Application of LDH assay for therapeutic efficacy evaluation of ex vivo tumor models

Funding Information: This work was supported by AbbVie and by iNOVA4Health – UIDB/04462/2020, a program financially supported by Fundação para a Ciência e Tecnologia (FCT) / Ministério da Educação e Ciência, through national funds. RM and TFM were recipients of PhD fellowships funded by FCT (SFRH/BD/132163/2017 and PD/BD/128377/2017, respectively) and CB was funded by “The Discoveries Centre for Regenerative and Precision Medicine” (European Commission Horizon 2020 Research and Innovation programme, under the Grant Agreement 739572). Publisher Copyright: © 2021, The Author(s).The current standard preclinical oncology models are not able to fully recapitulate therapeutic targets and clinically relevant disease biology, evidenced by the 90% attrition rate of new therapies in clinical trials. Three-dimensional (3D) culture systems have the potential to enhance the relevance of preclinical models. However, the limitations of currently available cellular assays to accurately evaluate therapeutic efficacy in these models are hindering their widespread adoption. We assessed the compatibility of the lactate dehydrogenase (LDH) assay in 3D spheroid cultures against other commercially available readout methods. We developed a standardized protocol to apply the LDH assay to ex vivo cultures, considering the impact of culture growth dynamics. We show that accounting for growth rates and background release levels of LDH are sufficient to make the LDH assay a suitable methodology for longitudinal monitoring and endpoint assessment of therapeutic efficacy in both cell line-derived xenografts (xenospheres) and patient-derived explant cultures. This method has the added value of being non-destructive and not dependent on reagent penetration or manipulation of the parent material. The establishment of reliable readout methods for complex 3D culture systems will further the utility of these tumor models in preclinical and co-clinical drug development studies.publishersversionpublishe

DEVELOPMENT OF THE PORTUGUESE VERSIONS OF FRAIL SCALE AND SARC-F: SCREENING TOOLS FOR PHYSICAL FRAILTY AND SARCOPENIA

A FRAIL Scale e a SARC-F são ferramentas válidas desenhadas respetivamente para o rastreio da fragilidade física e da sarcopenia. O presente trabalho teve como objetivo produzir versões validadas e adaptadas linguística e culturalmente para a língua portuguesa, mantendo equivalência conceitual às ferramentas originais. Para tal, foram seguidas as normas do Patient Reported Outcome (PRO) Consortium e aplicadas as doze etapas preconizadas para a obtenção do resultado final. As ferramentas foram aplicadas a uma amostra de conveniência da população alvo constituída por nove indivíduos, tendo se verificado que tanto a FRAIL Scale como o SARC-F mostraram ser de fácil compreensão e aplicação, ter boa aceitabilidade e validade aparente. As ferramentas produzidas neste trabalho poderão ser aplicadas tanto em contexto clínico como comunitário, no rastreio da fragilidade e da sarcopenia.info:eu-repo/semantics/publishedVersio

Investigação no ensino de engenharia biomédica: dois casos de estudo

Desde os anos oitenta que os avanços das ciências básicas e das ciências da engenharia têm dado um impulso sem precedentes à investigação médica, o que criou as condições para a afirmação da Engenharia Biomédica como ramo autónomo da Engenharia. Este novo ramo da Engenharia impôs-se em domínios que incluem a imagiologia biomédica, a bioinformática, a biotecnologia, a biomecânica, os biomateriais, a engenharia de tecidos, próteses e orgãos artificiais, a análise e modelação de sinais fisiológicos e a gestão de sistemas de saúde. Já no século XXI, o volume de saber acumulado e a motivação para acelerar desenvolvimentos científicos e tecnológicos, levou à criação e organização nas universidades de uma oferta de cursos de graduação e pós-graduação em Engenharia Biomédica. Apesar do espectro muito largo da Engenharia Biomédica e dos cursos criados, é possível, mesmo ao nível de Mestrado pós-Bologna (licenciatura pré-Bologna), dar uma formação avançada que permita aos alunos realizar trabalhos de investigação complexos e com impacto clínico. Neste artigo apresentamos dois destes casos de sucesso, representativos do papel da investigação ao longo do ensino bem actual da Engenharia Biomédica. No primeiro, intitulado “Caracterização Tridimensional da Placa de Ateroma da Bifurcação Carotídea com Ultrasonografia 3D”, é apresentado um método inovador de diagnóstico da Aterosclerose, baseado na reconstrução e caracterização tridimensional da lesão aterosclerótica ao nível da bifurcação carotídea. O segundo trabalho, intitulado “Reconstrução da Frente de Onda e Simulação da Acuidade Visual no Estudo do Impacto das Aberrações Ópticas em Olhos Submetidos a Cirurgia”, apresenta uma ferramenta computacional que gera informação adicional sobre a medição da frente de onda obtida com aparelhos comerciais, permitindo estudar o impacto de aberrações ópticas na acuidade visual de diferentes olhos. Este artigo está organizado em quatro partes. A primeira parte é uma nota histórica introdutória à Engenharia Biomédica em geral. A segunda parte apresenta o que do ponto de vista de ensino superior em Engenharia Biomédica, ao nível de Mestrado Integrado de 1º e 2º ciclos, pode em geral viabilizar a fácil inserção dos estudantes na Investigação, conduzindo-os de imediato a resultados de investigação concretos. A terceira parte constitui a parte central deste artigo onde se apresentam dois exemplos ilustrativos do sucesso da formação de 5 anos em Engenharia Biomédica e da inserção da investigação, desde muito cedo, nessa formação. Por fim, na quarta secção apresentam-se as conclusões

Algae as food in Europe: an overview of species diversity and their application

ABSTRACT: Algae have been consumed for millennia in several parts of the world as food, food supplements, and additives, due to their unique organoleptic properties and nutritional and health benefits. Algae are sustainable sources of proteins, minerals, and fiber, with well-balanced essential amino acids, pigments, and fatty acids, among other relevant metabolites for human nutrition. This review covers the historical consumption of algae in Europe, developments in the current European market, challenges when introducing new species to the market, bottlenecks in production technology, consumer acceptance, and legislation. The current algae species that are consumed and commercialized in Europe were investigated, according to their status under the European Union (EU) Novel Food legislation, along with the market perspectives in terms of the current research and development initiatives, while evaluating the interest and potential in the European market. The regular consumption of more than 150 algae species was identified, of which only 20% are approved under the EU Novel Food legislation, which demonstrates that the current legislation is not broad enough and requires an urgent update. Finally, the potential of the European algae market growth was indicated by the analysis of the trends in research, technological advances, and market initiatives to promote algae commercialization and consumption.info:eu-repo/semantics/publishedVersio

Feeding in advanced dementia: consensus report from the Portuguese Society of Internal Medicine and Portuguese Enteral and Parenteral Nutrition Society

A demência é uma síndrome neurológica de agravamento progressivo, sem cura, cuja prevalência tem vindo a aumentar devido ao envelhecimento da população. Existe um grande desconhecimento entre profissionais de saúde e cuidadores relativamente à melhor abordagem da alimentação nos doentes com demência avançada. Dado não existirem recomendações nacionais acerca deste tema, foi elaborado um documento de consenso da Sociedade Portuguesa de Medicina Interna e da Associação Portuguesa de Nutrição Entérica e Parentérica que explicita as orientações existentes relativas à abordagem dos problemas alimentares nos doentes com demência avançada. A demência avançada é uma condição terminal em que deve ser privilegiado o conforto do doente, frequentemente acamado, incapaz de comunicar verbalmente e com dificuldade na alimentação. Nesta população, a literatura atual não recomenda o uso de alimentação por sonda (nasogástrica, nasojejunal, gastrostomia percutânea ou jejunostomia percutânea), que está associada a maior risco de infeção, maior utilização de meios de contenção e desenvolvimento de úlceras de decúbito. Como alternativa, recomenda-se a alimentação por via oral de acordo com a tolerância e vontade do doente (alimentação de conforto). Do ponto de vista ético e legal, é legítimo não proceder à artificialização da alimentação na fase terminal da demência caso este procedimento seja contrário aos valores da pessoa e não se objetivem benefícios. Esta decisão deve ser tomada após discussão multidisciplinar incluindo o doente (se possível), representante legal, cuidadores, família e equipa de profissionais de saúde envolvidos, elaborando um plano individual de cuidados que permita a tomada de decisões no melhor interesse do doente.ABSTRACT - Dementia is a progressive neurological syndrome without cure whose prevalence is increasing due to population aging. There is a lack of knowledge among healthcare professionals and caregivers regarding the best feeding approach in patients with advanced dementia. As there are no national recommendations on this subject, a consensus report from the Portuguese Society of Internal Medicine and the Portuguese Enteral and Parental Nutrition Society was made, clarifying existing guidelines regarding the approach of eating difficulties in these patients. Advanced dementia is a terminal condition where patient comfort should be the goal. These patients are usually bedbound, have limited ability to communicate verbally and have difficulty eating. In this population, current literature does not support tube feeding (nasogastric tube, nasojejunal tube, percutaneous gastrostomy or percutaneous jejunostomy feeding), which is associated with higher rates of infection, greater use of chemical and physical restraints and development of pressure ulcers. As an alternative, careful hand feeding should be offered (comfort feeding). From an ethical and legal standpoint, it is acceptable not to use tube feeding in the terminal phase of dementia if it is against patient values and offers no benefits. This decision should be made using a multidisciplinary approach including the patient (if possible), legal representative, caregivers, family and healthcare professionals, in order to establish an individual care plan allowing decision making in the patient’s best interest.info:eu-repo/semantics/publishedVersio

Therapeutic angiogenesis induced by human umbilical cord tissue-derived mesenchymal stromal cells in a murine model of hindlimb ischemia

© 2016 The Author(s). Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.Background: Mesenchymal stem cells derived from human umbilical cord tissue, termed UCX®, have the potential to promote a full range of events leading to tissue regeneration and homeostasis. The main goal of this work was to investigate UCX® action in experimentally induced hindlimb ischemia (HLI). Methods: UCX®, obtained by using a proprietary technology developed by ECBio (Amadora, Portugal), were delivered via intramuscular injection to C57BL/6 females after unilateral HLI induction. Perfusion recovery, capillary and collateral density increase were evaluated by laser doppler, CD31 immunohistochemistry and diaphonisation, respectively. The activation state of endothelial cells (ECs) was analysed after EC isolation by laser capture microdissection microscopy followed by RNA extraction, cDNA synthesis and quantitative RT-PCR analysis. The UCX®-conditioned medium was analysed on Gallios flow cytometer. The capacity of UCX® in promoting tubulogenesis and EC migration was assessed by matrigel tubule formation and wound-healing assay, respectively. Results: We demonstrated that UCX® enhance angiogenesis in vitro via a paracrine effect. Importantly, after HLI induction, UCX® improve blood perfusion by stimulating angiogenesis and arteriogenesis. This is achieved through a new mechanism in which durable and simultaneous upregulation of transforming growth factor β2, angiopoietin 2, fibroblast growth factor 2, and hepatocyte growth factor, in endothelial cells is induced by UCX®. Conclusions: In conclusion, our data demonstrate that UCX® improve the angiogenic potency of endothelial cells in the murine ischemic limb suggesting the potential of UCX® as a new therapeutic tool for critical limb ischemia.This study was partially supported by PORLisboa-FEDER (project QREN 2013/30196 – ClinUCX).info:eu-repo/semantics/publishedVersio

OBSERVATORIO TERRITORIAL Y AMBIENTAL ALENTEJO, EXTREMADURA, CENTRO (OTALEX C): DE GIS A IDE.

In the scope of the Spain-Portugal INTERREG projects and FEDER funded POCTEP program, OTALEX C (Territorial and Environmental Monitoring Alentejo Extremadura Center) project aims at studying of various territorial, socioeconomic and environmental indicators. It is the fundamental objective of this project, to develop a geo-portal accessible via internet, for anyone, so that the information will be useful in making decisions related to land use and therefore sustainable development of the environment. Under this general framework over the past fifteen years, we have developed different projects that have set the standardization of data between Portugal and Spain, also was designed GIS systems, and developed regional models and indicator systems, culminating in the current Spatial Data Infrastructure SDI-OTALEX C

The Mossbauer effect using Fe-57-ferrabisdicarbollide ([o-(57)FESAN](-)): a glance into the potential of a low-dose approach for glioblastoma radiotherapy

Although a variety of cancers are initially susceptible to chemotherapy, they eventually develop multi-drug resistance. To overcome this situation, more effective and selective treatments are necessary using anti-tumour agents that act in two or more ways and offer greater therapeutic benefits over single-mechanism entities. In this study, we report on treating cancer with Na[3,3′-57Fe(1,2-C2B9H11)2], which offers the possibility of dual action (radiation-drug combinations) to improve the clinical benefits and reduce healthy tissue toxicity. An approach to evaluating the potential of [o-57FESAN]− to treat glioblastoma using the Mössbauer effect is presented. As the therapeutic outcomes rely on the amount and distribution of [o-57FESAN]− inside the cells, several studies, using magnetization, Mössbauer spectroscopy and nuclear microscopy techniques, were performed to ascertain the uptake of [o-57FESAN]− in U87 glioblastoma cells. [o-57FESAN]− was found to be within the cells; 29% of its uptake was in the nuclear fraction, which is a particularly desirable target, because the nucleus is the cell's control centre where DNA and the transcription machinery reside. Irradiation studies with 2D and 3D cellular models of U87 cells showed that the growth inhibition effect observed was more pronounced when [o-57FESAN]− was used in combination with the Mössbauer effect in low total dose regimens, suggesting that this procedure either alone or as adjuvant may be useful for glioblastoma treatment.This work was supported by the Spanish Ministerio de Economía y Competitividad (PID2019-106832RB-100) and the Generalitat de Catalunya (2017SGR1720) and by the Fundação para a Ciência e Tecnologia (FCT/MEC) for projects UID/MULTI/04349/2020, PTDC/BTM-TEC/29256/2017, UIDB/04565/2020, UIDP/04565/2020 (iBB/IST), LA/P/0140/2020 (Associate Laboratory Institute for Health and Bioeconomy – i4HB), PTDC/QUI-QIN/32240/2017, LISBOA-01-0145-FEDER-022096 (National Infrastructure Roadmap, LTHMFL-NECL) and GCT grant to A. C. Cerdeira (BL156/2019_IST-ID). A. B. Buades was enrolled in the PhD program of the UAB. C. I. G. Pinto is enrolled in the PhD scholarship 689 DFA/BD/07119/2020. The authors thank Dr Moulay Sougrati, Charles Gerhardt Institute, ICGM UMR 5253, Montpellier, France for a kind gift of 57FeCl2. The LMRI (Metrology Laboratory of Ionizing Radiation) team is acknowledged for their support in the X-ray irradiation setup.With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000917-S).Peer reviewe