https://hdl.handle.net/10216/26159 2026-07-10T14:32:58Z 2026-07-10T14:32:58Z https://hdl.handle.net/10216/171868 2026-01-10T01:58:37Z 2025-12-22T00:00:00Z

Title: N-Glycans as fundamental immune-checkpoints at the frontiers of inflammation, autoimmunity and cancer Abstract: Glycans cover the surface of essentially all cells, giving rise to the so-called glycocalyx. Glycans, and particularly N-glycans are key players in defining the fate and the nature of the immune response, participating in both pro-inflammatory and anti-inflammatory pathways occurring in inflammation, autoimmunity and cancer. Glycans and glycan-binding proteins (GBPs) are instrumental molecules that integrate the immunological landscape in homeostasis, being involved in how the host immune system perceives the self and non-self. Perturbations in these immune regulatory circuits created by glycans and GBPs may result in the loss of immune tolerance associated with autoimmunity and inflammatory conditions, but also in malignant transformation. This chapter will discuss the regulatory role played by N-glycans as fundamental immune-checkpoints in the interplay between the immune cells and their surrounding microenvironment associated with homeostasis and disease. Furthermore, we will also explore how N-glycans can act as promising diagnostic and prognostic biomarkers in disease, as well as appealing targets for new therapeutic approaches in inflammation, autoimmunity and cancer.

2025-12-22T00:00:00Z https://hdl.handle.net/10216/126851 2020-04-04T09:39:28Z 2019-08-31T00:00:00Z

Title: Delivery of Antisense Oligonucleotides Mediated by a Hydrogel System: In Vitro and In Vivo Application in the Context of Spinal Cord Injur Abstract: Biomaterials-based hydrogels are attractive drug-eluting vehicles in the context of RNA therapeutics, such as those utilizing antisense oligonucleotide or RNA interference based drugs, as they can potentially reduce systemic toxicity and enhance in vivo efficacy by increasing in situ concentrations. Here we describe the preparation of antisense oligonucleotide-loaded fibrin hydrogels exploring their applications in the context of the nervous system utilizing an organotypic dorsal root ganglion explant in vitro system and an in vivo model of spinal cord injury.

2019-08-31T00:00:00Z https://hdl.handle.net/10216/126850 2020-04-06T16:51:10Z 2018-10-30T00:00:00Z

Title: Molecular Recognition Force Spectroscopy for Probing Cell Targeted Nanoparticles In Vitro Description: In the development and design of cell targeted nanoparticle-based systems the density of targeting moieties plays a fundamental role in allowing maximal cell-specific interaction. Here, we describe the use of molecular recognition force spectroscopy as a valuable tool for the characterization and optimization of targeted nanoparticles toward attaining cell-specific interaction. By tailoring the density of targeting moieties at the nanoparticle surface, one can correlate the unbinding event probability between nanoparticles tethered to an atomic force microscopy tip and cells to the nanoparticle vectoring capacity. This novel approach allows for a rapid and cost-effective design of targeted nanomedicines reducing the need for long and tedious in vitro tests.

2018-10-30T00:00:00Z https://hdl.handle.net/10216/118214 2020-01-27T09:56:22Z 2016-01-01T00:00:00Z

Title: Studying T Cells N-Glycosylation by Imaging Flow Cytometry Abstract: Imaging flow cytometry is an emerging imaging technology that combines features of both conventional flow cytometry and fluorescence microscopy allowing quantification of the imaging parameters. The analysis of protein posttranslational modifications by glycosylation using imaging flow cytometry constitutes an important bioimaging tool in the glycobiology field. This technique allows quantification of the glycan fluorescence intensity, co-localization with proteins, and evaluation of the membrane/cytoplasmic expression. In this chapter we provide the guidelines to analyze glycan expression, particularly the ß1,6 GlcNAc branched N-glycans, on the membrane of intestinal T cells from inflammatory bowel disease patients.

2016-01-01T00:00:00Z https://hdl.handle.net/10216/117925 2020-01-27T09:56:22Z 2014-01-01T00:00:00Z

Title: Factors involved in ubiquitination and deubiquitination of PEX5, the peroxisomal shuttling receptor Abstract: Peroxisomal matrix proteins are synthesized on cytosolic ribosomes and post-translationally targeted to the organelle by the soluble factor PEX5. Besides a role as a receptor, and probably as a chaperone, PEX5 also holds the key to the matrix of the organelle. Indeed, the available data suggest that PEX5 itself pushes these proteins across the peroxisomal membrane using as driving force the strong protein-protein interactions that it establishes with components of the peroxisomal membrane docking/translocation module (DTM). In recent years, much has been learned on how this transport system is reset and kept fine-tuned. Notably, this involves covalent modification of PEX5 with ubiquitin. Two types of PEX5 ubiquitination have been characterized: monoubiquitination at a conserved cysteine, a mandatory event for the extraction of PEX5 from the DTM; and polyubiquitination, probably the result of a quality control mechanism aiming at clearing the DTM from entangled PEX5 molecules. Monoubiquitination of PEX5 is transient in nature and the factors that reverse this modification have recently been identified.

2014-01-01T00:00:00Z https://hdl.handle.net/10216/117911 2020-01-27T09:56:22Z 2017-01-01T00:00:00Z

Title: Determining the topology of peroxisomal proteins using protease protection assays Abstract: Protease protection assays are powerful tools to determine the topology of organelle proteins. Their simplicity, together with the fact that they are particularly suited to characterize endogenous proteins, are their major advantages and the reason why these assays have been in use for so many years. Here, we provide a detailed protocol to use with mammalian peroxisomes. Suggestions on how these assays can be controlled, and how to identify some technical pitfalls, are also presented.

2017-01-01T00:00:00Z https://hdl.handle.net/10216/110887 2020-01-27T09:56:22Z 2017-01-01T00:00:00Z

Title: Emerging Mechanisms and Roles for Asymmetric Cytokinesis Abstract: Cytokinesis completes cell division by physically separating the contents of the mother cell between the two daughter cells. This event requires the highly coordinated reorganization of the cytoskeleton within a precise window of time to ensure faithful genomic segregation. In addition, recent progress in the field highlighted the importance of cytokinesis in providing particularly important cues in the context of multicellular tissues. The organization of the cytokinetic machinery and the asymmetric localization or inheritance of the midbody remnants is critical to define the spatial distribution of mechanical and biochemical signals. After a brief overview of the conserved steps of animal cytokinesis, we review the mechanisms controlling polarized cytokinesis focusing on the challenges of epithelial cytokinesis. Finally, we discuss the significance of these asymmetries in defining embryonic body axes, determining cell fate, and ensuring the correct propagation of epithelial organization during proliferation.

2017-01-01T00:00:00Z https://hdl.handle.net/10216/82533 2021-05-04T09:21:58Z 2016-01-01T00:00:00Z

Title: Killing animals as a necessary evil? The case of animal research Abstract: This chapter addresses the question of killing animals in research, primarily from a moral perspective, but also taking into account some of the practical and scientific considerations with moral consequences in this context. We start by exploring in which situations animals are killed in research and whether these are always inevitable, analysing re-use and re-homing of animals as potential alternatives. We then discuss for whom – and under what circumstances -killing matters, considering situations where there may be a conflict between the wish to avoid killing and that to avoid suffering, and further take human-animal interactions into account. We argue that, although there are relevant practical, scientific and ethical arguments favouring the euthanasia of animals in most research contexts, there is a potential for rehabilitating more animals than is currently the practice.

2016-01-01T00:00:00Z https://hdl.handle.net/10216/81366 2021-05-04T09:16:50Z 2014-01-01T00:00:00Z

Title: Does the goal justify the methods? Harm and benefit in neuroscience research using animals Abstract: The goal of the present chapter is to open up for discussion some of the major ethical issues involved in animal-based neuroscience research. We begin by approaching the question of the moral acceptability of the use of animals in research at all, exploring the implications of three different ethical theories: contractarianism, utilitarianism and animal rights. In the remainder of the chapter we discuss more specific issues of neuroscience research within what we argue is the mainstream framework for research animal ethics, namely one based on harm-benefit analysis. We explore issues of harms and benefits and how to balance them as well as how to reduce harm and increase benefit within neuroscience research.

2014-01-01T00:00:00Z https://hdl.handle.net/10216/78733 2021-05-04T09:15:53Z 2012-01-01T00:00:00Z

Title: Bringing animal ethics teaching into the public domain: the Animalogos experience Abstract: An increasing number of academic researchers blog – most as a form of science communication to a wider public. This tool is particularly interesting as it provides a way for scientists to communicate directly with a public rather than depending on journalists. When we launched Animalogos (animalogos.blogspot.com) in December 2009, our main aim was to establish a forum of communication for people working professionally with animal ethics and animal welfare, in order to provide a Portuguese-language professional perspective on issues previously almost exclusively commented on by animal rights and animal protection NGOs. Increasingly, we have also integrated the blog into animal ethics teaching, where we now use it as the main examination tool in the animal ethics discipline of two courses: a postgraduate course in animal welfare and an undergraduate course in veterinary medicine. Individually, students have to write a comment to one of the existing blog posts, a comment which is published by the students themselves. In groups, they have to write a post on a topic of their own choice, which is published after revision. For examination purposes, the texts thus produced by the students are evaluated as a short written essay. Whether to be anonymous or not is the students’ own choice. Using the blog as a resource for teaching and examination started as a pilot project during the academic year 2010/11, during which the method was evaluated from the teachers’ perspective. The main advantages with involving students in the blog activity is that firstly students get to make a real contribution to the public debate and secondly that this draws attention to the blog. The main disadvantage we observed was that most posts written by students require substantial revision before they can be published, sometimes resulting in negative reactions from the students. Continuing the project in 2011/12, we have asked feedback from students. Overall, they view the experience as positive and consider examination through writing for the blog preferable to traditional exams (written or oral), essays or oral presentations. More than half of the students would like this examination approach for more disciplines within the course they are taking. All respondents think that anonymity should be an option when they are writing for the blog.

2012-01-01T00:00:00Z https://hdl.handle.net/10216/78436 2021-05-04T09:28:26Z 2014-01-01T00:00:00Z

Title: Welfare and Quantity of Life Abstract: The Animal welfare science is mostly focused on evaluating and improving the quality of life of animals that actually exist. This leaves out a range of ethically relevant issues regarding the quantity of life – in terms of number of animals living and the longevity of each animal. In many cases quantity and quality are related, and often there is a tension between the two. In this chapter, we develop a discussion around four practical cases presenting quality-quantity dilemmas: a) the issue of dairy cow longevity, b) the early slaughter of male dairy calves, c) the killing of newly-hatched male layer chicks and d) the conflict between reduction and refinement in animal research. The practical, economic and animal welfare aspects characterizing each case are presented together with relevant stakeholders’ perspective. We discuss the cases in light of the most relevant currents of thought in animal ethics, highlighting the main values at stake and which possible solutions may be sought according to each perspective.

2014-01-01T00:00:00Z