Supporting childbirth knowledge acquisition and decision-making through digital communication technology: the research design of an ongoing study following a mixed-method approach

This paper describes the research design of an ongoing study that overlaps three main fields: technology, health, and social science. This transdisciplinarity approach naturally brings challenges to the methodological plan, which this paper presents, and aims to guide the creation, validation and evaluation of a digital decision aid, and its comparison to a paper-based solution. Through the data collection from different natures, it is expected to be possible to understand the different sources, channels and formats of content that can contribute for childbirth knowledge acquisition; if communication can be facilitated between expectant parents, health care professionals, and childbirth educators; and ultimately, if the tool could provide a mean to create a document regarding birth preferences.publishe

Digital inclusion of nursing home residents: a usability evaluation of the digital kiosk siosLIFE™

The fast demographic ageing and technological progress are leading to a greater demand to develop digital solutions that can foster communication, information, and socialization of the elderly population. In the past decade, kiosks have been used to prevent digital exclusion and to promote the quality of life of this age group. This paper analyses siosLIFE™, a digital kiosk that is gradually getting attention from the public. The methodology adopted consisted in a usability test with guided tasks, using a cognitive walkthrough and think aloud protocol, with participants being residents from a nursing home. The results show that siosLIFE™ complies with some usability recommendations, but there are several improvements regarding the interface, contents, integration of support systems, and assistive technologies that can be made.publishe

Virtual reaction chambers as a tool for DNA sequencing

The topic of this thesis refers to the contribution of genetic variations, in which single nucleotide polymorphisms (SNP) can be included, to the manifestation and cause of disease. The facilitation of the treatment and cure of such conditions is done through identification and study of the cascade of biological mechanisms and processes; however the achievement of such goals is hampered by the formidable complexity and scale of the data sets involved in nucleic acid and protein analysis and thus the development of faster and cheaper methods is desirable. An intuitive trajectory for this development is the miniaturization of analytical methods with the incorporation of microfluidics which entails the manipulation of fluid analytes through channels and chambers on the microscale. This young field is projected to be instrumental in the realization of low temporal and financial cost analytical techniques. This thesis demonstrates the application of open-surface microfluidics to sequence DNA with the use of pyrosequencing. This method facilitates a reduction in instrument complexity and size and thus allows for functional integration or device disposability. Following incubation of the DNA with superparamagnetic particles, it was placed on a hydrophobic glass substrate. The DNA was then moved through microliter-volumes of mineral oil-coated water droplets via manipulation of the nanoparticles using a magnetic field; thus, these droplets could then be used either for pyrosequencing or for washing of the DNA strands. The reaction performance was determined, using the resequencing protocol with a 34 base pair (bps) of single-stranded DNA, to be highly linear for all 4 homopolymers events tested. De novo sequencing was performed with 51 and 81 bps while it was also verified that up to 7 homopolymers could be determined. This method displays full compatibility with previously demonstrated open surface steps for sample preparation and so confirms PCR on a flat glass substrate as an integrated sample-to-answer protocol. All assays were based on primer extension via DNA polymerase. A microfluidic device consisting of microliter-volume droplet-to-droplet DNA transport via manipulation of magnetic particles was used in this work. The VI difference in reaction kinetics for matched and mismatched configurations at the 3’-ends of the primer-template complex was employed in sample differentiation. The assay combined pyrosequencing technology with a sequence-by-synthesis bioluminometric DNA sequencing probe on one common microfluidic platform. Base-by-base sequencing was performed to obtain accurate single nucleotide polymorphism (SNP) scoring data with microliter volumes. The application of magnetically actuated beads to facilitate virtual chamber reactions for chip based DNA analysis was presented. Single base incorporation was seen to be detectable with the use of this pyrosequencing assay.Mit Abschluss des Humangenomprojekts verlagert sich der Fokus darauf, zu verstehen, wie genetische Variationen, wie z. B. Einzelnukleotid-Polymorphismus (SNP, single nucleotide polymorphism), zu Erkrankungen führen. Die enorme Menge an Sequenzinformationen müssen mit schnelleren, wirtschaftlicheren und leistungsfähigeren Technologien für die Analyse von RNA, DNA und Proteinen analysiert werden, um die die biologischen Mechanismen lebender Organismen zu verstehen und zu kontrollieren. Ein Ansatz ist die Miniaturisierung analytischer Methoden durch die Anwendung der Mikrofluidik, wobei Ströme in Kanälen auf der Mikrometer-Skala manipuliert werden. Es wird erwartet, dass Fortschritte in der Mikrofluidik-Chip-Technologie eine wichtige Rolle bei der Entwicklung wirtschaftlicher und schneller DNA-Analysemethoden spielen werden. In dieser Doktorarbeit wird die Anwendung von Mikrofluidik an offenen Oberfläche für die Sequenzierung von DNA mittels Pyrosequenzierung demonstriert. Dies bietet Vorteile bezüglich der Instrumentengröße, der Einfachheit, Verfügbarkeit und Funktionsintegration, insbesondere in Kombination mit den vielfältigen und flexiblen Möglichkeiten der Mikrofluidik an offenen Oberflächen. Die DNA wurde auf superparamagnetischen Partikeln inkubiert und auf ein Glassubstrat mit hydrophobischer Schicht platziert. Die Partikel mit gebundener DNA wurden mithilfe von Magnetkraft über Tropfen in Mikroliter-Größe, beschichtet mit Mineralöl zur Verhinderung von Verdunstung, bewegt. Diese Tropfen dienten als Reaktionsstationen für die Pyrosequenzierung sowie als Waschstationen. Das Sequenzierungsprotokoll mit 34 Basenpaaren (bps) einzelsträngiger DNA wurde zur Bestimmung der Reaktionsleistung verwendet und zeigte eine ausgezeichnete Linearität für alle 4 Homopolymere. Diese De-novo-Sequenzierung wurde mit 51 und 81 bps durchgeführt. Wir haben außerdem geprüft, dass bis zu 7 Homopolymere bestimmt werden können. Dieses Verfahren ist vollständig mit bisher verwendeten Schritten zur Probenvorbereitung an offenen Oberflächen kompatibel. Daher kann eine PCR als vollständig integriertes System von der VIII Probe bis zum Ergebnis auf einem flachen Glassubstrat zusammengestellt werden. In dieser Doktorarbeit werden Mikrofluidik-Anwendungen für verschiedene Genotypisierungsassay vorgestellt. Das übergeordnete Ziel ist die Kombination des Potentials des Chiplabor-Konzepts der Mikrofluidik mit Biochemie, um Verfahren für die DNA-Analyse zu entwickeln uns derzeit verfügbare Verfahren zu verbessern. Drei Genotypisierungsassays werden hier mithilfe von miniaturisierten Mikrofluidik-Methoden überprüft. Alle Assays basieren auf der Verlängerung von Primeren durch DNA-Polymerase. Ein Mikrofluidik-Instrument mit Handhabung von Tropfen zu Tropfen für Magnetpartikel für Volumen im Mikroliterbereich wurde in diesen Studien verwendet. Das Mikrofluidik-Verfahren nutzt die Vorteile der unterschiedlichen Reaktionskinetik für komplementäre und nicht-komplementäre Konfigurationen am 3‘-Ende des Primer-Template-Komplexes. Insgesamt beinhalteten die Assays die Anpassung der Pyrosequenzierungstechnologie, eines bioluminometrischen DNA-Sequenzierungsassays basierend auf Sequenzierung durch Synthese, an dieselbe Mikrofluidik-Plattform. Basenweise Sequenzierung wurde in einem Mikrofluidik-Instrument durchgeführt, um genaue SNP-Scoring-Daten für Volumina im Mikroliterbereich zu erzielen. In dieser Doktorarbeit wird die Anwendung virtueller Reaktionskammern von Partikel, die magnetisch ausgelöst werden, für die Chip-basierte DNA-Analyse präsentiert. Die Inkorporation von einzelnen Basen mithilfe der Pyrosequenzierungsreaktion wurde auf diesen Partikel beobachtet.KIST-europ

Do difficulties in emotion regulation impact self-esteem and adult attachment? – the role of trauma

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

Adverse childhood experiences and empathy: the role of interparental conflict

Communication abstract: Proceedings of the 5th International Congress of CiiEM - Reducing inequalities in Health and Society, held at Egas Moniz’ University Campus in Monte de Caparica, Almada, from June 16th to 18th, 2021.This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.The literature shows that adverse life experiences may harm individuals. The main objectives of this research were to study the relationship between adverse childhood experiences and empathy in adulthood and analyse differences between victims and nonvictims of interparental conflict. Our research evidenced that adverse childhood experiences affect individuals’ empathy in adulthood, and victims of interparental violence experienced other childhood victimization.info:eu-repo/semantics/publishedVersio

An Overview on the Recent Advances in Alternative Solvents as Stabilizers of Proteins and Enzymes

Currently, the use of alternative solvents is increasing, namely ionic liquids (ILs) and deep eutectic solvents (DESs) in diverse fields of knowledge, such as biochemistry, chemistry, chemical engineering, biotechnology and biomedicine. Particularly, when compared to traditional solvents, these alternative solvents have great importance for biomolecules due to the enhanced solubility, structure stability and the biological activity of biomolecules, such as protein and enzymes. Thus, in this review article, the recent developments and efforts on the technological developments carried out with ILs and DESs for the stabilization and activation of proteins and enzymes are provided. The most studied IL-and DES-based formulations for proteins and enzymes are discussed and the molecular mechanisms and interactions related to the increased stability promoted by these alternative solvents are disclosed, while emphasizing their main advantagespublishe

A Novel Function

Funding Information: This work was supported by Fundação para a Ciência e Tecnologia, Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES), through the grant number PTDC/QUI/64248/2006 (to A.S.P.), the Radiation Biology and Biophysics Doctoral Training Programme—RaBBiT (PD/00193/2012), Applied Molecular Biosciences Unit—UCIBIO (UIDP/04378/2020, i4HB—Institute for Health and Bioeconomy (LA/P/0140/2020) and CEFITEC (UIDB/00068/2020). A.V.A. (PD/BD/135477/2017 and COVID/BD/152498/2022) is supported by the RaBBiT programme. This work benefited from STSM funding by COST Action (CA15126 MOBIEU) and by the project CALIPSOplus under the Grant Agreement 730872 from the EU Framework Programme for Research and Innovation HORIZON 2020. Publisher Copyright: © 2022 by the authors.Encapsulins are protein nanocages capable of harboring smaller proteins (cargo proteins) within their cavity. The function of the encapsulin systems is related to the encapsulated cargo proteins. The Myxococcus xanthus encapsulin (EncA) naturally encapsulates ferritin-like proteins EncB and EncC as cargo, resulting in a large iron storage nanocompartment, able to accommodate up to 30,000 iron atoms per shell. In the present manuscript we describe the binding and protection of circular double stranded DNA (pUC19) by EncA using electrophoretic mobility shift assays (EMSA), atomic force microscopy (AFM), and DNase protection assays. EncA binds pUC19 with an apparent dissociation constant of 0.3 ± 0.1 µM and a Hill coefficient of 1.4 ± 0.1, while EncC alone showed no interaction with DNA. Accordingly, the EncAC complex displayed a similar DNA binding capacity as the EncA protein. The data suggest that initially, EncA converts the plasmid DNA from a supercoiled to a more relaxed form with a beads-on-a-string morphology. At higher concentrations, EncA self-aggregates, condensing the DNA. This process physically protects DNA from enzymatic digestion by DNase I. The secondary structure and thermal stability of EncA and the EncA−pUC19 complex were evaluated using synchrotron radiation circular dichroism (SRCD) spectroscopy. The overall secondary structure of EncA is maintained upon interaction with pUC19 while the melting temperature of the protein (Tm) slightly increased from 76 ± 1 °C to 79 ± 1 °C. Our work reports, for the first time, the in vitro capacity of an encapsulin shell to interact and protect plasmid DNA similarly to other protein nanocages that may be relevant in vivo.publishersversionpublishe

Efficient Protein Trans-Splicing at Low Vector Doses

Funding Information: Author Mariana V. Ferreira acknowledges Fundação para a Ciência e Tecnologia for PH.D. fellowship UI/BD/151256/2021 within the scope of the Ph.D. Program in Bioengineering—Cell Therapies and Regenerative Medicine. This work was funded by Fundação para a Ciência e Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through national funds to iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). Publisher Copyright: © 2023 by the authors.Adeno-associated viral (AAV) vectors represent one of the leading platforms for gene delivery. Nevertheless, their small packaging capacity restricts their use for diseases requiring large-gene delivery. To overcome this, dual-AAV vector systems that rely on protein trans-splicing were developed, with the split-intein Npu DnaE among the most-used. However, the reconstitution efficiency of Npu DnaE is still insufficient, requiring higher vector doses. In this work, two split-inteins, Cfa and Gp41-1, with reportedly superior trans-splicing were evaluated in comparison with Npu DnaE by transient transfections and dual-AAV in vitro co-transductions. Both Cfa and Gp41-1 split-inteins enabled reconstitution rates that were over two-fold higher than Npu DnaE and 100% of protein reconstitution. The impact of different vector preparation qualities in split-intein performances was also evaluated in co-transduction assays. Higher-quality preparations increased split-inteins’ performances by three-fold when compared to low-quality preparations (60–75% vs. 20–30% full particles, respectively). Low-quality vector preparations were observed to limit split-gene reconstitutions by inhibiting co-transduction. We show that combining superior split-inteins with higher-quality vector preparations allowed vector doses to be decreased while maintaining high trans-splicing rates. These results show the potential of more-efficient protein-trans-splicing strategies in dual-AAV vector co-transduction, allowing the extension of its use to the delivery of larger therapeutic genes.publishersversionpublishe

Bioinspired cyclic dipeptide functionalized nanofibers for thermal sensing and energy harvesting

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/ma16062477/s1, Figure S1: Optical microscopy; Figure S2: Output voltage as a function of time from Cyclo (L-Trp-L-Trp)@PLLA electrospun nanofibers; Figure S3: Piezoelectric current versus applied force for Cyclo (L-Trp-L-Trp)@PLLA and PLLA neat fibers, with the respective linear fits; Figure S4: Output voltage for low frequencies up to 10 Hz and output voltage as a function of time, from Cyclo (L-Trp-L-Trp)@PCL electrospun nanofibers.Nanostructured dipeptide self-assemblies exhibiting quantum confinement are of great interest due to their potential applications in the field of materials science as optoelectronic materials for energy harvesting devices. Cyclic dipeptides are an emerging outstanding group of ring-shaped dipeptides, which, because of multiple interactions, self-assemble in supramolecular structures with different morphologies showing quantum confinement and photoluminescence. Chiral cyclic dipeptides may also display piezoelectricity and pyroelectricity properties with potential applications in new sources of nano energy. Among those, aromatic cyclo-dipeptides containing the amino acid tryptophan are wide-band gap semiconductors displaying the high mechanical rigidity, photoluminescence and piezoelectric properties to be used in power generation. In this work, we report the fabrication of hybrid systems based on chiral cyclo-dipeptide L-Tryptophan-L-Tryptophan incorporated into biopolymer electrospun fibers. The micro/nanofibers contain self-assembled nano-spheres embedded into the polymer matrix, are wide-band gap semiconductors with 4.0 eV band gap energy, and display blue photoluminescence as well as relevant piezoelectric and pyroelectric properties with coefficients as high as 57 CN−1 and  35×10−6 Cm−2K−1, respectively. Therefore, the fabricated hybrid mats are promising systems for future thermal sensing and energy harvesting applications.This research was funded by Fundação para a Ciência e Tecnologia through FEDER (European Fund for Regional Development)-COMPETE-QREN-EU (ref. UID/FIS/04650/2013 and UID/FIS/04650/2019) and E-Field“Electric-Field Engineered Lattice Distortions (E-FiELD) for optoelectronic devices, ref. PTDC/NAN-MAT/0098/2020