No induction of antimicrobial resistance in Staphylococcus aureus and Listeria monocytogenes during continuous exposure to eugenol and citral

The aim of this study was to evaluate the adaptation response of Staphylococcus aureus, methicillin-resistant S.aureus (MRSA), and Listeria monocytogenes to the essential oil (EO), eugenol, and citral. The minimum inhibitory concentration of eugenol and citral was determined by agar dilution and microdilution. Adaptation to eugenol and citral was done by sequential exposure of the pathogens to increasing concentrations of the essential oils. The M2-A9 standard was used to determine the antibiotic susceptibility. The effect of eugenol and citral on the adherence ability was evaluated by the crystal violet assay. The impact of adaptation to eugenol on virulence was estimated using the Galleria mellonella model. No development of resistance to the components and antibiotics was observed in the adapted cells of S.aureus, MRSA, and L.monocytogenes. Eugenol and citral at subinhibitory concentration reduced the bacterial adherence. Adaptation to subinhibitory concentration of eugenol affected the virulence potential of S.aureus, MRSA, and L.monocytogenes. Eugenol and citral do not pose a risk of resistance development in a continuous mode of use. These EO components showed a high efficacy as antistaphylococcal and antilisterial biofilm agents. Adaptation at subinhibitory concentration of eugenol protected the larvae against listerial and staphylococcal infection.FCT - Fundacao para a Ciencia e Tecnologia [PEst-OE/EQB/LA0023/2013]info:eu-repo/semantics/publishedVersio

Biological properties of latex, aqueous extracts and bee products of Euphorbia Officinarum L.: a short review

Euphorbia officinarum L. is a Moroccan endemic plant known as “Tikiout” and “Daghmus” that can also be found in Mauritania, Western Sahara, and Algeria. In the present review, “Euphorbia officinarum”, “metabolites” “hemisynthesis” were the keywords used for the research in the Web search engine Google Scholar and in the database Web of Science. Triterpenes, phytosterols and ingol diterpenes were isolated and identified in the latex of Moroccan E. officinarum. More than sixty triterpenes were obtained by hemisynthesis from natural triterpenes. Some of these derivatives had insecticidal and antimicrobial activity (phytopathogenic bacteria). The total phenol content and the antioxidant and anti-α-glucosidase activities were dependent on the time and temperature of extractions and also on the plant solvent ratio. The antioxidant activity of monofloral honey of E. officinarum origin was attributed to the phenol fraction (this fraction, previously isolated from honey samples, had better activity than the entire honey).info:eu-repo/semantics/publishedVersio

Growth, photosynthetic pigments, phenolic content and biological activities of Foeniculum vulgare Mill., Anethum graveolens L. and Pimpinella anisum L. (Apiaceae) in response to zinc

The effect of zinc (Zn) on phenols, antioxidant activities (free radicals' scavenging activities, inhibition of lipid peroxidation, chelating activity and reducing power), and enzyme inhibition activities of acetylcholinesterase, lipoxygenase and tyrosinase of anise, dill and two cultivars of fennel [Latina" (F-L) and "Doux de Florence" (F-DF)] aqueous extracts was evaluated for the first time. At the same time, growth and photosynthetic pigment contents were also estimated. A significant decrease in all growth parameters, such as plant height, fresh and dry weights of aerial parts and roots was statistically proved in the presence of 2 mM Zn. An enhancement of Zn accumulation was observed, particularly in aerial part. Exposure to 2 mM Zn induced quantitative changes in the phenols of all tested extracts. In fennel extracts, a decrease of phenol content was observed when treated with Zn, whereas an increase was found in dill and anise extracts submitted to Zn treatment. All samples, either in the presence or absence of Zn, showed antioxidant activity, independent on the tested assay, with the exception of the cultivar F-DF (treated and non-treated), which was unable to scavenge NO radicals. Zn-treated dill and anise presented usually higher antioxidant activity. Concerning inhibitory activities against acetylcholinesterase, lipoxygenase and tyrosinase enzymes, both treated and non-treated F-DF did not inhibit acetylcholinesterase activity. Zinc-treated F-DF was also unable to inhibit lipoxygenase activity. The remaining species presented capacity for inhibiting those enzymes and tyrosinase as well, but their abilities varied in response to Zn excess.Portuguese National Funding Agency for Science, Research and Technology (Fundacao para a Ciencia e a Tecnologia - FCTPortugal) [UID/BIA/04325/2013-MEDTBIO, UID/Multi/04326/2013]info:eu-repo/semantics/publishedVersio

Bio-oils/FCC co-processing : Insights into the adsorption of guaiacol on Y zeolites with distinct acidity and textural properties

Acknowledgments The authors thank Fundação para a Ciência e a Tecnologia (FCT, Portugal) for financial funding (UIDB/00100/2020 and UIDP/00100/2020). Fernandes also thanks FCT for researcher contract hiring (DL No. 57/2016 regulation).Peer reviewedPostprin

Targeted gene disruption coupled with metabolic screen approach to uncover the LEAFY COTYLEDON1-LIKE4 (L1L4) function in tomato fruit metabolism

Key message Functional analysis of tomato L1L4 master transcription factor resulted in important metabolic changes affecting tomato fruit quality. Abstract Tomato fruits from mutant lines bearing targeted disruption of the heterotrimeric nuclear transcription factor Y (NF-Y) transcription factor (TF) gene LEAFY-COTYLEDON1-LIKE4 (L1L4, NF-YB6), a master regulator of biosynthesis for seed storage proteins and fatty acids, were evaluated for metabolites content and morphology. Metabolic screens using LC-MS/MS-based analysis and physico-chemical methods in different L1L4 mutants of the fourth generation allowed a comparative assessment of the effects of the TF disruption. Mutagenesis resulted in fruits phenotypically similar to wild-type with subtle shape differences in the distal end protrusion and symmetry. Conversely, mutant fruits from independent lines had significant variation in moisture content, titratable acidity and overall metabolite profiles including oxalic and citric acid, fructose, beta-carotene, total polyphenols and antioxidants. Lines 6, 7 and 9 were the richest in beta-carotene and antioxidant activity, line 4 in ascorbic acid and lines 4 and 8 in succinic acid. The reduced content of the anti-nutrient oxalic acid in several mutant fruits suggests that L1L4 gene may regulate the accumulation of this compound during fruit development. Detailed LC-MS/MS analysis of mutant seeds showed substantial differences in bioactive compounds compared to wild-type seeds. Taken together, the results suggest that the L1L4 TF is a significant regulator of metabolites both in tomato fruit and seeds providing a molecular target for crop improvement. Elucidation of the candidate genes encoding key enzymes in the affected metabolic pathways aimed to facilitate the L1L4 gene network exploration and eventually lead to systems biology approaches in tomato fruit quality.European funded COST ACTION [FA1106

Days out of role due to common physical and mental conditions in Portugal: results from the WHO World Mental Health Survey

Funding: The Portuguese Mental Health Study was carried out by the Department of Mental Health, NOVA Medical School, NOVA University of Lisbon, with collaboration of the CESOP – Portuguese Catholic University and was funded by the Champalimaud Foundation, the Gulbenkian Foundation, the Foundation for Science and Technology (FCT) and the Ministry of Health. The Portuguese Mental Health Study was carried out in conjunction with the World Health Organization WMH Survey Initiative which is supported by the National Institute of Mental Health (NIMH; R01 MH070884), the John D. and Catherine T. MacArthur Foundation, the Pfizer Foundation, the U.S. Public Health Service (R13-MH066849, R01-MH069864 and R01 DA016558), the Fogarty International Center (FIRCA R03-TW006481), the Pan American Health Organization, Eli Lilly and Company, OrthoMcNeil Pharmaceutical, GlaxoSmithKline and Bristol-Myers Squibb.publishersversionpublishe

Quick decline and stem pitting Citrus tristeza virus isolates induce a distinct metabolomic profile and antioxidant enzyme activity in the phloem sap of two Citrus species

Susceptibility to the severe Citrus tristeza virus (CTV), T36, is higher for Citrus macrophylla (CM) than for C. aurantium (CA). How host-virus interactions are reflected in host physiology is largely unknown. In this study, the profile of metabolites and the antioxidant activity in the phloem sap of healthy and infected CA and CM plants were evaluated. The phloem sap of quick decline (T36) and stem pitting (T318A) infected citrus, and control plants was collected by centrifugation, and the enzymes and metabolites analyzed. The activity of the antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), in infected plants increased significantly in CM and decreased in CA, compared to the healthy controls. Using LC-HRMS2 a metabolic profile rich in secondary metabolites was assigned to healthy CA, compared to healthy CM. CTV infection of CA caused a drastic reduction in secondary metabolites, but not in CM. In conclusion, CA and CM have a different response to severe CTV isolates and we propose that the low susceptibility of CA to T36 may be related to the interaction of the virus with the host’s metabolism, which reduces significantly the synthesis of flavonoids and antioxidant enzyme activity.ALG-01-0145-FEDER-30957; LA/P/0101/2020; ALG-01-0145-FEDER-022121info:eu-repo/semantics/publishedVersio

Nutritional characterization and storage ability of Salicornia ramosissima and Sarcocornia perennis for fresh vegetable salads

The aim of this work was to study the potential of two halophytes for fresh salads. Sarcocornia perennis (SAR) and Salicornia ramosissima (SAL) plant tips were harvested in May and July, and stored at 4 °C for 14 days. At harvest, mineral analyses (Na, K, Mg, Ca, P, N, Fe, Zn, Cu, Pb and Cd), chlorophyll content, protein, β-carotene and ascorbic acid were performed. Color (CIELab), phenolics, antioxidant activity and microbial contamination were measured at harvest and after 14 days storage at 4 °C, and a taste panel evaluated the products. SAL had generally slightly higher values of vitamin C, phenols and carotenoids than SAR. In SAR after 14 days of storage, the levels of phenols and the antioxidant activity were higher than at harvest, both in May and July. The opposite was observed for SAL. In this case, only the antioxidant activity was higher after 14 days of storage. Microbial contamination was very low, not reaching even half of critical limits. Both halophytes had good nutritional value, being suitable for use as fresh salads which can be stored for up to 14 days at 4 °C. SAR seems to be a better source of antioxidants, Ca and Fe than SAL and has less general quality and nutritional value changes from harvest time through cold storage. Fresh tips of both halophytes showed better appearance and nutritional quality when harvested in the early season (May) than at the end of the season (July).This research was funded by the Fundação para a Ciência e Tecnologia-FCT, Portugal, under the projects UIDB/05183/2020 MED and UID/BIA/04325/2013-MEDTBIO.info:eu-repo/semantics/publishedVersio

Effect of essential oils on the release of TNF-α and CCL2 by LPS-stimulated THP‑1 Cells

Plants and their constituents have been used to treat diverse ailments since time immemorial. Many plants are used in diverse external and internal formulations (infusions, alcoholic extracts, essential oils (EOs), etc.) in the treatment of inflammation-associated diseases, such as those affecting the respiratory tract or causing gastrointestinal or joint problems, among others. To support the traditional uses of plant extracts, EOs have been assessed for their alleged anti-inflammatory properties. However, the effect of EOs on the release of cytokines and chemokines has been much less reported. Considering their traditional use and commercial relevance in Portugal and Angola, this study evaluated the effect of EOs on the in vitro inhibition of the cytokine tumor necrosis factor-α (TNF-α) and the chemokine (C-C motif) ligand 2 (CCL2) by lipopolysaccharide (LPS)-stimulated human acute monocytic leukemia cells (THP-1 cells). Twenty EOs extracted from eighteen species from seven families, namely from Amaranthaceae (Dysphania ambrosioides), Apiaceae (Foeniculum vulgare), Asteraceae (Brachylaena huillensis, Solidago virgaurea), Euphorbiaceae (Spirostachys africana), Lamiaceae (Lavandula luisieri, Mentha cervina, Origanum majorana, Satureja montana, Thymbra capitata, Thymus mastichina, Thymus vulgaris, Thymus zygis subsp. zygis), Myrtaceae (Eucalyptus globulus subsp. maidenii, Eucalyptus radiata, Eucalyptus viminalis) and Pinaceae (Pinus pinaster) were assayed for the release of CCL2 and TNF-α by LPS-stimulated THP-1 cells. B. huillensis, S. africana, S. montana, Th. mastichina and Th. vulgaris EOs showed toxicity to THP-1 cells, at the lowest concentration tested (10 μg/mL), using the tetrazolium dye assay. The most active EOs in reducing TNF-α release by LPS-stimulated THP-1 cells were those of T. capitata (51% inhibition at 20 μg/mL) and L. luisieri (15–23% inhibition at 30 μg/mL and 78–83% inhibition at 90 μg/mL). L. luisieri EO induced a concentration-dependent inhibition of CCL2 release by LPS‑stimulated THP-1 cells (23%, 54% and 82% inhibition at 10, 30 and 90 μg/mL, respectively). These EOs are potentially useful in the management of inflammatory diseases mediated by CCL2 and TNF‑α, such as atherosclerosis and arthritis.The European Commission under the Seventh Framework Programme (FP7) of the European Union, Marie Curie International Research Staff Exchange Scheme (MC-IRSES). Project PEOPLE MC-IRSES, FP7-PEOPLE-2011-IRSES, PIRSES-GA-2011-295251 Fundação para a Ciência e Tecnologia (FCT/MCTES), FEDER, PT2020 PA, Compete 2020. Projects MED UIDB/05183/2020 and CESAM UIDP/50017/2020 + UIDB/50017/2020.info:eu-repo/semantics/publishedVersio

Molecular cloning and functional characterization of a monoterpene synthase isolated from the aromatic wild shrub Thymus albicans

The essential oil of Thymus albicans Hoffmanns. & Link, a native shrub from the Iberian Peninsula, is mainly composed of monoterpenes. In this study, a 1,8-cineole synthase was isolated from the 1,8-cineole chemotype. A partial sequence that lacked the complete plastid transit peptide but contained an extended C-terminal when compared to other related terpene synthases was generated by PCR and Rapid Amplification of cDNA Ends (RACE). The predicted mature polypeptide was 593 amino acids in length and shared 78% and 77% sequence similarity with the homologue 1,8-cineole synthase from Rosmarinus officinalis and Salvia officinalis, respectively. The putative protein possessed the characteristic conserved motifs of plant monoterpene synthases including the RRx(8)W and DDxxD motifs and phylogenetic analysis indicated that the amplified 1,8-cineole synthase bears greater sequence similarity with other 1,8-cineole synthases from Lamiaceae family relative to the terpene synthases from the genus Thymus. Functional expression of the recombinant protein in Escherichia coli revealed that in the presence of geranyl diphosphate (GPP) 1,8-cineole was the major product but that its production was too low for robust quantification. Other minor conversion products included a-pinene, beta-pinene, sabinene and beta-myrcene suggesting the isolated 1,8-cineole synthase may be a multi-product enzyme. To our knowledge, this is the first report of a functionally characterized monoterpene synthase from Thymus albicans.Ceratonia Project MONOTHYMUS, Universidade do Algarve, PortugalMinistry of Science, PortugalFCT under FEDER PT2020-Compete[FCT/UID/Multi/00631/2013/CEOT