Mycorrhizal induction of phenolic compounds and antioxidant properties of fungi and seedlings during the early steps of symbiosis.

Ectomycorrhizal (ECM) symbiosis plays a major role in nutrient cycling and the functioning of forest ecosystems. Trees with well developed ectomycorrhizas are more resistant to environmental stresses such as drought and to biotic stresses such as root pathogens. The establishment of ECM symbiosis is triggered by signals produced by both partners. These signals lead to morphological changes and a complex development of specific structures in both the plant root and the fungus. The development of the ECM fungi, Paxillus involutus and Pisolithus arhizus, in presence and absence of the symbiont – Pinus pinaster- was evaluated, as well as their antioxidant properties and phenolic compounds composition in response to the symbiotic association. ECM fungi grew less in the presence of Pinus pinaster, with Pisolithus arhizus being less affected in growth and thus being more adapted to this association. Protocatechuic acid was found only in Paxillus involutus, while Pinus pinaster roots, both in association and isolated, proved to have other phenolic acids, such as p-hydroxybenzoic and p-coumaric acids. The symbiosis between Paxillus involutus and Pinus pinaster had no major effects on the symbionts, while the association between Pisolithus arhizus and Pinus pinaster seems to generally decrease the antioxidant effects of both symbionts, despite the increase in p-coumaric and cinnamic acids in the ECM fungi

Influence of mycorrhizal symbiosis in antioxidant potential of fungi and seedlings.

Ectomycorrhizal (ECM) symbiosis plays a major role in nutrient cycling and the functioning of forest ecosystems. Trees with well developed ectomycorrhizas are more resistant to environmental stresses such as drought and to biotic stresses such as root pathogens [1]. The establishment of ECM symbiosis is triggered by signals produced by both partners. These signals lead to morphological changes and a complex development of specific structures in both the plant root and the fungus [2]

Experimental continuation of periodic orbits through a fold

We present a continuation method that enables one to track or continue branches of periodic orbits directly in an experiment when a parameter is changed. A control-based setup in combination with Newton iterations ensures that the periodic orbit can be continued even when it is unstable. This is demonstrated with the continuation of initially stable rotations of a vertically forced pendulum experiment through a fold bifurcation to find the unstable part of the branch.Comment: 4 page

Bioactive and chemical properties of edible mycorrhizal mushrooms from Northeast Portugal

Mushrooms are widely appreciated all over the world for their nutritional properties1 and pharmacological value as sources of important bioactive compounds.2,3 Mycorrhizal macrofungi associate with plant roots constituting a symbiotic relationship, beneficial for both plant and fungus. In the present work five edible mycorrhizal species (Amanita caesarea, Cortinarius anomalus, Cortinarius violaceus, Lactarius volemus and Suillus luteus) from Northeast Portugal were studied for their bioactive and chemical properties. Antioxidant activity was accessed by in vitro assays: radical scavenging activity, reducing power, and lipid peroxidation inhibition. Bioactive compounds such as phenolic compounds, vitamins (ascorbic acid and tocopherols), sugars and fatty acids were obtained by high performance liquid chromatography (HPLC) or gas-chromatography (GC) coupled to different detectors. The main compounds identified in each class were: Phenolic compounds- protocatechuic, p-hydroxybenzoic and p-coumaric acids; Tocopherols- the four isoforms ,, and ; Sugars- mannitol and trehalose; Fatty acids- oleic acid (C18:1n9), linoleic acid (C18:2n6) and palmitic acid (C16:0). A similar profile of metabolites was observed in the studied mushroom species with the order sugars > fat > ascorbic acid > phenolic compounds > tocopherols. Nevertheless, the samples revealed different compositions: prevalence of sugars in Lactarius volemus (28.22 1.81 g/100 g dw), ascorbic acid (343.96 ± 30.98 mg/100 g) and phenolic compounds (8.68 ± 0.38 mg/100 g) in Cortinarius anomalus and fat (3.45 ± 0.65 g/100 g), tocopherols (449.76 ± 52.96 g/100 g) and antioxidant activity in Suillus luteus (EC50 values 1.92 ± 0.08, 0.75 ± 0.03, 0.61 ± 0.02 mg/ml for radical scavenging activity, reducing power, and lipid peroxidation inhibition, respectively)

Towards the antioxidant and chemical characterization of mycorrhizal mushrooms from Northeast Portugal

Mushrooms are widely appreciated all over the world for their nutritional properties and pharmacological value as sources of important bioactive compounds. Mycorrhizal macrofungi associate with plant roots constituting a symbiotic relationship. This symbiosis could influence the production of secondary metabolites, including bioactive compounds. We focused on the evaluation of antioxidant potential and chemical composition of mycorrhizal mushrooms species from Northeast Portugal: Amanita caesarea, Amanita muscaria, Amanita pantherina, Chroogomphus fulmineus, Cortinarius anomalus, Cortinarius collinitus, Cortinarius violaceus, Lactarius quietus, Lactarius volemus, Russula sardonia, Suillus luteus and Tricholoma ustale. A similar profile of metabolites was observed in the studied species with the order sugars > fat > ascorbic acid > phenolic compounds > tocopherols. Nevertheless, the samples revealed different compositions: prevalence of sugars in L. volemus, fat and ascorbic acid in A. muscaria, phenolic compounds in C. anomalus and tocopherols and antioxidant activity in S. luteus

Chemical composition and biological activity of different residues obtained from the wine industry

In recent years, the bioactive compounds present in agri-food sub-products have attracted increased attention due to their health benefits and advantages within a circular economy context. Annually, wine production is responsible for the production of large amounts of phytotoxic waste, which elimination is considered challenging since these residues can be hazardous to the environment when used as fertilizers or just discarded. Phenolic compounds are secondary metabolites processed by plants that have shown several health benefits, acting as antioxidants, antimicrobials, anticarcinogenic, or antidiabetics, among others. Bioactive phenolic molecules have attracted considerable attention from the pharmaceutical, cosmetic, and food industries. So far, several studies have been developed on the characterization of grape pomace and its components (seeds, skins, and stems), especially directed to the residues of red varieties.1 However, less attention has been paid to other by-products generated during winemaking, such as waste from the production of white wine, wine lees and diatomaceous earth. The latter is used in the filtration of wine and constitutes about 250 tons/year of waste from the wine sector in Portugal alone, making it a very pertinent residue with still scarce information being found in the literature. In this context, within the framework of the BacchusTech project that seeks to develop new innovative processes, comprising the extraction, purification, and concentration of bioactive compounds present in winemaking residues, different residues including pomace, lees and diatomaceous earth were evaluated for their chemical composition and bioactivities. Residues were extracted using an hydroalcoholic solvent (80%, v/v), total phenolic compounds were estimated using the Folin- Ciocalteu reagent and individual phenolic compounds were identified and quantified by liquid chromatography coupled to mass spectrophotometry (HPLC-DAD-ESI-MSn). Additionally, the biological activity was assessed through TBARS, DPPH, and reducing power assays to determine the antioxidant activity, and the antimicrobial activity was evaluated by broth microdilution against eight bacteria and two fungi. The phenolic composition was in accordance with the previously reported in red wines.2,3 Fifteen non-anthocyanin phenolic compounds were found, five phenolic acids (gallic acid and derivatives, p-hydroxybenzoic and p-coumaric acid), four flavan-3-ols (procyanidin dimers), two O-glycosylated flavanols (isorhamnetin and quercetin derivatives), three flavanol aglycones (quercetin, kaempferol, and myricetin), and one unknown compound. Regarding anthocyanins, five compounds were found, namely malvidin derivatives linked to acyl groups. Wine lees and white grape pomace before distillation presented the highest contents of phenolic compounds; however, only diatomaceous earth sample reveal the presence of O-glycosylated flavonoids. All samples showed antibacterial and antifungal activity against most of the tested microorganisms. The best bacteriostatic activity was evidenced by the red and white grape pomace before distillation and diatomaceous earth, while the wine lees stood out for their fungistatic activity. In general, all samples showed promising antioxidant capacity, with very good results being obtained on TBARS assay, particularly for the white pomace after distillation (EC50 = 0.016±0.002 mg/mL), diatomaceous earth (EC50 = 0.063±0.001 mg/mL) and red pomace before distillation (EC50 = 0.08±0.04 mg/mL). Overall, the results obtained showed that the residues analyzed are good sources of bioactive compounds, namely anthocyanins and other phenolic compounds, which can be used as raw materials for the steps of concentration, purification and/or isolation of compounds of added value.To Campelo for supplying the winemaking residues. To project “BacchusTech - Integrated Approach for the Valorization of Winemaking Residues” (POCI-01-0247-FEDER-069583). L. Barros, M.I. Dias, and S. Heleno (CEECIND/00831/2018) thank the national funding by FCT, P.I., through the institutional and individual scientific employment program-contract for their contract. Funding: This work was financially supported by the Competitiveness and Internationalization Operational Program (COMPETE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and Foundation for Science and Technology (FCT, Portugal) through national funds FCT/MCTES to the CIMO (UIDB/00690/2020). The GIP-USAL is financially supported by the Spanish “Ministerio de Ciencia and Innovación” (PID2019-106167RB-I00) and “Junta de Castilla y León” (SA093P20 and CLU-2018-04).info:eu-repo/semantics/publishedVersio

Chemical characterization, antioxidant and antimicrobial activities of winemaking industry by-products

The exploitation of bioactive compounds trom agri-food by-products has been attracting an increasing interest from different industries within a circular economy context since several of compounds may have health properties or be used as natural colorants or preservatives. So far, several studies have been developed on the characterization of red wine grape pomace and its components (seeds, skins, and stems). However, less attention has been paid to other by-products, such as grape pomace from white wine, the residues after distillation, wine lees, and diatomaceous earth, the latter used in the filtration of wine and corresponding to about 250 tons/year of waste in Portugal. In this work, samples of these by-products were evaluated for their chemical composition by HPLC-DAD-ESI-Msn and bioactivities (antioxidant activity by TBARS, DPPH, and reducing power assays, and antimicrobial activity against eight bacteria and two fungi). Fifteen non-anthocyanin phenolic compounds were found, including 5 phenolic acids, 4 flavan-3-0Is, 2 0- glycosylated flavanols, 3 flavanol aglycones, and one unknown. In the red wine byproducts 12 anthocyanins were detected, most being malvidin derivatives. Wine lees and white grape pomace before distillation presented the highest contents of phenolic compounds. Ali samples showed antibacterial and antifunga! activity against most of the tested microorganisms, with red and white grape pomace and diatomaceous earth having the best bacteriostatic activity, while the lees stood out against fungi. Ali samples showed promising antioxidant capacity, with very good results obtained on TBARS for the white pomace after distillation and diatomaceous earth. Overall, the results show that besides grape pomace other wine industry byproducts are also good sources of bioactive compounds with high potential for exploitation.To Campelo for the winemaking residues. To projecl "BacchusTech - Inlegrated Approach for lhe Valorization of Winemaking Residues" (POCI-01-0247-FEDER-069583) and national funds FCT/MCTES to CIMO (UIDB/00690/2020). L. Barros, M. Dias, S. Heleno Ihank FCT, P.L, for Iheir scienlific employmenl program-contractinfo:eu-repo/semantics/publishedVersio

The contribution of phenolic acids to the anti-inflammatory activity of mushrooms: Screening in phenolic extracts, individual parent molecules and synthesized glucuronated and methylated derivatives

In the present study, the ethanolic extracts of fourteen edible mushrooms were investigated for their anti-inflammatory potential in LPS (lipopolysaccharide) activated RAW 264.7 macrophages. Furthermore the extracts were chemically characterized in terms of phenolic acids and related compounds. The identified molecules (p-hydroxybenzoic, p-coumaric and cinnamic acids) and their glucuronated and methylated derivatives obtained by chemical synthesis were also evaluated for the same bioactivity, in order to establish structure-activity relationships and to comprehend the effects of in vivo metabolism reactions in the activity of the compounds. The extracts of Pleurotus ostreatus, Macrolepiota procera, Boletus impolitus and Agaricus bisporus revealed the strongest anti-inflammatory potential (EC50 values 96 ± 1 to 190 ± 6 µg/mL, and also the highest concentration of cinnamic acid (656 to 156 µg/g), which was also the individual compound with the highest anti-inflammatory activity. The derivatives of p-coumaric acid revealed the strongest properties, specially the derivative methylated in the carboxylic group (CoA-M1) that exhibited similar activity to the one showed by dexamethaxone used as anti-inflammatory standard; by contrast, the derivatives of p-hydroxybenzoic revealed the lowest inhibition of NO production. All in all, whereas the conjugation reactions change the chemical structure of phenolic acids and may increase or decrease their activity, the glucuronated and methylated derivatives of the studied compounds are still displaying anti-inflammatory activity.The authors are grateful to Fundação para a Ciência e a Tecnologia (FCT, Portugal) and FEDER-COMPETE/QREN/EU for the financial support through the research projects PEst-OE/AGR/UI0690/2011 and PEst-C/QUI/UI0686/2011. S.A. Heleno (BD/70304/2010) and R.C. Calhelha (BPD/68344/2010) also thank FCT, POPH-QREN and FSE. C. Santos-Buelga is also thankful to the Spanish MINECO for financial support through the project BFU2012-35228

Narrativas dos profissionais dos ensinos pré-escolar e primeiro ciclo face às crianças com necessidades educativas especiais: o outro lado da história

[Resumo] As debilidades na formação dos educadores e professores quanto as NEE sao uma importante variável influenciadora na e para a construção de urna equidade educativa. Registe-se a afirmação proferida na Declaras;ao de Salamanca: "a preparação adequada de todo o pessoal educativo é o factor chave na promosção das escolas inclusivas." Toma-se imprescindível conhecermos as formas de pensar, de actuar, a diversidade de sentimentos e estratégias que os profissionais poderao demonstrar como resultado de lidarem com uma criança com NEE, para que possamos melhor compreender e interpretar a direcs;ao para a construção de um diálogo efectivo de parceria educacional. Pretende-se, assim, através de urna metodologia qualitativa - análise de narrativas - o desenvolvimento de um programa de forma<;ao que promova desenvolvimento de atitudes positivas face a escola inclusiva, a aquisis;ao de novas competencias de ensino, que permita aos profissionais serem mais responsivos as necessidades educativas das criança

Comparison between different extraction methods in the recovery of bioactive molecules from Melissa officinalis L. under sustainable cultivation: chemical and bioactive characterization

Melissa officinalis L., from the Lamiaceae family, is one of the most important medicinal and aromatic plants with potential in the market. With the passing of time, the use of medicinal plants in the treatment of some illness has gone from the simplest forms of local treatment to the industrial manufacture of phytotherapics. In addition to their medicinal effect, they can also be used in the form of infusions and decoctions and in various food preparations. In this sense, the objective of this work was to compare three different extraction methods: infusion (100% water), maceration (80:20 ethanol: water v:v) and ultrasound-assisted extraction (UAE) under previous optimized extraction conditions (33.0 +- 3.2 min, 371.7 +- 19.3Wand 39.9 +- 1.4% ethanol), in plants grown under sustainable cultivation under full irrigation in June. The parameters studied included bioactive evaluation by antioxidant (thiobarbituric acid reactive substances—TBARS), cytotoxicity (sulforhodamine B) and anti-inflammatory (RAWcells) assays. The composition of phenolic compounds and organic acids was also studied by GC-MS and HPLC-DAD, respectively. According to the obtained results, eight phenolic compounds were identified and quantified, being rosmarinic acid the main one for the three extraction methods (infusion: 107.1 +- 0.9 mg/g extract; maceration: 155.7 +- 0.3 mg/g extract; UAE: 118.7 0.6 mg/g extract). For Lithospermic acid A isomer (25.25 +- 0.01 mg/g) and Hydroxylsalvianolic E (111.70 +- 2.20 mg/g), the UAE revealed the lowest content of individual polyphenols, whereas maceration recorded the highest extractability. On the other hand, the content of six of the eight polyphenols detected for the ultrasound-assisted extraction was similar to the infusion and maceration methods. In terms of antioxidant activity, the infusions showed the highest capacity (3.00 +- 0.14 +-g/mL), followed by maceration (5.33 +- 0.30 +-g/mL) and UAE (12 +- 0.15 +-g/mL). The highest anti-inflammatory activity was verified for the infusion (244 +- 11 +-g/mL), followed by UAE (305 +- 9 +-g/mL), with no activity recorded for the maceration extract (>400 +-g/mL). The antitumor properties were evaluated in five cell lines, with the best results being recorded for infusion, except AGS (24 +- 1 +-g/mL) whereThe authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). L. Barros and M.I. Dias thank the national funding by FCT through the institutional scientific employment program-contract for her contract, while M. Carocho and S. Heleno thank FCT through the indiviual scientific employment program-contracts (CEECIND/00831/2018 and CEECIND/03040/2017). I. Oliveira thanks FCT for her PhD grant (BD/06017/2020). To FEDER-Interreg España-Portugal programme for financial support through the project TRANSCoLAB 0612_TRANS_CO_LAB_2_P; to ERDF through the Regional Operational ProgramNorth 2020, within the scope of Project GreenHealth— Norte-01-0145-FEDER-000042.info:eu-repo/semantics/publishedVersio