Epitope Mapping by NMR of a Novel Anti-Aβ Antibody (STAB-MAb)

FP7-SP3-People-606950 POCI-01-0145-FEDER-007728 Project No 022161Alzheimer´s Disease (AD) is one of the most common neurodegenerative disorders worldwide. Excess of β-amyloid (Aβ), a peptide with a high propensity to misfold and self-aggregate, is believed to be the major contributor to the observed neuronal degeneration and cognitive decline in AD. Here, we characterize the epitope of a novel anti-Aβ monoclonal antibody, the STAB-MAb, which has previously demonstrated picomolar affinities for both monomers (KD = 80 pM) and fibrils (KD = 130 pM) of Aβ(1–42) and has shown therapeutic efficacy in preclinical mouse models of AD. Our findings reveal a widespread epitope that embraces several key Aβ residues that have been previously described as important in the Aβ fibrillation process. Of note, STAB-MAb exhibits a stronger affinity for the N-terminus of Aβ and stabilizes an α-helix conformation in the central to N-terminal region of the peptide, in addition to disrupting a characteristic salt-bridge of a hairpin structure present in fibrils. The NMR derived epitope supports the observed results from ThT-monitored fluorescence and electron microscopy experiments, in which STAB-MAb was shown to inhibit the formation of aggregates and promote disruption of pre-formed fibrils. In combination with the published in vitro and in vivo assays, our study highlights STAB-MAb as a rare and versatile antibody with analytical, diagnostic and therapeutic efficacy.publishersversionpublishe

The role of DNA concentrations in forensic casework results : regression models application

Póster apresentado no 28th Congress of International Society For Forensic Genetics (ISFG 2019), Praga, República Checa, 9-13 de Setembro de 2019In forensic DNA typing, short tandem repeats (STRs) are the most frequently genotyped markers in order to distinguish between individuals and to relate them to a crime or to exonerate the innocent. In recent years, new controversies have arisen with the advent of more sensitive techniques, allowing profiles to be recovered from minimum amounts of DNA, hence, bringing challenges to weight of evidence evaluation for forensic DNA profiles obtained from low template DNA samples. Introduction of interpretation models, or even new weight of evidence software should be accompanied by a measure of uncertainty that is part of any biological analysis. Specially, due to stochastic effects, the reliability of the obtained profiles might differ between machinery, workflow and also PCR settings in use in different laboratories. In this work we try to understand the relation between Peak Area, DNA concentration and also size marker, using adequate regression models. Buccal swabs from 180 individuals, with unknown identity, were selected for this study. DNA was extracted with prep-n-go™ buffer and quantified using Quantifiler® Trio DNA Quantification kit in a 7500 Real-Time PCR System (Applied Biosystems). STR amplification was performed with Powerplex Fusion 6C amplification kit (Promega). Amplified PCR products were separated and detected in an Applied Biosystems® 3500 Genetic Analyzer using manufacturer’s conditions. Electrophoresis results were analysed with GeneMapper® ID-X v1.4. Statistical analysis was performed with R Studio. Our results allow having an important overview about the relation between DNA concentrations, peak area, and size of the studied genetic markers.N/

Forensic genetic analysis of South Portuguese population with the six dye Powerplex® Fusion 6C

Poster apresentado no 28th Congress of International Society For Forensic Genetics (ISFG 2019), Praga, República Checa, 9-13 de Setembro de 2019As an improvement in efficiency and in Human Discrimination Power, the new six dye multiplex kit PowerPlex® Fusion 6C System, by Promega, available for human identification can co-amplify 27 loci, in a single reaction, have been introduced in the last years with great success. This kit allows the amplification and detection of autosomal loci included in the expanded Combined DNA Index System CODIS, plus the loci Penta D, PENTA E and SE33 as well as Amelogenin for gender determination. Furthermore, this kit includes three Y –STRs (DYS391, DYS576 and DYS570), allowing allelic attribution in a total of 27 loci. This genetic markers extension satisfies not only CODIS but also European Standard Set recommendations. Thinking about continuous human migration movements, especially in a very cosmopolitan region like Lisbon and south of Portugal, and also, in keeping population studies and actualized STR databases we decided to update our previous studies. Our sample is composed of 600 unrelated individuals, from paternity testing with laboratory identity anonymised. DNA was extracted by Prep-n-go BufferTM(Thermo-Fisher Scientific). PCR amplification was performed with PowerPlex® Fusion 6C System, according to manufacturer’s guidelines. Fragment analysis was carried out in an Applied Biosystems® 3500 Genetic Analyser. Electrophoresis results were analysed with GeneMapper® ID-X v1.4. Allele frequencies and population statistics, including Hardy-Weinberg equilibrium p-values from exact test probabilities and forensic parameters were calculated with adequate software. In conclusion, our population information was updated in order to apply most recent data in our casework weight of evidence.N/

Estratégias educacionais do projeto Nutrition up 65: curso de qualificação de profissionais de saúde

INTRODUCTION: Nutrition knowledge among health professionals has a major impact on older adults’ healthcare. However, there is a lack of specialized education offer in this area. Within the Nutrition UP 65 Project, a health professional’s qualification nutrition course was created and implemented from April 2015 to April 2017. We aim to describe this process here. METHODOLOGY: At Faculdade de Ciências da Nutrição e Alimentação da Universidade do Porto, a course of twenty-seven hours was created by six professors. The course comprises eight hours of direct contact and nineteen hours of individual study and was accredited by the University of Porto and credited with one European Credit Transfer System (ECTS). Educational materials and presentations for each session were produced including posters and flyers. A final test was developed to evaluate attendee’s knowledge. Approximately 10% of national health units were identified, randomly selected in each country regional area (NUTS II – “Nomenclatura Comum das Unidades Territoriais para Fins Estatísticos”) and invited to participate. RESULTS: Thirty-five course editions were carried out by twenty-three Porto University accredited instructors for 784 professionals from 241 different health units. Concerning the knowledge evaluation, a 99% approval rate was achieved and the average mark score was of 16.8 out of 20. Course satisfaction survey revealed that 45.7% of the participants classified the course as “good” and 41.1% as “very good”. CONCLUSIONS: The overall balance of this branch of the Nutrition UP 65 project is very positive. The results on health professional’s adherence demonstrate that the interest in this area of nutrition care is high and that this nutrition course is a good opportunity to expand access to specialized education. This highlights the need for further initiatives promoting educational sessions targeting nutrition topics. These course sessions must be acknowledged as a starting point to a new approach in older adult’s nutrition knowledge in Portugal.INTRODUÇÃO: O conhecimento sobre nutrição dos profissionais de saúde tem um grande impacto na saúde dos indivíduos idosos. No entanto, há escassez de oferta de educação especializada nessa área. O Projeto Nutrition UP 65 foi criado e implementado a nível nacional um curso de qualificação em nutrição para profissionais de saúde, que decorreu entre abril de 2015 e abril de 2017. Pretende-se descrever aqui este processo. METODOLOGIA: Na Faculdade de Ciências da Nutrição e Alimentação da Universidade do Porto, seis professores elaboraram um curso de vinte e sete horas. Este curso é composto por oito horas de contacto direto e dezanove horas de estudo individual e foi acreditado e creditado pela Universidade do Porto com um European Credit Transfer System (ECTS). Foram produzidos materiais didáticos e apresentações, bem como cartazes e desdobráveis informativos. Desenvolveu-se uma prova final para avaliar os conhecimentos dos participantes. Cerca de 10% das unidades de saúde nacionais foram identificadas, selecionadas aleatoriamente em cada área regional (NUTS II) e convidadas a participar. RESULTADOS: Realizaram-se 35 edições do curso para 784 profissionais de 241 unidades de saúde diferentes por 23 instrutores credenciados pela Universidade do Porto. No teste de avaliação obteve-se uma taxa de aprovação de 99% e uma pontuação média de 16,8 em 20 valores. 45,7% dos participantes classificaram o curso como “bom” e 41,1% como “muito bom”. CONCLUSÕES: O balanço global desta vertente do Projeto Nutrition UP 65 é muito positivo. Os resultados da adesão dos profissionais de saúde demonstram que o interesse por esta área da intervenção nutricional é elevado e que este curso é uma boa oportunidade para expandir o acesso à formação especializada. Assim, destaca-se a necessidade de outras iniciativas que promovam sessões de formação voltadas para tópicos nutricionais. Estas sessões de formação devem ser reconhecidas como um ponto de partida para uma nova abordagem no conhecimento nutricional dos idosos em Portugal.info:eu-repo/semantics/publishedVersio

Chemical characterization, cytotoxic evaluation and anti-SARS-CoV2 activity of plant extracts rich in hydrolysable tannins

info:eu-repo/semantics/publishedVersio

Investigação de parentesco biológico: a importância de marcadores adicionais em casos de especial complexidade

Poster apresentado no 18º Congresso Nacional de Medicina Legal e Ciências Forenses. Coimbra, Portugal, 21-23 de Novembro de 2019A grande maioria das perícias de investigação de parentesco biológico realizadas pelo Instituto Nacional de Medicina Legal e Ciências Forenses (INMLCF) têm inicio com a ordem do Tribunal para realização da mesma. O mais frequente é o Tribunal dar ordem para nos serem presentes, como intervenientes, um trio constituído por um suposto pai, uma mãe e um(a) filho(a), havendo, no entanto, variações quanto ao número ou tipo de intervenientes, o que pode resultar em maior dificuldade em apresentar resultados com a robustez desejada. Em qualquer dos casos, genericamente, as conclusões possíveis de um estudo de parentesco biológico e mais concretamente de um estudo de paternidade são a exclusão ou não exclusão de paternidade relativamente ao suposto pai em estudo. A conclusão pela não exclusão é sempre acompanhada pela valorização estatística dos resultados, designadamente através do cálculo e apresentação do Índice de Parentesco (IP) e da Probabilidade de Parentesco (W). No caso de uma perícia de investigação de parentesco em que nos seja presente unicamente um filho biológico do suposto pai e um suposto filho biológico, sem possibilidade de estudo das amostras das respetivas mães biológicas nem da amostra biológica do suposto pai, e nos é pedido o estudo sobre a possibilidade de ambos serem filhos biológicos do mesmo pai, a impossibilidade de exclusão da paternidade pode estar associada a valores calculados de IP que podem ser pouco robustos. O ensaio Investigator® HDplex permite o estudo de marcadores genéticos adicionais aos habitualmente estudados na rotina pericial do INMLCF.N/

Dichloro Butenediamides as Irreversible Site‐Selective Protein Conjugation Reagent

We describe maleic-acid derivatives as robust cysteine-selective reagents for protein labelling with comparable kinetics and superior stability relative to maleimides. Diamide and amido-ester derivatives proved to be efficient protein-labelling species with a common mechanism in which a spontaneous cyclization occurs upon addition to cysteine. Introduction of chlorine atoms in their structures triggers ring hydrolysis or further conjugation with adjacent residues, which results in conjugates that are completely resistant to retro-Michael reactions in the presence of biological thiols and human plasma. By controlling the microenvironment of the reactive site, we can control selectivity towards the hydrolytic pathway, forming homogeneous conjugates. The method is applicable to several scaffolds and enables conjugation of different payloads. The synthetic accessibility of these reagents and the mild conditions required for fast and complete conjugation together with the superior stability of the conjugates make this strategy an important alternative to maleimides in bioconjugation