Ser treinador no futebol de formação - construção e aplicação de um sistema de observação e análise do talento no futebol

Dissertação para obtenção do grau de mestre em Desporto - Atividades desportivas para criançasO presente documento é referente ao estágio realizado na época desportiva 2022/2023, na modalidade de futebol, no escalão sub13, num clube de elite em Portugal. O documento está divido em duas partes, uma primeira parte referente ao contexto de estágio, com apresentação do clube, das funções e tarefas desempenhas enquanto treinador estagiário, e a descrição e reflexão de todo o processo de trabalho desenvolvido, nomeadamente nas áreas de planeamento, de intervenção no treino, de intervenção na competição, e na observação e analise. Uma segunda parte, onde se apresenta e desenvolve o estudo de aplicação prática na entidade de estágio. O objetivo do estudo passou pela criação de um sistema de observação e análise de talento no futebol de formação (SOTnFF), na componente técnica, tática, física e mental. O Sistema de Observação foi aplicado na entidade acolhedora, com a observação de 3 atletas, com níveis de potencial e rendimento diferentes, em três momentos competitivos distintos. O estudo permitiu caracterizar e avaliar os jovens jogadores, em função de um conjunto de indicadores, refletindo sobre as expetativas e análises internas do clube sobre o talento dos mesmos. Futuramente, espera-se a continua utilização desta ferramenta de análise e auxílio aos treinadores e coordenadores, com vista a definir e identificar um padrão de atleta talentoso para o escalão etário.This document relates to the internship carried out during the 2022/2023 sports season, in the sport of soccer, specifically in the under-13 age group, at an elite club in Portugal. The document is divided into two parts: the first part refers to the internship context, presenting the club, the functions, and tasks performed as a trainee coach. The second part presents and develops the study of practical application within the internship organization. The study aimed to create a Talent Observation and Analysis System in youth soccer (TOASyS), focusing on technical, tactical, physical, and mental aspects. The observation system was applied to three athletes in the hosting organization, each with different potential and performance levels, during three distinct competitive moments. The study allowed for evaluating and characterizing the players, comparing them based on a set of indicators, reflecting on their talent, as well as the club's internal expectations and analyses. It is expected that this analysis tool will continue to be utilized in the future by coaches and coordinators, aiming to define and identify a pattern of talented athletes for the age group.N/

Mechanistic insights into glycoside 3-oxidases involved in C-glycoside metabolism in soil microorganisms

Funding Information: We thank Diana Santos for preliminary data, Teresa Catarino with stopped-flow analysis, Tiago N. Cordeiro for help with Rosetta, Philippe Carpentier for support with krypton high-pressure experiments, Pedro Matias and Maximino Manzanera for valuable discussions. We thank the beamline staff at ESRF (Grenoble, France) and ALBA (Barcelona, Spain) for their support during the synchrotron data collection and Teresa Silva and Cristina Timóteo (Research Facilities, ITQB-NOVA) for technical assistance. The NMR data were acquired at CERMAX, ITQB-NOVA, Oeiras, Portugal, with equipment funded by FCT, project AAC 01/SAICT/2016. This work was supported by the Fundação para a Ciência e Tecnologia, Portugal, grants, 2022.02027.PTDC (L.O.M.), MOSTMICRO-ITQB (UIDB/04612/2020 and UIDP/04612/2020) (L.O.M. and M.R.V.), LS4FUTURE Associated Laboratory (LA/P/0087/2020) (L.O.M. and M.R.V.), PTDC/BII-BBF/29564/2017 (L.O.M.), UIDB/04326/2020, UIDP/043226/2020 and LA/P/0101/2020 (EPM) and FCT PhD fellowships 2020.07928 (A.T.), 2022.13872 (T.F.), and 2022.09426 (M.V.R.). B-Ligzymes (GA 824017) from the European Union’s Horizon 2020 Research and Innovation Program is also acknowledged for funding T.F. secondment at Zymvol and F.S. secondment at ITQB NOVA. L.M. and X.F.L. acknowledge PID2021-126897NB-I00 project and PRE2019-088412 fellowship, funded by MCIN/AEI/10.13039/501100011033/ FEDER, EU. Funding Information: We thank Diana Santos for preliminary data, Teresa Catarino with stopped-flow analysis, Tiago N. Cordeiro for help with Rosetta, Philippe Carpentier for support with krypton high-pressure experiments, Pedro Matias and Maximino Manzanera for valuable discussions. We thank the beamline staff at ESRF (Grenoble, France) and ALBA (Barcelona, Spain) for their support during the synchrotron data collection and Teresa Silva and Cristina Timóteo (Research Facilities, ITQB-NOVA) for technical assistance. The NMR data were acquired at CERMAX, ITQB-NOVA, Oeiras, Portugal, with equipment funded by FCT, project AAC 01/SAICT/2016. This work was supported by the Fundação para a Ciência e Tecnologia, Portugal, grants, 2022.02027.PTDC (L.O.M.), MOSTMICRO-ITQB (UIDB/04612/2020 and UIDP/04612/2020) (L.O.M. and M.R.V.), LS4FUTURE Associated Laboratory (LA/P/0087/2020) (L.O.M. and M.R.V.), PTDC/BII-BBF/29564/2017 (L.O.M.), UIDB/04326/2020, UIDP/043226/2020 and LA/P/0101/2020 (EPM) and FCT PhD fellowships 2020.07928 (A.T.), 2022.13872 (T.F.), and 2022.09426 (M.V.R.). B-Ligzymes (GA 824017) from the European Union’s Horizon 2020 Research and Innovation Program is also acknowledged for funding T.F. secondment at Zymvol and F.S. secondment at ITQB NOVA. L.M. and X.F.L. acknowledge PID2021-126897NB-I00 project and PRE2019-088412 fellowship, funded by MCIN/AEI/10.13039/501100011033/ FEDER, EU. Publisher Copyright: © 2023, The Author(s).C-glycosides are natural products with important biological activities but are recalcitrant to degradation. Glycoside 3-oxidases (G3Oxs) are recently identified bacterial flavo-oxidases from the glucose-methanol-coline (GMC) superfamily that catalyze the oxidation of C-glycosides with the concomitant reduction of O2 to H2O2. This oxidation is followed by C-C acid/base-assisted bond cleavage in two-step C-deglycosylation pathways. Soil and gut microorganisms have different oxidative enzymes, but the details of their catalytic mechanisms are largely unknown. Here, we report that PsG3Ox oxidizes at 50,000-fold higher specificity (k cat/Km) the glucose moiety of mangiferin to 3-keto-mangiferin than free D-glucose to 2-keto-glucose. Analysis of PsG3Ox X-ray crystal structures and PsG3Ox in complex with glucose and mangiferin, combined with mutagenesis and molecular dynamics simulations, reveal distinctive features in the topology surrounding the active site that favor catalytically competent conformational states suitable for recognition, stabilization, and oxidation of the glucose moiety of mangiferin. Furthermore, their distinction to pyranose 2-oxidases (P2Oxs) involved in wood decay and recycling is discussed from an evolutionary, structural, and functional viewpoint.publishersversionpublishe

Unveiling molecular details behind improved activity at neutral to alkaline pH of an engineered DyP-type peroxidase

DyP-type peroxidases (DyPs) are microbial enzymes that catalyze the oxidation of a wide range of substrates, including synthetic dyes, lignin-derived compounds, and metals, such as Mn2+ and Fe2+, and have enormous biotechnological potential in biorefineries. However, many questions on the molecular basis of enzyme function and stability remain unanswered. In this work, high-resolution structures of PpDyP wild-type and two engineered variants (6E10 and 29E4) generated by directed evolution were obtained. The X-ray crystal structures revealed the typical ferredoxin-like folds, with three heme access pathways, two tunnels, and one cavity, limited by three long loops including catalytic residues. Variant 6E10 displays significantly increased loops’ flexibility that favors function over stability: despite the considerably higher catalytic efficiency, this variant shows poorer protein stability compared to wild-type and 29E4 variants. Constant-pH MD simulations revealed a more positively charged microenvironment near the heme pocket of variant 6E10, particularly in the neutral to alkaline pH range. This microenvironment affects enzyme activity by modulating the pKa of essential residues in the heme vicinity and should account for variant 6E10 improved activity at pH 7–8 compared to the wild-type and 29E4 that show optimal enzymatic activity close to pH 4. Our findings shed light on the structure–function relationships of DyPs at the molecular level, including their pH-dependent conformational plasticity. These are essential for understanding and engineering the catalytic properties of DyPs for future biotechnological applications

Corrigendum to “Unveiling molecular details behind improved activity at neutral to alkaline pH of an engineered DyP-type peroxidase” Comput Struct Biotechnol J, 20 (2022), 3899–3910

Funding Information: This work was supported by the Fundação para a Ciência e Tecnologia, Portugal , grants CEECIND/02300/2017 , PTDC/BBBEBB/0122/2014 , PTDC/BII-BBF/29564/2017 , EXPL/BIA-BQM/0473/2021 , MOSTMICRO-ITQB ( UIDB/04612/2020 and UIDP/04612/2020 ), LS4FUTURE Associated Laboratory ( LA/P/0087/2020 ), BioISI ( UIDB/04046/2020 and UIDP/04046/2020 ), and projects UIDB/04326/2020 , UIDP/04326/2020 , LA/P/0101/2020 , and the operational programs CRESC Algarve 2020 and COMPETE through project EMBRC.PT ALG-01–0145-FEDER-022121 . Publisher Copyright: © 2022 The Author(s)The authors would like to add in the Funding section the project reference EXPL/BIA-BQM/0473/2021 in order to be like this: This work was supported by the Fundação para a Ciência e Tecnologia, Portugal, grants CEECIND/02300/2017, PTDC/BBBEBB/0122/2014, PTDC/BII-BBF/29564/2017, EXPL/BIA-BQM/0473/2021, MOSTMICRO-ITQB (UIDB/04612/2020 and UIDP/04612/2020), LS4FUTURE Associated Laboratory (LA/P/0087/2020), BioISI (UIDB/04046/2020 and UIDP/04046/2020), and projects UIDB/04326/2020, UIDP/04326/2020, LA/P/0101/2020, and the operational programs CRESC Algarve 2020 and COMPETE through project EMBRC.PT ALG-01–0145-FEDER-022121. The authors would like to apologise for any inconvenience caused.publishersversionpublishe