Repositório RCAAP

Macrofungi produce remarkable biomedical products that can make a significant contribution to health. Pisolithus arhizus is a fungal species of the phylum Basidiomycota. It is mainly found in association with the roots of some trees species, establishing a mutualistic relationship with them, facilitating the absorption of nutrients from the soil in exchange for carbohydrates produced by the host plant. As recently reviewed by us, this macrofungus has aroused great interest due to its promising therapeutic properties and bioactive effects. Crude extracts were prepared from the mycelium and culture medium of P.arhizus using a mixture of dichloromethane:methanol (2:1. The extracts were then separated into nine fractions using vacuum liquid chromatography. The fractions were tested for cytotoxicity against the RKO colon adenocarcinoma cell line and 3T3 fibroblasts cell line. Cell viability was assessed using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) reduction assay. Preliminary results revealed a dose dependent cytotoxicity against RKO cell line with no significant cytotoxicity to 3T3 cell lines, in fractions B and C. Those fractions corresponded to extraction with 70% Hex:30% EtOAc and 60% Hex:40% EtOAc, respectively. The results indicate that P.arhizus mycelium is a source of compounds with anticancer activity. However, more in-depth studies are needed to analyze the chemical compounds from the various parts of P. arhizus and investigate their biological and toxicological activities. Exploring the bioactive potential of P. arhizus is key to developing a complete understanding of its therapeutic benefits.

Medicinal plants and their components have been utilized in traditional medicine for centuries and have significantly influenced the development of modern medicine. Ruta graveolens, a Rutaceae medical plant, is known for its antibacterial, anti-inflammatory and cytotoxic properties. Actinobacteria are a rich source of compounds exhibiting diverse biological activities and potential therapeutic applications. The aim of this study was to perform the phylogenetic identification of a collection of actinobacterial strains previously isolated from R. graveolens and to investigate their bioactive potential. Actinobacterial strains previously isolated from stem, roots and leaves of R. graveolens were grown in Actinomycete Isolation Agar (AIA) or Starch-Casein-Nitrate-Agar (SCN). DNA from grown cultures was extracted and phylogenetically identified through 16S rRNA gene sequencing. For each strain, organic extracts were performed and used for the screening of antimicrobial activity, using the disk diffusion test, against four reference bacteria (Staphylococcus aureus, Bacillus subtilis, Salmonella typhimurium, Escherichia coli) and one yeast (Candida albicans). Thirty-two actinobacterial isolates were so far identified. Most of the strains was identified as Tsukamurella tyrosinosolvens, constituting 13 out of 32 isolates, followed by 8 Streptomyces sp., 7 Brevibacterium sediminis, 3 Microbacterium ginsengiterrae, and one Gordonia hydrophobica. The organic extracts obtained from each isolate were tested for their antimicrobial activity. Up to moment, no significant bioactivity was detected in the reference strains screened in this study. A collection of 32 actinobacterial strains was obtained from various parts of the medicinal plant R. graveolens. Though no relevant antimicrobial activity was yet found, extracts of these actinobacteria open new opportunities to explore their bioactive potentials with therapeutic applications.

Melanoma is the most aggressive and lethal form of skin cancer, with a high risk of metastatic spread. Obesity is recognized as a risk factor for various types of cancer. However, regarding melanoma, this association remains controversial. Obesity might act as a double-edged sword in melanoma, promoting primary tumour growth but at the same time limiting metastatic spread - the "obesity paradox”. Herein, we aimed to create a stable murine B16F10 melanoma cell line expressing both firefly luciferase (Luc) and green fluorescent protein (GFP), which will later be engrafted into diet induced-obesity animal model for future in vivo studies. B16F10-Luc-GFP cells were generated by transfection with premade lentiviral particles, featuring a construct with Luc and GFP under a cytomegalovirus promoter and mediated by a F2A element. The antibiotic selection marker (puromycin) is expressed under a Rous sarcoma virus promoter. Afterwards, the transfected cells were selected with 1 μg/ml of puromycin. The clones with the highest levels of GFP-positive cells and GFP fluorescence were purified by two rounds of cell sorting and submitted to fluorescence and bioluminescence quantification, morphology, injury, BrdU incorporation, 7-AAD, and PI cell cycle assays and compared to the parental cell line. B16F10-Luc-GFP were successfully generated, and both GFP fluorescence and D-luciferin bioluminescence are present and proportional to cell density. As expected, the parental cell line didn’t display GFP or Luc activities. Moreover, transduced cells exhibit similar morphology, motility, proliferation, viability, and cell cycle progression as B16F10 cells. Conclusions: Altogether, the future engraftment of B16F10-LucGFP in obese mice, will improve melanoma research models, enabling the in vivo and ex vivo visualization of primary tumours and metastasis, providing a better understanding of the underlying molecular mechanisms, to clarify the “obesity paradox” in melanoma.

Excessive alcohol consumption continues to pose a significant global health challenge, with detrimental effects on millions of individuals. Our laboratory has shown that alcohol exposure triggers reactive changes in astrocytes, including alterations in gene expression, activity, and proliferation, while also affecting microglial morphology and immune responses. We are to characterizing the effects of chronic alcohol consumption using a wellestablished voluntary alcohol drinking model in adult mice, to investigate the impact of chronic alcohol exposure on the prefrontal cortex (PFC), focusing on glial cell morphology, synaptic density, and behaviour. Mice are exposed to intermittent “every-other-day” (EOD) access to alcohol 15% (v/v) for 3 weeks, and behaviourally tested for anxiety, depression and memory, before sacrifice at 21 days of alcohol, or at 7 days of withdrawal. Brains were processed for glial cell analysis. Our preliminary findings revealed sex-specific responses following chronic alcohol exposure. Male mice exhibited increased astrocyte volume in the ventromedial PFC (vmPFC) and hyper-ramification in the ventrolateral PFC (vlPFC), whereas females displayed reductions in astrocyte size and complexity. Microglia morphology also differed between sexes, with females showing increased cell volume and males displaying reduced microglial volume in the vlPFC. These results suggest distinctive immune and synaptic responses to ethanol in males and females. Of note, we observed heightened inhibitory synapse density in the male PFC, while females exhibited increased excitatory synapse density. We are now conducting a proteomic analysis of PFC synaptosomes to identify important molecular targets in the crosstalk between neuros and glial cells. With this work we expect to clarify the complex interplay between chronic ethanol exposure, sex, and PFC function, find also new targets for innovative therapeutic approaches.

The A2 score is an 8-question patient-reported outcome measure that has been validated for ruling in (score ≥4) and ruling out (score 0-1) asthma. However, this screening tool has been validated in a cohort similar to the derivation cohort used. This study aims to validate the predictive accuracy of the A2 score in a primary care population against general practitioner (GP) clinical assessment and to determine whether the proposed cut-offs are the most appropriate. This accuracy study is a secondary analysis of the EPI-ASTHMA population-based study. Random adult participants recruited from primary healthcare centers in Portugal were analyzed. Participants answered the A2 score by telephone interview. Those with an A2 score ≥1 (plus 5% with an A2 score of 0) were invited to a diagnostic visit carried out by a GP to confirm or not a diagnosis of asthma. Diagnostic accuracy was assessed using receiver operating characteristic (ROC) curves. A total of 1283 participants (median 54[p25-p75 43-66] years; 60% female) were analyzed. The A2 score showed high discriminatory power in identifying asthma, with an area under the ROC curve of 82.9(95%CI 80.4-85.4)%. The proposed cut-off ≥4 was the most appropriate to rule in asthma (specificity 83.1%, positive predictive value 62.4%, accuracy 78%). Similarly, the proposed cut-off <2 was the most suitable for excluding asthma (sensitivity 92.7%, negative predictive value 93.7%, accuracy 60.5%). The A2 score is a useful tool to identify patients with asthma in a primary care population.

Phenol is a toxic compound present in a wide variety of foundry resins. Its quantification is important for the characterization of the resins as well as for the evaluation of free contaminants present in foundry wastes. Two chromatographic methods, liquid chromatography with ultraviolet detection (LC-UV) and gas chromatography with flame ionization detection (GC-FID), for the analysis of free phenol in several foundry resins, after a simple extraction procedure (30 min), were developed. Both chromatographic methods were suitable for the determination of phenol in the studied furanic and phenolic resins, showing good selectivity, accuracy (recovery 99–100%; relative deviations <5%), and precision (coefficients of variation <6%). The used ASTM reference method was only found to be useful in the analysis of phenolic resins, while the LC and GC methods were applicable for all the studied resins. The developed methods reduce the time of analysis from 3.5 hours to about 30 min and can readily be used in routine quality control laboratories.

The agricultural sector is continuously exploring sustainable and innovative approaches to enhance crop productivity, driving the quest for greener alternatives over synthetic fertilizers. Cyanobacteria have been reported as a promising agricultural fertilizer, with positive effects on crop yield. However, little is known about its potential effects on the produced vegetables' nutritional quality. Studies have documented that exposure of plants to cyanobacterial crude extracts stimulates the production of enzymatic and non-enzymatic antioxidants. Vitamin C, a non-enzymatic antioxidant, plays a crucial role in protecting cells from oxidative stress and spinach is known to be relatively high in vitamin C compared to other vegetables. The purpose of this study was to determine the content of vitamin C in spinach plants grown under the use of cyanobacterial biomass as fertilizer. Spinach plants were tested in soil, under six different conditions: (i) control, (ii) commercial chemical fertilizer, cyanobacterial biomass (iii) non-cyanotoxin producer, and cyanotoxin producer of (iv) Anatoxina (ANA), (v) Microcystin (MC), and (vi) Cylindrospermopsin (CYN). The analysis of vitamin C in spinach plants was performed according to EN 14130:2003. The results showed an increased content of vitamin C in all exposed groups when compared to the control (37 mg/Kg), with statistical differences in the conditions exposed to MC (56 mg/kg), ANA (76 mg/kg), and CYN (114 mg/kg). The results suggest that the rise in vitamin C may be related to a plant defence mechanism against oxidative stress potentially generated by exposure to cyanotoxins. Although the antioxidant content in spinach plants can be enhanced as a mechanism to cope with oxidative stress, prolonged exposure may increase plants' susceptibility to accumulate cyanotoxins, which may pose a risk to human health. Additional studies are needed to understand all the positive and negative effects of using cyanobacteria biomass as a biofertilizer.

Cellulose, a complex polysaccharide, is one of the main components of plant and algae biomass and therefore the most abundant polymer found in nature, serving as a significant carbon source in various ecosystems. Bioprospecting for cellulolytic microbes is a promising strategy for discovering potential biocatalysts for the hydrolysis of lignocellulosic materials, including residues. This work aims to assess the cellulolytic potential of microbial isolates (bacteria and fungi) sourced from diverse environments and to establish a synthetic microbial consortium capable to enhance the management of cellulosic waste. The experimental approach encompasses biodegradation assays coupled with optimized colorimetric methods, to estimate cellulose consumption (Congo Red assay) and the concomitant production of oligomeric/monomeric sugars (Dinitrosalicylic acid assay). For bacterial strains, these biodegradation assays involve batch incubations in minimal liquid medium supplied with cellulose as the sole carbon source, while solid medium is being used for fungi. Both assays are being run under mesophilic temperatures (28-30 ºC) and aerobic conditions. Upon achieving a final selection, strains exhibiting the best cellulose degradation performances will be assembled into a synthetic consortium that will be evaluated for their cellulose degradation capability. Future steps will also include phylogenetic identification of the cellulose degrading isolates through 16S and ITS rRNA gene sequence analysis. This study has the potential to identify novel cellulolytic microbes, highlighting the substantial role microorganisms play in ecosystem processes and their diverse applications in biotechnology.

Here we report the molecular and antimicrobial susceptibility profile of extended-spectrum β-lactamase (ESBL)-producing strains found in the Portuguese northern occidental coast region (Minho). For this purpose, bacteria isolated from clinical hospitalised and non-hospitalised patients over a period of 2 years were identified and minimal inhibitory concentrations (MICs) were determined by microdilution methods according to the Clinical and Laboratory Standards Institute (formerly the National Committee for Clinical Laboratory Standards) guidelines on Enterobacteriaceae. Additionally, ESBL phenotypic identification was confirmed by the Etest (AB BIODISK, Solna, Sweden). Various methods of molecular identification of the β-lactamase (bla) genes, involving polymerase chain reaction (PCR) and sequencing strategies, were used in this study. The ESBL-producing strains (n = 193) were isolated from urine (n = 127), sputum (n = 42), bronchoalveolar lavage (n = 14), blood (n = 7) and ascitic fluid (n = 3). The most frequent ESBL-producing organism isolated in the present study was Escherichia coli (67.9%; n = 131), followed by Klebsiella pneumoniae (30.6%; n = 59), Klebsiella oxytoca (0.5%; n = 1), Enterobacter aerogenes (0.5%; n = 1) and Citrobacter freundii (0.5%; n = 1). The ESBL detected in the present studywere the TEM type (40.4%), CTX-M type (36.8%) and SHV type (22.8%). TEM-52 and TEM-24 were the most frequent TEM types (20.2% and 12.9%, respectively). Members of TEM-10 (4.1%) and TEM-116 (2.1%) were also detected. Within the CTX-M family, CTX-M-9 group was represented by CTX-M-9 (13.5%) and CTX-M-14 (8.4%). In the CTX-M-1 group, CTXM-15 was the most frequent type (12.4 %), followed by CTX-M-1 (2.1%), CTX-M-3 (0.5%) and CTX-M-32 (0.5%). Regarding CTX-M types, it appears that CTX-M-14 is widespread among the northwestern Iberian Peninsula [1]. Klebsiella pneumoniae harbouring a CTX-M-15 enzyme was described for the first time in Portugal in 2005 [2] in the Lisbon area, but CTX-M-15 enzyme has also recently been found by us in the north of Portugal in another Enterobacteriaceae member, isolated from bloodstream infections [3] among seven patients in two different hospitals. Other ESBL-producing species (not E. coli or K. pneumoniae)were also found. This is the first time that C. freundii has been described as a producer of CTX-M-32 in this country. The SHV enzymes occurred only in 23.3% of all ESBL-producing organisms. Within this type, the most frequent type was SHV-12 (12.4%), followed by SHV-5 (8.8%) and finally SHV-2 (2.1%). Some isolates co-produced more than one ESBL type: TEM-52/CTX-M-14 (0.5%); TEM-116/CTX-M-14 (0.5%); and TEM-116/CTX-M-15 (0.5%). MIC testing showed that isolates producing ESBLs were mostly susceptible to carbapenems (100%) and amikacin (99.5%). In contrast, ESBL-producing strains presented low susceptibility rates to cefepime and quinolones. Indeed, 98.9% of the ESBL-producing strains were cefepime-resistant and 85.4% were resistant to quinolones (ciprofloxacin and norfloxacin). In the generality, these high levels of resistance to quinolones were more conspicuous in members of the CTX-M family (98.1%) than TEM and SHV types (80.8% and 72.1%, respectively). In this study, cefepime presented a surprisingly low activity against ESBL-producing microorganisms. Recent literature refers to the inoculum effect exhibited by cefepime [4]. Nevertheless, we believe that this should not be pointed out as a single explanation once MIC determination is performed using inoculum concentrations of 0.5 McFarland standard. In our sample, only two K. pneumoniae harbouring SHV-2 ESBL were susceptible to cefepime. All the other clinical isolates (98.9%) expressing the ESBL phenotype were resistant to cefepime. It seems interesting that a recent study showed that cefepimewas successfully administered to three patients (two females and one male) aged between 47 years and 87 years carrying a Gram-negative ESBL-positive strain [5]. Nevertheless, other studies worldwide have begun to describe the emergence of high resistance to cefepime among Gram-negative ESBL-producers [6]. The present work showed a high diversity of ESBL enzymes occurring in the north of Portugal. In this country, the most prevalent type is still the TEM type, but CTX-M is growing rapidly [7]. The emergence of ESBL-producers resistant to cefepime in Portugal is a matter of concern. We believe that the uncontrolled use of cephalosporins may have an important role in the acquisition of resistance mechanisms, particularly the production of ESBL enzymes. Establishment of policies to monitor drug delivery in hospital and ambulatory pharmacies as well as implementation of public health defence strategies towards health promotion and drug resistance prevention appear to be urgent.

The increasing prevalence of antimicrobial resistances is now a worldwide problem. Investigating the mechanisms by which pets harboring resistant strains may receive and/or transfer resistance determinants is essential to better understanding how owners and pets can interact safely. Here, we characterized the genetic determinants conferring resistance to β-lactams and quinolones in 38 multidrug-resistant Escherichia coli isolated from fecal samples of dogs, through PCR and sequencing. The most frequent genotype included the β-lactamase groups TEM (n = 5), and both TEM + CTX-M-1 (n = 5). Within the CTX-M group, we identified the genes CTX-M-32, CTX-M-1, CTX-M-15, CTX-M-55/79, CTX-M-14 and CTX-M-2/44. Thirty isolates resistant to ciprofloxacin presented two mutations in the gyrA gene and one or two mutations in the parC gene. A mutation in gyrA (reported here for the first time), due to a transversion and transition (TCG → GTG) originating a substitution of a serine by a valine in position 83 was also detected. The plasmid-encoded quinolone resistance gene, qnrs1, was detected in three isolates. Dogs can be a reservoir of genetic determinants conferring antimicrobial resistance and thus may play an important role in the spread of antimicrobial resistance to humans and other co-habitant animals.

Animal nutrition has been severely challenged by the ban on antimicrobials as growth promoters. This has fostered the study of alternative methods to avoid colonisation by pathogenic bacteria as well as to improve the growth of animals and feed conversion efficiency. These new options should not alter the normal intestinal microbiota, or affect it as little as possible. The use of probiotics, which are live microorganisms that beneficially affect the host by improving its intestinal microbial balance, can be seen as a promising way to achieve that goal. In this study, New Zealand White rabbits were fed diets containing an autochthonous probiotic of Enterococcus spp., with the strains EaI, EfaI and EfaD, and Escherichia coli, with the strains ECI 1, ECI 2 and ECD, during a 25-d trial, to evaluate the impact of the probiotic on the faecal microbiota, including population dynamics and antimicrobial resistance profiles. A control group of rabbits, which was fed a diet containing a commonly used mixture of antimicrobials (colistin, oxytetracycline, and valnemulin), was also studied. To assess the colonisation ability of the mentioned probiotic, the faecal microbiota of the rabbits was characterised up to 10 d after the administration had ended. Isolates of enterococci and E. coli were studied for phylogenetic relationships using enterobacterial repetitive intergenic consensus (ERIC-PCR) and pulse-field gel electrophoresis (PFGE), respectively. Although partially affected by an unexpected clinical impairment suffered by the rabbits in the experimental group, our results showed the following. The difference between the growth rate of the animals treated with antimicrobials and those fed the probiotic was not statistically significant (P> 0.05). The competitive exclusion product was present in the faecal samples in a large proportion, but stopped being recovered by culture as soon as the administration ended and the housing conditions were changed. Multidrug-resistant strains of enterococci and E. coli were more commonly recovered from faecal samples of animals fed diets containing antimicrobials, than from rabbits fed diets with our probiotic formula. The use of E. coli probiotics to prevent infection by enteropathogenic strains must be carefully considered due to the possible occurrence of gastrointestinal signs. On the other hand, enterococci strains may be more effective, but lack the long-term colonisation ability.

No desenvolvimento fisiopatológico da diabetes tipo 2 (DM2) ocorrem variados desequilíbrios metabólicos. Há evidências crescentes de que a disfunção metabólica precede em muito as manifestações clínicas, sendo a pré-diabetes a condição determinante. A modulação da pré-diabetes com intervenções no estilo de vida pode diminuir o risco de desenvolver DM2 em 40 a 70%. O principal problema associado à pré-diabetes é o sobrediagnóstico e o sobretratamento. Conhecer e compreender os desequilíbrios metabólicos nesta fase inicial da evolução não clínica da doença é fundamental para identificar biomarcadores que permitirão identificar pacientes com alto risco de progressão diabética e desta forma atuar nos casos de maior risco de desenvolvimento da DM2. Este estudo tem como principal objetivo o levantamento dos mais recentes biomarcadores relevantes em pré-diabetes. Foi realizada uma pesquisa de artigos publicados entre 2011 e 2021 sobre palavras-chave: ‘Biomarcadores’, ‘Pré--diabetes’, ‘Glicemia de jejum alterada’ e ‘Tolerância à Glicose Diminuída’. Artigos duplicados foram excluídos e procedeu-se à aplicação dos critérios de exclusão que envolviam a associação com outras comorbilidades ou com a dieta. Dos 29 artigos incluídos procedeu-se à separação dos biomarcadores por metabolitos e microRNAs. Os metabolitos mais citados e com resultados mais promissores foram: 2-Hidroxibutirato, aminoácidos aromáticos e de cadeia ramificada, adiponectina, acilcarnitina, proteína C reativa, ferritina, albumina glicada, glicina, Linoleilglicerofosfocolina e triglicerídeos. Os microRNAs identificados mais promissores são: miRNA-15a, miRNA-23a, miRNA--29a, miRNA-126, miRNA-150, miRNA-192, miRNA-320, miRNA-375. Em conclusão, a existência de um painel de biomarcadores permitirá identificar com precisão os indivíduos com maior risco de desenvolverem DM2, promovendo significativamente a saúde dos pacientes e reduzindo a despesa para o serviço nacional de saúde.

O presente Relatório de Estágio reflete o percurso e as atividades realizadas como estagiária, desde fevereiro até ao mês de junho. Surge no âmbito da unidade curricular de Prática Pedagógica Supervisionada, do 2º semestre do 1º ano de Mestrado em Educação Pré-Escolar e Ensino do 1º Ciclo do Ensino Básico. Pretende-se desta forma demonstrar o processo de formação desenvolvido com o intuito de adquirir as competências definidas no programa da unidade curricular em consonância com o pefil do educador estabelecido pelo Decreto-Lei nº241/2001. Este processo teve consequências não só na formação pessoal e profissional da estagiária mas também no desenvolvimento das crianças. O estágio foi realizado em díade num Jardim de Infância da rede pública, no concelho de Ramalde. A recolha de dados sobre a turma e sobre o meio envolvente para a caracterização devida e para a elaboração das reflexões, foi feita através da observação direta e da interação com as crianças, bem como das suas fichas pessoais e de inquéritos. Para um exercício da prática profissional devidamente fundamentado, privilegiaram-se documentos orientadores como as Orientações Curriculares para a Educação Pré-Escolar e as Metas de Aprendizagem para o Pré-Escolar. Ao longo de todo este percurso, procurou-se adequar as estratégias e modelos às características específicas das crianças e do grupo, articular saberes teóricos e desenvolver um papel ativo e um perfil de constante reflexão. Através do trabalho em díade e do trabalho em cooperação com a professora supervisora e a educadora cooperante, houve a preocupação de avaliar o desempenho da estagiária e o crescimento pessoal e profissional, com o objetivo de formar um profissional competente e capaz de tomar decisões favoráveis e assegurar o bem-estar das crianças.

O presente relatório de estágio de qualificação profissional, realizado no âmbito da Unidade Curricular Prática Pedagógica Supervisionada no 1.º Ciclo do Ensino Básico, inserida no Mestrado em Educação Pré-Escolar e Ensino do 1.º Ciclo do Ensino Básico, intenta apresentar a intervenção desenvolvida em contexto de estágio, evidenciando a sua dimensão fulcral no processo de formação da mestranda, nomeadamente, na construção do seu perfil profissional e na consolidação das suas convicções sobre a conceção educativa que se lhe afigura pertinente para com as exigências de um mundo em permanente mudança. Alicerçando a sua conceção educativa em quatro pilares estruturais – aprender a conhecer, aprender a fazer, aprender a viver em sociedade e aprender a ser (Delors et al., 1998) – a mestranda desenvolveu a sua prática pedagógica sustentada em pressupostos teóricos e legais que a fundamentaram e numa atitude crítica e reflexiva que favoreceu a prossecução de ciclos sucessivos de observação, planificação, ação e reflexão, inerentes à metodologia de investigação-ação (Latorre, 2008). Num comprometimento com a melhoria do processo de ensino e aprendizagem e com o seu aperfeiçoamento profissional, edificou a sua intervenção sobre perspetivas pedagógicas construtivistas, valorizando a consecução da integração curricular, a diversificação de estratégias e a diferenciação pedagógica. No final do percurso de estágio, evidencia-se o desenvolvimento significativo de competências inerentes ao perfil do docente do 1.º Ciclo do Ensino Básico, que não termina, contudo, no momento da profissionalização, mas se prolonga, de forma inequívoca e essencial, ao longo da vida (Decretos- Lei n.º 240/2001 e n.º 241/2001, de 30 de agosto).

O presente relatório de estágio (RE) insere-se na unidade curricular Prática Educativa Supervisionada em Educação Pré-Escolar (EPE), no âmbito do Mestrado em Educação Pré- Escolar. Este apresenta e analisa a intervenção da mestranda nos contextos de educação em creche e educação pré-escolar, especificamente numa sala com crianças de dois anos e numa sala com crianças de quatro anos. Analisar-se-á tanto os fundamentos legais que sustentam a prática educativa como os referenciais teóricos que orientaram a ação pedagógica. Reconhecendo a relevância do meio envolvente no processo de aprendizagem da criança, serão caracterizados elementos como as instituições de estágio, a organização do ambiente educativo das salas de atividades e os grupos de crianças. Além disso, será apresentada a metodologia adotada, nomeadamente a metodologia de investigação-ação, que integra processos de observação, planificação, avaliação e reflexão, com o objetivo de aperfeiçoar o processo educativo. Este documento inclui, ainda, a descrição e reflexão das ações desenvolvidas nos diferentes contextos, destacando a importância da pintura como instrumento para o desenvolvimento integral das crianças e articulando conhecimentos teóricos com experiências práticas. Assim, este relatório pretende evidenciar as aprendizagens adquiridas pelas crianças, bem como os aspetos que contribuíram para o crescimento profissional da mestranda.

O presente relatório de estágio descreve o percurso formativo desenvolvido pela mestranda no âmbito da Práticas Educativas Supervisionadas, nas valências de Educação em Creche e Educação Pré-Escolar, integradas no Mestrado em Educação Pré-Escolar da Escola Superior de Educação do Porto. Este documento centra-se nas aprendizagens profissionais construídas ao longo do estágio, articulando os fundamentos teóricos, pedagógicos e legais adquiridos neste percurso. Para tal, o mesmo está estruturado em três grandes capítulos, sendo que o primeiro capítulo apresenta o enquadramento teórico-legal, o segundo capítulo caracteriza os contextos da PES e detalha a metodologia de investigação adotada e o terceiro capítulo reflete sobre as ações desenvolvidas no âmbito da prática. Merece destaque o caráter reflexivo do relatório de estágio, firmado pela metodologia de Investigação-Ação, através da qual a mestranda analisou criticamente as problemáticas emergentes da prática educativa, focando-se no tópico do papel do adulto enquanto facilitador e interlocutor nas interações. Neste sentido, todas as ações educativas desenvolvidas foram sustentadas por um processo interativo de observação, planificação, ação e reflexão, estabelecendo a intencionalidade pedagógica centrada nos interesses e necessidades de aprendizagem das crianças. A investigação baseou-se em referenciais teóricos de orientação socioconstrutivista e em pedagogias participativas, que fundamentaram uma práxis pedagógica centrada na criança, valorizando a escuta ativa e a construção partilhada do conhecimento. Assim, o processo vivido permitiu à mestranda transformar práticas, consolidar competências pessoais e profissionais, bem como desenvolver uma intervenção educativa intencional, sensível e ética, reconhecendo a criança como agente ativo do seu próprio desenvolvimento e aprendizagens.

Air pollution represents a serious risk not only to environment and human health, but also to historical heritage. In this study, air pollution of the Oporto Metropolitan Area and its main impacts were characterized. The results showed that levels of CO, PM10 and SO2 have been continuously decreasing in the respective metropolitan area while levels of NOx and NO2 have not changed significantly. Traffic emissions were the main source of the determined polycyclic aromatic hydrocarbons (PAHs; 16 PAHs considered by U.S. EPA as priority pollutants, dibenzo[a,l]pyrene and benzo[j]fluoranthene) in air of the respective metropolitan area. The mean concentration of 18 PAHs in air was 69.9±39.7 ng m−3 with 3–4 rings PAHs accounting for 75% of the total ΣPAHs. The health risk analysis of PAHs in air showed that the estimated values of lifetime lung cancer risks considerably exceeded the health-based guideline level. Analytical results also confirm that historical monuments in urban areas act as passive repositories for air pollutants present in the surrounding atmosphere. FTIR and EDX analyses showed that gypsum was the most important constituent of black crusts of the characterized historical monument Monastery of Serra do Pilar classified as “UNESCO World Cultural Heritage”. In black crusts, 4–6 rings compounds accounted approximately for 85% of ΣPAHs. The diagnostic ratios confirmed that traffic emissions were the major source of PAHs in black crusts; PAH composition profiles were very similar for crusts and PM10 and PM2.5.

Este relatório examina a relação entre a memória, a plasticidade cerebral e o processo de aprendizagem de inglês como língua estrangeira no contexto do primeiro ciclo do ensino básico. O estudo investiga de que forma os diferentes tipos de memória, como a de curto e longo prazo, influenciam a aquisição linguística, bem como o impacto da plasticidade cerebral no desenvolvimento cognitivo ao longo desse processo. A componente empírica deste estudo é fundamentada com observações em contexto de sala de aula, testes de memória e entrevistas realizadas com alunos e professores durante o período de estágio, de outubro de 2024 a janeiro de 2025, abrangendo quatro turmas. Embora os resultados ofereçam dados relevantes, é importante destacar algumas limitações do estudo, como o tamanho da amostra e a duração relativamente curta do estágio, o que pode ter influenciado a profundidade das conclusões. Os resultados sugerem que memória e plasticidade cerebral estão interligadas, sendo que estratégias de ensino adequadas potenciam ambos os processos e favorecem a retenção linguística. O estudo realça a relevância de abordagens pedagógicas multimodais que envolvam diferentes tipos de memória, promovendo a neuroplasticidade e otimizando os resultados da aprendizagem. Além disso, os resultados contribuem para a compreensão de como as práticas pedagógicas podem apoiar a consolidação da memória e estimular o desenvolvimento cognitivo, fornecendo dados valiosos para o aperfeiçoamento das metodologias de ensino de línguas a crianças.

Este estudo tem como objetivo avaliar o impacto da gamificação e dos jogos na motivação e no desempenho dos alunos do 1º Ciclo do Ensino Básico na aprendizagem de Inglês. O seu principal objetivo é avaliar de que forma estas estratégias podem estimular o envolvimento dos alunos e contribuir para a eficácia do processo de ensino-aprendizagem. O trabalho de investigação-ação, realizado no âmbito de um estágio curricular, tem por base uma abordagem teórica e empírica, integrando observações de aulas, análise de planificações e entrevistas com os alunos. Através de uma metodologia qualitativa, procurou-se compreender as perceções dos intervenientes e identificar práticas eficazes para aumentar a motivação e o envolvimento dos alunos, contribuindo para um ensino mais dinâmico e eficaz. Os resultados obtidos evidenciaram que a introdução de jogos e de atividades gamificadas contribuiu positivamente para o aumento da motivação e da participação dos alunos, especialmente quando estas estratégias foram adaptadas aos diferentes perfis de aprendizagem. Verificou-se também que os alunos sentiram-se mais felizes e envolvidos nas aulas com recurso a estas metodologias, no entanto também ficou clara a necessidade de equilibrar estes recursos com abordagens não lúdicas para garantir a eficácia do processo educativo.

The growing complexity and unpredictability of the external environment in which Non-Profit Organizations (NPOs) operate have increasingly highlighted the need for formal management processes to ensure their effectiveness and long-term viability. The aim of this study is to assess the extent of strategic planning implementation at the Confraria of Santo António, a Private Institution of Social Solidarity (IPSS) in Portugal, evaluate the impact of this implementation on organizational effectiveness, and propose strategies to formalize the organization's strategic management process. Non-profit organizations, especially those in the social sector, play a critical role in addressing pressing social challenges. However, these organizations often face significant barriers, such as limited financial resources, environmental instability, and heightened competition. Among the most pressing challenges is the need for a sustainable management model that guarantees the continued delivery of services while adapting to external changes. Strategic planning has proven to be an effective tool in addressing these issues, as it enables organizations to define long-term goals, allocate resources efficiently, and measure success (Serra et al., 2010; Tucci & Roy, 2022). Despite its recognized value, many NPOs, including the Confraria of Santo António, fail to implement formal strategic planning, which weakens their capacity to respond effectively to these challenges (Fonseca, 2014). This study follows a case study methodology, which allows for an in-depth examination of the practices and challenges faced by the organization (Yin, 2014). Given the research objectives, the approach is primarily qualitative, employing a combination of documentary analysis and semi-structured interviews. The documentary analysis involved a review of key institutional documents, including the organization's annual report and accounts, management reports, and any available strategic planning documents. This analysis helped establish the extent to which strategic planning was present in the organization and provided a baseline for the interviews that followed. The interviews were conducted with four key internal stakeholders at Confraria of Santo António: the Vice-President, the General Director, the Accountant, and the Technical Director. These individuals were selected due to their direct involvement in decision-making processes and their in-depth knowledge of the organization's operations. The semi-structured interview format allowed for flexibility in exploring key themes, such as the organization's approach to strategic planning, its sustainability, and its efforts to create social value. The data collected from the interviews was transcribed and analyzed using the Iramuteq software, which employs Descending Hierarchical Classification (DHC) to identify patterns and categorize responses based on their lexical similarities (Sousa et al., 2020). The analysis revealed several key findings regarding the implementation of strategic planning at Confraria of Santo António. First, it was determined that the organization does not have a formal, documented strategic plan. While the organization does engage in some informal planning and decision-making, these efforts are not systematically structured or aligned with long-term goals. As a result, the organization faces difficulties in responding to environmental changes, securing diverse funding sources, and managing its resources efficiently. Second, the study identified several strengths in the organization’s current practices. These included a strong commitment to its social mission, a dedicated staff, and a long history of service provision. The organization has successfully addressed short-term challenges and provided quality services to its beneficiaries. However, the lack of a formal strategic plan means that these strengths are not always leveraged to their full potential, and the organization struggles to maintain long-term sustainability. Third, the research highlighted several weaknesses in the organization’s current management practices. These include a reactive approach to problem-solving, limited capacity for long-term planning, and a lack of clear performance evaluation metrics. The organization’s reliance on external funding sources, such as government subsidies, further exacerbates its vulnerability to external shocks. Additionally, the absence of a formal strategic plan limits the organization's ability to adapt to changes in the social and economic environment, making it difficult to achieve its long-term goals. The data also revealed a strong recognition among stakeholders of the need for strategic planning to improve organizational effectiveness. Key interviewees expressed support for the development of a Strategic Plan that would guide the organization’s actions, help align its resources with its mission and ensure long-term sustainability. In light of the above, this study makes several contributions to the field of strategic management in the third sector, with significant implications for theory, practice, and policy. From a theoretical point of view, the study provides empirical evidence of the challenges faced by NPOs that lack formal strategic planning, particularly within the social sector in Portugal. The findings highlight management deficits in many NPOs and the resulting negative impact on organizational performance, confirming earlier studies by Fonseca (2014) and Meneses (2012). Furthermore, the study emphasizes the critical role of strategic planning in enhancing organizational sustainability and effectiveness, reinforcing its significance as a strategic management tool, in alignment with the work of Tucci and Roy (2022). Additionally, this research provides a case study of Confraria of Santo António, offering practical implications into how NPOs can overcome the barriers to strategic planning. The proposed Strategic Plan, which is presented as part of the study, serves as a framework for formalizing the organization’s management processes. The plan outlines specific steps for improving resource allocation, diversifying funding sources, and establishing performance evaluation mechanisms. The study also offers recommendations for fostering a culture of strategic thinking within the organization and ensuring that long-term goals are aligned with the organization's mission. From a policy perspective, the study highlights the critical need for greater adoption of formal management tools in NPOs, especially in environments that demand flexibility and long-term vision. The findings suggest that policymakers should encourage and support the implementation of strategic planning in the third sector, as it is essential for ensuring the sustainability and effectiveness of NPOs in meeting social needs. By providing a strategic framework, this research also advocates for policy initiatives aimed at enhancing the capacity of NPOs to deal with dynamic external conditions effectively.