ABC2-Artigos (em revistas ou actas indexadas)
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- 1-Methylcyclopropene and lemongrass essential oil nanocoatings effect on the preservation of cold stored ‘Rocha’ pearPublication . Gago, Custódia; Guerreiro, Adriana; Cruz, Sandra; Martins, Nuno; Cabrita, Maria João; Miguel, Maria; Faleiro, Maria Leonor; Antunes, Maria DulceThe effects of coating 'Rocha' pear with alginate-based nanoemulsions enriched with lemongrass essential oil (LG) was evaluated and compared to the usual 1-MCP treatment. Fruit were treated with 1-MCP (312 nL L-1) or coated with nanoemulsions: sodium alginate 2 % (w/w) + lemongrass essential oil 1.25 % (w/w) (LG 1.25 %) or lemongrass essential oil 2.5 % (w/w) (LG 2.5 %). Then, fruit were stored at 0 degrees C and 90-95 % relative humidity (RH), for eight months. Fruit samples were collected at harvest and after two, four, six and eight months of cold storage, and then transferred to shelf-life at 22 degrees C. Upon removal and after 7 d shelf-life, fruit symptoms of superficial scald and internal browning, ethylene production, color CIE (L*, hue), firmness, soluble solids content (SSC), titratable acidity (TA), weight loss, electrolytic leakage (EL), antioxidant activity and fatty acids of pear peel, microbial growth and sensory analyses were evaluated. Coatings and 1-MCP reduced fruit color evolution and preserved better firmness than control. Coatings and 1-MCP did not affect SSC and TA. Treatments did not influence the sensory quality. Microbial growth was within the safety limits in all treatments. Treatments with 1-MCP and LG-nanoemulsions were similarly efficient to reduce superficial scald, nevertheless the LG-nanoemulsions showed higher internal disorders after 8 months of storage and LG 2.5 % had higher decay at the same period, similar to control. 1-MCP treated fruit had the lowest softening rate after shelf-life up to 4 months and LG 2.5 % showed higher weight loss. Also, ethylene production was higher in control and LG 1.25 % up to 6 months plus shelf-life, while after 8 months there was no difference among treatments. This study suggests that 1-MCP is the most efficient for preserving quality of 'Rocha' pear for 8 months, while up to 6 months the best effect is obtained with LG 1.25 % nanocoatings.
- Advanced therapy medicinal products development - from guidelines to medicines in the marketPublication . Frederico, Catarina; Vieira da Conceição, André Filipe; Nóbrega, Clévio; Mendonça, Liliana S.In Europe, Advanced Therapy Medicinal Products (ATMPs) include medicines based on gene therapy, somaticcell therapy, tissue-engineered products, and combined ATMPs. ATMPs constitute an emerging and innovative class of medicines used to treat multiple pathologies and are particularly relevant in pathologies where therapeutic options are limited and require high medical needs. These therapies act, among others, through the insertion of recombinant nucleic acids, including genes, to promote a therapeutic effect and through the restoration of cell functions, and repairing or replacing damaged cells and tissues impaired in pathological conditions. Despite their unique potential, these therapies face challenges related to scientific complexity, production processes, regulatory approval, and market access that hinder their development and availability. Based on official European guidelines, the present review explores the current regulatory framework for the non-clinical and clinical development of advanced therapies. We aimed to discuss the regulations applied to the different types of ATMPs, as well as the challenges associated with their development until these therapies reach the market. Accordingly, topics such as the implementation of proof-of-concept studies to provide evidence supporting the potential clinical effect; biodistribution studies to evaluate tissue distribution and persistence; and toxicology studies to assess potential undesirable effects, integration potential of viral vectors, tumorigenicity, and germline transmission, are discussed. This work also covers some of the ATMPs available to patients on the EU market.
- Advancing glioblastoma research with innovative brain organoid-based modelsPublication . Dias Correia, Cátia; Calado, Sofia; Matos, Alexandra; Oleiro Esteves, Filipa Alexandra; De Sousa-Coelho, Ana Luísa; Campinho, Marco António; Teotónio Fernandes, Mónica AlexandraGlioblastoma (GBM) is a relatively rare but highly aggressive form of brain cancer characterized by rapid growth, invasiveness, and resistance to standard therapies. Despite significant progress in understanding its molecular and cellular mechanisms, GBM remains one of the most challenging cancers to treat due to its high heterogeneity and complex tumor microenvironment. To address these obstacles, researchers have employed a range of models, including in vitro cell cultures and in vivo animal models, but these often fail to replicate the complexity of GBM. As a result, there has been a growing focus on refining these models by incorporating human-origin cells, along with advanced genetic techniques and stem cell-based bioengineering approaches. In this context, a variety of GBM models based on brain organoids were developed and confirmed to be clinically relevant and are contributing to the advancement of GBM research at the preclinical level. This review explores the preparation and use of brain organoid-based models to deepen our understanding of GBM biology and to explore novel therapeutic approaches. These innovative models hold significant promise for improving our ability to study this deadly cancer and for advancing the development of more effective treatments.
- Automedicação: hábitos durante um período de pandemia - estudo transversalPublication . Espírito-Santo, Margarida; Estêvão, M. Dulce; Campos, EricaA automedicação é uma prática muito comum e a pandemia de COVID-19 poderá ter criado condições para haver um maior recurso às farmácias comunitárias, particularmente em situações de doença ligeira. Esse estudo teve como objetivo analisar os hábitos de automedicação de uma amostra da população Portuguesa durante o período inicial da pandemia (março-novembro 2020), identificar os medicamentos não sujeitos a receita médica (MNSRM) e suplementos alimentares mais consumidos, bem como as situações que levaram os indivíduos a recorrer à automedicação e a informação sobre estes produtos. Foi realizado um estudo transversal, observacional, através da aplicação de um questionário com recurso a uma plataforma da internet, durante o mês de dezembro 2020. Os dados foram obtidos de forma anónima, e analisados com o programa IBM SPSS v26. A amostra em análise era composta por 170 indivíduos, tendo-se verificado que a pandemia parece não ter afetado a frequência de ida à farmácia. As situações que mais levaram as pessoas a recorrer à automedicação foram dores de cabeça e dores musculares. Os medicamentos mais utilizados neste período pelos participantes neste estudo foram os analgésicos e os anti-inflamatórios. Apesar de todas as restrições impostas pela situação pandémica, parece não ter havido um impacto negativo no recurso às farmácias comunitárias, nem um aumento significativo da prática de automedicação. Foi também assinalado o papel relevante dos profissionais de Farmácia, em particular nas situações de automedicação, como fonte de informação sobre o uso dos medicamentos e dos suplementos alimentares.
- Autophagy in Spinocerebellar Ataxia Type 3: From pathogenesis to therapeuticsPublication . Paulino, Rodrigo; Nóbrega, ClévioMachado–Joseph disease (MJD) or spinocerebellar ataxia 3 (SCA3) is a rare, inherited, monogenic, neurodegenerative disease, and the most common SCA worldwide. MJD/SCA3 causative mutation is an abnormal expansion of the triplet CAG at exon 10 within the ATXN3 gene. The gene encodes for ataxin-3, which is a deubiquitinating protein that is also involved in transcriptional regulation. In normal conditions, the ataxin-3 protein polyglutamine stretch has between 13 and 49 glutamines. However, in MJD/SCA3 patients, the size of the stretch increases from 55 to 87, contributing to abnormal protein conformation, insolubility, and aggregation. The formation of aggregates, which is a hallmark of MJD/SCA3, compromises different cell pathways, leading to an impairment of cell clearance mechanisms, such as autophagy. MJD/SCA3 patients display several signals and symptoms in which the most prominent is ataxia. Neuropathologically, the regions most affected are the cerebellum and the pons. Currently, there are no disease-modifying therapies, and patients rely only on supportive and symptomatic treatments. Due to these facts, there is a huge research effort to develop therapeutic strategies for this incurable disease. This review aims to bring together current state-of-the-art strategies regarding the autophagy pathway in MJD/SCA3, focusing on evidence for its impairment in the disease context and, importantly, its targeting for the development of pharmacological and gene-based therapies.
- BARX2/FOXA1/HK2 axis promotes lung adenocarcinoma progression and energy metabolism reprogrammingPublication . Xie, Kai; Feng, Jian; Fan, Dingwei; Wang, Shi; Luo, Jing; Ren, Zhijian; Zheng, Chao; Diao, Yifei; De Mello, Ramon Andrade; Tavolari, Simona; Brandi, Giovanni; Roden, Anja C.; Ren, Binhui; Shen, Yi; Xu, LinBackground: Metabolic reprogramming is an emerging cancer feature that has recently drawn special attention since it promotes tumor cell growth and proliferation. However, the mechanism of the Warburg effect is still largely unknown. This research aimed to reveal the effects of BarH-like homeobox 2 (BARX2) in regulating tumor progression and glucose metabolism in lung adenocarcinoma (LUAD).Methods: Expression of BARX2 was measured by quantitative real-time polymerase chain reaction (qRTPCR) in LUAD cell line and tissues, and the tumor-promoting function of BARX2 in LUAD cells was detected in vitro and in vivo xenograft models. The metabolic effects of BARX2 were examined by detecting glucose uptake, the production levels of lactate and pyruvate, and the extracellular acidification rate (ECAR). Chromatin immunoprecipitation (ChIP) assay and luciferase reporter gene assay were used to identify the underlying molecular mechanism of BARX2 regulation of HK2. Further studies showed that transcription factor FOXA1 directly interacts with BARX2 and promotes the transcriptional activity of BARX2.Results: BARX2 was remarkably up-regulated in LUAD tissues and positively linked to advanced clinical stage and poor prognosis. In vitro and in vivo data indicated ectopic expression of BARX2 enhanced cell proliferation and tumorigenesis, whereas BARX2 knockdown suppressed these effects. Metabolic-related experiments showed BARX2 promoted the reprogramming of glucose metabolism. Mechanistically, the BARX2/FOXA1/HK2 axis promoted LUAD progression and energy metabolism reprogramming.Conclusions: In summary, our research first defined BARX2 as a tumor-promoting factor in LUAD andthat it may act as a novel prognostic biomarker and new therapeutic target for the disease.
- Biological consequences of Vanadium effects on formation of reactive oxygen species and lipid peroxidationPublication . Aureliano, Manuel; De Sousa-Coelho, Ana Luísa; Dolan, Connor C.; Roess, Deborah A.; Crans, Debbie C.Lipid peroxidation (LPO), a process that affects human health, can be induced by exposure to vanadium salts and compounds. LPO is often exacerbated by oxidation stress, with some forms of vanadium providing protective effects. The LPO reaction involves the oxidation of the alkene bonds, primarily in polyunsaturated fatty acids, in a chain reaction to form radical and reactive oxygen species (ROS). LPO reactions typically affect cellular membranes through direct effects on membrane structure and function as well as impacting other cellular functions due to increases in ROS. Although LPO effects on mitochondrial function have been studied in detail, other cellular components and organelles are affected. Because vanadium salts and complexes can induce ROS formation both directly and indirectly, the study of LPO arising from increased ROS should include investigations of both processes. This is made more challenging by the range of vanadium species that exist under physiological conditions and the diverse effects of these species. Thus, complex vanadium chemistry requires speciation studies of vanadium to evaluate the direct and indirect effects of the various species that are present during vanadium exposure. Undoubtedly, speciation is important in assessing how vanadium exerts effects in biological systems and is likely the underlying cause for some of the beneficial effects reported in cancerous, diabetic, neurodegenerative conditions and other diseased tissues impacted by LPO processes. Speciation of vanadium, together with investigations of ROS and LPO, should be considered in future biological studies evaluating vanadium effects on the formation of ROS and on LPO in cells, tissues, and organisms as discussed in this review.
- Biphasic Npas4 expression promotes inhibitory plasticity and suppression of fear memory consolidation in micePublication . Brito, David V.C.; Kupke, Janina; Sokolov, Rostilav; Cambridge, Sidney; Both, Martin; Bengtson, C. Peter; Rozov, Andrei; Oliveira, Ana M. M.Long-term memories are believed to be encoded by unique transcriptional signatures in the brain. The expression of immediate early genes (IEG) promotes structural and molecular changes required for memory consolidation. Recent evidence has shown that the brain is equipped with mechanisms that not only promote, but actively constrict memory formation. However, it remains unknown whether IEG expression may play a role in memory suppression. Here we uncovered a novel function of the IEG neuronal PAS domain protein 4 (Npas4), as an inducible memory suppressor gene of highly salient aversive experiences. Using a contextual fear conditioning paradigm, we found that low stimulus salience leads to monophasic Npas4 expression, while highly salient learning induces a biphasic expression of Npas4 in the hippocampus. The later phase requires N-methyl-D-aspartate (NMDA) receptor activity and is independent of dopaminergic neurotransmission. Our in vivo pharmacological and genetic manipulation experiments suggested that the later phase of Npas4 expression restricts the consolidation of a fear memory and promote behavioral flexibility, by facilitating fear extinction and the contextual specificity of fear responses. Moreover, immunofluorescence and electrophysiological analysis revealed a concomitant increase in synaptic input from cholecystokinin (CCK)-expressing interneurons. Our results demonstrate how salient experiences evoke unique temporal patterns of IEG expression that fine-tune memory consolidation. Moreover, our study provides evidence for inducible gene expression associated with memory suppression as a possible mechanism to balance the consolidation of highly salient memories, and thereby to evade the formation of maladaptive behavior.
- Blood management in total knee arthroplasty: an exploratory study regarding the use of drain and tranexamic acid in two types of instrumentationPublication . Dias, Pedro Alexandre; Caldeira-Dantas, Sofia; Carvalho, Maria Miguel; Marreiros, Ana; de Almeida Fontes, Ana Paula; Sousa, João PauloBackground: This study investigates how presence or absence of drain or tranexamic acid (TXA) in 2 different types of instrumentation with invasion/sparing femoral canal affects hemoglobin drop after total knee arthroplasty (TKA). Methods: This was a retrospective observational study that included 736 individuals divided in five groups, regarding conventional instrumentation (CI) or patient specific instrumentation (PSI), presence or absence of drain and TXA. Mean percentage of hemoglobin drop after the first postoperative day was compared for each group. Results: Patients submitted to CI using drain and no TXA revealed the highest value (21,3%±9,1; p<0,001). Groups who received TXA and drain was not used, presented the lowest blood loss. Moreover, with the introduction of TXA PSI lost its advantage over CI. Conclusions: TKA using CI, without drain and with TXA administration showed better results, reducing mean percentage of hemoglobin drop after surgery.
- Brain-targeted drug delivery - nanovesicles directed to specific brain cells by brain-targeting ligandsPublication . Moreira, Ricardo; Nóbrega, Clévio; Almeida, Luís Pereira de; Mendonça, LilianaNeurodegenerative diseases are characterized by extensive loss of function or death of brain cells, hampering the life quality of patients. Brain-targeted drug delivery is challenging, with a low success rate this far. Therefore, the application of targeting ligands in drug vehicles, such as lipid-based and polymeric nanoparticles, holds the promise to overcome the blood-brain barrier (BBB) and direct therapies to the brain, in addition to protect their cargo from degradation and metabolization. In this review, we discuss the barriers to brain delivery and the different types of brain-targeting ligands currently in use in brain-targeted nanoparticles, such as peptides, proteins, aptamers, small molecules, and antibodies. Moreover, we present a detailed review of the different targeting ligands used to direct nanoparticles to specific brain cells, like neurons (C4-3 aptamer, neurotensin, Tet-1, RVG, and IKRG peptides), astrocytes (Aquaporin-4, D4, and Bradykinin B2 antibodies), oligodendrocytes (NG-2 antibody and the biotinylated DNA aptamer conjugated to a streptavidin core Myaptavin-3064), microglia (CD11b antibody), neural stem cells (QTRFLLH, VPTQSSG, and NFL-TBS.40–63 peptides), and to endothelial cells of the BBB (transferrin and insulin proteins, and choline). Reports demonstrated enhanced brain-targeted delivery with improved transport to the specific cell type targeted with the conjugation of these ligands to nanoparticles. Hence, this strategy allows the implementation of high-precision medicine, with reduced side effects or unwanted therapy clearance from the body. Nevertheless, the accumulation of some of these nanoparticles in peripheral organs has been reported indicating that there are still factors to be improved to achieve higher levels of brain targeting. This review is a collection of studies exploring targeting ligands for the delivery of nanoparticles to the brain and we highlight the advantages and limitations of this type of approach in precision therapies.