Browsing by Author "Silva, Ana Margarida"
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- Assessment of 83 pharmaceuticals in WWTP influent and effluent samples by UHPLC-MS/MS: Hourly variationPublication . Paíga, Paula; Correia, Manuela; Fernandes, Maria João; Silva, Ana Margarida; Martins de Carvalho, Maria Manuela; Vieira, Joana; Jorge, Sandra; Silva, Jaime Gabriel; Freire, Cristina; Delerue-Matos, CristinaThe removal efficiency of pharmaceuticals in wastewater treatment plants (WWTPs) is variable and some of these compounds pass these plants almost intact and others presenting a removal efficiency close to 100%. Their incomplete removal results in a continuous discharge of pharmaceuticals into the environment. To assess the profile of contamination of influents and effluents over a day, a set of 83 pharmaceuticals were evaluated hourly in a WWTP in Leiria, Portugal. The composite samples of the influent and effluent were also collected. Concentrations varied from 1 in WWTP influents, and carbamazepine, fluoxetine, sertraline the pharmaceuticals with an RQ > 1 in WWTP effluents.
- Assessment of a Formulation Containing a Castanea sativa Shells Extract on Skin Face Parameters: In Vivo EvaluationPublication . Silva, Ana Margarida; Costa, Paulo C.; Delerue-Matos, Cristina; Rodrigues, FranciscaIn the last years, the demand for new eco-friendly ingredients has increased on the cosmetic market. Consumers are more aware of sustainable principles and, simultaneously, more conscious regarding the skin aging process. Chestnut (Castanea sativa) shells are a food by-product produced in high quantities in Europe. This waste has been described as a rich source of phenolic compounds with skin biological effects, such as antioxidant, antiradical, and anti-inflammatory activities. Despite the huge number of assays reporting the richness of chestnut shell extracts in bioactive compounds as well as the development of cosmetic formulations containing these extracts, no in vivo assays have assessed their clinical efficacy in human volunteers. The aim of this study was to evaluate the effect on skin face parameters of a formulation containing a chestnut shell extract in human volunteers (n = 22) who applied the product twice per day, for 56 days. For that, biophysical techniques, including Corneometer®, Cutometer®, and PrimosPremium, were employed, allowing the quantification of skin hydration and firmness, as well as of wrinkles’ depth and volume and wrinkles’ skin roughness. The results demonstrated that the formulation led to a slight decrease in roughness and wrinkles’ depth, although no significant differences with respect to a placebo were observed. In addition, a clear improvement of skin hydration was achieved (t0 = 54.00 Arbitrary Units (A.U.) and t56 = 58.62 A.U.). In contrast to the placebo, the active formulation increased skin firmness up to 31.76% in 50.00% of the volunteers, but without significant differences with respect to the placebo, probably due to the short period of treatment. A long-term use of the product is recommended to possibly observe significant differences in all parameters.
- Chemical characterization and bioactive properties of a coffee-like beverage prepared from Quercus cerris kernelsPublication . Pinto, Diana; Diaz Franco, Santiago; Silva, Ana Margarida; Cupara, Snezana; Koskovac, Marijana; Kojicic, Ksenija; Soares, Sónia; Rodrigues, Francisca; Sut, Stefania; Dall'Acqua, Stefano; Oliveira, M Beatriz P PIn the present study, a coffee-like beverage was prepared from Quercus cerris seeds. Seeds presented 60.4% carbohydrates (of which 26.9% is fiber), 6.3% proteins and 4.0% fat. In the coffee-like beverage, the TPC and TFC were 2070.2 mg GAE per L and 285.3 mg CEQ per L, respectively. The IC50 values determined using FRAP and DPPH assays were, respectively, 203.1 mg L-1 and 271.6 mg L-1. Concerning the reactive species scavenging capacity, the extract was more effective against O2˙-, HClO and ˙NO (namely, IC50 = 17.24 ± 0.24 μg mL-1, IC50 = 9.25 ± 0.92 μg mL-1 and IC50 = 0.65 ± 0.06 μg mL-1). Qualitative and quantitative analyses by 1D- and 2D-NMR and LC-MS indicated high levels of ellagic acid and gallotannin or ellagitannin derivatives. The optimal range of non-cytotoxic concentrations in Caco-2 and HT29-MTX cell lines was between 0.1 and 1.0 μg mL-1. This is the first study that evaluates the Q. cerris seed extract as a beverage.
- Chemical composition and bioactivity of oilseed cake extracts obtained by subcritical and modified subcritical waterPublication . Švarc-Gajić, Jaroslava; Rodrigues, Francisca; Moreira, Manuela M.; Delerue-Matos, Cristina; Morais, Simone; Dorosh, Olena; Silva, Ana Margarida; Bassani, Andrea; Dzedik, Valentin; Spigno, GiorgiaRecovery of bioactive compounds from biowaste is gaining more and more interest in circular economy models. The oilseed cakes are usually insufficiently exploited by most technologies since they represent valuable matrices abundant in proteins, minerals, and phytochemicals, but their use is mostly limited to feed ingredients, fertilizers or biofuel production. This study was thus focused on the exploration of new valorization pathways of oilseed cakes by subcritical water, representing a safe and economic alternative in the creation of value chains. Pumpkin, hemp, and flax seed cakes were treated with subcritical water in nitrogen and carbon-dioxide atmospheres, as well as in nitrogen atmosphere with the addition of acid catalyst. The degradation of carbohydrate fraction was studied by quantifying sugars and sugar degradation products in the obtained extracts. The extracts obtained under different conditions were further compared chemically with respect to total phenols and flavonoids, as well as to the content of individual phenolic compounds. Furthermore, the effects of subcritical water treatment conditions on antioxidant, antiradical and cytotoxic properties of thus obtained extracts were defined and discussed.
- Development and Optimization of a Topical Formulation with Castanea sativa Shells Extract Based on the Concept “Quality by Design”Publication . Oliveira, Nágilla; Cádiz-Gurrea, María de La Luz; Silva, Ana Margarida; Macedo, Catarina; Rodrigues, Francisca; Costa, PauloThe proposed study aims to develop and optimize a topical formulation with Castanea sativa shells extract considering the concept of Quality by Design, focusing on a planned development that consider the vulnerabilities of the entire process through risk analysis tools and design of experiments (DoE). A Box–Behnken design with three factors and three levels was used as a statistical tool for the execution of the DoE and the analysis of the response surface methodology responses. The independent variables studied were the quantity of sodium lauryl sulfate (%) (X1), beeswax (%) (X2) and macadamia oil (%) (X3); the dependent variables were pH (Y1), viscosity (Y2) and adhesiveness (Y3). According to the mathematical model, the optimal formulation contains 0.93% of sodium lauryl sulfate, 5.00% of beeswax and 10.00% of macadamia oil. The optimal formulation with the extract was prepared and characterized over the time, regarding organoleptic and technological characteristics, allowing conclusions to be reached regarding its stability. The formulation presented a pleasant odor and was light brown in color, it also demonstrated pseudoplastic-thixotropic behavior and a small reduction in the formulation consistency after 30 days of storage. This study demonstrated the efficiency of the Quality by Design methodology to understand the product variability, supporting that this approach favors a better understanding of the whole process and enables to design a robust development stage, reducing costs and generating high-quality products
- Eco-friendly insights on kiwiberry leaves valorization through in-vitro and in-vivo studiesPublication . Silva, Ana Margarida; Garcia, Juliana; Dall'Acqua, Stefano; Costa, Paulo; Delerue-Matos, Cristina; Rodrigues, FranciscaOn the last decade, the demand for novel active skin ingredients based on sustainable principles arise as a trend. These new concerns transformed the consumer preferences, which are not actually solved by the industry portfolio. Regulatory aspects establish guidelines to assess the safety and efficacy of new cosmetic ingredients, being this step imperative to introduce a new ingredient on the market. Our last studies reported the richness of Actinidia arguta (kiwiberry) leaves in bioactive compounds with potential skin applications, but further studies are necessary to validate this extract. Therefore, the current study aimed to screen the hydroalcoholic extract of A. arguta leaves regarding elastase and hyaluronidase inhibition, achieving excellent results (65.62 ± 2.09% and 54.64 ± 4.17%, respectively). The cell viability was assessed in keratinocytes and fibroblasts (0.1–1000 μg/mL), ensuring the absence of cytotoxic effects. Afterwards, 3D models, namely a skin (EpiSkin™) and an ocular (SkinEthic™ HCE, respectively) model, were used to estimate the irritant potential. The viability achieved in both models after exposure to the extract was, respectively, 55.18 ± 10.60% and 101.15 ± 5.96%. The IL-1α released for the skin and ocular models were, respectively, 0 pg/mL and 35.60 ± 2.61 pg/mL, being the extract classified as non-irritant for both models. Lastly, in-vivo studies performed through a patch test in 10 human volunteers proved the absence of allergic or irritative reactions. Overall, the results attained highlighted that the hydroalcoholic kiwiberry leaves extract is an excellent cosmetic ingredient, being not irritant to skin application
- Evaluation of the potential of marine cyanobacteria towards Alzheimer’s diseasePublication . Moreira, Ana Rita; Silva, Ana Margarida; Vieira, Mónica; Martins, RosárioAlzheimer’s disease (AD) is one of the most prevalent neurodegenerative diseases (NDs) and the major cause of dementia. Despite the constant efforts to achieve cure, no treatment is currently effective and new approaches are urgently needed. Marine biotechnology has been at the frontline for the bioprospection of natural sources to treat ND. Several marine compounds have already provided evidence for anti-AD drugs by acting in mechanisms such as the inhibition of acetylcholinesterase (AChE). Among marine organisms, cyanobacteria have been recognized as a major source of bioactive compounds and the potential towards AD has been highlighted, namely AChE inhibitors. This project is included in a screening program that aims to evaluate the potential of cyanobacteria from the Culture Collection of CIIMAR -LEGE-CC against AD. In this work, 80 fractions of marine cyanobacteria were screened for the potential to inhibit AChE since this is a proven enzyme target to ameliorate the loss of cholinergic signal and to induce cytotoxicity in the neuroblastoma cell line SHY5Y, in the normal fibroblasts cell line 3T3L1, and in the endothelial cells from central nervous system capillaries hCMEC/D3. Results considering the enzymatic assays and cytotoxic assays revealed that the fractions are not toxic or with low toxicity. The strains Phormidium sp; an unidentified filamentous Synechococcales and Nodosilinea nodulosa revealed moderate toxicity towards AChE.
- Exploring the bioactive potential of Pisolithus (Basidiomycota): comprehensive insights into antimicrobial, anticancer, and antioxidant properties for innovative applicationsPublication . S. Oliveira, Rui; Preto, Marco; Santos, Germana; Silva, Ana Margarida; Vasconcelos, Vitor; Martins, RosárioAddressing pressing health concerns, modern medical research seeks to identify new antimicrobials to combat drug resistance, novel molecules for cancer treatment, and antioxidants for inflammation-related diseases. Pisolithus (Basidiomycota) is a ubiquitous and widely distributed fungal genus in forest ecosystems, known for establishing ectomycorrhizal associations with a range of host plants, enhancing their growth, and conferring protection against biotic and abiotic stresses. Beyond ecological applications, Pisolithus yields bioactive compounds with medicinal potential. This comprehensive review explores the transversal biological activity of Pisolithus fungi, aiming to provide a thorough overview of their antimicrobial, anticancer, and antioxidant potential. The focus is on elucidating bioactive compounds within Pisolithus to trigger further research for innovative applications. Compounds from Pisolithus displayed antimicrobial activity against a broad spectrum of microorganisms, including antibiotic-resistant bacteria. The efficacy of Pisolithus-derived compounds matched established medications, emphasizing their therapeutic potential. In anticancer research, the triterpene pisosterol stood out with documented cytotoxicity against various cancer cell lines, showcasing promise for novel anticancer therapies. Pisolithus was also recognized as a potential source of antioxidants, with basidiocarps exhibiting high antioxidant activity. In vivo validation and comprehensive studies on a broader range of compounds, together with mechanistic insights into the mode of action of Pisolithus-derived compounds, are compelling areas for future research.
- Formulation, Characterization, and Cytotoxicity Evaluation of Lactoferrin Functionalized Lipid Nanoparticles for Riluzole Delivery to the BrainPublication . Teixeira, Maria Inês; Lopes, Carla Martins; Gonçalves, Hugo; Catita, José; Silva, Ana Margarida; Rodrigues, Francisca; Amaral, Maria Helena; Costa, Paulo C.Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with a very poor prognosis. Its treatment is hindered by a lack of new therapeutic alternatives and the existence of the blood–brain barrier (BBB), which restricts the access of drugs commonly used in ALS, such as riluzole, to the brain. To overcome these limitations and increase brain targeting, riluzole-loaded nanostructured lipid carriers (NLC) were prepared and functionalized with lactoferrin (Lf), facilitating transport across the BBB by interacting with Lf receptors expressed in the brain endothelium. NLC were characterized with respect to their physicochemical properties (size, zeta potential, polydispersity index) as well as their stability, encapsulation efficiency, morphology, in vitro release profile, and biocompatibility. Moreover, crystallinity and melting behavior were assessed by DSC and PXRD. Nanoparticles exhibited initial mean diameters between 180 and 220 nm and a polydispersity index below 0.3, indicating a narrow size distribution. NLC remained stable over at least 3 months. Riluzole encapsulation efficiency was very high, around 94–98%. FTIR and protein quantification studies confirmed the conjugation of Lf on the surface of the nanocarriers, with TEM images showing that the functionalized NLC presented a smooth surface and uniform spherical shape. An MTT assay revealed that the nanocarriers developed in this study did not cause a substantial reduction in the viability of NSC-34 and hCMEC/D3 cells at a riluzole concentration up to 10 μM, being therefore biocompatible. The results suggest that Lf-functionalized NLC are a suitable and promising delivery system to target riluzole to the brain
- Influence of temperature on the subcritical water extraction of Actinidia arguta leaves: A screening of pro-healthy compoundsPublication . Silva, Ana Margarida; Luís, Ana Sofia; Moreira, Manuela M.; Ferraz, Ricardo; Brezo-Borjan, Tanja; Svarc-Gajic, Jaroslava; Costa, Paulo C.; Delerue-Matos, Cristina; Rodrigues, FranciscaActinidia arguta is a species disseminated in Europe and classified by the Chinese Herbal Medicine as a medicinal plant. The fruit (kiwiberry) has been extensively exploited for multiple purposes, while leaves where discarded. The objective of this study was to evaluate the optimal Subcritical Water Extraction (SWE) temperature (110 °C - 160 °C) of antioxidants and polyphenols from A. arguta leaves. The optimal temperature of extraction was 123 °C, revealing the highest phenolic and flavonoid contents and good scavenging efficiencies against HOCl (IC50 = 17.06 μg/mL) and O2●- (IC50 = 335.2 μg/mL), without toxicity on intestinal cells. The phenolic profile was characterized by high amounts of phenolic acids (e.g., gallic acids), flavanols (catechin) and flavonols (e.g., quercetin-3-O-galactoside). This work allows to conclude that SWE can be a useful extraction technique for the recovery of polyphenolics from A. arguta leaves.
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