Does water addition during the industrial milling phase affect the chemical-sensory quality of olive oils? The case of cv. Arbequina oils.

At olive oil industrial extraction, water addition is a practice overcoming the formation of thick pastes. The effect of water addition (0 to 6.2%, kgadded water/kgolives), during the industrial milling of cv. Arbequina olives, on the oils' chemical-sensory quality, was evaluated. Despite the extra virgin olive oil classification, compared with the water incorporation (1.2-6.2%), extraction without water addition resulted into oils that showed less primary oxidation (lower peroxide values and K232), greater total phenolic content (+12-22%) and higher oxidative stability (+22-31%). No water addition increased the oils secoiridoids content (+5-13%), mainly oleacein (+27-79%). Oils extracted without water addition had a more intense ripe fruity sensation (≥ +11%) but lower fruit intensities (at least -4%). Thus, the quality and stability of the cv. Arbequina oils can be favoured if extracted without adding water during the olives industrial milling.

Impact of fresh olive leaves addition during the extraction of Arbequina virgin olive oils on the phenolic and volatile profiles.

Leaves incorporation during the extraction of olive oils can enhance their chemical-sensory quality. Thus, leaves from cvs. Arbequina or Santulhana were added (1%, w/w) during the extraction of Arbequina oils using an Abencor system, being discussed the impacts on the phenolics and volatiles formation enzymatic pathways. Leaves addition contributed to a significant decrease (P-value < 0.05) of the contents of secoiridoids (-11%), C6-aldehydes (-16%), and ester compounds (-22%). This could be tentatively related to a reduction of the enzymatic activity of secoiridoids biosynthesis and lipoxygenase pathways, promoted by the leaves' addition. Moreover, in the presence of leaves, the oils' total contents of phenolics and volatiles were significantly reduced (-7 and -17%, respectively). Contrary, the incorporation of leaves significantly increased (P-value < 0.05) the contents of C6-alcohols (+37%) and the intensities of the green fruity (+25%) and apple (+30%) sensations.

Anthraquinones, Diphenyl Ethers, and Their Derivatives from the Culture of the Marine Sponge-Associated Fungus Neosartorya spinosa KUFA 1047.

Previously unreported anthraquinone, acetylpenipurdin A (4), biphenyl ether, neospinosic acid (6), dibenzodioxepinone, and spinolactone (7) were isolated, together with (R)-6-hydroxymellein (1), penipurdin A (2), acetylquestinol (3), tenellic acid C (5), and vermixocin A (8) from the culture of a marine sponge-associated fungus Neosartorya spinosa KUFA1047. The structures of the previously unreported compounds were established based on an extensive analysis of 1D and 2D NMR spectra as well as HRMS data. The absolute configurations of the stereogenic centers of 5 and 7 were established unambiguously by comparing their calculated and experimental electronic circular dichroism (ECD) spectra. Compounds 2 and 5-8 were tested for their in vitro acetylcholinesterase and tyrosinase inhibitory activities as well as their antibacterial activity against Gram-positive and Gram-negative reference, and multidrug-resistant strains isolated from the environment. The tested compounds were also evaluated for their capacity to inhibit biofilm formation in the reference strains.

Plasmodium stage-selective antimalarials from Lophira lanceolata stem bark.

Targeting the transmissible stages of the Plasmodium parasite that develop in the human and mosquito host is a crucial strategy for malaria control and elimination. Medicinal plants offer a prolific source for the discovery of new antimalarial compounds. The recent identification of the gametocytocidal activity of lophirone E, obtained from the African plant Lophira lanceolata (Ochnaceae), inspired the evaluation of the plant also against early sporogonic stages of the parasite development. The bioassay-guided phytochemical study led to the isolation of two known lanceolins and of a new glycosylated bichalcone, named glucolophirone C. Its stereostructure, including absolute configuration of the bichalcone moiety, was elucidated by means of NMR, HRMS, ECD and computational calculations. Lanceolin B proved to be a potent inhibitor of the development of Plasmodium early sporogonic stages indicating that the plant produces two different stage-specific antimalarial agents acting on transmissible stages in the human and mosquito host.

Secondary Metabolites from the Culture of the Marine Sponge-Associated Fungi Talaromyces tratensis and Sporidesmium circinophorum.

Wortmin (1), meso-1,4-bis(4-methoxybenzyl)-2,3-butanediol (2), and a new isocoumarin derivative tratenopyrone (3) were isolated from the marine sponge-associated fungus Talaromyces tratensis KUFA 0091. A new diphenyl ether derivative, circinophoric acid (4), was isolated, together with the previously reported anthraquinones catenarin and physcion, the benzophenone monomethylsoluchrin, and ß-ergosterol-5,8-endoperoxide, from the marine sponge-associated fungus Sporidesmium circinophorum KUFA 0043. The structures of the new compounds were established based on an extensive analysis of 1D and 2D NMR spectra, and, in the case of compounds 2-4, also by X-ray analysis. All of the isolated compounds were tested for their antibacterial activity against Gram-positive and Gram-negative bacteria, and multidrug-resistant isolates from the environment, as well as for their anti-quorum sensing based on the pigment production of Chromobacterium violaceum ATCC 31523. None of the compounds exhibited either antibacterial (MIC > 256 µg/mL) or anti-quorum sensing activities. The compounds were also inactive in the antifungal (MIC > 512 µg/mL) and cancer cell line (GI50 > 150 µM) assays.

Single-cultivar extra virgin olive oil classification using a potentiometric electronic tongue.

Label authentication of monovarietal extra virgin olive oils is of great importance. A novel approach based on a potentiometric electronic tongue is proposed to classify oils obtained from single olive cultivars (Portuguese cvs. Cobrançosa, Madural, Verdeal Transmontana; Spanish cvs. Arbequina, Hojiblanca, Picual). A meta-heuristic simulated annealing algorithm was applied to select the most informative sets of sensors to establish predictive linear discriminant models. Olive oils were correctly classified according to olive cultivar (sensitivities greater than 97%) and each Spanish olive oil was satisfactorily discriminated from the Portuguese ones with the exception of cv. Arbequina (sensitivities from 61% to 98%). Also, the discriminant ability was related to the polar compounds contents of olive oils and so, indirectly, with organoleptic properties like bitterness, astringency or pungency. Therefore the proposed E-tongue can be foreseen as a useful auxiliary tool for trained sensory panels for the classification of monovarietal extra virgin olive oils.

Improvement of vegetables elemental quality by espresso coffee residues.

Spent coffee grounds (SCG) are usually disposed as common garbage, without specific reuse strategies implemented so far. Due to its recognised richness in bioactive compounds, the effect of SCG on lettuce's macro- and micro-elements was assessed to define its effectiveness for agro industrial reuse. A greenhouse pot experiment was conducted with different amounts of fresh and composted spent coffee, and potassium, magnesium, phosphorous, calcium, sodium, iron, manganese, zinc and copper were analysed. A progressive decrease on all lettuce mineral elements was verified with the increase of fresh spent coffee, except for potassium. In opposition, an increment of lettuce's essential macro-elements was verified when low amounts of composted spent coffee were applied (5%, v/v), increasing potassium content by 40%, manganese by 30%, magnesium by 20%, and sodium by 10%, of nutritional relevance This practical approach offers an alternative reuse for this by-product, extendable to other crops, providing value-added vegetable products.

Inhibition of quorum sensing in Pseudomonas aeruginosa by sesquiterpene lactones.

Six sesquiterpene lactones (SLs) of the goyazensolide and isogoyazensolide-type isolated from the Argentine herb Centratherum punctatum were evaluated on their ability to inhibit virulence factors of Pseudomonas aeruginosa ATCC 27853. Although compounds were not able to completely inhibit bacterial growth at 200µg/ml, the SLs do altered biofilm formation, elastase activity, and production of N-acyl-homoserinelactones (AHLs) which are known quorum sensing autoinducers at lower concentration. Compounds 2, 3, and 5 displayed significant inhibitory effects on P. aeruginosa biofilm formation at 0.5µg/ml being compound 3 (1.32µM) the most potent (42%). Compounds 2, 3, 4, 5 and 6, inhibited 39, 44, 42, 32 and 35% the production of AHLs at 100µg/ml and inhibited by more than 50% the elastase activity at 0.5µg/ml. Our results clearly indicated that sesquiterpene lactones are good candidates for the development of new antimicrobial agents acting not as bactericidal but as antipathogenic agents.

Kale extract increases glutathione levels in V79 cells, but does not protect them against acute toxicity induced by hydrogen peroxide.

This study aims to evaluate the antioxidant potential of extracts of Brassica oleracea L. var. acephala DC. (kale) and several materials of Pieris brassicae L., a common pest of Brassica cultures using a cellular model with hamster lung fibroblast (V79 cells) under quiescent conditions and subjected to H2O2 induced oxidative stress. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and glutathione was determined by the 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB)-oxidized glutathione (GSSG) reductase recycling assay. The phenolic composition of the extracts was also established by HPLC-DAD. They presented acylated and non acylated flavonoid glycosides, some of them sulfated, and hydroxycinnamic acyl gentiobiosides. All extracts were cytotoxic by themselves at high concentrations and failed to protect V79 cells against H2O2 acute toxicity. No relationship between phenolic composition and cytotoxicity of the extracts was found. Rather, a significant increase in glutathione was observed in cells exposed to kale extract, which contained the highest amount and variety of flavonoids. It can be concluded that although flavonoids-rich extracts have the ability to increase cellular antioxidant defenses, the use of extracts of kale and P. brassicae materials by pharmaceutical or food industries, may constitute an insult to health, especially to debilitated individuals, if high doses are consumed.

Further insights into chemical characterization through GC-MS and evaluation for anticancer potential of Dracaena draco leaf and fruit extracts.

The present study reports for the first time the amino acid and fatty acid compositions and the antitumoral activity of aqueous extracts obtained from Dracaena draco L. leaf and fruit. Metabolite profiles were determined by gas chromatography-ion trap-mass spectrometry (GC-IT-MS), with several amino acids, palmitic, linolenic and stearic acid being identified in the leaf extract, and only proline, oleic and stearic acid in the fruit extract. The in vitro antiproliferative activities of the extracts were tested against human colon (Caco-2), kidney (A-498), and liver (HepG2) cancer cell lines. In addition, primary cultures of normal and cancerous renal cells derived from kidney cancer patients were treated with D. draco extracts (0-400 µg/mL). Antiproliferative and cytotoxic effects were determined by the MTT assay. D. draco extracts inhibited proliferation of human colon and renal tumor cells in vitro, whereas no or weak effect was observed in HepG2 cells. Compared to the fruit extract, D. draco leaf extract exhibited stronger antiproliferative activity against all cancer cells. Our results indicate that D. draco, particularly the leaf, may be useful as a cancer chemopreventive and/or chemotherapeutic agent for colon and kidney cancers.

Chemical assessment and in vitro antioxidant capacity of Ficus carica latex.

Ficus species possess latex-like material within their vasculatures, affording protection and self-healing from physical attacks. In this work, metabolite profiling was performed on Ficus carica latex. Volatiles profile was determined by HS-SPME/GC-IT-MS, with 34 compounds being identified, distributed by distinct chemical classes: 5 aldehydes, 7 alcohols, 1 ketone, 9 monoterpenes, 9 sesquiterpenes and 3 other compounds. Sesquiterpenes constituted the most abundant class in latex (ca. 91% of total identified compounds). Organic acids composition was also characterized, by HPLC-UV, and oxalic, citric, malic, quinic, shikimic and fumaric acids were determined. Malic and shikimic acids were present in higher amounts (ca. 26%, each). The antioxidant potential of this material was checked by distinct in vitro chemical assays. A concentration-dependent activity was noticed against DPPH, nitric oxide and superoxide radicals. Additionally, acetylcholinesterase inhibitory capacity was evaluated, but a weak effect was found.

Human cancer cell antiproliferative and antioxidant activities of Juglans regia L.

Several studies suggest that regular consumption of nuts, mostly walnuts, may have beneficial effects against oxidative stress mediated diseases such as cardiovascular disease and cancer. Walnuts contain several phenolic compounds which are thought to contribute to their biological properties. The present study reports the total phenolic contents and antioxidant properties of methanolic and petroleum ether extracts obtained from walnut (Juglans regia L.) seed, green husk and leaf. The total phenolic contents were determined by the Folin-Ciocalteu method and the antioxidant activities assessed by the ability to quench the stable free radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes. Methanolic seed extract presented the highest total phenolic content (116 mg GAE/g of extract) and DPPH scavenging activity (EC(50) of 0.143 mg/mL), followed by leaf and green husk. In petroleum ether extracts, antioxidant action was much lower or absent. Under the oxidative action of AAPH, all methanolic extracts significantly protected the erythrocyte membrane from hemolysis in a time- and concentration-dependent manner, although leaf extract inhibitory efficiency was much stronger (IC(50) of 0.060 mg/mL) than that observed for green husks and seeds (IC(50) of 0.127 and 0.121 mg/mL, respectively). Walnut methanolic extracts were also assayed for their antiproliferative effectiveness using human renal cancer cell lines A-498 and 769-P and the colon cancer cell line Caco-2. All extracts showed concentration-dependent growth inhibition toward human kidney and colon cancer cells. Concerning A-498 renal cancer cells, all extracts exhibited similar growth inhibition activity (IC(50) values between 0.226 and 0.291 mg/mL), while for both 769-P renal and Caco-2 colon cancer cells, walnut leaf extract showed a higher antiproliferative efficiency (IC(50) values of 0.352 and 0.229 mg/mL, respectively) than green husk or seed extracts. The results obtained herein strongly indicate that walnut tree constitute an excellent source of effective natural antioxidants and chemopreventive agents.

Metabolic and bioactivity insights into Brassica oleracea var. acephala.

Seeds of Brassica oleracea var. acephala (kale) were analyzed by HPLC/UV-PAD/MSn-ESI. Several phenolic acids and flavonol derivatives were identified. The seeds of this B. oleracea variety exhibited more flavonol derivatives than those of tronchuda cabbage (Brassica oleracea var. costata), also characterized in this paper. Quercetin and isorhamnetin derivatives were found only in kale seeds. Oxalic, aconitic, citric, pyruvic, malic, quinic, shikimic, and fumaric acids were the organic acids present in these matrices, malic acid being predominant in kale and citric acid in tronchuda cabbage seeds. Acetylcholinesterase (AChE) inhibitory activity was determined in aqueous extracts from both seeds. Kale leaves and butterflies, larvae, and excrements of Pieris brassicae reared on kale were also evaluated. Kale seeds were the most effective AChE inhibitor, followed by tronchuda cabbage seeds and kale leaves. With regard to P. brassicae material, excrements exhibited stronger inhibitory capacity. These results may be explained by the presence of sinapine, an analogue of acetylcholine, only in seed materials. A strong concentration-dependent antioxidant capacity against DPPH, nitric oxide, and superoxide radicals was observed for kale seeds.

Volatile constituents throughout Brassica oleracea L. Var. acephala germination.

In this work, the volatile composition of kale ( Brassica oleracea L. var. acephala) and its variation during germination were monitored during the first 9 days of seedling development by headspace solid-phase microextraction (HS-SPME) combined with gas chromatography/ion trap-mass spectrometry (GC/IT-MS). Differences were found among the materials in the distinct analyzed periods. A total of 66 volatile compounds, distributed in several chemical classes, were determined: alcohols, carbonyl compounds (ketones, aldehydes, and esters), norisoprenoids, and terpenes, among others, sulfur compounds being the most abundant group in seeds and sprouts that exhibited allyl isothiocyanate as the major compound. Leaves of fully developed ground plant had the highest content of norisoprenoids, alcohols, and carbonyl compounds; in opposition, they showed lower levels of sulfur compounds, suggesting that these are important molecules for the development of kale, whereas the others are produced mainly during its growth.

Protective effect of quince (Cydonia oblonga Miller) fruit against oxidative hemolysis of human erythrocytes.

The aim of this study was to determine the phenolic content and evaluate the antioxidant activity of quince (Cydonia oblonga) fruit. For this purpose, fruits were separated into pulps, peels and seeds and methanolic extracts were prepared. The phenolic profiles were determined by HPLC/UV and antioxidant properties were studied for their ability to quench the stable free radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes. The main phenolic compounds were 5-O-caffeoylquinic acid for pulp and peel (57% and 29%, respectively) and stellarin-2 for seed (18%). Total phenolics content was 2.5, 6.3 and 0.4g/kg of methanolic extract for pulp, peel and seed, respectively. Pulp and peel extracts showed similar DPPH free radical scavenging activities (EC(50) of 0.6 and 0.8 mg/ml, respectively), while seed extract presented much lower antioxidant potential (EC(50) of 12.2mg/ml). Under the oxidative action of AAPH, pulp and peel extracts showed significant protection of the erythrocyte membrane from hemolysis, in a time- and concentration-dependent manner. Seed extracts by themselves induced extensive hemolysis. These results indicate higher antioxidant activity for certain parts of quince fruit, namely pulp and peel, that may therefore represent accessible sources of natural antioxidants with potential application in nutritional/pharmaceutical fields, as preventive or therapeutic agents in diseases in which free radicals are implicated.

Screening of antioxidant phenolic compounds produced by in vitro shoots of Brassica oleracea L. var. costata DC.

The phenolic compounds produced by in vitro shoots of Brassica oleracea L. var. costata DC were screened by HPLC-DAD-MS/MS. Thirty seven compounds were characterized, which included chlorogenic acids, flavonoids (the majority of them were hydroxycinnamic acid esters of kaempferol and quercetin glycosides) and hydroxycinnamic acyl glycosides (with predominance of synapoyl gentiobiosides). The antioxidant capacity of the shoots was assessed against DPPH radical and two reactive oxygen species (superoxide radical and hypochlorous acid). A strong concentration-dependent antioxidative capacity was verified in the DPPH and superoxide radicals assays, but a reduced effect was noticed against hypochlorous acid. The results obtained indicate that the in vitro production of B. oleracea var. costata shoots can become important in the obtention of a noticeable dietary source of compounds with health protective potential.

Evaluation of free radical-scavenging and antihemolytic activities of quince (Cydonia oblonga) leaf: a comparative study with green tea (Camellia sinensis).

This study aimed to determine the phenolic profile and to investigate the antioxidant potential of quince (Cydonia oblonga) leaf, comparing it with green tea (Camellia sinensis). For these purposes, methanolic extracts were prepared and phenolics content of quince leaf was determined by HPLC/UV. The antioxidant properties were assessed by Folin-Ciocalteu reducing capacity assay and by the ability to quench the stable free radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes.5-O-Caffeoylquinic acid was found to be the major phenolic compound in quince leaf extract. Quince leaf exhibited a significantly higher reducing power than green tea (mean value of 227.8 +/- 34.9 and 112.5 +/- 1.5 g/kg dry leaf, respectively). Quince leaf extracts showed similar DPPH radical-scavenging activities (EC50 mean value of 21.6 +/- 3.5 microg/ml) but significantly lower than that presented by green tea extract (EC50 mean value of 12.7 +/- 0.1 microg/ml). Under the oxidative action of AAPH, quince leaf methanolic extract significantly protected the erythrocyte membrane from hemolysis in a similar manner to that found for green tea (IC50 mean value of 30.7 +/- 6.7 and 24.3 +/- 9.6 microg/ml, respectively, P > 0.05). These results point that quince leaf may have application as preventive or therapeutic agent in diseases in which free radicals are involved.

Targeted metabolite analysis and biological activity of Pieris brassicae fed with Brassica rapa var. rapa.

For the first time, an insect-plant system, Pieris brassicae fed with Brassica rapa var. rapa, was tested for its biological capacity, namely, antioxidant (DPPH*, *NO, and O(2)*- radicals) and antimicrobial (bacteria and fungi) activities. Samples from the insect's life cycle (larvae, excrements, exuviae, and butterfly) were always found to be more efficient than the host plant. Also, P. brassicae materials, as well as its host plant, were screened for phenolics and organic acids. The host plant revealed higher amounts of both compounds. Two phenolic acids, ferulic and sinapic, as well as kaempferol 3-Osophoroside, were common to insect (larvae and excrements) and plant materials, with excrements being considerably richer. Detection of sulfated compounds in excrements, absent in host plant, revealed that metabolic processes in this species involved sulfation. Additionally, deacylation and deglycosilation were observed. All matrices presented the same organic acids qualitative profile, with the exception of excrements.

Triacylglycerol composition of walnut (Juglans regia L.) cultivars: characterization by HPLC-ELSD and chemometrics.

A total of 26 walnut (Juglans regia L.) samples from 9 cultivars (Arco, Franquette, Hartley, Lara, Marbot, Mayette, Mellanaise, Parisienne, and Rego) harvested in the 2001, 2002, and 2003 crop years and grown in two geographical origins (Braganca and Coimbra, Portugal) were evaluated with regard to their triacylglycerol composition. The methodology employed was reversed-phase high-performance liquid chromatography coupled to an evaporative light-scattering detector (RP-HPLC-ELSD) after extraction of the lipidic fraction of the nuts. Nine compounds were separated, identified, and quantified. All samples presented an identical qualitative profile composed by LLnLn, LLLn, LLL, OLLn, OLL, PLL, OOL, and PLO (P = palmitoyl; O = oleoyl; L = linoleoyl; Ln = linonenoyl). Trilinolein (LLL) was the major triglyceride, followed by dilinoeoyl-oleoyl-glycerol (OLL) and dilinoleoyl-linolenoyl-glycerol (LLLn), with mean values of 37.7, 18.5, and 18.4%, respectively. Significant differences in composition were found between cultivars, and these differences were also significant when cultivars were grouped by year of production, showing that besides genetic factors, the triacylglycerol composition can be strongly influenced by environmental factors.

Determination of sterol and fatty acid compositions, oxidative stability, and nutritional value of six walnut (Juglans regia L.) cultivars grown in Portugal.

Six cultivars (Franquette, Marbot, Mayette, Mellanaise, Lara, and Parisienne) of walnuts (Juglans regia L.) were collected during the 2001 crop, from Bragança, Portugal. Chemical composition, including moisture, total oil content, crude protein, ash, carbohydrates, and nutritional value, was evaluated. Fat was the predominant component, ranging from 62.3 to 66.5%. Total oil was extracted and analyzed for fatty acids, sterols, oxidative stability, and peroxide value. Fatty acids and sterols were determined by gas-liquid chromatography coupled to a flame ionization detector. Eighteen fatty acids were quantified. Polyunsaturated fatty acids and, in particular, linoleic acid were predominant. Beta-Sitosterol, delta(5)-avenasterol, and campesterol were the major sterols found. Differences were observed among the studied cultivars, especially in peroxide values and in the sterol profile.