Dand5 is involved in zebrafish tailbud cell movement

Funding Information: This work was supported by the Fundação para a Ciência e a Tecnologia (FCT-ANR/BEX-BID/0153/2012 research grant and by the Project LysoCil funded by the European Union Horizon 2020 research and innovation under grant agreement No. 811087. SSL was funded by an FCT CEEC contract for Principal Investigator reference 2018CEECIND/02170/2018 and CB was funded by a FCT SFRH/BD/141034/2018 PhD fellowship. NMS and IGC Fish Facilities were supported from the research infrastructure Congento, co-financed by Lisboa Regional Operational Programme (Lisboa 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and FCT under the project LISBOA-01-0145-FEDER-022170. Publisher Copyright: Copyright © 2023 Bota, Martins and Lopes.During vertebrate development, symmetry breaking occurs in the left-right organizer (LRO). The transfer of asymmetric molecular information to the lateral plate mesoderm is essential for the precise patterning of asymmetric internal organs, such as the heart. However, at the same developmental time, it is crucial to maintain symmetry at the somite level for correct musculature and vertebrae specification. We demonstrate how left-right signals affect the behavior of zebrafish somite cell precursors by using live imaging and fate mapping studies in dand5 homozygous mutants compared to wildtype embryos. We describe a population of cells in the vicinity of the LRO, named Non-KV Sox17:GFP+ Tailbud Cells (NKSTCs), which migrate anteriorly and contribute to future somites. We show that NKSTCs originate in a cluster of cells aligned with the midline, posterior to the LRO, and leave that cluster in a left-right alternating manner, primarily from the left side. Fate mapping revealed that more NKSTCs integrated somites on the left side of the embryo. We then abolished the asymmetric cues from the LRO using dand5−/− mutant embryos and verified that NKSTCs no longer displayed asymmetric patterns. Cell exit from the posterior cluster became bilaterally synchronous in dand5−/− mutants. Our study revealed a new link between somite specification and Dand5 function. The gene dand5 is well known as the first asymmetric gene involved in vertebrate LR development. This study revealed a new link for Dand5 as a player in cell exit from the maturation zone into the presomitic mesoderm, affecting the expression patterns of myogenic factors and tail size.publishersversionpublishe

Cracking the breast cancer glyco-code through glycan-lectin interactions: Targeting immunosuppressive macrophages

UIDB/04462/2020 UIDP/04378/2020 UIDB/04378/2020 PhD fellowships 2020.07623.BDThe immune microenvironment of breast cancer (BC) is composed by high macrophage infiltrates, correlated with the most aggressive subtypes. Tumour-associated macrophages (TAM) within the BC microenvironment are key regulators of immune suppression and BC progression. Nevertheless, several key questions regarding TAM polarisation by BC are still not fully understood. Recently, the modulation of the immune microenvironment has been described via the recognition of abnormal glycosylation patterns at BC cell surface. These patterns rise as a resource to identify potential targets on TAM in the BC context, leading to the development of novel immunotherapies. Herein, we will summarize recent studies describing advances in identifying altered glycan structures in BC cells. We will focus on BC-specific glycosylation patterns known to modulate the phenotype and function of macrophages recruited to the tumour site, such as structures with sialylated or N-acetylgalactosamine epitopes. Moreover, the lectins present at the surface of macrophages reported to bind to such antigens, inducing tumour-prone TAM phenotypes, will also be highlighted. Finally, we will discuss and give our view on the potential and current challenges of targeting these glycan-lectin interactions to reshape the immunosuppressive landscape of BC.publishersversionpublishe

Molecular biomarkers associated with respiratory insufficiency in amyotrophic lateral sclerosis

Abstract in proceedings of the Fourth International Congress of CiiEM: Health, Well-Being and Ageing in the 21st Century, held at Egas Moniz’ University Campus in Monte de Caparica, Almada, from 3–5 June 2019.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.info:eu-repo/semantics/publishedVersio

Tubulin post-translational modifications: the elusive roles of acetylation

This work was funded by Instituto Politécnico de Lisboa IPL/2021/ObeCil_ESTeSL and IPL/2022/WintCilGlu_ESTeSL.Microtubules (MTs), dynamic polymers of α/β-tubulin heterodimers found in all eukaryotes, are involved in cytoplasm spatial organization, intracellular transport, cell polarity, migration, division, and cilia biology. MTs functional diversity depends on the differential expression of distinct tubulin isotypes and is amplified by a vast number of different post-translational modifications (PTMs). The addition/removal of PTMs to α- or β-tubulins is catalyzed by specific enzymes and allows combinatory patterns largely enriching the distinct biochemical and biophysical properties of MTs, creating a code read by distinct proteins, including microtubule-associated proteins (MAPs), which allow cellular responses. This review is focused on tubulin-acetylation, whose cellular roles continue to generate debate. We travel through the experimental data pointing to α-tubulin Lys40 acetylation role as being an MT stabilizer and a typical PTM of long-lived MTs, to the most recent data, suggesting that Lys40 acetylation enhances MT flexibility and alters the mechanical properties of MTs, preventing MTs from mechanical aging characterized by structural damage. Additionally, we discuss the regulation of tubulin acetyltransferases/desacetylases and their impacts on cell physiology. Finally, we analyze how changes in MT acetylation levels have been found to be a general response to stress and how they are associated with several human pathologies.info:eu-repo/semantics/publishedVersio

Development of photocatalytic poly(vinylidenefluoride-trifluoroethylene)/TiO₂ porous membranes

Photocatalysis has become an attractive process to remove contaminants from aquatic environments1. Photocatalysis consists on the use of solar light to generate highly oxidizing species, most often the hydroxyl radical (•OH), to destroy harmful compounds 2. Catalysts are required in this process and, due to its significant oxidizing properties under UV irradiation, TiO2 is the most widely used photocatalyst [ref?=6]. One disadvantage of the UV/photocatalyst process is the fast recombination rate of the electron-hole pairs3. Doping the photocatalysts with other elements is an effective approach to overcome this problem[ref]. Nevertheless, one of the largest drawbacks of the use of photocatalyst particles is the recycling and reutilization of the nanoparticles, which is time consuming and requires expensive processes2. In order to overcome this drawback, TiO2 immobilization in several substrates (e.g. glass, zeolite, silica and ceramic) has been explored. This work reports the optimization and photocatalytic activity of poly(vinylidenefluoride - trifluoroethylene) (PVDF-TrFE)/TiO2 nanocomposites.Work supported by FEDER (FCT), project PEST-C/FIS/UI607/2011 and PEST-C/QUI/UIO686/2011 and Matepro – Optimizing Materials and Processes”, ref. NORTE-07-0124-FEDER-00003

Zebrafish motile cilia as a model for primary ciliary dyskinesia

Funding Information: Funding: This work was a product of the Project LYSOCIL funded by the European Union Horizon 2020 research and innovation under grant agreement No 811087. It received funding from Fundação para a Ciencia e tecnologia, grant PTDC/BEX-BID/1411/2014; M.R. was funded by the fellowship PD/BD/136927/2018. P.S was funded by the fellowship SFRH/BD/111611/2015; C.B. was funded by the fellowship SFRH/BD/141034/2018; SSL was funded by FCT CEEC-IND 2018. Funding Information: Acknowledgments: The authors want to thank the Fish Facility from NMS. This work was developed with the support of the Fish Facility NMS|FCM that is part of CONGENTO, a Research Infrastructure co-financed by Lisboa Regional Operational Programme (Lisboa2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and Fundação para a Ciência e Tecnologia (Portugal) LISBOA-01-0145-FEDER-022170. A.P. wants to thank the Royal Brompton and Harefield hospital, part of the Guy’s and St Thomas’ foundation trust, London. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Zebrafish is a vertebrate teleost widely used in many areas of research. As embryos, they develop quickly and provide unique opportunities for research studies owing to their transparency for at least 48 h post fertilization. Zebrafish have many ciliated organs that include primary cilia as well as motile cilia. Using zebrafish as an animal model helps to better understand human diseases such as Primary Ciliary Dyskinesia (PCD), an autosomal recessive disorder that affects cilia motility, currently associated with more than 50 genes. The aim of this study was to validate zebrafish motile cilia, both in mono and multiciliated cells, as organelles for PCD research. For this purpose, we obtained systematic high-resolution data in both the olfactory pit (OP) and the left–right organizer (LRO), a superficial organ and a deep organ embedded in the tail of the embryo, respectively. For the analysis of their axonemal ciliary structure, we used conventional transmission electron microscopy (TEM) and electron tomography (ET). We characterised the wild-type OP cilia and showed, for the first time in zebrafish, the presence of motile cilia (9 + 2) in the periphery of the pit and the presence of immotile cilia (still 9 + 2), with absent outer dynein arms, in the centre of the pit. In addition, we reported that a central pair of microtubules in the LRO motile cilia is common in zebrafish, contrary to mouse embryos, but it is not observed in all LRO cilia from the same embryo. We further showed that the outer dynein arms of the microtubular doublet of both the OP and LRO cilia are structurally similar in dimensions to the human respiratory cilia at the resolution of TEM and ET. We conclude that zebrafish is a good model organism for PCD research but investigators need to be aware of the specific physical differences to correctly interpret their results.publishersversionpublishe

Potential of mannan or dextrin nanogels as vaccine carrier/adjuvant systems

Polymeric nanogels have been sophisticatedly designed promising a new generation of vaccine delivery/adjuvant systems capable of boosting immune response, a strategic priority in vaccine design. Here, nanogels made of mannan or dextrin were evaluated for their potential as carriers/adjuvants in vaccine formulations. Since lymph nodes are preferential target organs for vaccine delivery systems, nanogels were biotin-labeled, injected in the footpad of rats, and their presence in draining lymph nodes was assessed by immunofluorescence. Nanogels were detected in the popliteal and inguinal lymph nodes by 24h upon subcutaneous administration, indicating entrapment in lymphatic organs. Moreover, the model antigen ovalbumin was physically encapsulated within nanogels and physicochemically characterized concerning size, zeta potential, ovalbumin loading, and entrapment efficiency. The immunogenicity of these formulations was assessed in mice intradermally immunized with ovalbuminmannan or ovalbumindextrin by determining ovalbumin-specific antibody serum titers. Intradermal vaccination using ovalbuminmannan elicited a humoral immune response in which ovalbumin-specific IgG1 levels were significantly higher than those obtained with ovalbumin alone, indicating a TH2-type response. In contrast, dextrin nanogel did not show adjuvant potential. Altogether, these results indicate that mannan nanogel is a material that should be explored as a future antigen delivery system.The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work is supported by the Fundação para a Ciência e a Tecnologia (FCT) Portugal, post-doc grant SFRH/BPD/70524/2010 and the International Iberian Nanotechnology Laboratory (INL), PhD grant. The authors thank the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684). The authors also acknowledge the Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462)

Classificação Geotécnica do maciço rochoso da Pedreira do Monte das Flores – Évora (Portugal)

No âmbito da atualização do processo de homologação da Pedreira do Monte das Flores - Évora, como fornecedora de balastro ferroviário, procedeu-se à classificação geotécnica do maciço rochoso existente no local. Esta exploração possui 96,90 ha de área arrendada estando 70,70 ha concessionados à exploração. Geologicamente, a área estudada pertence à Zona de Ossa-Morena (Maciço de Évora), enquanto parte integrante do Maciço Ibérico que constitui o setor mais ocidental e contínuo da Cadeia Orogénica Varisca na Europa. Também os eventos tectonotérmicos alpinos se fizeram sentir neste maciço de forma atenuada, permitindo a preservação da história geológica mesozóica [Moita, 2008]. A unidade geológica onde se insere a exploração é constituída por litótipos ígneos e metamórficos de idade precâmbrica e paleozóica [Andrade et al., 1976], denotando-se um domínio das formações de rochas eruptivas. Na área estudada ocorre um afloramento de quartzodiorito e granodiorito de grão médio, não porfiróide, onde os minerais mais representativos da rocha são o feldspato potássico, a plagioclase e o quartzo, sem evidenciarem qualquer orientação à vista desarmada [Moita, 2008]. Neste estudo fez-se a descrição geotécnica da qualidade do maciço rochoso que ocorre na Pedreira do Monte das Flores. Por se tratar de um sistema de classificação generalista e correntemente utilizado na avaliação do comportamento geomecânico dos maciços rochosos, utilizou-se a Descrição Geotécnica Básica (“Basic Geotechnical Description”- BGD), proposta pela Sociedade Internacional de Mecânica das Rochas [ISRM, 1981]. O principal objetivo da aplicação desta classificação foi o de efetuar um zonamento geotécnico do maciço rochoso, quer do local atualmente em exploração, quer da área contígua para onde se prevê o alargamento da corta da exploração, com base no reconhecimento geológico e, na amostragem efetuada em locais selecionados para posterior realização de ensaios laboratoriais de caracterização mecânica. Este reconhecimento geológico de superfície permitiu identificar o tipo de rocha presente, determinar as características estruturais e caracterizar a alteração do maciço rochoso, assim como, definir as diferentes famílias de fraturas presentes e quantificar o espaçamento entre as descontinuidades nestas famílias. Com o objetivo de determinar as características mecânicas, tais como, a resistência à compressão uniaxial e o ângulo de atrito das fraturas, foi realizada uma campanha de ensaios laboratoriais e de campo, nomeadamente, o ensaio de resistência à compressão uniaxial, complementado pelo ensaio de carga pontual, de modo a determinar o valor daquela resistência. Realizou-se também o ensaio com o martelo de Schmidt dado ser um ensaio simples e rápido na caracterização de materiais [Pinho, 2003], que permite estimar o valor da resistência à compressão uniaxial dos planos das descontinuidades do maciço rochoso (JCS), nos diferentes locais de amostragem. O parâmetro JCS, o coeficiente de rugosidade da descontinuidade (JRC) e a tensão efectiva normal (’n ), foram necessários para obter o ângulo de atrito das diaclases (Øpico), de acordo com o método proposto pela Sociedade Internacional de Mecânica das Rochas [ISRM, 1978]. O estudo realizado permitiu concluir que o maciço apresenta boa qualidade, em regra, homogénea relativamente às suas características geológicas e geotécnicas. No entanto, distinguem-se duas zonas, ZG1 e ZG2, com base em pequenas diferenças nos valores da resistência à compressão uniaxial da rocha e do ângulo de atrito das fraturas

2,3-Diphosphoglycerate and the Protective Effect of Pyruvate Kinase Deficiency against Malaria Infection—Exploring the Role of the Red Blood Cell Membrane

Malaria remains a major world public health problem, contributing to poverty and inequality. It is urgent to find new efficacious tools with few adverse effects. Malaria has selected red blood cell (RBC) alterations linked to resistance against infection, and understanding the protective mechanisms involved may be useful for developing host-directed tools to control Plasmodium infection. Pyruvate kinase deficiency has been associated with resistance to malaria. Pyruvate kinase-deficient RBCs display an increased concentration of 2,3-diphosphoglycerate (2,3-DPG).We recently showed that 2,3-DPG impacts in vitro intraerythrocytic parasite growth, induces a shift of the metabolic profile of infected cells (iRBCs), making it closer to that of noninfected ones (niRBCs), and decreases the number of parasite progenies that invade new RBCs. As an increase of 2,3-DPG content may also have an adverse effect on RBC membrane and, consequently, on the parasite invasion, in this study, we explored modifications of the RBC morphology, biomechanical properties, and RBC membrane on Plasmodium falciparum in vitro cultures treated with 2,3-DPG, using atomic force microscopy (AFM)-based force spectroscopy and other experimental approaches. The presence of infection by P. falciparum significantly increased the rigidity of parasitized cells and influenced the morphology of RBCs, as parasitized cells showed a decrease of the area-to-volume ratio. The extracellular addition of 2,3-DPG also slightly affected the stiffness of niRBCs, making it more similar to that of infected cells. It also changed the niRBC height, making the cells appear more elongated. Moreover, 2,3-DPG treatment influenced the cell surface charge, becoming more negative in treated RBCs than in untreated ones. The results indicate that treatment with 2,3-DPG has only a mild effect on RBCs in comparison with the effect of the presence of the parasite on the host cell. 2,3-DPG is an endogenous host metabolite, which may, in the future, originate a new antimalarial tool with few adverse effects on noninfected cells.publishersversionpublishe

Impact of phlorotannin extracts from fucus vesiculosus on human gut microbiota

Recent studies indicate that plant polyphenols could be pointed as potential prebiotic candidates since they may interact with the gut microbiota, stimulating its growth and the production of metabolites. However, little is known about the fate of brown seaweeds’ phlorotannins during their passage throughout the gastrointestinal tract. This work aimed to evaluate the stability and bioaccessibility of Fucus vesiculosus phlorotannins after being submitted to a simulated digestive process, as well as their possible modulatory effects on gut microbiota and short-chain fatty acids production following a fermentation procedure using fecal inoculates to mimic the conditions of the large intestine. The stability of phlorotannins throughout the gastrointestinal tract was reduced, with a bioaccessibility index between 2 and 14%. Moreover, slight alterations in the growth of certain commensal bacteria were noticed, with Enterococcus spp. being the most enhanced group. Likewise, F. vesiculosus phlorotannins displayed striking capacity to enhance the levels of propionate and butyrate, which are two important short-chain fatty acids known for their role in intestinal homeostasis. In summary, this work provides valuable information regarding the behavior of F. vesiculosus phlorotannins along the gastrointestinal tract, presenting clear evidence that these compounds can positively contribute to the maintenance of a healthy gastrointestinal condition.info:eu-repo/semantics/publishedVersio