this phytocomplex was reached by partial dehydration of the OMWW, followed by liquid-liquid extraction with ethyl acetate and middle pressure liquid chromatography (MPLC) on a Sephadex LH-20 column. The phytocomplex accounted for 6% of the total phenolic content of the OMWW. The phytocomplex and individual compounds were tested for antioxidant capacity by the oxygen radical absorbance capacity (ORAC) method. The ORAC phytocomplex produced 10,000 ORAC units/g dry weight, whereas the cellular antioxidant activity, measured by the cellular antioxidant activity in red blood cell (CAA-RBC) method, demonstrated that the phytocomplex and all of the components are able to permeate the cell membrane thus exhibiting antioxidant activity inside the red blood cells. Our phytocomplex could be employed in the formulation of fortified foods and nutraceuticals, with the goal to obtain substantial health protective effects due to the suitable combination of the component molecules.