Development of a new approach based on NADH dehydrogenase subunit 1(ND1) marker and high resolution melting (HRM) analysis towards the authentication of the geographical origin of honey

In the present study, an approach for verifying the geographical origin of honey based on its entomological origin is proposed. The method was developed based on the polymerase chain reaction (PCR) amplification of a new markercontaining different single nucleotide polymorphisms characteristic of honeybees of different mitochondrial (mtDNA) lineages, therefore generating different fluorescent curves on HRM analysis. The method was successfully applied to honeys from Portugal and Italy, expected to be from lineages A and C, respectively, demonstrating their origin compliance.The authors are grateful to Programa Nacional Apícola 2020-2022 for funding the project “AUTENT+ Desenvolvimento de abordagens inovadoras com vista à valorização e exploração do potencial de mercado do mel Português”, to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO UIDB/00690/2020, to Fenapícola and Capemel for supplying the Portuguese honeys and to Dr. Antonio Nanetti (CREA-AA) for the Italian honeys. D. Henriques is supported by the project Bee Happy (POCI-01-0145-FEDER-029871) funded by FEDER, COMPETE 2020-POCIand FCT and A.R. Lopes by the PhD scholarship funded by the FCT SFRH/BD/143627/2019.info:eu-repo/semantics/publishedVersio

Synthetic approaches to a challenging and unusual structure—an amino-pyrrolidine guanine core

Associate Laboratory for Green Chemistry-LAQV, which is financed by national funds from FCT/MCTES (UIDB/50006/2020). Grant number SFRH/BD/136692/2018 from FCT/MCTES.The synthesis of an unreported 2-aminopyrrolidine-1-carboxamidine unit is here described for the first time. This unusual and promising structure was attained through the oxidative decarboxylation of amino acids using the pair of reagents, silver(I)/peroxydisulfate (Ag(I)/S2O82−) followed by intermolecular (in the case of L-proline derivative) and intramolecular trapping (in the case of acyl L-arginine) by N-nucleophiles. The L-proline approach has a broader scope for the synthesis of 2-aminopyrrolidine-1-carboxamidine derivatives, whereas the intramolecular cyclization afforded by the L-acylarginines, when applied, results in higher yields. The former allowed the first synthesis of cernumidine, a natural alkaloid isolated in 2011 from Solanum cernuum Vell, as its racemic form.publishersversionpublishe

High resolution melting analysis of a COI mini-barcode as a simple approach for the entomological authentication of honey

Honey is highly valued for its taste, aroma, content in bioactive compounds and for being a natural food. In the European Union (EU), market demand for honey is higher than the domestic production and therefore a substantial amount of honey is imported. According to a 2014 European Parliament report on fraud in the food chain, honey was ranked as the 6th food product most prone to adulterateration.1 Up until now, honey authenticity addressed mainly sugars addition and botanical origin. However, an increased attention has recently been paid to honey entomological origin as it also relates to its geographical origin since honeybees carrying mitochondrial DNA (mtDNA) of distinct ancestries can be found across Europe. While in Portugal the predominant mtDNA of the autochthonous subspecies Apis mellifera iberiensis belongs to the A-lineage, when moving towards the northeastern part of the Iberian Peninsula this lineage is gradually replaced by the M-lineage. The native distribution of the M-lineage A. m. mellifera expands from the Pyrenees to Scandinavia and from the British Isles to the Ural Mountains while the C-lineage A. m. ligustica and A. m. carnica are naturally found in the Apennine and Balkan peninsulas, respectivelyThis work was funded by the project-2022, financed by IFAP. The authors are also grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020), to Fenapícola and Capemel for supplying the Portuguese honeys, to Dr. Antonio Nanetti (CREA-AA) for the Italian honeys and to António Pajuelo (Pajuelo Consultores Apícolas S.L.) for the Spanish honeys. D. Henriques is supported by the project BeeHappy (POCI-01-0145-FEDER-029871) funded by FEDER, COMPETE 2020-POCI and FCT. A. R. Lopes and A. Quaresma acknowledge the PhD scholarship funded by the FCT (SFRH/BD/143627/2019 and DFA/BD/5155/2020, respectively).info:eu-repo/semantics/publishedVersio

Climate adaptation, transitions, and socially innovative action-research approaches

Climate change may be a game-changer for scientific research by promoting a science that is grounded in linking the production of knowledge and societal action in a transition toward more sustainable development pathways. Here, we discuss participatory action-research (PAR) as a way of thinking and leading investigations that may promote incremental and transformative changes in the context of climate change adaptation research. Our exploration is addressed in the Portuguese context, where PAR and sustainable transition studies are still marginal, and adaptation processes are a recent topic on political agendas. We describe the characteristics of PAR and use two studies of adaptation to illustrate how research and practice co-evolve through interactive cycles. The two studies are works in progress, rather than completed PAR processes. Climate change adaptation is an ongoing and long-term process. Moreover, in Portugal, as in many regions of the world, climate change adaptation is a fairly new topic. Thus, both case studies are now initiating a long-term process of change and adaptation. The completion of one research cycle is a realistic expectation that we have achieved in the two case study experiences. In our discussion of the case studies, we consider how these experiences provide insights into the role of PAR for long-term regime changes. We conclude by pointing to the societal needs addressed by PAR, as a pragmatically oriented and context-specific research design. The approach can be complementary to other frameworks in sustainable transition studies such as transition management. Being more pragmatically oriented, PAR cycles may influence incrementally transformative changes that can be guided by transition management’s long-term design for governing sustainable transitions

Low Serum Levels of DKK2 Predict Incident Low-Impact Fracture in Older Women

There are currently no robust noninvasive markers of fragility fractures. Secreted frizzled related protein-1 (sFRP-1), dickkopf-related protein 1 (DKK1) and DKK2, and sclerostin (SOST) inhibit Wnt signaling and interfere with osteoblast-mediated bone formation. We evaluated associations of serum levels of sFRP-1, DKK1, DKK2, and SOST with incident low-impact fracture and BMD in 828 women aged ≥65 years from EpiDoC, a longitudinal population-based cohort. A structured questionnaire during a baseline clinical appointment assessed prevalent fragility fractures and clinical risk factors (CRFs) for fracture. Blood was collected to measure serum levels of bone turnover markers and Wnt regulators. Lumbar spine and hip BMD were determined by DXA scanning. Follow-up assessment was performed through a phone interview; incident fragility fracture was defined by any new self-reported low-impact fracture. Multivariate Cox proportional hazard models were used to analyze fracture risk adjusted for CRFs and BMD. During a mean follow-up of 2.3 ± 1.0 years, 62 low-impact fractures were sustained in 58 women. A low serum DKK2 level (per 1 SD decrease) was associated with a 1.5-fold increase in fracture risk independently of BMD and CRFs. Women in the two lowest DKK2 quartiles had a fracture incidence rate of 32 per 1000 person-years, whereas women in the two highest quartiles had 14 fragility fractures per 1000 person-years. A high serum sFRP1 level was associated with a 1.6-fold increase in fracture risk adjusted for CRFs, but not independently of BMD. Serum levels of SOST (r = 0.191; p = 0.0025) and DKK1(r = -0.1725; p = 0.011) were correlated with hip BMD, but not with incident fragility fracture. These results indicate that serum DKK2 and sFRP1 may predict low-impact fracture. The low number of incident fractures recorded is a limitation and serum levels of Wnt regulators should be further studied in other populations as potential noninvasive markers of fragility fractures. © 2019 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.publishersversionpublishe

Molecular Interactions in Ionic Liquids: The NMR Contribution towards Tailored Solvents

Ionic liquids have been on the spotlight of chemical research field in the last decades. Their physical properties (low vapor pressure, thermal stability, and conductivity) and the possibility of fine tuning make them a versatile class of compounds for a wide range of applications, such as catalysis, energy, and material sciences. Ionic liquids can establish multiple intermolecular interactions with solutes such as electrostatic, van der Waals, or hydrogen bonds. The prospect of designing ionic liquid structures toward specific applications has attracted the attention to these alternative solvents. However, their rational design demands a molecular detailed view, and Nuclear Magnetic Resonance is a unique and privileged technique for this purpose, as it provides atomic resolution and at the same time enables the study of dynamic information. In this chapter, we provide an overview about the application of Nuclear Magnetic Resonance spectroscopy techniques as a methodology for the rational design of ionic liquids as solvents for small organic compounds, CO2 capture, and polymers such as cellulose focusing mainly in the last 10 years

A robust assay to monitor ataxin-3 amyloid fibril assembly

Spinocerebellar ataxia type 3 (SCA3) is caused by the expansion of a glutamine repeat in the protein ataxin-3, which is deposited as intracellular aggregates in affected brain regions. Despite the controversial role of ataxin-3 amyloid structures in SCA3 pathology, the identification of molecules with the capacity to prevent aberrant self-assembly and stabilize functional conformation(s) of ataxin-3 is a key to the development of therapeutic solutions. Amyloid-specific kinetic assays are routinely used to measure rates of protein self-assembly in vitro and are employed during screening for fibrillation inhibitors. The high tendency of ataxin-3 to assemble into oligomeric structures implies that minor changes in experimental conditions can modify ataxin-3 amyloid assembly kinetics. Here, we determine the self-association rates of ataxin-3 and present a detailed study of the aggregation of normal and pathogenic ataxin-3, highlighting the experimental conditions that should be considered when implementing and validating ataxin-3 amyloid progress curves in different settings and in the presence of ataxin-3 interactors. This assay provides a unique and robust platform to screen for modulators of the first steps of ataxin-3 aggregation—a starting point for further studies with cell and animal models of SCA3.This study was supported by FEDER funds through the COMPETE 2020—Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020; Portuguese funds through FCT in the framework of the projects “PQTools: Molecular tools for Machado-Joseph Disease” (POCI-01-0145-FEDER-031173), “NAPPIT-MJD:Nuclear ataxin-3 protein-protein interactions as therapeutic targets in Machado-Joseph disease” (POCI-01-0145-FEDER-029056), “AggreGATE: Targeting diffusible oligomers of alpha-synuclein and ataxin-3: a drug repurposing opportunity for the treatment of neurodegenerative diseases” (POCI-01-0145-FEDER-031323), and “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274); the European Union’s Horizon 2020 Research and Innovation programme under grant agreement ID 952334 “PhasAGE”. The work was also supported by a research grant from National Ataxia Foundation to A.S. F.F. is the recipient of an FCT PhD fellowship (SFRH/BD/133009/2017)

Essential genetic findings in neurodevelopmental disorders

Neurodevelopmental disorders (NDDs) represent a growing medical challenge in modern societies. Ever-increasing sophisticated diagnostic tools have been continuously revealing a remarkably complex architecture that embraces genetic mutations of distinct types (chromosomal rearrangements, copy number variants, small indels, and nucleotide substitutions) with distinct frequencies in the population (common, rare, de novo). Such a network of interacting players creates difficulties in establishing rigorous genotype-phenotype correlations. Furthermore, individual lifestyles may also contribute to the severity of the symptoms fueling a large spectrum of gene-environment interactions that have a key role on the relationships between genotypes and phenotypes.Herein, a review of the genetic discoveries related to NDDs is presented with the aim to provide useful general information for the medical community.info:eu-repo/semantics/publishedVersio

Carcinogenic Ability of Schistosoma Haematobium Possibly through Oncogenic Mutation of KRAS Gene

Schistosoma haematobium is a parasitic flatworm that infects millions of people, mostly in the developing world, and is associated with high incidence of bladder cancer, although why is not clear. Previously, we have used CD-1 mice to show that Schistosoma haematobium total antigen (Sh) has a carcinogenic ability. Sh intravesically instillation induced the development of several urothelial lesions, namely nodular hyperplasia and dysplasia (LGIUN—Low Grade Intra-Urothelial Neoplasia) after 40 weeks of treatment. These results suggested that Sh induce urothelium malignization. Bladder carcinoma frequently harbours gene mutations that constitutively activate the receptor tyrosine kinase-Ras pathway for this reason we studied activating mutations in KRAS gene. Twenty percent of the bladders with dysplasia presented a KRAS mutation in codon 12 of exon 2. We concluded from these results that the parasite extract of S. haematobium has carcinogenic ability possibly through oncogenic mutation of KRAS gene

Avaliação da Vulnerabilidade das Águas Subterrâneas no Município de Santa Cruz do Sul, RS/Brasil

Nesta pesquisa realizada no município de Santa Cruz do Sul, RS-Brasil, utilizou-se dados pré-existentes em banco de dados de empresas privadas, estatais, municipais e federais com o objetivo de espacializar as informações através da metodologia GOD e do aplicativo SURFER™ versão 6.0. Com essa metodologia GOD utilizada foi possível prever e avaliar a vulnerabilidade natural dos aqüíferos através dos parâmetros: grau de confinamento hidráulico (G); ocorrência do suprajacente (O) e distância do nível da água (D). Foi utilizada uma planilha eletrônica, MS Excel™, para dispor os parâmetros hidrodinâmicos e físico-químicos das águas subterrâneas. Ilustra-se a superfície potenciométrica indicativa do fluxo subterrâneo, as concentrações de flúor e o índice de vulnerabilidade. A faixa de variação do flúor foi de 0,0 até 3,6 mgL-1, e o índice de vulnerabilidade obtido, apresentou dos 25 poços simulados; 4 poços na classe desprezível, 7 poços na classe baixa, á poços na classe média e apenas 1 na classe alta. Discutem-se alguns aspectos relativos á legislação das águas subterrâneas e seu uso para abastecimento público