A Critical Perspective on 3D Liver Models for Drug Metabolism and Toxicology Studies

The poor predictability of human liver toxicity is still causing high attrition rates of drug candidates in the pharmaceutical industry at the non-clinical, clinical, and post-marketing authorization stages. This is in part caused by animal models that fail to predict various human adverse drug reactions (ADRs), resulting in undetected hepatotoxicity at the non-clinical phase of drug development. In an effort to increase the prediction of human hepatotoxicity, different approaches to enhance the physiological relevance of hepatic in vitro systems are being pursued. Three-dimensional (3D) or microfluidic technologies allow to better recapitulate hepatocyte organization and cell-matrix contacts, to include additional cell types, to incorporate fluid flow and to create gradients of oxygen and nutrients, which have led to improved differentiated cell phenotype and functionality. This comprehensive review addresses the drug-induced hepatotoxicity mechanisms and the currently available 3D liver in vitro models, their characteristics, as well as their advantages and limitations for human hepatotoxicity assessment. In addition, since toxic responses are greatly dependent on the culture model, a comparative analysis of the toxicity studies performed using two-dimensional (2D) and 3D in vitro strategies with recognized hepatotoxic compounds, such as paracetamol, diclofenac, and troglitazone is performed, further highlighting the need for harmonization of the respective characterization methods. Finally, taking a step forward, we propose a roadmap for the assessment of drugs hepatotoxicity based on fully characterized fit-for-purpose in vitro models, taking advantage of the best of each model, which will ultimately contribute to more informed decision-making in the drug development and risk assessment fields.This research has been supported by FCT (Portugal) through the research grants and scholarship PTDC/MED-TOX/29183/2017, UIDB/04138/2020, UIDP/04138/2020, and SFRH/BD/144130/2019 to JR and by the H2020, European Commission, though the MSCA-IF-EF-ST – Standard EF to MC (GA-845147-LIV-AD-ON-A-CHIP). Conflict of Interestinfo:eu-repo/semantics/publishedVersio

A neural network based fall detector

In this project we present an intelligent fall detector system based on a 3-axis accelerometer and a neural network model that allows recognizing several possible motion situations and performing an emergency call only when a fall situation occurs, with low false negatives rate and low false positives rate. The system is based on a two module platform. The first one is a Mobile Station (MS) and should be carried always by the person. An accelerometer is implemented in this module and its information is transferred via a radio-frequency channel (RF) to the Base Station (BS). The BS is fixed and is connected to a GSM (Global System for Mobile communication) module. A neural network model was built into the BS and is able to identify falls from other possible motion situations, based on the received information. According to the neural network response the system sends a SMS (Short Message Service) to a destination number requesting for assistance

Otimização de uma metodologia para a determinação da composição em ácidos gordos da membrana de eritrócitos por GC-FID

Atualmente é consensual a ideia de que a composição lipídica tem influência determinante no risco de algumas doenças crónicas. A ingestão de grandes quantidades de ácidos gordos saturados e ácidos gordos trans tem sido associado a dislipidémias e aumento do risco de doenças cardiovasculares, enquanto que o consumo de ácidos gordos polinsaturados, especialmente ómega-3, tem sido associado a diversos benefícios para a saúde. Pelo facto de os eritrócitos apresentarem um elevado tempo de vida, tem sido sugerido por diversos autores que o perfil em ácidos gordos das membranas destas células poderá ser usado não apenas como um biomarcador que reflita a ingestão de macronutrientes da dieta, mas também como um biomarcador associado a diferentes patologias como diabetes, cancro e doenças cardiovasculares. Por este motivo, o presente trabalho teve como objetivo desenvolver um método simples e rápido para a identificação e quantificação de ácidos gordos presentes na membrana de eritrócitos utilizando cromatografia gasosa com detetor de ionização em chama (GC-FID). Para tal, foram obtidas amostras de sangue, a partir das quais se procedeu ao isolamento da fração contendo eritrócitos. Seguidamente foram testados diferentes protocolos para a preparação da amostra a ser analisada por GC-FID, incluindo um método clássico modificado (método de Folch) e quatro métodos rápidos, para os quais a realização da extração lipídica e reação de derivatização decorrem num passo único. No que concerne aos métodos rápidos, foi avaliada a influência de diferentes parâmetros, nomeadamente diferentes tempos de metilação e a realização, ou não, de reação de saponificação. Com base nos resultados obtidos, selecionou-se o método rápido com saponificação e com tempo de metilação de 60 minutos como sendo o mais adequado para o objetivo pretendido, permitindo obter um maior número de ácidos gordos identificados. Agradecimentos: Ao apoio financeiro concedido pela FCT no âmbito dos projetos PTDC/SAU-ENB/116929/2010 e EXPL/EMS-SIS/2215/2013, COMPETE, QREN e União Europeia (FEDER).info:eu-repo/semantics/publishedVersio

Development of a methodology using GC-FID for the quantitativo analysis of fatty acids from red blood cells

In the last years, there has been an increasing interest in evaluating possible relations between fatty acid patterns and the risk for chronic diseases. Currently, it is generally accepted that higher intakes of saturated and trans fatty acids are related with dyslipidemia and increased risk for cardiovascular diseases, while the consumption of polyunsaturated fatty acids, especially omega-3, has been positively associated with several health benefits. So far, most studies concerning the analysis of blood fatty acids (FA) composition have been performed using plasma or serum, with red blood cells (RBCs) usually being discarded [l]. However, because of the long half-life (120 days) of these cells, the FA profile of RBCs membrane may reflect longer-term markers of nutritional intake compared with plasma or urine [l].This work received financiai support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e Tecnologia) through project projetos PTDC/SAU-ENB/116929/2010 andEXPL/EMS-SIS/2215/2013info:eu-repo/semantics/publishedVersio

Bovine milk lactoferrin selectively kills highly metastatic prostate cancer PC-3 and osteosarcoma MG-63 cells in vitro

Prostate cancer and osteosarcoma are the second most common type of cancer affecting men and the fifth most common malignancy among adolescents, respectively. The use of non-toxic natural or natural-derived products has been one of the current strategies for cancer therapy, owing to the reduced risks of induced-chemoresistance development and absence of secondary effects. In this perspective, lactoferrin (Lf), a natural protein derived from milk, emerges as a promising anticancer agent due to its well-recognized cytotoxicity and anti-metastatic activity. Here, we aimed to ascertain the potential activity of bovine Lf (bLf) against highly metastatic cancer cells. The bLf effect on prostate PC-3 and osteosarcoma MG-63 cell lines, both displaying plasmalemmal V-ATPase, was studied and compared with the breast cancer MDA-MB-231 and the non-tumorigenic BJ-5ta cell lines. Cell proliferation, cell death, intracellular pH, lysosomal acidification and extracellular acidification rate were evaluated. Results show that bLf inhibits proliferation, induces apoptosis, intracellular acidification and perturbs lysosomal acidification only in highly metastatic cancer cell lines. In contrast, BJ-5ta cells are insensitive to bLf. Overall, our results establish a common mechanism of action of bLf against highly metastatic cancer cells exhibiting plasmalemmal V-ATPase. This study opens promising perspectives for further research on the anticancer role of Lf, which ultimately will contribute to its safer and more rational application in the human therapy of these life-threatening cancers.This study was supported by national funds through Fundação para a Ciência e Tecnologia (FCT) under the scope of the projects: UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569), UID/ BIO/04469/2013 (POCI-01-0145-FEDER-006684), FCT-ANR/ BEX-BCM/0175/2012, PEstOE/BIA/UI4050/2014, RECI/BBBEBI/0179/2012 (FCOMP-01-0124-FEDER-027462), and PTDC/ SAU-BMA/121028/2010.info:eu-repo/semantics/publishedVersio

Educação para o Desenvolvimento Sustentável: O Caso de Estudo dos Cursos Técnicos Superiores Profissionais do Instituto Politécnico da Guarda

O Ensino Superior (ES) defronta-se com um novo contexto de atuação, decorrente da globalização, de uma sociedade cada vez mais baseada no conhecimento, assim como da rápida evolução tecnológica que exigem uma constante (re)adaptação do saber e das competências do capital humano. Perante este cenário de mudança e os desafios que dele advêm, um dos objetivos da Estratégia Europa 2020 (EE2020), para a década 2010-2020, traduz-se em investir na qualificação superior da população europeia. Como tal, foi formulada a meta de, até 2020, os Estados-Membros ampliarem a percentagem da população com diploma de ES, na faixa etária compreendida entre os 30 e os 34 anos. A criação dos Cursos Técnicos Superiores Profissionais (CTeSP) foi a solução encontrada por Portugal para dar resposta às necessidades do país ao nível das qualificações do seu capital humano e, de igual forma, alcançar a meta europeia estabelecida. Neste contexto e, no âmbito dos processos de submissão de candidaturas a apoios financeiros provenientes de fundos europeus e nacionais para financiamento dos CTeSP, surge a necessidade de desenvolvimento do presente estudo - Educação para o Desenvolvimento Sustentável: o Caso de Estudo dos Cursos Técnicos Superiores Profissionais do Instituto Politécnico da Guarda (IPG). Assim, o presente relatório de estágio profissionalizante tem como objetivo principal apresentar uma proposta de Educação para o Desenvolvimento Sustentável do IPG, através de um Sistema de Monitorização do Percurso Académico e/ou Profissional dos Estudantes de CTeSP. A metodologia de investigação subdivide-se, primeiramente, numa análise teórica, que através da revisão de literatura sistematiza a evolução das políticas europeias e nacionais da educação e da formação no ES, bem como da qualidade e avaliação ao nível deste sector. Posteriormente, segue-se uma análise empírica, com base no método de caso de estudo (Yin, 2018) e à aplicação de uma análise estatística descritiva exploratória, aos dados obtidos através do questionário empregue aos estudantes de CTeSP do IPG. Os resultados obtidos permitem concluir que a maioria dos estudantes se encontram a trabalhar em atividades profissionais diretamente relacionadas com a área de formação obtida no CTeSP (66,8% do total) e que se encontram satisfeitos relativamente ao CTeSP frequentado (71,8% do total)

Dual antibiotherapy of tuberculosis mediated by inhalable locust bean gum microparticles

Despite the existence of effective oral therapy, tuberculosis remains a deadly pathology, namely because of bacterial resistance and incompliance with treatments. Establishing alternative therapeutic approaches is urgently needed and inhalable therapy has a great potential in this regard. As pathogenic bacteria are hosted by alveolar macrophages, the co-localisation of antitubercular drugs and pathogens is thus potentiated by this strategy. This work proposes inhalable therapy of pulmonary tuberculosis mediated by a single locust bean gum (LBG) formulation of microparticles associating both isoniazid and rifabutin, complying with requisites of the World Health Organisation of combined therapy. Microparticles were produced by spray-drying, at LBG/INH/RFB mass ratio of 10/1/0.5. The aerodynamic characterisation of microparticles revealed emitted doses of more than 90% and fine particle fraction of 38%, thus indicating the adequacy of the system to reach the respiratory lung area, thus partially the alveolar region. Cytotoxicity results indicate moderate toxicity (cell viability around 60%), with a concentration-dependent effect. Additionally, rat alveolar macrophages evidenced preferential capture of LBG microparticles, possibly due to chemical composition comprising mannose and galactose units that are specifically recognised by macrophage surface receptors. (C) 2017 Elsevier B.V. All rights reserved.National Portuguese funding through FCT - Fundacao para a Ciencia e a Tecnologia [PTDC/DTP-FTO/0094/2012, UID/BIM/04773/2013, UID/Multi/04326/2013, UID/QUI/00100/2013, PEst-OE/QUI/UI4023/2011

A review on the laser-assisted flow deposition method: growth of ZnO micro and nanostructures

Zinc oxide (ZnO) is a widely versatile semiconductor with major importance from the technological point of view, presenting the advantage to be grown by a large number of techniques and with one of the richest varieties of morphologies. Due to the special interest of this semiconductor, several methods have been developed to control the production of its nanostructures. Herein, we report the development of a vapour-based method, designated by laser-assisted flow deposition (LAFD), able of producing ZnO micro and nanocrystals with different morphologies, with a high crystalline and optical quality. This new process allows high yield of ZnO production, showing great prospects for scalable applications. In the present work, we review in detail the main growth parameters and their relationship with the produced morphologies, in addition to their influence in the structural and optical properties. Furthermore, an assessment of the possible growth mechanisms that may be involved in this new method is reported. Some potential applications of the ZnO structures produced by LAFD were also evaluated, with focus on the photocatalysis and photovoltaic fields. Additionally, the possibility of synthesizing ZnO composite nanostructures, as well as the growth of other metal oxides using this technique was explored.publishe

An antibiogram classificattion system based on an hybrid hough transform and gradient approach

The antibiogram performed by the disc diffusion method is a test frequently used in clinical microbiology. In this test, the result is given by the diameter of the inhibition zone formed around the antibiotic disco In general, this measurement is performed manually. The main objective of this work was to develop an automatic image analysis system to assess the susceptibility of microorganisms to different antibiotics. As a first step, several images of antibiograms were obtained. Subsequently the images were subjected to image processing techniques. After the elimination of noise, the Hough transform was used to detect the antibiotic discs. Next, from the center of each identified disc, the inhibition zone was detected recurring to a gradient method. According to the diameter of the inhibition zone a susceptibility classification was made. The results prove the validity of the developed tool to detect the antibiotics discs and to segment the inhibition zones used in microorganism's susceptibilfty evaluation

Psoriatic arthritis and ankylosing spondylitis impact on health-related quality of life and working life : a comparative population-based study

© 2001-2020 Sociedade Portuguesa de ReumatologiaIntroduction: Psoriatic arthritis (PsA) and ankylosing spondylitis (AS) are chronic disorders that significantly impact patients’ quality of life (QoL), health care systems and society. There is very limited data on the epidemiology and the impact of PsA and AS in Portugal, so in this study we aim to: 1) estimate the prevalence of PsA and AS in the adult Portuguese population; 2) compare health-related quality of life (QoL) of PsA and AS with the one of other rheumatic and musculoskeletal diseases (RMD) and with subjects with no rheumatic diseases; 3) compare early retirement and productivity loss among PsA and AS with other RMD. Methods: We used data from EpiReumaPt, a population-based survey, conducted from 2011 to 2013, in which 10661 subjects, over 18 years old, were screened for RMD. Spondyloarthritis (SpA) was defined by a positive expert opinion combined with the fulfillment of the Assessment of Spondyloarthritis International Society (ASAS) criteria for axial and peripheral SpA. Estimates were computed as weighted proportions considering the study design. Logistic regressions were used to compare AS/PsA subjects with other RMD and the adult Portuguese population without rheumatic diseases. Results: Prevalence rate of SpA was of 1.6% (95% CI 1.2% to 2.1%). Subjects with AS or PsA had worse QoL, reflected by EQ5D score when compared with the adult Portuguese population without rheumatic diseases (b=- 0.08; p=0.031). AS and PsA also had worse QoL when compared with participants with other RMD (b=-0.22; p>0.001). AS and in comparison to patients with other RMD, PsA subjects retired early due to their illness (OR=4.95; 95% CI 1.54% to 15.93%). A significant proportion of patients with SpA (13.6%) referred absenteeism in the previous 12 months to the interview. Conclusions: AS and PsA were found to be associated with poor QoL and a high rate of disease-related early retirement, emphasizing the burden of such rheumatic conditions in Portugal.info:eu-repo/semantics/publishedVersio