Efeito da salinidade da água do mar no rendimento, composição e atividades biológicas de frações polissacarídicas da Chlorophyta Caulerpa cupressoides var. flabellata

Seaweeds sulfated polysaccharides have been described as having various pharmacological activities. However, nothing is known about the influence of salinity on the structure of sulfated polysaccharides from green seaweed and pharmacological activities they perform. Therefore, the main aim of this study was to evaluate the effect of salinity of seawater on yield and composition of polysaccharides-rich fractions from green seaweed Caulerpa cupressoides var. flabellata, collected in two different salinities beaches of the coast of Rio Grande do Norte, and to verify the influence of salinity on their biological activities. We extracted four sulfated polysaccharides-rich fractions from C. cupressoides collected in Camapum beach (denominated CCM F0.3; F0.5; F1.0; F2.0), which the seawater has higher salinity, and Buzios beach (denominated CCB F0.3; F0.5; F1.0; F2.0). Different from that observed for other seaweeds, the proximate composition of C. cupressoides did not change with increased salinity. Moreover, interestingly, the C. cupresoides have high amounts of protein, greater even than other edible seaweeds. There was no significant difference (p>0.05) between the yield of polysaccharide fractions of CCM and its CCB counterparts, which indicates that salinity does not interfere with the yield of polysaccharide fractions. However, there was a significant difference in the sulfate/sugar ratio of F0.3 (p<0.05) and F0.5 (p<0.01) (CCM F0.3 and CCB F0.5 was higher than those determined for their counterparts), while the sulfate/sugar ratio the F1.0 and F2.0 did not change significantly (p>0.05) with salinity. This result suggested that the observed difference in the sulfate/sugar ratio between the fractions from CCM and CCB, is not merely a function of salinity, but probably also is related to the biological function of these biopolymers in seaweed. In addition, the salinity variation between collection sites did not influence algal monosaccharide composition, eletrophoretic mobility or the infrared spectrum of polysaccharides, demonstrating that the salinity does not change the composition of sulfated polysaccharides of C. cupressoides. There were differences in antioxidant and anticoagulant fractions between CCM and CCB. CCB F0.3 (more sulfated) had higher total antioxidant capacity that CCM F0.3, since the chelating ability the CCM F0.5 was more potent than CCB F0.5 (more sulfated). These data indicate that the activities of sulfated polysaccharides from CCM and CCB depend on the spatial patterns of sulfate groups and that it is unlikely to be merely a charge density effect. C. cupressoides polysaccharides also exhibited anticoagulant activity in the intrinsic (aPTT test) and extrinsic pathway (PT test). CCB F1.0 and CCM F1.0 showed different (p<0,001) aPTT activity, although F0.3 and F0.5 showed no difference (p>0,05) between CCM and CCB, corroborating the fact that the sulfate/sugar ratio is not a determining factor for biological activity, but rather for sulfate distribution along the sugar chain. Moreover, F0.3 and F0.5 activity in aPTT test was similar to that of clexane®, anticoagulant drug. In addition, F0.5 showed PT activity. These results suggest that salinity may have created subtle differences in the structure of sulfated polysaccharides, such as the distribution of sulfate groups, which would cause differences in biological activities between the fractions of the CCM and the CCB

Monitoramento sazonal da atividade antioxidante das folhas de Erythroxylum suberosum A.St.-Hil. : correlação com o teor de hiperosídeo e isoquercitrina

Dissertação (mestrado em Ciências Farmacêuticas)—Universidade de Brasília, Faculdade de Ciências da Saúde, Pós-graduação em Ciências Farmacêuticas, 2015.

Desenvolvimento de um aditivo natural à base de catequina: otimização da extração e estabilização a partir de frutos de Arbutus unedo L.

The common Mediterranean ornamental strawberry-tree (Arbutus unedo L.) produces an edible reddish sweet berry that is found to be bland and tasteless unless it is consumed overripe, otherwise it is discarded or used as basic agricultural sub residue. The bioactive properties of this fruit have been reported and related with phenolic compounds, mainly flavan-3-ols, such as catechin and procyanidins, which has opened the opportunity to exploit their extraction from alternative sources.The common Mediterranean ornamental strawberry-tree (Arbutus unedo L.) produces an edible reddish sweet berry that is found to be bland and tasteless unless it is consumed overripe, otherwise it is discarded or used as basic agricultural sub residue. The bioactive properties of this fruit have been reported and related with phenolic compounds, mainly flavan-3-ols, such as catechin and procyanidins, which has opened the opportunity to exploit their extraction from alternative sources. This study compares and optimizes the maceration, microwave and ultrasound extraction techniques in the recovery of a catechin extract from Arbutus unedo L. fruits and evaluate the stability of flavan-3-ols during storage and application processes. To obtain conditions that maximize the catechin extraction yield, a response surface methodology was used. Maceration and microwave extractions were found to be the most effective methods, capable of yielding 1.38±0.1 and 1.70±0.3 mg of catechin/g dry weight (dw) in the corresponding optimal extraction conditions. The optimal conditions for maceration were 93.2±3.7 min, 79.6±5.2 ºC and 23.1±3.7 % of ethanol, while for the microwave extraction were 42.2±4.1 min, 137.1±8.1 ºC and 12.1±1.1 % of ethanol. The microwave system was a quicker solution, conducting to slightly higher yields of catechin than maceration, but this one needed lower temperatures to reach similar yields. The ultrasound method was the least effective solution in terms of catechin yield extraction (0.71±0.1 mg/g at 42.4±3.6 min, 314.9±21.2 W and 40.3±3.8 %. ethanol). The stability was tested with of the catechin-enriched extract (60% flavan-3-ols and 22% catechin), obtained under the best maceration conditions, was tested. Therefore, catechin-enriched extracts were submitted to physical and chemical stability studies, considering the main affecting variables (time, temperature and pH): i) a stability study of the extracts during storage as powder system; and ii) a stability study of the extracts in simulated food environment (aqueous solution system). The measured responses were the flavan-3-ols and catechin contents, determined by HPLC-DAD, and the antioxidant activity of the extracts evaluated by hydrophilic assays. Mechanistic and phenomenological equations were used to describe the responses, and the optimal conditions for flavan-3-ols (including catechin) stability as powder extract during a month were pH= 5.4 and T= -20ºC; while its stability in aqueous solution remained during the 24 h of application at pH<4 and T<30ºC. This study compares and optimizes the maceration, microwave and ultrasound extraction techniques in the recovery of a catechin extract from Arbutus unedo L. fruits and evaluate the stability of flavan-3-ols during storage and application processes. To obtain conditions that maximize the catechin extraction yield, a response surface methodology was used. Maceration and microwave extractions were found to be the most effective methods, capable of yielding 1.38±0.1 and 1.70±0.3 mg of catechin/g dry weight (dw) in the corresponding optimal extraction conditions. The optimal conditions for maceration were 93.2±3.7 min, 79.6±5.2 ºC and 23.1±3.7 % of ethanol, while for the microwave extraction were 42.2±4.1 min, 137.1±8.1 ºC and 12.1±1.1 % of ethanol. The microwave system was a quicker solution, conducting to slightly higher yields of catechin than maceration, but this one needed lower temperatures to reach similar yields. The ultrasound method was the least effective solution in terms of catechin yield extraction (0.71±0.1 mg/g at 42.4±3.6 min, 314.9±21.2 W and 40.3±3.8 %. ethanol). The stability was tested with of the catechin-enriched extract (60% flavan-3-ols and 22% catechin), obtained under the best maceration conditions, was tested. Therefore, catechin-enriched extracts were submitted to physical and chemical stability studies, considering the main affecting variables (time, temperature and pH): i) a stability study of the extracts during storage as powder system; and ii) a stability study of the extracts in simulated food environment (aqueous solution system). The measured responses were the flavan-3-ols and catechin contents, determined by HPLC-DAD, and the antioxidant activity of the extracts evaluated by hydrophilic assays. Mechanistic and phenomenological equations were used to describe the responses, and the optimal conditions for flavan-3-ols (including catechin) stability as powder extract during a month were pH= 5.4 and T= -20ºC; while its stability in aqueous solution remained during the 24 h of application at pH<4 and T<30ºC.

Investigation on possibility of survival and growth enhancing by selected seaweeds from Persian Gulf in post larvae shrimp Litopenaeus vannamei

The use of antibiotics in aquaculture has been limited. Scientifics seeking for natural substitutes to prevent of aquatic animals diseases. Considering seaweeds are rich of nutritions and bioactive compounds, the purpose of this study is: investigation the potential and use possibility of native seaweeds from Persian Gulf in shrimp aquculture industry to improve growth, survival of postlarvae and to resistance against pathogens such as vibriosis. For this propose 7 macroalgae species from Bushehr province coast, inclouding: green algae (C. iyengarii), brown algae (S. angutifolium and S. ilicifolium) and red algae (L. snyderiae, K. alvarezii and G. corticata) were collected and identified. Then seaweed extracts abtained by Water, Ethanol, Methanol and Chloroform solvents by soaking method. In vitro antibacterial activity of extracts against Gr+ bacteria (S. aureus and B. subtilis) and Gr- bacteria (V. harveyi, V. alginolyticus and E. coli) was conducted by Agar diffusion, MIC and MBC methods. Antioxidant activity also by DPPH and EC50 methods was investigated. According to results of these two tests four seaweeds species (S. angutifolium, L. snyderiae, K. alvarezii and G. corticata) were selected for use in shrimp postlarvae (PL22) diets by Bio-Encapsulation (Artemia enrichment). Before of enrichment, toxicity effect of extracts to Artemia nauplii were evaluated by determination of LC50 24 h method. From results of this section Ethanol extracts were selected to bioencapsulation. After encapsulation shrimp postlarvae divided to 12 groups in triplicate, namely: C-, C+, S (200), S (400), S (600), L(200), L(400), L(600), G(300), G(600), K(300) and K(600). During 30 days of reared period C- and C+ use of basal diet and unenriched Artemia, but the other groups use of basal diet and enriched Artemia. Except C-, the shrimps in first day of culture put in 107 cfu/ml v. harveyi suspension for 30 minutes, and after water exchange 10 ml of this dose was added to reared aquaria. After 30 days survival percentage, obtained weight and SGR% were investigated. To evaluate vibrio loading, every 10 days 5 postlarvae were sampled randomly for vibrio count. Results showed that vibrio count in C- was less than the others and in C+ was more than the others. In treatments vibrio count in L(200) was the most and L(600) was the less. Survival rate in C- was the most and after that G(600) with 79.4±6.6% and then S(300) and K(600) were 73.3±7.3% and 70.6±6.6% respectively that were significantly compare the other (P < 0.01). Also the C+ was the less with 33.3±6.6% that difference was significant (P< 0.01). In this study growth parameters of all groups that fed by enriched Artemia were better than C+ (P<0.05). After cultre period 10 shrimp of every aquarium disinfected and reared for 10 days like before treatment. After 10 days the shrimps were challenged by 3×108 cfu/ml V. harveyi and mortality was recorded for 7 days. The all of animals in C- were survive but more than 90% of C+ were dead. And survival in all of treatments were better the C+ (P<0.05). The study showed the ethanol extracts of selected seaweed from Persian Gulf is a good source for growth, Survival and disease control in shrimp larviculture.

Viability of probiotic bacteria and some chemical and sensory characteristics in cornelian cherry juice during cold storage

Background: Increased popularity of vegetarianism, lactose intolerance, and the high cholesterol content in dairy products, are all factors that have recently increased the demand for nondairy probiotic products. The objective of this study is to evaluate the effect of refrigeration on the viability of probiotics and asses someof the chemical and sensory characteristics in cornelian cherry juice. Results: The Iranian native probiotic strain (L. casei T4) showed greater viability compared to industrial types (viable count of 8.67 log cfu/mL versus <6.0 log cfu/mL at d 28). However, this most tolerant Iranian strain, could not withstand the conditions of ‘Natural juice’ at pH 2.6 for more than 7 d. Following a pH adjusted treatment (to pH ~3.5), the viability of the strain was improved to 28 d with some evidence of increased growth of the probiotic. However, the level of antioxidant activity, anthocyanin and phenolic compounds, revealed a slight decrease during cold storage. The changes in the chemical profile of the sample containing L. casei T4 indicated fermentation activity during cold storage. Sensory evaluation results showed significant differences between samples containing L. casei TD4 and other samples in taste, odor and overall acceptance in a complimentary way. Conclusions: The results showed that low pH and presence of inhibitor phenolic compounds of cornelian cherry juice have negative effect on viability of probiotics, especially industrial strains during refrigerated storage.

Caracterização físico-química e efeitos de frações polissacarídicas da alga amansia multifida na coagulação, inflamação, radicais livres e viabilidade celular

Galactans are polysaccharides sulfated present in the cell wall of red algae. Carrageenans are galactans well known in the food industry as gelling polysaccharides and for induce inflammatory process in rodents as animal model. The extraction of polysaccharides from A. multifida has been carried out by proteolysis and precipitation in different volumes of acetone, which produced three fractions (F1, F2, and FT). Chemical and physical analyses revealed that these fractions are sulfated galactan predominantly. Results of the antioxidant activity assays showed that all of these fractions have antioxidant activity and that was associated with sulfate content of the analysis of reducing power and total antioxidant capacity. However, these fractions were not effective against lipid peroxidation. The fraction FT presented higher activity on the APTT test at 200 μg (> 240 s). The assessment of the hemolytic activity showed that the FT fraction has the best activity, increasing lyses by the complement system to 42.3% (50 μg) (p< 0,001). The fraction FT showed the best yield, anticoagulant and hemolytic activity between the three fractions and therefore it was choose for the in vivo studies. The Inflammation assessment using the FT fraction (50 mg / kg MB) showed that the cellular migration and the IL-6 production increased 670.1% (p< 0,001) and 531.8% (p< 0,001), respectively. These results confirmed its use as an inflammation inducer in animal model. Cytotoxicity assay results showed that all fractions have toxic effects on 3T3 and HeLa cells after exposition of 48 hours, except when 100 μg for both F1 and FT were used. These results arise the discussion whether these polysaccharides it should be used as additive in foods, cosmetics and medicines.

Atividades anticoagulante e antioxidante de extratos brutos ricos em polissacarídeos sulfatados das macroalgas marinhas marrons Canistrocarpus cervicornis, Dictyota mertensii e Dictyopteris delicatula e de Heterofucanas de Canistrocarpus cervicornis

In the present study, extracts rich-sulfated polysaccharides were obtained from three different species of Dictyotales (a class of brown macroalgae): Canistrocarpus cervicornis, Dictyota mertensii and Dictyopteris delicatula and their anticoagulant and antioxidant activities were evaluated. All extracts showed anticoagulant activity on aPTT assay, but not on PT assay. Extracts also exhibited total antioxidant activity, superoxide radical scavenging capacity and ferric chelating property. The extract from C. cervicornis showed the best results and was choose to have their sulfated polysaccharides fractioned and subsequently analysed. Thus, six fractions (CC-0.3, CC-0.5, CC-0.7, CC-1.0, CC-1.2 and CC-2.0) were obtained by proteolysis followed by sequential acetone precipitation. Agarose gel eletrophoresis stained with blue toluidine, confirmed the presence of sulfated polysaccharides in all fractions. Chemical analyses showed that all fractions presented heterofucans mainly constitued by fucose, galactose, glucuronic acid and sulfate. Any fraction changed the PT. However, all fractions were able to double the aPTT on a dose-dependent manner. CC- 0.3, CC-0.5, CC-0.7 and CC-1.0 needed only 0.100 mg/mL to double the aPTT, result only 1.25 times higher than the Clexane® (0.080 mg/mL), a commercial low molecular heparin. The heterofucans presented appreciable total antioxidant capacity, low capacity on scavenging hydroxyl radical and good efficiency on scavenging superoxide radicals (except CC-1.0). CC-1.2 showed 43.1 % on superoxide radical scavenging. This result was higher than that showed by the same concentration of gallic acid (41.8 %), a known antioxidant. Furthermore, the heterofucans showed excelent activity on ferrous chelating activity (except CC-0.3). CC-0.5, CC-0.7 and CC-1.0 showed the highest activities with 47.0 % of ferrous chelating activity, a result 2.0 times lesser than that exhibited by the same concentration of EDTA. These results clearly indicated the beneficial effects of heterofucans extracted from C. cervicornis as potential anticoagulant and antioxidant agents. However additional steps of purification, structural studies, besides in vivo experiments are needed for these fucans may be used as therapeutic agents

Composição química e atividade antioxidante de maracujá-roxo (Passiflora edulis Sims edulis)em diferentes graus de maturação

O maracujá-roxo é um fruto tropical que tem ganho destaque devido ao seu valor nutricional, organolético e às emergentes descobertas acerca das suas propriedades farmacêuticas e antioxidantes. Durante o processo de maturação decorrem várias reações de ordem física e química, e no intuito de enriquecer o conhecimento acerca de como a composição química e as propriedades do maracujá-roxo evoluem ao longo da maturação, este trabalho consistiu na caracterização física do fruto inteiro e na caracterização química geral das sementes, casca e polpa em frutos separados em cinco graus de maturação diferentes (G1 a G5). Foi também avaliada a atividade antioxidante (atividade antiradicalar e poder redutor), e o teor em compostos fenólicos (fenóis totais, derivados do ácido hidroxicinâmico e flavonóis) das diferentes matrizes que compõem o fruto ao longo da maturação. A partir dos resultados obtidos verificou-se na casca que o teor em cinza bruta aumenta essencialmente entre G1 e G2 enquanto se verificou uma diminuição da proteína. Em relação à semente, o teor em cinza bruta aumenta gradualmente, e o teor em proteína aumenta de G1 para G2, estabilizando posteriormente nos 8% (base seca). O teor em gordura aumenta gradualmente ao longo da maturação, verificando-se um maior acumulo entre G1 e G2 (9,9 e 19,1% respetivamente). No que respeita à polpa, há uma diminuição nos teores de cinza e proteína e aumento dos sólidos solúveis totais.Verificou-se um aumento ligeiro do pH ao longo da maturação (entre 2,8 a 3,1) e uma diminuição da acidez (entre 12,1 e 6,7 g de ácido cítrico 100 mL-1). Constatou-se que os açúcares predominantes na polpa foram a sacarose, frutose e glucose. Quanto aos ácidos orgánicos, o ácido cítrico foi o maioritário em todos os graus de maturação e teores mais baixos foram quantificados para os ácidos málico e ascórbico. Entre as diferentes partes do fruto estudadas, as cascas foram a matriz mais antioxidante, aumentando o seu potencial bioativo durante a maturação. As sementes apresentaram valores mais elevados de fenóis totais, derivados do ácido hidroxicinâmico e flavonóis. Foi verificado que a atividade antioxidante esteve correlacionada com os valores de fenóis totais presentes nas diferentes partes do fruto ao longo da maturação. De acordo com o conhecimento dos autores, este é o primeiro estudo que toma em consideração as alterações sofridas pelas diferentes partes do maracujá-roxo produzido em Portugal, ao longo da maturação.

Dietary supplements based on borututu: wich is the most suitable choice for bioactive purposes?

Borututu ( Cochlospermum angolensis Welw.) is a widespread tree in Angola used since antiquity by traditional healers for the prevention and treatment of hepatic diseases and for the prophylaxis of malaria [1]. This plant is mostly consumed as infusions but is also available as dietary supplements, such as piiis, capsules, and syrups, among others. In the present study, the aim was to evaluate the proximate composition and energetic contribution of borututu as weii as its composition in hydrophilic (sugars and organic acids) and lipophilic (fatty acids and tocopherols) compounds, given the fact that this plant is directly introduced in some dietary supplements. Furthermore, the bioactivity (antioxidant, hepatoprotective and antimicrobial activities) of three different formulations of borututu (infusion, pills, and syrup) was assessed and compared, and since plant beneficial properties are often ascribed to phenolic compounds [2], the phenolic profile of the formulations was also analysed. Carbohydrates (88 g/100 g) and fat (2.5 g/100 g) were the major and tl1e minor components of the studied borututu dry barks, respectively, with an energetic contribution of 384 kcal/100 g. Fructose was the most abundant sugar (1.3 g/100 g), foilowed by sucrose, trehalose and glucose (1.1, 0.98 and 0.79 g/100 g, respectively). Oxalic (0.70 g/100 g), malic (0.63 g/100 g) and citric (0.57 g/100 g) acids were present in higher amounts but shikimic and fumaric acids were also detected. Among the fatty acids found in borututu, a prevalence of saturated fatty acids (SF A; 48.2%) was observed, whereas polyunsaturated (PUFA) and monounsaturated (MUFA) fatty acids were detected in relative percentages of 30.9% and 20.8%, respectively. P-tocopherol was the most abundant of the four isoforms found in the sample, foiiowed by o-, a- and y-tocopherol, present in concentrations of 597,43, 3.7 and 2.0 g/100 g, respectively. Borututu infusion revealed the highest antioxidant activity, with EC50 values ranging from 20 to 600 J.lg/mL and was the only formulation inhibiting the growth of an HepG2 ceii line, with a Gl5o value of 146 J.lg/mL. This formulation.also revealed the best antimicrobial capacity and proved to be able to inhibit the growth of Escherichia coli, E. coli ESBL, Staphylococcus aureus and Pseudomonas aeruginosa, with MIC values of 50, 6.2, 1.6 and 25 mg!mL, respectively. Pills revealed activity against some of the studied bacterial strains and the syrup did not reveal antimicrobial activity at the studied concentration. Eilagic acids, methyl ellagic acids, eucaglobulinlglobulusin B and (epi)gaiiocatechin-0-gallate were the compounds present in all the different formulations. The highest concentration of phenolic compounds was found in the infusion extract. Protocatechuic acid was the most abundant phenolic compound in the infusions, the only preparation where it was detected, whereas ( epi)gaiiocatechin- 0-gallate was the main phenolic in the pills and eucaglobulinlglobulusin in the syrup. In a general way, borututu proved to be a good source of phytochemicals such as phenolic compounds, with the infusions revealing the best bioactive properties.

Crocin and β-Carotene bleaching assays as analytical tools in the optimization of the extraction of hydrophilic and lipophilic antioxidants from tomato

Tomato (Lycopersicon esculentum Mill.) is the second most important vegetable crop worldwide and a rich source of hydrophilic (H) and lipophilic (L) antioxidants. The H fraction is constituted mainly by ascorbic acid and soluble phenolic compounds, while the L fraction contains carotenoids (mostly lycopene), tocopherols, sterols and lipophilic phenolics [1,2]. To obtain these antioxidants it is necessary to follow appropriate extraction methods and processing conditions. In this regard, this study aimed at determining the optimal extraction conditions for H and L antioxidants from a tomato surplus. A 5-level full factorial design with 4 factors (extraction time (I, 0-20 min), temperature (T, 60-180 •c), ethanol percentage (Et, 0-100%) and solid/liquid ratio (S/L, 5-45 g!L)) was implemented and the response surface methodology used for analysis. Extractions were carried out in a Biotage Initiator Microwave apparatus. The concentration-time response methods of crocin and P-carotene bleaching were applied (using 96-well microplates), since they are suitable in vitro assays to evaluate the antioxidant activity of H and L matrices, respectively [3]. Measurements were carried out at intervals of 3, 5 and 10 min (initiation, propagation and asymptotic phases), during a time frame of 200 min. The parameters Pm (maximum protected substrate) and V m (amount of protected substrate per g of extract) and the so called IC50 were used to quantify the response. The optimum extraction conditions were as follows: r~2.25 min, 7'=149.2 •c, Et=99.1 %and SIL=l5.0 giL for H antioxidants; and t=l5.4 min, 7'=60.0 •c, Et=33.0% and S/L~l5.0 g/L for L antioxidants. The proposed model was validated based on the high values of the adjusted coefficient of determination (R2.wi>0.91) and on the non-siguificant differences between predicted and experimental values. It was also found that the antioxidant capacity of the H fraction was much higher than the L one.

From the field to the table: ionizing radiation as a feasible postharvest treatment for fresh and dried plant foods

Food irradiation is a treatment that involves subjecting in-bulk or packaged food to a controlled dose of ionizing radiation, with a clearly defined goal. It has been used for disinfestation and sanitization of food commodities and to retard postharvest ripening and senescence processes, being a sustainable alternative to chemical agents 1 . Doses up to 10 kGy are approved by several international authorities for not offering negative effects to food from a nutrition and toxicology point of view 2 . However, the adoption of this technology for food applications has been a slow process due to some misunderstandings by the consumer who often chooses non-irradiated foods. In this study, the effects of the ionizing radiation treatment on physical, chemical and bioactive properties of dried herbs and its suitability for preserving quality attributes of fresh vegetables during cold storage were evaluated. The studied herbs, perennial spotted rockrose (Tuberaria lignosa (Sweet) Samp.) and common mallow (Malva neglecta Wallr.) were freeze-dried and then irradiated up to 10 kGy in a Cobalt-60 chamber. The selected vegetables, watercress (Nasturtium officinale R. Br.) and buckler sorrel (Rumex induratus Boiss. Reut.) were rinsed in tap water, packaged in polyethylene bags, submitted to irradiation doses up to 6 kGy and then were stored at 4 C for a period of up to 12 days. Physical, chemical and bioactive parameters of irradiated and non-irradiated samples were evaluated using different methodologies the colour was measured with a colorimeter, individual chemical compounds were analyzed by chromatographic techniques, antioxidant properties were evaluated using in vitro assays based on different reaction mechanisms, and other quality analyses were performed following official methods of analysis. The irradiation treatment did not significantly affect the colour of the perennial spotted rockrose samples, or its phenolic composition and antioxidant activity 3 . Medium doses preserved the colour of common mallow and a low dose did not induce any adverse effect in the organic acids profile. The green colour of the irradiated vegetables was maintained during cold storage but the treatment had pros and cons in other quality attributes. The 2 kGy dose preserved free sugars and favoured polyunsaturated fatty acids (PUFA) while the 5 kGy dose favoured tocopherols and preserved the antioxidant properties in watercress samples. The 6 kGy dose was a suitable option for preserving PUFA and the ω-6 ω-3 fatty acids ratio in buckler sorrel samples. This comprehensive experimental work allowed selecting appropriate processing doses for the studied plant foods in order to preserve its quality attributes and edibility.

A prospective study on bioactive properties of wild mushrooms mycelium grown in vitro under different conditions

Wild mushrooms have been extensively studied for their value as sources of high quality nutrients and of powerful physiologically bioactive compounds [1,2]. The present study was designed to evaluate the in vitro development of two wild edible mushroom species: Pleurotus eryngii (DC.) Quél. and Suillus belinii (Inzenga) Watling, by testing different solid (Potato Dextrose Agar medium –PDA and Melin-Norkans medium- MMN) and liquid culture media (Potato dextrose broth- PDB and Melin-Norkans medium- MMN). Each strain of mushroom produces a special type of mycelium and this range of characteristics varies in form, color and growth rate. S. bellinii presents a pigmented and rhizomorphic mycelia, whereas, P. eryngii has depigmented and cottony mycelia. The mycelium isolated and grown in PDA showed a faster radial growth compared to the mycelium isolated and grown in both solid and liquid incomplete MMN medium. P. eryngii exhibited a rapid growth and a higher mycelia biomass in both medium compared to S. belinii. Moreover, the obtained mycelia will be characterized in terms of well-recognized bioactive compounds namely, phenolic acids and mycosterols (mainly ergosterol), by using high performance liquid chromatography coupled to diode array and ultraviolet detectors, respectively. These compounds will be correlated to mycelia bioactivity: i) antioxidant activity, evaluated through free radicals scavenging activity, reducing power and lipid peroxidation inhibition in vitro assays; ii) anti-inflammatory activity, assessed through nitric oxide production inhibition in murine macrophages (RAW 264.7 cell line); iii) cytotoxic activity, evaluated either in human tumor cell lines (MCF-7- breast adenocarcinoma, NCIH460- non-small cell lung cancer, HeLa- cervical carcinoma and HepG2- hepatocellular carcinoma) as also in a non-tumor porcine primary liver cells culture established in-house (PLP2). Overall, our expectation is that the bioactive formulations obtained by in vitro culture can be applied as nutraceuticals or incorporated in functional foods.

Utilization of free and microencapsulated Rosmarinus oficinalis L. extracts as bioactives ingredientes for new functional foods developmen

Currently, many consumers search for food with functional characteristics beyond their nutritional properties. Thus, the concept of functional food becomes a hot topic, allowing the obtaining of health benefits, including disease prevention. In this context, plants are recognized as sources of a wide range of bioactives, mainly phenolic compounds. In particular, the Rosmarinus officina/is L., commonly referred as rosemary, has several phenolic compounds with different bioactive properties such as antioxidant, antiinflammatory and antimicrobial activities, among others [!]. Hence, this plant has great potential for incorporation into foods in order to confer bioactivity to the final products. However, it should be highlighted that the bioactive compounds if exposed to adverse environments, for example: light, moisture, extreme pH, storage, food processing conditions, can be degraded leading to the consequent loss of bioactivity [2]. The microencapsulation is an alternative to overcome this problematic of bioactive compounds, as also to ensure controlled release, or target deliver to a specific site [3]. In this work, lyophilized rosemary aqueous extract prepared by in:'usion was used as a functional ingredient for cottage cheeses, after proving that it possesses, both higher content in phenolic compounds and higher antioxidant activity, comparatively with the corresponding hydroethanolic extract. The rosemary aqueous extract revealed, for example, a DPPH scavenging activity with an EC50 value of 73.44±0.54j!g/mL and presented as main phenolic compound the caffeic acid dimer, commonly named as rosmarinic acid. For the functionalized cottage cheeses, a decrease of bioactivity was observed after seven days under storage in fridge, when the extracts were incorporated in its free form. Therefore, to preserve the antioxidant activity, the rosemary aqueous extract was efficiently microencapsulated by using an atomization/coagulation technique and alginate as the matrix material and thereafter incorporated into the cottage cheeses. The final microspheres showed a size, estimated by OM using a magnification of I OOx, ranging between 51.1 and 122.6 J!m and an encapsulation efficiency, estimated through an indirect method, approaching 100%. Overall, the introduction of both free and microencapsulated extracts did not change the nutritional value of cottage cheeses, providing bioactivity that was more preserved with microencapsulated extracts putting in evidence the importance of using microencapsulation to develop effective functional foods.

Micoencapsulation of bioactive plant extracts: a tool to functionalize cottage cheeses

Foeniculum vulgare Mill. (fennel) and Matricaria recutita L. (chamomile) are two examples of plants with reported antioxidant and antimicrobial properties, which can be related with their composition in phenolic compounds [1,2]. Furthermore, according to previous results of our research group, the direct incorporation of the aqueous extracts showed capacity to maintain the nutritional properties of the cottage cheeses, up to 7 days of storage, while improving the antioxidant potential. However, after 14 days, a decrease in the antioxidant properties was observed [1,2], which can be related with factors such as light, moisture, temperature and pH, that can cause bioactive compounds degradation. Therefore, the aim of the present study was to prepare microcapsules with the aqueous extracts of fennel and chamomile for incorporation in cottage cheese samples, in order to protect the bioactive molecules present in the extracts, such as phenolic compounds, and prevent the decrease of the antioxidant activity observed after the 14 days period. The microspheres were prepared using an atomization/coagulation technique. Sodium alginate was used as the matrix material to produce the microspheres that were characterized through optical microscopy (OM), during and after atomization, for inspecting morphology. The encapsulation efficiency (EE) was determined by HPLC-DAD by an indirect method by analysing the coagulation solution. FTIR was also used to attest the presence of the extract inside of the alginate matrix. These microencapsulated extracts were incorporated in cottage cheese samples that were further characterized in terms of nutritional properties and antioxidant potential right after incorporation, and after 7 and 14 days of storage at 4•c. The EE was estimated as -100% and the FTIR analysis confirmed the presence of the extracts inside the microspheres. The results showed that the incorporation of the microencapsulated extracts did not cause changes in the nutritional value of cottage cheeses (through a comparison with control samples without extracts). The predominant fatty acids were palmitic (C16:0) and oleic (CI8:0) acids. The order of abundance of fatty acids was as follows: saturated fatty acids (SF A)> monounsaturatcd fatty acids (MUF A)> polyunsaturated fatty acids (PUF A). Regarding free sugars, lactose was the only sugar identified and quantified in all samples. Regarding the antioxidant activity, the samples functionalized with the microencapsulated extracts showed a higher preservation of this property even after the 7th day of storage. Overall, the incorporation of the protected plant extracts in dairy foods can be a strategy to provide health benefits to consumers.

Propriedades antioxidante, anti-hemostástica e antiproliferativa de galactanas sulfatadas da alga vermelha hypnea musciformis (wulfen) j. V. Lamouroux

Marine algae are one of the major sources of biologic compounds. In extracellular matrix of these organisms there are sulfated polysaccharides that functions as structural components and provides protection against dehydration. The fraction 1.0 (F1.0) rich in sulfated galactans obtained from red seaweed Hypnea musciformis was physicochemical characterized and evaluated for pharmacologic activity through antioxidant activity, cytotoxic action on erythrocytes, anticoagulant, stimulatory action under antithrombotic heparan sulfate synthesis and their effects on cell proliferation and cycle cell progression. The main components of F1.0 were carbohydrates (49.70 ± 0.10%) and sulfate (44.59 ± 0.015%), presenting phenolic compounds (4.79 ± 0.016%) and low protein contamination (0.92 ± 0.001%). Fraction 1.0 showed polidisperse profile and signs in infrared analysis in 1262, 1074 and 930, 900 and 850 attributed to sulfate esters S=O bond, presence of a 3,6- anidrogalactose C-O bond, non-sulfated β-D-galactose and a C-O-SO4 bond in galactose C4, respectively. The fraction rich in sulfated galactans exhibited strong antioxidant action under lipid peroxidation assay with IC50 of 0.003 mg/mL. Besides the inhibition of hemolysis induced by H2O2 in erythrocytes treated with F1.0, this fraction did not promote significant cytotoxity under erythrocytes membranes. F1.0 exhibited low anticoagulant activity causing moderate direct inhibition of enzimatic activity of thrombin. This fraction promoted stimulation around of 4.6 times on this synthesis of heparan sulfate (HS) by rabbit aortic endothelial cells (RAEC) in culture when was compared with non treated cells. The fraction of this algae displayed antiproliferative action under RAEC cells causing incresing on cell number on S fase, blocking the cycle cell progression. Thus F1.0 presented cytostatic and no cytotoxic action under this cell lineage. These results suggest that F1.0 from H. musciformis have antioxidant potential which is a great effect for a compound used as food and in food industry which could be an alternative to food industry to prevent quality decay of lipid containing food due to lipid peroxidation. These polysaccharides prevent the lipid peroxidation once the fraction in study exhibited strong inhibitory action of this process. Furthermore that F1.0 present strong antithrombotic action promoting the stimulation of antithrombotic HS synthesis by endothelial cells, being important for thrombosis preventing, by its inhibitory action under reactive oxygen species (ROS) in some in vitro methods, being involved in promotion of hypercoagulability state.