Maria João Rodrigues

Characterization and identification of microflora from soaked cod and respective salted raw materials

The microflora of soaked cod products from salted and dried salted cod were identified to the species level by the Vitek® automated identification system using the Gram-positive identification (GPI) and Gram-negative identification plus (GNI+) cards, and a cluster analysis was performed to determine the similarity among the strains. Within the Gram-positive bacteria, Staphylococcus epidermidis, S. hominis, S. warneri, S. haemolyticus, S. simulans and S. saprophyticus were found. Regarding Gram-negative, Stenotrophomonas maltophilia, Vibrio alginolyticus, Pasteurella haemolytica, Enterobacter agglomerans, E. asburiae, E. cloacae, Pseudomonas putida, P. fluorescens, Shewanella putrefaciens, Morganella morganii, Actinobacillus urea, Acinetobacter lwoffi/junii and non-fermentative bacilli were identified. Several species found in the soaked products were also isolated from the salted and dried salted cod. Thus, despite of the low water activity of these products, the mentioned strains were able to survive. Some of these, had the ability to produce H2S and/or presented decarboxylase activity, therefore it is probable that the growth of these bacteria during soaking might contribute to the spoilage of soaked cod. Aeromonas hydrophila and A. caviae were not isolated from the soaked food, but from the raw material (dried salted cod). Their presence might constitute a potential hazard, especially considering that these foods may be eaten raw.

Maritime Halophyte Species from Southern Portugal as Sources of Bioactive Molecules

Extracts of five halophytes from southern Portugal (Arthrocnemum macrostachyum, Mesembryanthemum edule, Juncus acutus, Plantago coronopus and Halimione portulacoides), were studied for antioxidant, anti-inflammatory and in vitro antitumor properties. The most active extracts towards the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical were the methanol extracts of M. edule (IC50 = 0.1 mg/mL) and J. acutus (IC50 = 0.4 mg/mL), and the ether extracts of J. acutus (IC50 = 0.2 mg/mL) and A. macrostachyum (IC50 = 0.3 mg/mL). The highest radical scavenging activity (RSA) against the 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical was obtained in the ether extract of J. acutus (IC50 = 0.4 mg/mL) and H. portulacoides (IC50 = 0.9 mg/mL). The maximum total phenolic content (TPC) was found in the methanol extract of M. edule (147 mg gallic acid equivalents (GAE)/g) and in the ether extract of J. acutus (94 mg GAE/g). Significant decreases in nitric oxide (NO) production were observed after incubation of macrophages with lipopolysaccharide (LPS) and the chloroform extract of H. portulacoides (IC50 = 109 µg/mL) and the hexane extract of P. coronopus (IC50 = 98.0 µg/mL). High in vitro cytotoxic activity and selectivity was obtained with the ether extract of J. acutus. Juncunol was identified as the active compound and for the first time was shown to display selective in vitro cytotoxicity towards various human cancer cells.

In vitro antioxidant and anti-inflammatory properties of Limonium algarvense flowers' infusions and decoctions: A comparison with green tea (Camellia sinensis).

This work reports the in vitro antioxidant and anti-inflammatory activities and toxicity of infusions and decoctions of Limonium algarvense flowers, and green tea. The total contents in different phenolic groups and the quantification of individual phenolics by HPLC are also reported. L. algarvense and green tea had similar antioxidant properties, except for hydroxyl radical-scavenging activity, higher on green tea, and iron chelating potential, higher on L. algarvense. The later species also had the uppermost anti-inflammatory potential. Green tea decoction had the highest content of phenolic groups, but the infusion of L. algarvense had higher amounts of salicylic, gallic and coumaric acids. L. algarvense was not toxic, whereas green tea was toxic for S17 cells. Under our experimental conditions, infusions and decoctions of L. algarvense flowers had similar or higher antioxidant and anti-inflammatory properties than green tea, and thus, may be useful for alleviating symptoms associated with oxidative and inflammatory-related diseases.

Botryococcus braunii and Nannochloropsis oculata extracts inhibit cholinesterases and protect human dopaminergic SH-SY5Y cells from H2O2-induced cytotoxicity

Extracts of Botryococcus braunii and Nannochloropsis oculata were evaluated for inhibitory activity against acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and tyrosinase (TYRO) and capacity to attenuate hydrogen peroxide (H2O2)-induced injury in the human dopaminergic cell line SH-SY5Y. We also report the antioxidant activity, the total phenolic content (TPC) and the fatty acid (FA) profile of these microalgae. Both species had low levels of TPC and considerable amounts of polyunsaturated fatty acids (PUFA). The highest radical scavenging activity (RSA) against 1,1-diphenyl-2-picrylhydrazyl (DPPH) was observed in the acetone extract of B. braunii and in the diethyl ether extracts of both strains. The acetone extract of B. braunii had the highest RSA against 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid). The extracts had a higher capacity to chelate iron than copper, and the highest iron chelation was achieved with the hexane extract of N. oculata. The diethyl ether and water extracts of the latter species also displayed the highest copper chelation. Except for the acetone extract of B. braunii and the water extract of N. oculata, all samples inhibited AChE, especially the hexane extract of N. oculata. Samples had moderate BChE inhibition and no effect towards TYRO. Almost all samples effectively protected neuronal cells against oxidative stress induced by H2O2. These results suggest possible novel applications of biomass from those microalgae in the pharmaceutical industry and/or as functional foods.

Novel approach to bis(indolyl)methanes: de novo synthesis of 1-hydroxyiminomethyl derivatives with anti-cancer properties.

A versatile and broad range approach to previously unknown bis(indolyl)methane oximes based on two consecutive hetero Diels-Alder cycloaddition reactions of electrophilic conjugated nitrosoalkenes with indoles is disclosed. The cytotoxic properties and selectivity of some adducts against several human cancer cell lines pointing to a promising role in the development of anti-tumoural drugs, in particular for leukaemia and lymphoma.

Methanol extracts from Cystoseira tamariscifolia and Cystoseira nodicaulis are able to inhibit cholinesterases and protect a human dopaminergic cell line from hydrogen peroxide-induced cytotoxicity

Abstract Context Marine macroalgae contain several bioactive molecules that may be developed as functional foods, but information about their neuroprotective potential is scarce. Objective The objective of this study is to determine the in vitro antioxidant and neuroprotective features of marine algae from the southern coast of Portugal and to assess the total content of different types of bioactives. Materials and methods Methanol extracts from 21 macroalgal species from the southern Portugal were evaluated for in vitro antioxidant and acetylcholinesterase (AChE) inhibition. Active extracts were further evaluated for inhibitory activity against butyrylcholinesterase (BuChE) and tyrosinase (TYRO), and for their ability to attenuate hydrogen peroxide (H2O2)-induced toxicity in SH-SY5Y cells. The total contents of different phenolic groups were determined for the most active extracts. Results Cystoseira tamariscifolia (Hudson) Papenfuss (Sargassaceae) had the highest antiradical activity (92%, 1 mg/mL). Cystoseira nodicaulis (Withering) M. Roberts (Sargassaceae) (75%) and Cystoseira humilis Schousboe ex Kützing (Sargassaceae) (70%) had the highest iron-chelating activity at 10 mg/mL. Cystoseira baccata (S.G. Gmelin) P.C. Silva (Sargassaceae) was more active towards copper (66%, 10 mg/mL). Cystoseira tamariscifolia had the highest AChE inhibitory capacity (85%, 10 mg/mL). Cystoseira tamariscifolia and C. nodicaulis were also active against BuChE and TYRO, and were able to protect SH-SY5Y cells against oxidative stress induced by H2O2. Cystoseira tamariscifolia had the highest content of all the groups of phenolics, and was particularly enriched in hydroxycinnamic acids (106 mg CAE/g DW). Discussion and conclusion Results indicate that C. tamariscifolia and C. nodicaulis are important sources of nutraceutical compounds and may be considered functional foods that could improve cognitive functions.

Biological Activities and Chemical Composition of Methanolic Extracts of Selected Autochthonous Microalgae Strains from the Red Sea

Four lipid-rich microalgal species from the Red Sea belonging to three different genera (Nannochloris, Picochlorum and Desmochloris), previously isolated as novel biodiesel feedstocks, were bioprospected for high-value, bioactive molecules. Methanol extracts were thus prepared from freeze-dried biomass and screened for different biological activities. Nannochloris sp. SBL1 and Desmochloris sp. SBL3 had the highest radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl, and the best copper and iron chelating activities. All species had potent butyrylcholinesterase inhibitory activity (>50%) and mildly inhibited tyrosinase. Picochlorum sp. SBL2 and Nannochloris sp. SBL4 extracts significantly reduced the viability of tumoral (HepG2 and HeLa) cells with lower toxicity against the non-tumoral murine stromal (S17) cells. Nannochloris sp. SBL1 significantly reduced the viability of Leishmania infantum down to 62% (250 µg/mL). Picochlorum sp. SBL2 had the highest total phenolic content, the major phenolic compounds identified being salicylic, coumaric and gallic acids. Neoxanthin, violaxanthin, zeaxanthin, lutein and β-carotene were identified in the extracts of all strains, while canthaxanthin was only identified in Picochlorum sp. SBL2. Taken together, these results strongly suggest that the microalgae included in this work could be used as sources of added-value products that could be used to upgrade the final biomass value.

A comparative evaluation of biological activities and bioactive compounds of the seagrasses Zostera marina and Zostera noltei from southern Portugal.

This work assessed the antioxidant potential, acetylcholinesterase (AChE) inhibition and the in vitro cytotoxic activity of extracts of the seagrasses Zostera marina and Zostera noltei collected from southern Portugal. The total phenolic contents (TPCs), the rosmarinic acid (RA) concentration (HPLC/DAD) and the fatty acid (FA) profile (GC/MS) are also described. Z. marina had the highest TPC, radical scavenging activity against DPPH radicals and copper chelating activity. Z. noltei had metal chelation capacity to copper and iron ions. None of the species was able to inhibit AChE. Both seagrasses had high levels of polyunsaturated FAs. Z. marina significantly and selectively reduced the viability of tumorous neuronal cells. Z. noltei was highly toxic for the three cell lines tested and was selective against hepatocarcinoma cells at the concentration of 100 μg/mL. RA was the main compound identified in Z. marina, but not in Z. noltei.

First report of the nutritional profile and antioxidant potential of Holothuria arguinensis, a new resource for aquaculture in Europe

Abstract This work reports for the first time the nutritional profile and antioxidant potential of the edible sea cucumber Holothuria arguinensis from the North-eastern Atlantic. H. arguinensis has high levels of protein, with the amino acids profile dominated by alanine, glycine and proline and low lysine/arginine ratios. Its carbohydrate and energetic contents are also low as well as the total lipid levels, although its lipid profile is rich in polyunsaturated fatty acids (PUFA), especially arachidonic, eicosapentaenoic and docosahexaenoic acids. In addition, H. arguinensis has high levels of calcium. The water and ethanol extracts show ability to scavenge free radicals and to chelate copper and iron ions. Our results indicate that H. arguinensis has a balanced nutritional quality suitable for human consumption. In addition, it contains compounds with antioxidant potential; thus its intake can contribute for a healthy and well-balanced diet. Graphical abstract

Unlocking the in vitro anti-inflammatory and antidiabetic potential of Polygonum maritimum

Abstract Context: Several Polygonum species (Polygonaceae) are used in traditional medicine in Asia, Europe and Africa to treat inflammation and diabetes. Objective: Evaluate the in vitro antioxidant, anti-inflammatory and antidiabetic potential of methanol and dichloromethane extracts of leaves and roots of the halophyte Polygonum maritimum L. Material and methods: Antioxidant activity was determined (up to 1 mg/mL) as radical-scavenging activity (RSA) of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), copper (CCA) and iron (ICA) chelating activities and iron reducing power (FRAP). NO production was measured in lipopolysaccharide (LPS)-stimulated macrophages for 24 h at concentrations up to 100 μg/mL and antidiabetic potential was assessed by α-amylase and α-glucosidase inhibition (up to 10 mg/mL) assays. The phytochemical composition of the extracts was determined by gas chromatography-mass spectrometry (GC-MS). Results: The methanol leaf extract had the highest activity against DPPH• (IC50 = 26 μg/mL) and ABTS+• (IC50 = 140 μg/mL), FRAP (IC50 = 48 μg/mL) and CCA (IC50 = 770 μg/mL). Only the dichloromethane leaf extract (LDCM) showed anti-inflammatory activity (IC50 = 48 μg/mL). The methanol root (IC50 = 19 μg/mL) and leaf (IC50 = 29 μg/mL) extracts strongly inhibited baker’s yeast α-glucosidase, but LDCM had higher rat’s α-glucosidase inhibition (IC50 = 2527 μg/mL) than acarbose (IC50 = 4638 μg/mL). GC-MS analysis identified β-sitosterol, stigmasterol, 1-octacosanol and linolenic acid as possible molecules responsible for the observed bioactivities. Conclusions: Our findings suggest P. maritimum as a source of high-value health promoting commodities for alleviating symptoms associated with oxidative and inflammatory diseases, including diabetes.