Selective albumin-binding surfaces modified with a thrombin-inhibiting peptide

Blood-contacting medical devices have been associated with severe clinical complications, such as thrombus formation, triggered by the activation of the coagulation cascade due to the adsorption of certain plasma proteins on the surface of biomaterials. Hence, the coating of such surfaces with antithrombotic agents has been used to increase biomaterial haemocompatibility. Biomaterial-induced clotting may also be decreased by albumin adsorption from blood plasma in a selective and reversible way, since this protein is not involved in the coagulation cascade. In this context, this paper reports that the immobilization of the thrombin inhibitor D-Phe-Pro-D-Arg-D-Thr-CONH2 (fPrt) onto nanostructured surfaces induces selective and reversible adsorption of albumin, delaying the clotting time when compared to peptide-free surfaces. fPrt, synthesized with two glycine residues attached to the N-terminus (GGfPrt), was covalently immobilized onto self-assembled monolayers (SAMs) having different ratios of carboxylate-hexa(ethylene glycol)- and tri(ethylene glycol)-terminated thiols (EG6-COOH/EG3) that were specifically designed to control GGfPrt orientation, exposure and density at the molecular level. In solution, GGfPrt was able to inactivate the enzymatic activity of thrombin and to delay plasma clotting time in a concentration-dependent way. After surface immobilization, and independently of its concentration, GGfPrt lost its selectivity to thrombin and its capacity to inhibit thrombin enzymatic activity against the chromogenic substrate n-p-tosyl-Gly-Pro-Arg-p-nitroanilide. Nevertheless, surfaces with low concentrations of GGfPrt could delay the capacity of adsorbed thrombin to cleave fibrinogen. In contrast, GGfPrt immobilized in high concentrations was found to induce the procoagulant activity of the adsorbed thrombin. However, all surfaces containing GGfPrt have a plasma clotting time similar to the negative control (empty polystyrene wells), showing resistance to coagulation, which is explained by its capacity to adsorb albumin in a selective and reversible way. This work opens new perspectives to the improvement of the haemocompatibility of blood-contacting medical devices

Accelerated protein synthesis via one–pot ligation–deselenization chemistry

Peptide ligation chemistry has revolutionized protein science by facilitating access to synthetic proteins. Here, we describe the development of additive-free ligation-deselenization chemistry at β-selenoaspartate and γ-selenoglutamate that enables the generation of native polypeptide products on unprecedented timescales. The deselenization step is chemoselective in the presence of unprotected selenocysteine, which is highlighted in the synthesis of selenoprotein K. The power of the methodology is also showcased through the synthesis of three tick-derived thrombin-inhibiting proteins, each of which were assembled, purified, and isolated for biological assays within a few hours. The methodology described here should serve as a powerful means of accessing synthetic proteins, including therapeutic leads, in the future

iFloW: an integrated logistics software system for inbound supply chain traceability

Visibility plays an important role in supply chain management. Such visibility is not only important for better planning, but especially for real-time execution related with the traceability of goods. In inbound supply chain management, logistics planners need to trace raw materials from their requests in order to properly plan a plant’s production. The iFloW (Inbound Logistics Tracking System) integrates logistics providers IT applications and Global Positioning System (GPS) technology to track and trace incoming freights. The Estimated Time of Arrival (ETA) is updated in real-time allowing an improved materials planning process. This paper presents the iFloW project and describes how these issues are addressed and validated in a real pilot project.This research is sponsored by the Portugal Incentive System for Research and technological Development PEst-UID/CEC/00319/2013 and by project in co-promotion no 36265/2013 (Project HMIExcel—2013–2015)

Characterization of a new GmFAD3A allele in Brazilian CS303TNKCA soybean cultivar

Soybean is one of the most important crops cultivated worldwide. Soybean oil has 13% palmitic acid, 4% stearic acid, 20% oleic acid, 55% linoleic acid and 8% linolenic acid. Breeding programs are developing varieties with high oleic and low polyunsaturated fatty acids (linoleic and linolenic) to improve the oil oxidative stability and make the varieties more attractive for the soy industry. The main goal of this study was to characterize the low linoleic acid trait in CS303TNKCA cultivar. We sequenced CS303TNKCA GmFAD3A, GmFAD3B and GmFAD3C genes and identified an adenine point deletion in the GmFAD3A exon 5 (delA). This alteration creates a premature stop codon, leading to a truncated protein with just 207 residues that result in a non-functional enzyme. Analysis of enzymatic activity by heterologous expression in yeast support delA as the cause of low linolenic acid content in CS303TNKCA. Thus, we developed a TaqMan genotyping assay to associate delA with low linolenic acid content in segregating populations. Lines homozygous for delA had a linolenic acid content of 3.3 to 4.4%, and the variation at this locus accounted for 50.83 to 73.70% of the phenotypic variation. This molecular marker is a new tool to introgress the low linolenic acid trait into elite soybean cultivars and can be used to combine with high oleic trait markers to produce soybean with enhanced economic value. The advantage of using CS303TNKCA compared to other lines available in the literature is that this cultivar has good agronomic characteristics and is adapted to Brazilian conditions

The MOVE.TE Falls Prevention and Management Program: lessons learnt in the Portuguese context

MOVE.TE is a non-profit participatory physiotherapy platform that aims at translating knowledge in the field of physiotherapy and developing freely available evidence-based physiotherapy programmes targeting the primary care services of the Portuguese National Health service. A group of volunteer academics and clinicians collaborated at different stages and time points to create the first ever falls prevention and management programme and guidance for Physiotherapy in primary care, in Portugal. This report describes this seven-step process. In spite of many challenges, this project constitutes an example of advocacy in physiotherapy for the promotion of better healthcare for older adults.info:eu-repo/semantics/publishedVersio

Using scrum together with UML models: a collaborative university-industry R&D software project

Conducting research and development (R&D) software projects, in an environment where both industry and university collaborate, is challenging due to many factors. In fact, industrial companies and universities have generally different interests and objectives whenever they collaborate. For this reason, it is not easy to manage and negotiate the industrial companies’ interests, namely schedules and their expectations. Conducting such projects in an agile framework is expected to decrease these risks, since partners have the opportunity to frequently interact with the development team in short iterations and are constantly aware of the characteristics of the system under development. However, in this type of collaborative R&D projects, it is often advantageous to include some waterfall practices, like upfront requirements modeling using UML models, which are not commonly used in agile processes like Scrum, in order to better prepare the implementation phase of the project. This paper presents some lessons learned that result from experience of the authors in adopting some Scrum practices in a R&D project, like short iterations, backlogs, and product increments, and simultaneously using UML models, namely use cases and components.This research is sponsored by the Portugal Incentive System for Research and Technological Development PEst-UID/CEC/00319/2013 and by project in co–promotion nº 36265/2013 (Project HMIExcel - 2013-2015)

Accelerated protein synthesis via one–pot ligation–deselenization chemistry

Peptide ligation chemistry has revolutionized protein science by facilitating access to synthetic proteins. Here, we describe the development of additive-free ligation-deselenization chemistry at β-selenoaspartate and γ-selenoglutamate that enables the generation of native polypeptide products on unprecedented timescales. The deselenization step is chemoselective in the presence of unprotected selenocysteine, which is highlighted in the synthesis of selenoprotein K. The power of the methodology is also showcased through the synthesis of three tick-derived thrombin-inhibiting proteins, each of which were assembled, purified, and isolated for biological assays within a few hours. The methodology described here should serve as a powerful means of accessing synthetic proteins, including therapeutic leads, in the future

description of the methodological approach

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