Direct in vivo evidence of protective effects of grape seed procyanidin fractions and other antioxidants against ethanol-induced oxidative DNA damage in mouse brain cells

Ethanol is a principle ingredient of alcoholic beverages with potential neurotoxicity and carcinogenicity, and the ethanol-associated oxidative DNA damage in the central nervous system is well documented. The present work studied the possible protective effects of grape seed oligomer and polymer procyanidin fractions against ethanol-induced toxicity and compared these with resveratrol and other well-known antioxidants (ascorbic acid and vitamin E). By using the single cell gel electrophoresis (comet assay), a simple and sensitive technique for genotoxicity studies, the potential genotoxicity of acute and chronic ethanol administration in the different brain regions was investigated. Acute ethanol administration, at the dose of 2.5 or 5.0 g kg(-1) i.p., could induce significant DNA damage in cerebellum and hippocampus. Chronic administration of ethanol at the dose of 2.5 or 5.0 g kg-1 p.o. for 30 days could induce significant DNA damage in cerebellum, hippocampus, hypothalamus, and cortex, which could be auto-repaired at least 3 days after ethanol withdrawal. Oral administration of grape seed oligomer and polymer procyanidins and resveratrol (25, 50, and 100 mg kg(-1)) for 3 days before acute ethanol (5.0 g kg(-1), i.p.) or repeated administration of these substances together with ethanol (5.0 g kg(-1), p.o.) for 30 consecutive days could significantly inhibit DNA damage in brain cells induced by ethanol. As compared, ascorbic acid (50, 100, and 200 mg kg(-1)) and vitamin E (100, 200, and 400 mg kg(-1)) could also present protective effects on ethanol-induced DNA damage. Furthermore, the concentrations of ethanol and acetaldehyde in brain regions of the mice were detected by gas chromatography after administration of ethanol plus antioxidants. All of the results indicated that ethanol could induce region-specific oxidative DNA damage in which the cerebellum and hippocampus were more vulnerable, but intake of grape seed procyanidins or other natural antioxidants could protect the brain against ethanol-induced genotoxicity.     

Affinity purification of copper-chelating peptides from sunflower protein hydrolysates

Copper-chelating peptides were purified from sunflower protein hydrolysates by affinity chromatography using immobilized copper. A variety of protein hydrolysates were obtained by incubation with the proteases Alcalase and Flavourzyme for different periods of time. Chelating activity was indirectly determined by measuring the inhibitory effect of hydrolysates on the oxidation of beta-carotene by copper. Copper-binding peptides purified from the two hydrolysates that inhibited oxidation by copper the most contained 25.4 and 42.0% histidine and inhibited beta-carotene oxidation 8 and 3 times more than the original hydrolysates, which had 2.4 and 2.6% histidine, respectively. Thus, histidine content is not the only factor involved in antioxidant activity, and probably other factors such as peptide size and amino acid sequence are also important. This work shows that affinity chromatography can be used for the purification of copper-chelating peptides and probably other metals of nutritional interest such as calcium, iron, and zinc. In addition to their antioxidant potential, chelating peptides are of nutritional interest because they increase bioavailability of minerals.     

Cytotoxic and antioxidant activity of 4-methylthio-3-butenyl isothiocyanate from Raphanus sativus L. (Kaiware Daikon) sprouts

There is high current interest in the chemopreventive potential of Brassica vegetables (cruciferae), particularly due to their content in glucosinolates (GL), which upon myrosinase hydrolysis release the corresponding isythiocyanates (ITC). Some ITCs, such as sulforaphane (SFN) from broccoli ( Brassica oleacea italica), have been found to possess anticancer activity through induction of apoptosis in selected cell lines, as well as indirect antioxidant activity through induction of phase II detoxifying enzymes. Japanese daikon ( Raphanus sativus L.) is possibly the vegetable with the highest per capita consumption within the Brassicaceae family. Thanks to a recently improved gram scale production process, it was possible to prepare sufficient amounts of the GL glucoraphasatin (GRH) as well as the corresponding ITC 4-methylthio-3-butenyl isothiocyanate (GRH-ITC) from its sprouts. This paper reports a study on the cytotoxic and apoptotic activities of GRH-ITC compared with the oxidized counterpart 4-methylsulfinyl-3-butenyl isothiocyanate (GRE-ITC) on three human colon carcinoma cell lines (LoVo, HCT-116, and HT-29) together with a detailed kinetic investigation of the direct antioxidant/radical scavenging ability of GRH and GRH-ITC. Both GRH-ITC and GRE-ITC reduced cell proliferation in a dose-dependent manner and induced apoptosis in the three cancer cell lines. The compounds significantly ( p < 0.05) increased Bax and decreased Bcl2 protein expression, as well as producing caspase-9 and PARP-1 cleavage after 3 days of exposure in the three cancer cell lines. GRH-ITC treatment was shown to have no toxicity with regard to normal human lymphocytes (-15 +/- 5%) in comparison with SFN (complete growth inhibition). GRH and GRH-ITC were able to quench the 2,2-diphenyl-1-picrylhydrazyl radical, with second-order rate constants of 14.0 +/- 2.8 and 43.1 +/- 9.5 M(-1) s(-1), respectively (at 298 K in methanol), whereas the corresponding value measured here for the reference antioxidant alpha-tocopherol was 425 +/- 40 M (-1) s (-1). GRH reacted with H2O2 and tert-butyl hydroperoxide in water (pH 7.4) at 37 degrees C, with rate constants of 1.9 +/- 0.3 x 10(-2) and 9.5 +/- 0.3 x 10(-4) M(-1) s (-1) (paralleling recently developed synthetic antioxidants) being quantitatively (>97%) converted to GRE. It is demonstrated that GRH-ITC has interesting antioxidant/radical scavenging properties, associated with a selective cytotoxic/apoptotic activity toward three human colon carcinoma cell lines, and very limited toxicity on normal human T-lymphocytes.     

Analysis of the distribution of endemic and rare arthropods in high endemism areas of Algarve-South Portugal

The study of several arthropod groups in high endemism areas (HEA) of Algarve, the Southernmost province of Portugal, has revealed endemic species and species not yet recorded for the Portuguese fauna. The list includes 3 species of Isopoda endemic to Algarve, to Portugal or to the Iberian Peninsula, 2 species new to Portugal and 2 species new to the Iberian Peninsula; 2 species of Pseudoscorpiones not yet cited to Portugal; 11 species of Pauropoda new to Portugal, including one new species probably endemic; 15 species of Collembola endemic to the Portuguese or to the Iberian Peninsula faunas (Gama et al. 1997) and 37 species and 5 subspecies of Staphylinidae endemic to Algarve.With the exception of Collembola, which have been studied by us, the remaining groups have been submitted for study to several specialists.These findings heighten the biological richness of these areas of high endemism in terms of conservation of biological diversity. The authors draw attention to the necessity to preserve or restore the biological value of these areas, encouraging the adoption of protective measures and the creation of research programmes that include the implementation of taxonomic studies and co-operation between taxonomists and ecologists in order to preserve the biodiversity of these areas.     

Evaluation of the Influence of Natural and Antrhopogenic Processes on Water Quality in Karstic Region

This paper presents a comprehensive study on the quality of surface and groundwater in an environmentally sensitive karstic watershed strongly pressed by urban growth. The objective of the study was to assess the integrated effects of human activities and natural characteristics of karstic environments on the quality of surface and groundwater using multivariate statistical techniques. Data from 18 physical, chemical, and microbiological water quality variables obtained throughout a hydrological year were analyzed using principal components analysis (PCA) and cluster analysis. The PCA was carried out individually for surface water and groundwater. Our goal was to study the behavior of the water quality variables at each of these systems, as well as to infer the importance of these variables on the dynamics of the water resources in the region. Our results for surface water showed that the 18 original variables could be reduced to five principal components which together accounted for 69% of the total variation in the data, whereas for groundwater, 70% of the total variation in the data was explained by five principal components. In order to evaluate the impacts of anthropic activities on the water quality at the monitoring locations, the cluster analysis was applied to the ten sampling points. The analysis identified five clusters, two of which are formed by points with low contamination levels and three where the anthropic interference is noticeable. The results suggest the existence of specific contamination sources in many points, including in the groundwater, and highlight the natural vulnerability of the karstic environments.     

A critical assessment of soil amendments (slaked lime/acidic fertilizer) for the phytomanagement of moderately contaminated shooting range soils

Purpose  

The effects of the addition of an acidic fertilizer solution and/or slaked lime (5.5 g Ca(OH)₂ kg⁻¹) on a slightly acidic shooting range soil (pH 6.1, % organic carbon 5.4) with moderate metal (e.g., 620 mg kg⁻¹ Pb) and metalloid (17 mg kg⁻¹ Sb) concentrations on metal and Sb solubility and plant accumulation were investigated.     

Determination of nonprotein amino acids and betaines in vegetable oils by flow injection triple-quadrupole tandem mass spectrometry: a screening method for the detection of adulterations of olive oils

A novel screening method using an automated flow injection electrospray ionization tandem mass spectrometry system is proposed for the simultaneous determination of five nonprotein amino acids (β-alanine, alloisoleucine, ornithine, citrulline, pyroglutamic acid) and three betaines (glycine betaine, trigonelline, proline betaine) after derivatization with butanolic HCl. MS/MS experiments were carried out in a triple-quadrupole instrument using multiple reaction monitoring mode in <2 min. The proposed method provided high fingerprinting power to identify the presence of five of the studied compounds in different types of vegetable oils (soybean, sunflower, corn, olive) with LODs at parts per billion levels. The method was validated, and different mixtures of extra virgin olive oil with seed oils were analyzed, achieving the typification for the detection of adulterations in extra virgin olive oils up to 2% w/w. The nonprotein amino acid ornithine was confirmed as a marker for adulteration in the olive oils analyzed.     

Phenolic compounds and saponins in quinoa samples (Chenopodium quinoa Willd.) grown under different saline and nonsaline irrigation regimens

Quinoa is a pseudocereal from South America that has received increased interest around the world because it is a good source of different nutrients and rich in antioxidant compounds. Thus, this study has focused on the effects of different agronomic variables, such as irrigation and salinity, on the phenolic and saponin profiles of quinoa. It was observed that irrigation with 25% of full water restitution, with and without the addition of salt, was associated with increases in free phenolic compounds of 23.16 and 26.27%, respectively. In contrast, bound phenolic compounds were not affected by environmental stresses. Saponins decreased if samples were exposed to drought and saline regimens. In situations of severe water deficit, the saponins content decreased 45%, and 50% when a salt stress was added. The results suggest that irrigation and salinity may regulate the production of bioactive compounds in quinoa, influencing its nutritional and industrial values.     

Structural, physical, and chemical modifications induced by microwave heating on native agar-like galactans

Native agars from Gracilaria vermiculophylla produced in sustainable aquaculture systems (IMTA) were extracted under conventional (TWE) and microwave (MAE) heating. The optimal extracts from both processes were compared in terms of their properties. The agars' structure was further investigated through Fourier transform infrared and NMR spectroscopy. Both samples showed a regular structure with an identical backbone, β-d-galactose (G) and 3,6-anhydro-α-l-galactose (LA) units; a considerable degree of methylation was found at C6 of the G units and, to a lesser extent, at C2 of the LA residues. The methylation degree in the G units was lower for MAE(opt) agar; the sulfate content was also reduced. MAE led to higher agar recoveries with drastic extraction time and solvent volume reductions. Two times lower values of [η] and M(v) obtained for the MAE(opt) sample indicate substantial depolymerization of the polysaccharide backbone; this was reflected in its gelling properties; yet it was clearly appropriate for commercial application in soft-texture food products.     

Tocopherol composition and antioxidant activity of Spanish wild vegetables

Traditional use of noncultivated vegetables has decreased with the development of agriculture and global supply chains. However, some species are still consumed as part of our traditional Mediterranean diet. Plants are among the most important sources of natural antioxidants for retarding lipid oxidative rancidity in foods or for pharmaceutical applications against chronic diseases related to free radicals production. The present study reports tocopherols composition and antioxidant activity of eight wild greens traditionally used in Spain. According to the edible part consumed, two groups were differentiated. Leafy vegetables whose young stems with leaves are consumed (Apium nodiflorum (L.) Lag., Foeniculum vulgare Mill., Montia fontana L. and Silene vulgaris (Moench) Garcke), and wild asparagus whose young shoots with leaf buds scarcely developed are eaten (Asparagus acutifolius L., Bryonia dioica Jacq., Humulus lupulus L. and Tamus communis L.). Among the leafy vegetables, Silene vulgaris and Apium nodiflorum presented the highest antioxidant capacity and antioxidants contents. Among the wild asparagus, the highest antioxidant capacity was obtained in Humulus lupulus.