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Abstract:We report the determination of phenolic compounds in olive leaves by reversedphase HPLC/DAD, and the evaluation of their in vitro activity against several microorganisms that may be causal agents of human intestinal and respiratory tract infections, namely Gram positive (Bacillus cereus, B. subtilis and Staphylococcus aureus), Gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae) and fungi (Candida albicans and Cryptococcus neoformans). Seven phenolic compounds were identified and quantified: caffeic acid, verbascoside, oleuropein, luteolin 7-O-glucoside, rutin, apigenin 7-O-glucoside and luteolin 4'-O-glucoside. At low concentrations olive leafs extracts showed an unusual combined antibacterial and antifungal action, which suggest their great potential as nutraceuticals, particulalry as a source of phenolic compounds.

a b s t r a c tIn order to promote the use of mushrooms as source of nutrients and nutraceuticals, several experiments were performed in wild and commercial species. The analysis of nutrients included determination of proteins, fats, ash, and carbohydrates, particularly sugars by HPLC-RI. The analysis of nutraceuticals included determination of fatty acids by GC-FID, and other phytochemicals such as tocopherols, by HPLC-fluorescence, and phenolics, flavonoids, carotenoids and ascorbic acid, by spectrophotometer techniques. The antimicrobial properties of the mushrooms were also screened against fungi, Gram positive and Gram negative bacteria. The wild mushroom species proved to be less energetic than the commercial sp., containing higher contents of protein and lower fat concentrations. In general, commercial species seem to have higher concentrations of sugars, while wild sp. contained lower values of MUFA but also higher contents of PUFA. a-Tocopherol was detected in higher amounts in the wild species, while c-tocopherol was not found in these species. Wild mushrooms revealed a higher content of phenols but a lower content of ascorbic acid, than commercial mushrooms. There were no differences between the antimicrobial properties of wild and commercial species. The ongoing research will lead to a new generation of foods, and will certainly promote their nutritional and medicinal use.

Phenolic compounds of dark and clear honeys from Trás-os-Montes of Portugal were extracted with Amberlite XAD-2 and evaluated for their antioxidant and antimicrobial activities. The antioxidant effect was studied using the in vitro test capacity of scavenge the 2,2-diphenyl-1-picryhydrazyl (DPPH) free radical and of reducing power of iron (III)/ferricyanide complex. The antimicrobial activity was screened using three Gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus, Staphylococcus lentus) and three Gram-negative bacteria (Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli). The results obtained from the partial identification of honey phenolic compounds by high-performance liquid chromatography with a diode array detector showed that p-hydroxibenzoic acid, cinnamic acid, naringenin, pinocembrin and chrysin are the phenolic compounds present in most of the samples analyzed. Antioxidant potential was dependent of honey extract concentration and the results showed that dark honey phenolic compounds had higher activity than the obtained from clear honey. In the biological assays, results showed that S. aureus were the most sensitive microrganisms and B. subtilis, S. lentus, K. pneumoniae and E. coli were each moderately sensitive to the antimicrobial activity of honey extracts. Nevertheless, no antimicrobial activity was observed in the test with P. aeruginosa.

a b s t r a c tThe present study aimed to characterize five commercial honeys available in the Portuguese market in respect to their floral origins, physicochemical parameters and microbial safety and commercial quality assessment. Pollen profile, colour, moisture content, ash, acidity, electrical conductivity, pH, reducing sugars, apparent sucrose and HMF were the parameters analysed in each honey sample. Aerobic mesophiles, moulds and yeasts, fecal coliforms and sulphite-reducing clostridia were the microbial contaminants of interest studied. The antimicrobial effect against four fermentative yeasts was determined.Concerning the physicochemical parameters, all honey samples were found to meet European Legislation (EC Directive 2001/110) for all parameters, except for HMF and apparent sucrose. Microbiologically, the commercial quality was considered good and all samples showed to be negative in respect to safety parameters. We also verified that the presence of honey differentially affected the growth of fermentative yeasts under study, depending on the type of yeast, but this growth was not significantly influenced by the type of honey used.

A marked decrease in human cancers, including breast cancer, bone cancer, and cervical cancer, has been linked to the consumption of vegetable and fruit, and the corresponding chemoprotective effect has been associated with the presence of several active molecules, such as kaempferol. Kaempferol is a major flavonoid aglycone found in many natural products, such as beans, bee pollen, broccoli, cabbage, capers, cauliflower, chia seeds, chives, cumin, moringa leaves, endive, fennel, and garlic. Kaempferol displays several pharmacological properties, among them antimicrobial, anti-inflammatory, antioxidant, antitumor, cardioprotective, neuroprotective, and antidiabetic activities, and is being applied in cancer chemotherapy. Specifically, kaempferol-rich food has been linked to a decrease in the risk of developing some types of cancers, including skin, liver, and colon. The mechanisms of action include apoptosis, cell cycle arrest at the G2/M phase, downregulation of epithelial-mesenchymal transition (EMT)-related markers, and phosphoinositide 3-kinase/protein kinase B signaling pathways. In this sense, this article reviews data from experimental studies that investigated the links between kaempferol and kaempferol-rich food intake and cancer prevention. Even though growing evidence supports the use of kaempferol for cancer prevention, further preclinical and clinical investigations using kaempferol or kaempferol-rich foods are of pivotal importance before any public health recommendation or formulation using kaempferol.

Different cultivars of walnut (Juglans regia L.) leaves (Cv. Lara, Franquette, Mayette, Marbot, Mellanaise and Parisienne) grown in Portugal, were investigated in what concerns phenolic compounds and antimicrobial and antioxidant properties. Phenolics analysis was performed by reversed-phase HPLC/DAD and 10 compounds were identified and quantified: 3- and 5-caffeoylquinic acids, 3- and 4-p-coumaroylquinic acids, p-coumaric acid, quercetin 3-galactoside, quercetin 3-pentoside derivative, quercetin 3-arabinoside, quercetin 3-xyloside and quercetin 3-rhamnoside. The antimicrobial capacity was screened against Gram positive (Bacillus cereus, B. subtilis, Staphylococcus aureus) and Gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae) and fungi (Candida albicans, Cryptococcus neoformans). Walnut leaves selectively inhibited the growth of Gram positive bacteria, being B. cereus the most susceptible one (MIC 0.1mg/mL). Gram negative bacteria and fungi were resistant to the extracts at 100mg/mL. Lara walnut leaves were also submitted to antibacterial assays using 18 clinical isolates of Staphylococcus sp. Antioxidant activity was accessed by the reducing power assay, the scavenging effect on DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals and beta-carotene linoleate model system. In a general way, all of the studied walnut leaves cultivars presented high antioxidant activity (EC(50) values lower than 1mg/mL), being Cv. Lara the most effective one.

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