A Metabolomics-Inspired Strategy for the Identification of Protein Covalent Modifications

This work was supported by Fundação para a Ciência e a Tecnologia (FCT), Portugal, through projects UID/QUI/00100/2019, IF/01091/2013/CP1163/CT0001 and PTDC/QUIQAN/32242/2017 as well as doctoral fellowships SFRH/BD/102846/2014 (to CC) and SFRH/BD/140157/2018 (to JN);joint funding from FCT and the COMPETE Program is also acknowledge through RNEM-LISBOA-01-0145-FEDER-022125-funded postdoctoral fellowship (to JM).Identification of protein covalent modifications (adducts) is a challenging task mainly due to the lack of data processing approaches for adductomics studies. Despite the huge technological advances in mass spectrometry (MS) instrumentation and bioinformatics tools for proteomics studies, these methodologies have very limited success on the identification of low abundant protein adducts. Herein we report a novel strategy inspired on the metabolomics workflows for the identification of covalently-modified peptides that consists on LC-MS data preprocessing followed by statistical analysis. The usefulness of this strategy was evaluated using experimental LC-MS data of histones isolated from HepG2 and THLE2 cells exposed to the chemical carcinogen glycidamide. LC-MS data was preprocessed using the open-source software MZmine and potential adducts were selected based on the m/z increments corresponding to glycidamide incorporation. Then, statistical analysis was applied to reveal the potential adducts as those ions are differently present in cells exposed and not exposed to glycidamide. The results were compared with the ones obtained upon the standard proteomics methodology, which relies on producing comprehensive MS/MS data by data dependent acquisition and analysis with proteomics data search engines. Our novel strategy was able to differentiate HepG2 and THLE2 and to identify adducts that were not detected by the standard methodology of adductomics. Thus, this metabolomics driven approach in adductomics will not only open new opportunities for the identification of protein epigenetic modifications, but also adducts formed by endogenous and exogenous exposure to chemical agents.publishersversionpublishe

Mover – Activity Monitor and Fall Detector for Android

Mover is an exciting application that allows you to track your activity level and helps you become more active. Mover includes also an experimental fall detection system

Probable Person-to-Person Transmission of Legionnaires’ Disease

Correspondence to the Editor.Legionnaires’ disease is an often severe form of pneumonia that is typically acquired by susceptible persons (e.g., elderly persons and smokers) through inhalation of aerosols that contain legionella species.1-4 A cluster of cases of this disease occurred in Vila Franca de Xira, Portugal, in 2014

The compound event that triggered the destructive fires of October 2017 in Portugal

Portugal is regularly affected by destructive wildfires that have severe social, economic, and ecological impacts. The total burnt area in 2017 (∼540,000 ha) marked the all-time record value since 1980 with a tragic toll of 114 fatalities that occurred in June and October events. The local insurance sector declared it was the costliest natural disaster in Portugal with payouts exceeding USD295 million. Here, the 2017 October event, responsible for more than 200,000 ha of burnt area and 50 fatalities is analyzed from a compound perspective. A prolonged drought led to preconditioned cumulative hydric stress of vegetation in October 2017. In addition, on 15 October 2017, two other major drivers played a critical role: 1) the passage of hurricane Ophelia off the Coast of Portugal, responsible for exceptional meteorological conditions and 2) the human agent, responsible for an extremely elevated number of negligent ignitions. This disastrous combination of natural and anthropogenic drivers led to the uncontrolled wildfires observed on 15 October

Highly water soluble room temperature superionic liquids of APIs

Fundacao para a Ciencia e a Tecnologia through project (PEst-C/LA0006/2013). The authors also thank Prof. Madalena Dionisio and Dr Natalia Correia for their support with the DSC analyses.Herein a straightforward approach for the enhancement of the water solubility of common antibiotic and NSAID active pharmaceutical ingredients (APIs) is presented. The APIs are converted into ionic liquids (API-ILs) and molten salts by combination with the organic superbases TMG, DBU and DBN. The prepared superionic liquids were characterized by 1H and 13C NMR as well as FTIR spectroscopy and elemental analysis. Most products are amorphous non-polymorphic room temperature ionic liquids with very high solubility in water, which may enhance the bioavailability of the API-ILs in comparison with the parent drugs.authorsversionpublishe

Acetylcholine release and choline uptake by cuttlefish (Sepia officinalis) optic lobe synaptosomes

Acetylcholine (ACh), which is synthesized from choline (Ch), is believed to hold a central place in signaling mechanisms within the central nervous system (CNS) of cuttlefish (Sepia officinalis) and other coleoid cephalopods. Although the main elements required for cholinergic function have been identified in cephalopods, the transmembrane translocation events promoting the release of ACh and the uptake of Ch remain largely unsolved. The ACh release and Ch uptake were quantitatively studied through the use of in vitro chemiluminescence and isotopic methods on a subcellular fraction enriched in synaptic nerve endings (synaptosomes) isolated from cuttlefish optic lobe. The ACh release evoked by K+ depolarization was found to be very high (0.04 pmol ACh.s-(1).mg(-1) protein). In response to stimulation by veratridine, a secretagogue (a substance that induces secretion) that targets voltage-gated Na+ channels, the release rate and the total amount of ACh released were significantly lower, by 10-fold, than the response induced by KCl. The high-affinity uptake of choline was also very high (31 pmol Ch.min(-1).mg(-1) protein). The observed ACh release and Ch uptake patterns are in good agreement with published data on preparations characterized by high levels of ACh metabolism, adding further evidence that ACh acts as a neurotransmitter in cuttlefish optic lobe.FCTPOCTI/BSE/46721/2002SFRH/BPD/14677/2003SFRH/BD/1079/2000SFRH/BD/6403/2001SFRH/BD/18101/2004EC (to Y.D.) - LIPIDIET - QLK1-CT-2002-0017

Instalação de um ensaio de produção intensiva de biomassa lenhosa para energia

No âmbito de um projecto de investigação tendo como propósito o desenvolvimento de sistemas sustentáveis de produção de biomassa lenhosa para energia, foi instalado em Bragança, em 2008, um ensaio de produção intensiva de biomassa lenhosa com base em culturas de curta rotação. Este ensaio tem como objectivos específicos a avaliação dos efeitos de factores como material vegetal, densidade de plantação, rega e fertilização no crescimento e produção de biomassa lenhosa. Nesta comunicação descrevemos a instalação do ensaio relativamente ao desenho experimental seguido, estrutura interna das parcelas de estudo, material vegetal utilizado, operações de preparação do solo e de plantação e ainda gestão da cultura com respeito a fertilização, rega e protecção. Avaliamos também a sobrevivência das plantas no final da primeira estação de crescimento, bem como apresentamos os custos gerais associados à instalação. A apresentação deste trabalho permite partilhar com a comunidade científica e técnica nacional a experiência da instalação deste tipo de culturas, muito recentes no nosso país, e dessa forma contribuir para a definição de processos ajustados e eficientes de instalação e gestão de culturas lenhosas para produção de biomassa para energia em Portugal

Cryptic Prophages Contribution for Campylobacter jejuni and Campylobacter coli Introgression

Campylobacter coli and C. jejuni, the causing agents of campylobacteriosis, are described to be undergoing introgression events, i.e., the transference of genetic material between different species, with some isolates sharing almost a quarter of its genome. The participation of phages in introgression events and consequent impact on host ecology and evolution remain elusive. Three distinct prophages, named C. jejuni integrated elements 1, 2, and 4 (CJIE1, CJIE2, and CJIE4), are described in C. jejuni. Here, we identified two unreported prophages, Campylobacter coli integrated elements 1 and 2 (CCIE1 and CCIE2 prophages), which are C. coli homologues of CJIE1 and CJIE2, respectively. No induction was achieved for both prophages. Conversely, induction assays on CJIE1 and CJIE2 point towards the inducibility of these prophages. CCIE2-, CJIE1-, and CJIE4-like prophages were identified in a Campylobacter spp. population of 840 genomes, and phylogenetic analysis revealed clustering in three major groups: CJIE1-CCIE1, CJIE2-CCIE2, and CJIE4, clearly segregating prophages from C. jejuni and C. coli, but not from human- and nonhuman-derived isolates, corroborating the flowing between animals and humans in the agricultural context. Punctual bacteriophage host-jumps were observed in the context of C. jejuni and C. coli, and although random chance cannot be fully discarded, these observations seem to implicate prophages in evolutionary introgression events that are modulating the hybridization of C. jejuni and C. coli species.F.F.V. is funded by Fundação para a Ciência e a Tecnologia (FCT) through an Assistant Researcher grant CEECIND/03023/2017, and a project grant (PTDC/BTM-SAL/28978/2017) that supported this work. The work is partially supported by National funds from FCT, projects UIDB/04138/2020 and UIDP/04138/2020. Campylobacter strains were sequenced under the GenomePT project (POCI-01-0145-FEDER-022184), supported by COMPETE 2020—Operational Programme for Competitiveness and Internationalization (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by FCT. This work was also supported by Fundos FEDER through the Programa Operacional Factores de Competitividade—COMPETE and by Fundos Nacionais through the FCT within the scope of the project UID/BIM/00009/2019 (Centre for Toxicogenomics and Human Health-ToxOmics).info:eu-repo/semantics/publishedVersio

Recurrent Campylobacter jejuni Infections with In Vivo Selection of Resistance to Macrolides and Carbapenems: Molecular Characterization of Resistance Determinants

Erratum in: Microbiol Spectr. 2023 Dec 12;11(6):e0312123. doi: 10.1128/spectrum.03121-23. Epub 2023 Oct 10.Free PMC article: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10434052/We present two independent cases of recurrent multidrug-resistant Campylobacter jejuni infection in immunocompromised hosts and the clinical challenges encountered due to the development of high-level carbapenem resistance. The mechanisms associated with this unusual resistance for Campylobacters were characterized. Initial macrolide and carbapenem-susceptible strains acquired resistance to erythromycin (MIC . 256mg/L), ertapenem (MIC . 32mg/L), and meropenem (MIC . 32mg/L) during treatment. Carbapenem-resistant isolates developed an in-frame insertion resulting in an extra Asp residue in the major outer membrane protein PorA, within the extracellular loop L3 that connects b-strands 5 and 6 and forms a constriction zone involved in Ca21 binding. The isolates presenting the highest MIC to ertapenem exhibited an extra nonsynonymous mutation (G167AjGly56Asp) at PorA’s extracellular loop L1. IMPORTANCE Carbapenem susceptibility patterns suggest drug impermeability, related to either insertion and/or single nucleotide polymorphism (SNP) within porA. Similar molecular events occurring in two independent cases support the association of these mechanisms with carbapenem resistance in Campylobacter spp.Importance: Carbapenem susceptibility patterns suggest drug impermeability, related to either insertion and/or single nucleotide polymorphism (SNP) within porA. Similar molecular events occurring in two independent cases support the association of these mechanisms with carbapenem resistance in Campylobacter spp.This work was partially supported by GenomePT (ref. POCI-01-0145-FEDER-022184) from Fundação para a Ciência e Tecnologia, Portugal.info:eu-repo/semantics/publishedVersio