Eduardo Rosa

Natural Bioactive Compounds from Winery By-Products as Health Promoters: A Review

The relevance of food composition for human health has increased consumers’ interest in the consumption of fruits and vegetables, as well as foods enriched in bioactive compounds and nutraceuticals. This fact has led to a growing attention of suppliers on reuse of agro-industrial wastes rich in healthy plant ingredients. On this matter, grape has been pointed out as a rich source of bioactive compounds. Currently, up to 210 million tons of grapes (Vitis vinifera L.) are produced annually, being the 15% of the produced grapes addressed to the wine-making industry. This socio-economic activity generates a large amount of solid waste (up to 30%, w/w of the material used). Winery wastes include biodegradable solids namely stems, skins, and seeds. Bioactive compounds from winery by-products have disclosed interesting health promoting activities both in vitro and in vivo. This is a comprehensive review on the phytochemicals present in winery by-products, extraction techniques, industrial uses, and biological activities demonstrated by their bioactive compounds concerning potential for human health.

The antimicrobial effects of glucosinolates and their respective enzymatic hydrolysis products on bacteria isolated from the human intestinal tract

Aims:  The aim of the study was to evaluate the in vitro antibacterial activity of glucosinolates and their enzymatic hydrolysis product against bacteria isolated from the human intestinal tract.

Antimicrobial activity of phenolics and glucosinolate hydrolysis products and their synergy with streptomycin against pathogenic bacteria.

The purpose of the present study was to evaluate the in vitro antibacterial effects of different classes of important and common dietary phytochemicals (5 simple phenolics - tyrosol, gallic acid, caffeic acid, ferulic acid, and chlorogenic acid; chalcone - phloridzin; flavan-3-ol - (-) epicatechin; seco-iridoid - oleuropein glucoside; 3 glucosinolate hydrolysis products - allylisothiocyanate, benzylisothiocyanate and 2-phenylethylisothiocyanate) against Escherichia coli, Pseudomonas aeruginosa, Listeria monocytogenes and Staphylococcus aureus. Another objective of this study was to evaluate the effects of dual combinations of streptomycin with the different phytochemicals on antibacterial activity. A disc diffusion assay was used to evaluate the antibacterial activity of the phytochemicals and 3 standard antibiotics (ciprofloxacin, gentamicin and streptomycin) against the four bacteria. The antimicrobial activity of single compounds and dual combinations (streptomycin-phytochemicals) were quantitatively assessed by measuring the inhibitory halos. The results showed that all of the isothiocyanates had significant antimicrobial activities, while the phenolics were much less efficient. No antimicrobial activity was observed with phloridzin. In general P. aeruginosa was the most sensitive microorganism and L. monocytogenes the most resistant. The application of dual combinations demonstrated synergy between streptomycin and gallic acid, ferulic acid, chlorogenic acid, allylisothiocyanate and 2-phenylethylisothiocyanate against the Gram-negative bacteria. In conclusion, phytochemical products and more specifically the isothiocyanates were effective inhibitors of the in vitro growth of the Gram-negative and Gram-positive pathogenic bacteria. Moreover, they can act synergistically with less efficient antibiotics to control bacterial growth.

Daily Variation in Glucosinolate Concentrations in the Leaves and Roots of Cabbage Seedlings in Two Constant Temperature Regimes

Limited information is available on the glucosinolate variation within the Brassica plant and the relationship between the pattern and concentration of glucosinolates in the aerial parts and the roots has received little attention. Early studies carried out under -eld conditions have shown that glucosinolate levels may vary considerably throughout a 24 h period. The purpose of the present study was to show whether, under controlled conditions, temperature was a factor in glucosinolate variation and to determine whether such variation might be due to translocation of glucosinolates between the aerial parts of the plant and its roots. Cabbage seedlings were maintained at 20 and 30iC over 2 days and leaves and roots sampled at 02 : 00 h, 06 : 00 h, 10 : 00 h, 14 : 00 h, 18 : 00 h and 22 : 00 h. The glucosinolates 2-propenyl- and 3-methylsulphinylpropyl- with an average of 261 and 167 kmol 100 g~1 DW, respectively, were the two main glucosinolates in the aerial part of the plant whilst in the roots 1-methoxyindol- 3-ylmethyl-, 2-phenylethyl- and 3-methylsulphinylpropyl, with 495, 495 and 385 kmol 100 g~1 DW respectively, showed the highest average concentrations. Total and individual glucosinolates in the roots and in the aerial part of the plant showed the highest concentrations in the dark cycle, at 02 : 00 h and 22 : 00 h, respectively, whilst the lowest levels were during the light cycle, mainly at 18 : 00 h. The results suggest that temperature was not a major factor in the short-term variation in glucosinolate levels. Although there was a very high sig- ni-cant di†erence between the total glucosinolate levels in the aerial part of the plant (581 kmol 100 g~1 DW) and roots (2124 kmol 100 g~1 DW), the results of the present study do not support the concept of translocation between aerial part and roots, suggesting that other factors may be involved.

Seasonal effects on bioactive compounds and antioxidant capacity of six economically important brassica vegetables.

Research on natural and bioactive compounds is increasingly focused on their effects on human health, but there are unexpectedly few studies evaluating the relationship between climate and natural antioxidants. The aim of this study was analyze the biological role of six different Brassica vegetables (Brassica oleracea L. and Brassica rapa L.) as a natural source of antioxidant compounds. The antioxidant activity may be assigned to high levels of L-ascorbic acid, total phenolics and total flavonoids of each sample. The climate seasons affected directly the concentration of bioactive components and the antioxidant activity. Broccoli inflorescences and Portuguese kale showed high antioxidant activity in Spring-Summer whilst turnip leaves did so in Summer-Winter. The Brassica vegetables can provide considerable amounts of bioactive compounds and thus may constitute an important natural source of dietary antioxidants.

The effect of light and temperature on glucosinolate concentration in the leaves and roots of cabbage seedlings

In previous studies it was shown that the concentration of total and individual glucosinolates in brassicaceous plants can vary signi-cantly over a 24-h period grown either in the -eld or under controlled conditions. The present study shows total and individual glucosinolate variation during a single day. Seedlings of cabbage grown under controlled conditions and at 14 and 15 days after emergence were moved to 20iC (Exp A) and 30iC (Exp B), with a constant photosynthetic photon Nux density of 480 lmol m~2 s~1 and 75% relative humidity, over a 2-day period, during which time aerial parts and roots were sampled at regular intervals. Whilst the glucosinolate patterns of the aerial part of the plant and of the roots remained the same, the levels of major glucosinol- ates in the aerial part, averaged over all sampling times and 2 days, were 233 ^ 60 lmol 100 g~1 DW for 3-methylsulphinylpropyl and 72 ^ 22 for 2- propenyl; in the roots, 2-phenylethyl and 1-methoxyindol-3-ylmethyl showed the highest average concentrations, with 678 ^ 355 lmol 100 g~1 DW and 411 ^ 122, respectively. Total and individual glucosinolate levels showed very high signi-cant di†erences between the two plant parts. Despite the constant temperature, light and relative humidity, glucosinolates varied within a 24-h period, showing ultradian rhythms that are common to several metabolic pro- cesses in plants. The results con-rm previous observations that at a temperature of 20iC, close to the optimum for growth and development, the diurnal variation in glucosinolate concentration, was smaller than at 30iC. 1998 Society of

Phenolics and Antioxidant Properties of Fruit Pulp and Cell Wall Fractions of Postharvest Banana (Musa acuminata Juss.) Cultivars

Banana fruits are important foods, but there have been very few studies evaluating the phenolics associated with their cell walls. In the present study, (+) catechin, gallocatechin, and (-) epicatechin, as well as condensed tannins, were detected in the soluble extract of the fruit pulp; neither soluble anthocyanidins nor anthocyanins were present. In the soluble cell wall fraction, two hydroxycinnamic acid derivatives were predominant, whereas in the insoluble cell wall fraction, the anthocyanidin delphinidin, which is reported in banana cell walls for the first time, was predominant. Cell wall fractions showed remarkable antioxidant capacity, especially after acid and enzymatic hydrolysis, which was correlated with the total phenolic content released after the hydrolysis of the water-insoluble polymer, but not for the posthydrolysis water-soluble polymer. The acid hydrolysis released various monosaccharides, whereas enzymatic hydrolysis released one peak of oligosaccharides. These results indicate that banana cell walls could be a suitable source of natural antioxidants and that they could be bioaccessible in the human gut.

Production, purification and characterisation of polysaccharides from Pleurotus ostreatus with antitumour activity

BACKGROUND Mushroom polysaccharides play an important role in functional foods because they exhibit biological modulator properties such as antitumour, antiviral and antibacterial activities. The present study involved the production, purification and characterisation of intracellular and extracellular free and protein-bound polysaccharides from Pleurotus ostreatus and the investigation of their growth-inhibitory effect on human carcinoma cell lines. RESULTS Several fermentation parameters were obtained: batch polysaccharide productivities of 0.013 ± 8.12 × 10⁻⁵ and 0.037 ± 0.0005 g L⁻¹ day⁻¹ for intracellular and extracellular polysaccharides respectively, a maximum biomass concentration of 9.35 ± 0.18 g L⁻¹ , P(max) = 0.935 ± 0.018 g L⁻¹ day⁻¹, µ(max) = 0.218 ± 0.02 day⁻¹, Y(EP/X) = 0.040 ± 0.0015 g g⁻¹ and Y(IP/X) = 0.014 ± 0.0003 g g⁻¹ . Some polysaccharides exhibited superoxide dismutase (SOD)-like activity of 50-200 units. Fourier transform infrared analysis of the polysaccharides revealed absorption bands characteristic of such biological macromolecules. Cytotoxicity assays showed that both intracellular and extracellular polysaccharides exhibited antitumour activity towards several tested human carcinoma cell lines in a dose-dependent manner. CONCLUSION The polysaccharides of P. ostreatus exhibited high SOD-like activity, which strongly supports their biological effect on tumour cell lines. The extracellular polysaccharides presented the highest antitumour activity towards the RL95 carcinoma cell line and should be further investigated as an antitumour agent.

Phytochemical characterization and antioxidant properties of baby-leaf watercress produced under organic production system

The aim of this study was to evaluate the phytochemical composition and the antioxidant properties of watercress (Nasturtium officinale R. Br., Brassicaceae) produced under organic production system. Fresh baby-leaf watercress samples were collected from local organic farms. High performance liquid chromatography with diode array detection (HPLC-DAD) and high performance liquid chromatography with diode array detection mass spectrometry (HPLC-MS) were used to assess the phytochemical composition, and spectrophotometric methods were used to assess the antioxidant capacity. Two major classes of healthier secondary plant metabolites were identified: phenolics and glucosinolates. The major phenolics determined were chlorogenic acid, quercetin-3-O-rutinoside, dicaffeoyltartaric acid, and isorhamnetin. The glucosinolates were composed exclusively of gluconasturtiin, the precursor of the anticarcinogenic, and antimicrobial compound 2-phenylethyl isothiocyanate. The extracts of organic young baby-leaf watercress presented high antioxidant capacity and this property was highly related with caffeic acid, quercetin-3-O-rutinoside, isorhamnetin, and glucosnastrutiin. The results achieved showed that baby-leaf watercress can provide high contents of antioxidant compounds at levels even superior to similar adult plant material.

Glucosinolate assessment in Brassica oleracea leaves by near-infrared spectroscopy

Glucosinolates and their hydrolysis products are of great concern because they are responsible for many of both the beneficial and harmful properties of glucosinolate-containing plants. Cabbage ( Brassica oleracea L.) is a species highly consumed as a leaf vegetable in many countries showing high qualitative and quantitative differences in glucosinolate composition among cultivars. The standard analytical techniques for determining glucosinolate composition using conventional methods lead to high costs, labour input and delays, all of which affect both the availability of data and the taking of swift decisions. In contrast, near-infrared spectroscopy (NIRS) has emerged as a rapid and cost-effective technique of analysis for many agro-food products. In this work we test the potential of NIRS for screening the total glucosinolates (t-GSL), gluconapin (GNA), gluconasturtiin (GNAST) and neoglucobrassicin (NGBS) contents of cabbage leaf cultivars coming from Portugal and Spain. NIRS calibrations resulted in coefficients of determination and standard deviation to standard error of cross-validation ratio of 0·83 and 2·38 for t-GSL; 0·70 and 1·85 for GNA; 0·62 and 1·63 for GNAST; and 0·60 and 1·58 for NGBS, respectively. An examination of the loadings of the equation for t-GSL suggested that O-H groups of water, C-H combinations of the methylene group and also N-H groups of amides were the molecular associations most strongly used in modelling total glucosinolates. It was concluded that NIRS shows a high potential as an analytical method for total and individual glucosinolate routine analysis in cabbage.