One of the trends in downstream processing comprises the use of ?anything-but-chromatography? methods to overcome the current downfalls of standard packed-bed chromatography. Precipitation and magnetic separation are two techniques already proven to accomplish protein purification from complex media, yet never used in synergy. With the aim to capture antibodies directly from crude extracts, a new approach combining precipitation and magnetic separation was developed and named as affinity magnetic precipitation. A precipitation screening, based on the Hofmeister series, and a commercial precipitation kit were tested with affinity magnetic particles to assess the best condition for antibody capture from human serum plasma and clarified cell supernatant. The best conditions were obtained when using PEG3350 as precipitant at 4°C for 1h, reaching 80% purity and 50% recovery of polyclonal antibodies from plasma, and 99% purity with 97% recovery yield of anti-TNFα mAb from cell supernatants. These results show that the synergetic use of precipitation and magnetic separation can represent an alternative for the efficient capture of antibodies. This article is protected by copyright. All rights reserved

Reduction of N2O to N2 is catalysed by nitrous oxide reductase in the last step of the denitrification pathway. This multicopper enzyme has an electron transferring centre, CuA, and a tetranuclear copper-sulfide catalytic centre, “CuZ”, which exists as CuZ*(4Cu1S) or CuZ(4Cu2S). The redox behaviour of these metal centres in Marinobacter hydrocarbonoclasticus nitrous oxide reductase was investigated by potentiometry and for the first time by direct electrochemistry. The reduction potential of CuA and CuZ(4Cu2S) was estimated by potentiometry to be +275 ± 5 mV and +65 ± 5 mV vs SHE, respectively, at pH 7.6. A proton-coupled electron transfer mechanism governs CuZ(4Cu2S) reduction potential, due to the protonation/deprotonation of Lys397 with a pKox of 6.0 ± 0.1 and a pKred of 9.2 ± 0.1. The reduction potential of CuA, in enzyme samples with CuZ*(4Cu1S), is controlled by protonation of the coordinating histidine residues in a two-proton coupled electron transfer process. In the cyclic voltammograms, two redox pairs were identified corresponding to CuA and CuZ(4Cu2S), with no additional signals being detected that could be attributed to CuZ*(4Cu1S). However, an enhanced cathodic signal for the activated enzyme was observed under turnover conditions, which is explained by the binding of nitrous oxide to CuZ0(4Cu1S), an intermediate species in the catalytic cycle.

BackgroundHalf of human cancers harbour TP53 mutations that render p53 inactive as a tumor suppressor. As such, reactivation of mutant (mut)p53 through restoration of wild-type (wt)-like function represents one of the most promising therapeutic strategies in cancer treatment. Recently, we have reported the (S)-tryptophanol-derived oxazoloisoindolinone SLMP53-1 as a new reactivator of wt and mutp53 R280K with in vitro and in vivo p53-dependent antitumor activity. The present work aimed a mechanistic elucidation of mutp53 reactivation by SLMP53-1.
Methods and results
By cellular thermal shift assay (CETSA), it is shown that SLMP53-1 induces wt and mutp53 R280K thermal stabilization, which is indicative of intermolecular interactions with these proteins. Accordingly, in silico studies of wt and mutp53 R280K DNA-binding domain with SLMP53-1 unveiled that the compound binds at the interface of the p53 homodimer with the DNA minor groove. Additionally, using yeast and p53-null tumor cells ectopically expressing distinct highly prevalent mutp53, the ability of SLMP53-1 to reactivate multiple mutp53 is evidenced.
Conclusions
SLMP53-1 is a p53-activating agent with the ability to directly target wt and a set of hotspot mutp53.
General Significance
This work reinforces the encouraging application of SLMP53-1 in the personalized treatment of cancer patients harboring distinct p53 status.

Three new additional skull specimens of Talarurus plicatospineus have been recovered from the Upper Cretaceous (Cenomanian–Santonian) Bayanshiree Formation, of Bayan Shiree cliffs, eastern Gobi Desert, Mongolia. The skulls feature unique characters such as an anteriorly protruded single internarial caputegulum, around 20 flat or concave nasal-area caputegulae surrounded by a wide sulcus, a vertically oriented elongate loreal caputegulum with a pitted surface, an elongate lacrimal caputegulum positioned above the posterodorsal border of the maxilla, two longitudinally arranged large frontoparietal caputegulae surrounded by smaller rhomboid caputegulae, small but elongate medial supraorbital caputegulae, a posterior supraorbital caputegulum that is four times larger than the anterior one, up to three transverse parallel grooves on the dorsal surface of the posterior supraorbital caputegulum, postocular caputegulae along the ventral to posterior rim of the orbit that extend almost to the anteroventral margin of the squamosal horn, a longitudinal furrow tapering towards the apex of the squamosal horn, a lateral nuchal caputegulum four to five times larger than other nuchal caputegulae, and a pterygovomerine keel with a ventral margin that is dorsally positioned to the alveolar ridge. The phylogenetic analysis result showed that Talarurus is sister to the clade that includes the derived Asian ankylosaurines (Saichania chulsanensis, Tarchia kielanae, and Zaraapelta nomadis). It also shows that there was dispersal of ankylosaurines from Asia into western North America before the Cenomanian. Moreover, the rostral differences between Talarurus and Tsagantegia, another ankylosaur from the same formation, suggest possible niche partitioning between these taxa.

Polynomial interpolation or regression models are an important tool in Derivativefree Optimization, acting as surrogates of the real function. In this work, we propose the use of these models in the multiobjective framework of directional direct search, namely the one of Direct Multisearch. Previously evaluated points are used to build quadratic polynomial models, which are minimized in an attempt of generating nondominated points of the true function, defining a search step for the algorithm. Numerical results state the competitiveness of the proposed approach.

Abstract This paper presents a critical review of the state of the art of experimental research concerning the seismic response of reinforced concrete flat slab frames. After a summary of tests on connections, the paper examines tests carried out on frames with gravity and cyclic lateral loading, and shake table tests; scaled specimens and one real scale study are included. A discussion of the results reached so far is provided focusing on the global response, the different load types and effects; the ultimate rotations at failure in relation to the gravity shear and a classification of different failure modes for different types of connections. Based on this analysis, the research needs are highlighted. An experimental program launched to address these open questions is described. Further open topics are highlighted.

The lithostratigraphy of the Triassic deposits of the Jameson Land Basin in central East Greenland is revised. The new Scoresby Land Supergroup is now composed of the Wordie Creek, Pingo Dal, Gipsdalen and Fleming Fjord Groups. This paper only deals with the lithostratigraphy of the late Early-Late Triassic continental deposits of the latter three groups with emphasis on the vertebratebearing Fleming Fjord Group. The new Pingo Dal Group consists of three new formations, the Rødstaken, Paradigmabjerg and Klitdal Formations (all elevated from members), the new Gipsdalen Group consists of three new formations, the Kolledalen, Solfaldsdal (with the new Gråklint Member) and Kap Seaforth Formations (all elevated from members), and the new Fleming Fjord Group is subdivided into three new formations, the Edderfugledal, Malmros Klint and Ørsted Dal Formations (all elevated from members). The Edderfugledal Formation contains two cyclic bedded, lacustrine members, a lowermost Sporfjeld Member (elevated from beds), and an uppermost Pingel Dal Member (elevated from beds). The lacustrine red beds of the Malmros Klint Formation are not subdivided. The lacustrine and fluvial Ørsted Dal Formation contains three new members. In the eastern and central part of the basin, the formation is initiated by cyclic bedded, red lacustrine mudstones of the Carlsberg Fjord Member (elevated from beds), while in the northwestern part of the basin the lowermost part of the formation is composed of grey fluvial conglomerates and sandstones with subordinate red mudstones of the Bjergkronerne Member (elevated from beds). The uppermost part of the formations in most of the basin is composed of cyclic bedded, variegated lacustrine mudstones and grey to yellowish marlstones of the Tait Bjerg Member (elevated from beds). The sediments in the Fleming Fjord Group contain remains of a rich and diverse vertebrate fauna including dinosaurs, amphibians, turtles, aeotosaurs, pterosaurs, phytosaurs and mammaliaforms. Most vertebrate bones have been found in uppermost Malmros Klint Formation, and in the Carlsberg Fjord and Tait Bjerg Members. The Norian–early Rhaetian, lacustrine Fleming Fjord Group was deposited at about 41° N on the northern part of the supercontinent Pangaea. Lacustrine sedimentation was controlled by seasonal as well as longer-term (orbital) variation in precipitation. Precipitation was probably brought to the basin by southwesterly winds. The lacustrine sediments of the uppermost Fleming Fjord Group show deposition during increasingly humid conditions changing the lake environment from an ephemeral lake-steppe area to a perennial lake. This evolution of lake environment suggests a change from a winter-wet temperate climate to one with precipitation throughout the year.

No contexto do Observatório de Literacia Oceânica, o estudo da coconstrução de um museu virtual online investiga a aprendizagem dos alunos pertencentes às comunidades piscatórias, contribuindo para a literacia histórica. O envolvimento em tarefas concretas associadas à aprendizagem é considerado, consensualmente, um fator de motivação. No presente estudo trabalharam as artes da pesca artesanal, enquadradas por conteú dos da disciplina de História. Utilizando ambientes tecnologicamente enriquecidos, pretende-se compreender as interações entre o conhecimento formal e informal destes alunos, desenvolvendo-se o estudo de forma a que os conhecimentos sejam signi{\textbackslash}icativos e usáveis, promovendo a intervenção na sociedade, enquanto agentes de mudança. A metodologia assentou num estudo empı́rico qualitativo, com base na etnogra{\textbackslash}ia crı́tica, combinado com design-based research, para a construção do protótipo do museu virtual online. As estratégias de ação, técnicas de recolha e aná lise de dados desenvolveram-se num contexto de observação participativa, notas de campo e aplicação de vários instrumentos, bem como na coconstrução do protótipo por aproximações sucessivas estruturantes da plataforma digital selecionada. Constatou-se a ausência de interação entre os saberes formais e informais dos alunos, um desconhecimento da história das suas comunidades, mas também a valorização das histórias de famı́lia. Os alunos foram observadores crı́ticos das suas práticas reconhecendo, por exemplo, a desvalorização dos seus saberes informais e a sua falta de relação com os conteú dos da disciplina de História. Neste processo identi{\textbackslash}icou-se a necessidade de introduzir temáticas relacionadas com a Literacia Histórica, História das Comunidades Capariquenses e Sustentabilidade, considerando-o o meio privilegiado para a disseminação do seu património cultural.

The elastic parameters, strengths, and intralaminar fracture toughness are determined for an E-Glass polymer composite material system, statically and at high strain rate, adapting methodologies previously developed by the authors for different carbon composites. Dynamic experiments are conducted using tension and compression Split-Hopkinson Bars (SHBs). A unique set of experimental parameters is obtained, and reported together with the experimental set-up, in order to ensure reproducibility. While in-plane elastic and strength properties were obtained by testing one specimen geometry, intralaminar fracture properties required the testing of different sized notched specimens with scaled geometries. This allowed the use of the size-effect method for the determination of the dynamic R-curve. When comparing these results with those previously obtained for a carbon/epoxy material system, it is observed that the dynamic fracture toughness exhibits a much more significant increase in both tension and compression. The obtained results permit the identification of the softening law at different strain rates, allowing its use in any analytical or numerical strength predictive method.

This project is inserted in Observatório de Literacia Oceânica, and its aim is the co-construction of a virtual museum of the Fishery Arts, based on History concepts and contents related with formal and informal knowledge. In these communities, the oral and intergenerational transmission of life stories occurs mainly within the family circle. Our main purpose was to find ways that allowed students to acquire usable and significant pasts, and not only fixed histories. The acquisition of historical literacy can change the way they apprehend and make use of knowledge, turning them in to agents of change, disseminators of their cultural heritage, with the help of a technologically enriched environment that enables network communication. The access to digital education should promote students' autonomy and responsibility when constructing knowledge. In the present study, artisanal fishing was the object of study, related to the contents of the discipline of History. Making use of technologically enriched environments we intended to analyse how the interactions between the formal and informal knowledge of these students occurred, developing the study in such a way that their knowledge could be meaningful and usable. It is intended to promote their intervention in society as agents of change, identifying and resolving problems associated with the communities to which they belong. In a perspective of curricular innovation, and considering that a sustainable development implies a change of paradigm in education, the adoption of differentiated, transformative pedagogies aiming an active learning, involving direct participation, collaboration and problem solving for the promotion of school success, it is intended to create a space for critical reflection, generating new knowledge and knowledge shared and enriched so that students participate in the construction of knowledge in order to make it meaningful and usable. Participation and negotiation amongst participants, within a perspective of shared responsibility and active citizenship, promotes cohesion and equality. We have considered the context of the school community and the context of the community in which the project is inserted, so that the tasks / knowledges that students have, can become meaningful and promote the quality and sustainability of the environment in which they are inserted. Education faces challenges that are a reflection of society’s problems. In the search for solutions it is essential the participation and collaboration of students who, with their formal and informal knowledge, can address problems that have cultural, social, political, economic and environmental impacts in their communities, as well as a local, regional and global scope, both in present and past times. Using digital environments and instruments we will try to understand how the interactions between formal and informal knowledge take place and how the performance of students from the fishing communities can be improved. Keywords: historical literacy; formal and informal knowledge; virtual museum.