Geographical origin discrimination of the green coffee bean and analytical qualification of the roasting profilles

Tese de doutoramento, Química (Química Analítica), Universidade de Lisboa, Faculdade de Ciências, 2011ADI Agency (Agência de Inovação), project POCTI Med. 2.4 - Projecto ORITOCAFÉ (2004-2006); Fundação para a Ciência e a Tecnologia (FCT, SFRH/BD/28354/2006), project PTDC/AGR-AAM/104357/2008 (IsoGeoCoffee), (2010-2012

Experimental design and sample preparation in forest tree matabolomics

ReviewAppropriate experimental design and sample preparation are key steps in metabolomics experiments, highly influencing the biological interpretation of the results. The sample preparation workflow for plant metabolomics studies includes several steps before metabolite extraction and analysis. These include the optimization of laboratory procedures, which should be optimized for di erent plants and tissues. This is particularly the case for trees, whose tissues are complex matrices to work with due to the presence of several interferents, such as oleoresins, cellulose. A good experimental design, tree tissue harvest conditions, and sample preparation are crucial to ensure consistency and reproducibility of the metadata among datasets. In this review, we discuss the main challenges when setting up a forest tree metabolomics experiment for mass spectrometry (MS)-based analysis covering all technical aspects from the biological question formulation and experimental design to sample processing and metabolite extraction and data acquisition. We also highlight the importance of forest tree metadata standardization in metabolomics studiesinfo:eu-repo/semantics/publishedVersio

Ferulic acid exerts antidepressant-like effect in the tail suspension test in mice: Evidence for the involvement of the serotonergic system

AbstractFerulic acid (4-hydroxy-3-methoxycinnamic acid) is a phenolic compound present in several plants with claimed beneficial effects in prevention and treatment of disorders linked to oxidative stress and inflammation. In this study, we aimed to verify the possible antidepressant-like effect of acute oral administration of ferulic acid in the forced swimming test (FST) and tail suspension test (TST) in mice. Additionally, the mechanisms involved in the antidepressant-like action and the effects of the association of ferulic acid with the antidepressants fluoxetine, paroxetine, and sertraline in the TST were investigated. Ferulic acid produced an antidepressant-like effect in the FST and TST (0.01–10mg/kg, p.o.), without accompanying changes in ambulation. The pretreatment of mice with WAY100635 (0.1mg/kg, s.c., a selective 5-HT1A receptor antagonist) or ketanserin (5mg/kg, i.p., a 5-HT2A receptor antagonist) was able to reverse the anti-immobility effect of ferulic acid (0.01mg/kg, p.o.) in the TST. The combination of fluoxetine (5mg/kg, p.o.), paroxetine (0.1mg/kg, p.o.) or sertraline (1mg/kg, p.o.) with a sub-effective dose of ferulic acid (0.001mg/kg, p.o.) produced a synergistic antidepressant-like effect in the TST, without causing hyperlocomotion in the open-field test. Taken together, these results demonstrate that ferulic acid exerts antidepressant-like effect in the FST and TST in mice through modulation of the serotonergic system

Drought response of cowpea (Vigna unguiculata (L.) Walp.) landraces at leaf physiological and metabolite profile levels

Drought threatens the world’s food production, particularly in Sub Saharan Africa low external input and rain fed agricultural systems, where cowpea (Vigna unguiculata (L.) Walp.) is an important food crop. In the context of growing concerns regarding climate changes implications on water availability, this study aimed at 1) to evaluate the drought responses in cowpea landraces with contrasting drought tolerance levels (A55 – high sensitivity; A80 - mild sensitivity; A116 - tolerant), 2) using an integrated physiological (leaf gas exchanges; chlorophyll a fluorescence) and biochemical (photoprotective pigments; RuBisCO activity; primary metabolite profiling) analysis to identify drought tolerance probes, in plants submitted to three water availability levels (well-watered, WW; mild drought, MD; severe drought, SD). A116 plants maintained a better water status under drought, what could justify the higher Pn and Pnmax values in MD, as well as higher photochemical use of energy (reflected in the photochemical quenching (qL) and in the quantum yield of non-cyclic electron transport (Y(II))), and the lower need of photoprotective thermal dissipation mechanisms (given by the non-photochemical quenching (qN), and the quantum yield of regulated energy dissipation at photosystem PSII (Y(NPQ))), in MD and SD plants. Greater declines of net (Pn) and potential (Pnmax) photosynthesis were observed in A55 plants, which frequently showed significant impacts already under MD conditions in most parameters, whereas A80 usually displayed and intermediate behaviour. Still, even A55 showed some acclimation response, regarding photoprotective mechanisms associated with high contents of zeaxanthin, lutein, and carotenes, and high Y(NPQ), and qN values, supporting the absence of an increase in the non-regulated energy dissipation at PSII (Y(NO) did not increased) even in SD plants. Additionally, A55 was not significantly affected in RuBisCO activity, which showed to be quite resilient in cowpea. A primary metabolite profiling, complemented with a partial least square discrimination analysis (PLS-DA), allowed a better separation of A116 and A55 plants according to their degree of drought tolerance. In response to drought, A116 showed the greatest accumulation of most responsive metabolites, 14 in total, with sucrose, fucose, urea, alanine and putrescine being exclusively increased in this genotype, suggesting that they can be candidates as drought tolerance proxies. Other compounds, as proline, valine, isoleucine (among amino acids), and rhamnose and raffinose (among sugars) showed close increase patternsinfo:eu-repo/semantics/publishedVersio

Controlling Macrophage Polarization to Modulate Inflammatory Cues Using Immune-Switch Nanoparticles

The persistence of inflammatory mediators in tissue niches significantly impacts regenerative outcomes and contributes to chronic diseases. Interleukin-4 (IL4) boosts pro-healing phenotypes in macrophages (MÏ ) and triggers the activation of signal transducer and activator of transcription 6 (STAT6). Since the IL4/STAT6 pathway reduces MÏ responsiveness to inflammation in a targeted and precise manner, IL4 delivery offers personalized possibilities to overcome inflammatory events. Despite its therapeutic potential, the limited success of IL4-targeted delivery is hampered by inefficient vehicles. Magnetically assisted technologies offer precise and tunable nanodevices for the delivery of cytokines by combining contactless modulation, high tissue penetration, imaging features, and low interference with the biological environment. Although superparamagnetic iron oxide nanoparticles (SPION) have shown clinical applicability in imaging, SPION-based approaches have rarely been explored for targeted delivery and cell programming. Herein, we hypothesized that SPION-based carriers assist in efficient IL4 delivery to MÏ , favoring a pro-regenerative phenotype (M2Ï ). Our results confirmed the efficiency of SPION-IL4 and MÏ responsiveness to SPION-IL4 with evidence of STAT6-mediated polarization. SPION-IL4-treated MÏ showed increased expression of M2Ï associated-mediators (IL10, ARG1, CCL2, IL1Ra) when compared to the well-established soluble IL4. The ability of SPION-IL4 to direct MÏ polarization using sophisticated magnetic nanotools is valuable for resolving inflammation and assisting innovative strategies for chronic inflammatory conditions.This research was funded by the European Research Council, Consolidator Grant Magtendon, grant number 772817. FCT-Fundação para a Ciência e a Tecnologia, grant number SFDH/BD/144816/2019. FCT-Fundação para a Ciência e a Tecnologia under the Scientific Employment Stimulus—Individual Call: 2020.01157.CEECIND

Primary metabolite profile changes in coffea spp. Promoted by single and combined exposure to drought and elevated co2 concentration

info:eu-repo/grantAgreement/FCT/OE/PD%2FBD%2F113475%2F2015/PT PD/00035/2013 CRA-RED-00053-16Climate change scenarios pose major threats to many crops worldwide, including coffee. We explored the primary metabolite responses in two Coffea genotypes, C. canephora cv. Conilon Clone 153 and C. arabica cv. Icatu, grown at normal (aCO2) or elevated (eCO2) CO2 concentrations of 380 or 700 ppm, respectively, under well-watered (WW), moderate (MWD), or severe (SWD) water deficit conditions, in order to assess coffee responses to drought and how eCO2 can influence such responses. Primary metabolites were analyzed with a gas chromatography time-of-flight mass spectrometry metabolomics platform (GC-TOF-MS). A total of 48 primary metabolites were identified in both genotypes (23 amino acids and derivatives, 10 organic acids, 11 sugars, and 4 other metabolites), with differences recorded in both genotypes. Increased metabolite levels were observed in CL153 plants under single and combined conditions of aCO2 and drought (MWD and SWD), as opposed to the observed decreased levels under eCO2 in both drought conditions. In contrast, Icatu showed minor differences under MWD, and increased levels (especially amino acids) only under SWD at both CO2 concentration conditions, although with a tendency towards greater increases under eCO2. Altogether, CL153 demonstrated large impact under MWD, and seemed not to benefit from eCO2 in either MWD and SWD, in contrast with Icatu.publishersversionpublishe

Will Casuarina glauca Stress Resilience Be Maintained in the Face of Climate Change?

Actinorhizal plants have been regarded as promising species in the current climate change context due to their high tolerance to a multitude of abiotic stresses. While combined salt-heat stress effects have been studied in crop species, their impact on the model actinorhizal plant, Casuarina glauca, has not yet been fully addressed. The effect of single salt (400 mM NaCl) and heat (control at 26/22 C, supra optimal temperatures at 35/22 C and 45/22 C day/night) conditions on C. glauca branchlets was characterised at the physiological level, and stress-induced metabolite changes were characterised by mass spectrometry-based metabolomics. C. glauca could withstand single salt and heat conditions. However, the harshest stress condition (400 mM NaCl, 45 C) revealed photosynthetic impairments due to mesophyll and membrane permeability limitations as well as major stress-specific differential responses in C and N metabolism. The increased activity of enzymatic ROS scavengers was, however, revealed to be sufficient to control the plant oxidative status. Although C. glauca could tolerate single salt and heat stresses, their negative interaction enhanced the effects of salt stress. Results demonstrated that C. glauca responses to combined salt-heat stress could be explained as a sum of the responses from each single applied stressinfo:eu-repo/semantics/publishedVersio

Routine screening of harmful microorganisms in beach sands: implications to public health

Beaches worldwide provide recreational opportunities to hundreds of millions of people and serve as important components of coastal economies. Beach water is often monitored for microbiological quality to detect the presence of indicators of human sewage contamination so as to prevent public health outbreaks associated with water contact. However, growing evidence suggests that beach sand can harbor microbes harmful to human health, often in concentrations greater than the beach water. Currently, there are no standards for monitoring, sampling, analyzing, or managing beach sand quality. In addition to indicator microbes, growing evidence has identified pathogenic bacteria, viruses, and fungi in a variety of beach sands worldwide. The public health threat associated with these populations through direct and indirect contact is unknown because so little research has been conducted relating to health outcomes associated with sand quality. In this manuscript, we present the consensus findings of a workshop of experts convened in Lisbon, Portugal to discuss the current state of knowledge on beach sand microbiological quality and to develop suggestions for standardizing the evaluation of sand at coastal beaches. The expert group at the "Microareias 2012" workshop recommends that 1) beach sand should be screened for a variety of pathogens harmful to human health, and sand monitoring should then be initiated alongside regular water monitoring; 2) sampling and analysis protocols should be standardized to allow proper comparisons among beach locations; and 3) further studies are needed to estimate human health risk with exposure to contaminated beach sand. Much of the manuscript is focused on research specific to Portugal, but similar results have been found elsewhere, and the findings have worldwide implications

Routine screening of harmful microorganisms in beach sands: implications to public health

Beaches worldwide provide recreational opportunities to hundreds of millions of people and serve as important components of coastal economies. Beach water is often monitored for microbiological quality to detect the presence of indicators of human sewage contamination so as to prevent public health outbreaks associated with water contact. However, growing evidence suggests that beach sand can harbor microbes harmful to human health, often in concentrations greater than the beach water. Currently, there are no standards for monitoring, sampling, analyzing, or managing beach sand quality. In addition to indicator microbes, growing evidence has identified pathogenic bacteria, viruses, and fungi in a variety of beach sands worldwide. The public health threat associated with these populations through direct and indirect contact is unknown because so little research has been conducted relating to health outcomes associated with sand quality. In this manuscript, we present the consensus findings of a workshop of experts convened in Lisbon, Portugal to discuss the current state of knowledge on beach sand microbiological quality and to develop suggestions for standardizing the evaluation of sand at coastal beaches. The expert group at the "Microareias 2012" workshop recommends that 1) beach sand should be screened for a variety of pathogens harmful to human health, and sand monitoring should then be initiated alongside regular water monitoring; 2) sampling and analysis protocols should be standardized to allow proper comparisons among beach locations; and 3) further studies are needed to estimate human health risk with exposure to contaminated beach sand. Much of the manuscript is focused on research specific to Portugal, but similar results have been found elsewhere, and the findings have worldwide implications