Polysaccharides in plant cell walls serve as a rich carbon and energy source, yet their structural complexity presents a barrier to efficient degradation. To address this, anaerobic microorganisms like R. flavefaciens have developed sophisticated multi-enzyme complexes known as cellulosomes, which enable the efficient breakdown of these recalcitrant polysaccharides. These complexes are assembled through high-affinity interactions between cohesin (Coh) modules in scaffoldin proteins and dockerin (Doc) modules in cellulosomal enzymes. R. flavefaciens FD-1 harbors one of the most intricate cellulosomes described to date, comprising over 200 Doc-containing proteins encoded in its genome. Despite substantial research on this cellulosome, the role of a group of truncated but functional dockerins, known as group-2 Docs, remains unclear. In this study, we present a detailed structural and binding analysis of a Coh-Doc complex involving the cohesin from the cell-anchoring scaffoldin ScaE and a group-2 Doc that bears only one of the two Ca+2-coordinating loops that characterise the canonical Docs. Our findings reveal a novel tripartite binding mechanism, in which the cohesin can simultaneously bind two distinct dockerin units in three alternative conformations. This discovery provides new insights into the modular versatility of the R. flavefaciens cellulosome and sheds light on the mechanisms that enhance its efficiency in polysaccharide degradation.

The Upper Jurassic beds of the Lusitanian Basin in central Portugal yield diverse dinosaurian fauna, dated to the Kimmeridgian–Tithonian interval. Saurischian dinosaurs are, overall, more abundant than their ornithischian counterparts, in terms of both specimens collected and species recognized. Iguanodontians are so far represented by the styracosternan Draconyx loureiroi, the dryosaurid Eousdryosaurus nanohallucis and the enigmatic dryomorphan Hesperonyx martinhotomasorum. Here we aim to highlight the diversity of this clade in the Late Jurassic of Portugal, presenting evidence for yet another species of ankylopollexian iguanodontian dinosaur, represented by the specimen SHN.JJS.015, which is housed at the Sociedade de História Natural, Torres Vedras. Detailed comparisons rule out attribution to previously known taxa, and phylogenetic analyses that include SHN.JJS.015 indicate early-diverging ankylopollexian affinities for this specimen. As there is no robust diagnosis, we do not erect a new formal species for it at this stage. Nevertheless, this specimen represents a previously unreported taxon that highlights greater diversity than previously estimated among the iguanodontians of the Late Jurassic and highlights the importance of Europe in diversification and dispersal events of this clade. A series of smaller, isolated femora from the same sub-basin as SHN.JJS.015 may represent the same taxon, presenting evidence of thriving communities of ankylopollexians during the Kimmeridgian–Tithonian interval in Portugal.

The Late Triassic deposits in the Jameson Land Basin, central East Greenland, stand as a crucial fossil area, yielding a diverse Norian vertebrate fauna. This basin, situated at a palaeolatitude of 41° N on the northern rim of Pangea and bordered in the North by the Boreal Sea, was a hub of activity during the Late Triassic. A large ephemeral to perennial lake system developed in the central and eastern parts of the basin, with rivers transporting sediment from the uplands northwest of the basin. Our research focused on the microvertebrate remains recovered from an expedition to Jameson Land in 1991 which yielded significant findings. These specimens, meticulously photographed and listed in a catalogue, comprise 950 vertebrate remains. Notably, we have identified new taxa never described from the Late Triassic sediments of Greenland, including sharks (Lissodus, Rhomphaiodon), bony fish (Gyrolepis), and reptiles (Doswelliidae and Rhynchocephalia). The revision of two sphenodontians jaw fragments as Clevosauridae, in association with Lissodus lepagei, Lissodus cf. Lissodus minimus, Rhomphaiodon sp., Saurichthys sp., and Gyrolepis sp., which are taxa that are highly documented in Europe and Asia, confirms a relationship between faunas of Greenland and Eurasia during the Late Triassic, a statement previously based on macro vertebrate studies.

Viral proteins typically exist in the context of complex virions or in the even more complex host cells in which they replicate. Hence, meaningful insights into virus protein structure often need to account for this context. Various flavors of in situ cryo-electron microscopy (cryo-EM), such as cryo-electron tomography, are key methods for the contextual study of virus protein structure in pleomorphic virions and host cells. Here, we review recent in situ cryo-EM work on three selected phenomena in RNA virus replication: the maturation and nuclear entry of HIV-1, the membrane-bound replication organelles of positive-sense RNA viruses, and the membrane-less viral factories of negative-sense RNA viruses. We highlight cases where the imaged phenomena are targets of novel antiviral drugs (such as the recently approved antiretroviral Lenacapavir), drug candidates, and antiviral strategies. Finally, we discuss recent technical advances that extend the reach of in situ cryo-EM in virology.

Accurate blast models are essential for disaster management and emergency preparedness. Semi-empirical methods, which rely on free-field assumptions, struggle to provide precise data for complex building shapes because they ignore shock wave reflections. This research numerically investigates the effects of an explosion on a large building of complex geometrical configuration using blastFoam, estimating the façade damage and assessing risks to occupants. To evaluate the importance of accurately modelling the building’s geometry, four levels of detail were considered. Additionally, simulations were also performed using the Load Blast Enhanced (LBE) method from LS-DYNA to compare the estimates of this faster semi-empirical approach with those obtained through CFD analyses. The findings reveal that simpler CFD models are adequate for façade analysis and injury assessment around the building but fall short for predicting injury distribution within interior spaces or between buildings. The LBE underestimates both structural damage and human injury levels, while detailed CFD highlights the importance of accounting for interior walls and windows to enhance blast pressure predictions.

ABSTRACT The first mammaliaforms emerged in the Late Triassic, but their exact origins remain unclear due to the scarcity of fossils from this period. One of the earliest diverging mammaliaform groups, the order Docodonta, became unusually ecomorphologically diverse compared with other early mammals, and this may be connected to the possession of complex molar cusp morphology. The specimen described here, found in the Rhætelv Formation of the Kap Stewart Group (Rhaetian–Sinemurian) of central East Greenland, provides novel information on docodontan origins and evolution, as well as key biogeographic insights into early mammal dispersal. Nujalikodon cassiopeiae gen. et sp. nov. is the first mammaliaform found in the Rhætelv Formation, and is likely to be Early Jurassic (Hettangian) in age. Comprising an incomplete dentary with a single preserved molar, it was visualized using micro-computed tomography; the molar bears similarities to the putative early docodontan Delsatia, and docodontan Dobunnodon. Phylogenetic analysis places Nujalikodon cassiopeiae as a basal member of Docodonta or a close sister taxon, making it one of the oldest definitive docodontans and pushing the origin of the group back to at least the Early Jurassic. It provides insights into the development of docodontan dental complexity, a key factor in their ecological diversification during the Middle to Late Jurassic. Its presence in Greenland supports the hypothesis that docodontans originated in the region now comprising Europe and Greenland before dispersing across the rest of Laurasia.

Carbohydrate-binding-modules (CBMs) are discrete auxiliary protein modules with a non-catalytic carbohydrate-binding function and that exhibit a great diversity of binding specificities. CBMcarb-DB is a curated database that classifies the three-dimensional structures of CBM–carbohydrate complexes determined by single-crystal X-ray diffraction methods and solution NMR spectroscopy. We designed the database architecture and the navigation tools to query the database with the Protein Data Bank (PDB), UniProtKB, and GlyTouCan (universal glycan repository) identifiers. Special attention was devoted to describing the bound glycans using simple graphical representation and numerical format for cross-referencing to other glycosciences and functional data databases. CBMcarb-DB provides detailed information on CBMs and their bound oligosaccharides and features their interactions using several open-access applications. We also describe how the curated information provided by CBMcarb-DB can be integrated with AI algorithms of 3D structure prediction, facilitating structure–function studies. Also in this chapter, we discuss the exciting convergence of CBMcarb-DB with the glycan array repository, which serves as a valuable resource for investigating the specific binding interactions between glycans and various biomolecular targets. The interaction of the two fields represents a significant milestone in glycosciences. CBMcarb-DB is freely available at https://cbmdb.glycopedia.eu/ and https://cbmcarb.webhost.fct.unl.pt.

The structural and chemical diversity of β-glucans is reflected on the variety of essential biological roles tackled by these polysaccharides. This natural heterogeneity requires an elaborate assortment of enzymatic mechanisms to assemble, degrade or modify, as well as to extract their full biotechnological potential. Recent metagenomic efforts have provided an unprecedented growth in potential new biocatalysts, most of which remain unconfirmed or uncharacterized. Here we report the first biochemical and structural characterization of two bacterial β-glucanases from the recently created glycoside hydrolase family 157 (LaGH157 and BcGH157) and investigate their molecular basis for substrate hydrolysis. Structural analysis by X-ray crystallography revealed that GH157 enzymes belong to clan GH-A, possessing a (β/α)8-barrel fold catalytic domain, two β-sandwich accessory domains and two conserved catalytic glutamates residues, with relative positions compatible with a retaining mechanism of hydrolysis. Specificity screening and enzyme kinetics suggest that the enzymes prefer mixed-linkage glucans over β-1,3-glucans. Activity screening showed that both enzymes exhibit pH optimum at 6.5 and temperature optimum for LaGH157 and BcGH157 at 25 °C and 48 °C, respectively. Product analysis with HPAEC-PAD and LC-MS revealed that both enzymes are endo-1,3(4)-β-glucanases, capable of cleaving β-1,3 and β-1,4-linked glucoses, when preceded by a β-1,3 linkage. Moreover, BcGH157 needs a minimum of 4 subsites occupied for hydrolysis to occur, while LaGH157 only requires 3 subsites. Additionally, LaGH157 possesses exohydrolytic activity on β-1,3 and branching β-1,6 linkages. This unusual bifunctional endo-1,3(4)/exo-1,3–1,6 activity constitutes an expansion on our understanding of β-glucan deconstruction, with the potential to inspire future applications.

Este relatório pretende descrever e analisar o trabalho de campo realizado no âmbito do projeto CERP. O projeto pretende compreender o estado da arte em Portugal da negociação coletiva portuguesa com referências à transição sustentável e justa no setor dos transportes e no setor público.

Este relatório pretende fazer uma revisão da literatura sobre a negociação coletiva portuguesa com referências à transição sustentável e justa no setor dos transportes e no setor público.
Portugal tem sido dos países que iniciou já o seu caminho de transição e encerrou as centrais a carvão. Mas em países do Leste europeu, por exemplo, existe muita indústria associada à produção de electricidade a partir do carvão, e onde o encerramento levanta sérias questões sociais. Nestes casos, em particular, é necessário um processo gradual que inclua medidas de apoio à formação e reconversão dos trabalhadores envolvidos.

As mentioned in the introduction to this volume, artificial intelligence (AI) functionalities are swiftly becoming integral to knowledge work and digital services. They are, however, also applied in all areas of human work. That is making the public services in Portugal more accessible and efficient. Since mid-90s, with the national strategy towards the Information Society (Missão para a Sociedade da Informação), a wide program for the promotion and development of public digital services have started. In the last decades, and with the definition of this national strategy, successive governments and the national parliament have contributed to stimulate democratic participation, and support decision-making and policy development.

Shock absorbers have been widely used in the automotive and aeronautical industries for many years. Inspired on these devices, the paper presents an analytical and numerical assessment of a high performance protective system for building structures against blast loads, which is composed of a shielding element connected to the main structure, at the floor levels, through ductile Energy Absorbing Connectors (EACs). The EACs exploit the external tube inversion mechanism to absorb a significant part of the imparted kinetic energy from the blast wave. While the system prototype has been developed in laboratory, it was characterized and tested in a full-scale blast testing campaign. A validated finite element model was used next to analyze its performance in a more demanding design scenario. The introduction of EACs notably reduces the peak horizontal loads and the kinetic energy transferred to the protected structure, being expected a significant reduction of the stresses in the supporting vertical elements, in addition to the protection of structural and non-structural members. These results encourage further studies of the presented protective system that can be potentially employed for a large variety of blast threat scenarios, especially when increasing the stand-off is not a possible/viable option and sensitive facilities have to be protected.

The challenges of disciplinary integration in teaching practice, especially in the implementation of tasks that promote interdisciplinary approaches, highlight the importance of teachers' professional knowledge in this process. Using a naturalistic approach, this study aims to characterize the professional knowledge of a physics teacher when adopting an interdisciplinary approach using technology. The results of the research revealed that the teacher mobilizes different types of knowledge, showing a differentiated knowledge of the most appropriate mathematical application to teach a given piece of content, and knowledge of how pedagogical strategies can be aided through different applications using technology.

ABSTRACTVirtual palaeontology is a growing field, leading palaeontologists to better understand the microanatomy of many extinct species. The application of techniques such as CT and μCT-scanning allows the researchers to study micro-anatomical features in a non-invasive way and make inferences on the palaeobiology of animals. Dinosaurs have been extensively studied using these techniques, with particular focus on the microanatomy of the cranium, whereas relatively little is known of other cranial elements, such as the lower jaw. Here, we aim to fill this gap, describing the microanatomy of the specimen ML 768, an isolated dentary belonging to a dryosaurid iguanodontian dinosaur from the Upper Jurassic of Lourinhã Fm. The dentary ML 768 was subjected to μCT-scanning, and subsequently the data were segmented in Avizo and rendered in Blender. We identified functional and replacement teeth, recognising remnants of old replacement cycles. Furthermore, we mapped a rich neurovascular network present in the dentary and compared it with reference literature. We found that the high vascularisation is shared with other cerapodan dinosaurs with high tooth replacement rates, although homoeostasis may have also played a role in the development of this condition. Further evidence is needed to appreciate the macroevolutionary significance of these findings.

Abstract The Monastery of Alcobaça houses in reserve the ceramic tiles that adorned the floor of the church's apse. These tiles were removed during rehabilitation works and many preserve part of their original fixing mortars. A comprehensive analysis of 21 samples was conducted using a multi-analytical approach (X-ray fluorescence, X-ray diffraction, scanning electron microscopy–energy-dispersive X-ray spectroscopy, μ-Raman spectroscopy, thermogravimetry–differential thermal analysis, optical microscopy, and colorimetry). Results suggest compositional variations in the samples from the back and sides of the ceramic tiles; however, the mineralogy and general characteristics of the aggregates remained consistent between the samples and are coherent with the local geology, suggesting a shared historical origin.