Mesenchymal stem cells in the umbilical cord: phenotypic characterization, secretome and applications in central nervous system regenerative medicine

Mesenchymal Stem Cells (MSCs), have been defined and characterized by: 1) their ability to adhere to plastic culture flasks; 2) the positive expression of CD105, CD73, CD90 membrane antigens, and the lack of expression of others (e.g CD45 and CD34) and 3) the ability of differentiation under adequate conditions along the osteogenic, chondrogenic and adipogenic lineages. In recent years, cells with these characteristics have been isolated from the Wharton’s jelly of the Umbilical Cord (UC). Similarly to bone marrow MSCs, they have shown multilineage differentiation potential and to be able to provide trophic support to neighboring cells. According to the literature, there are two main populations of cells with a mesenchymal character within the human UC: Wharton's jelly Mesenchymal Stem Cells (WJ-MSCs) and Human Umbilical Cord Perivascular Cells (HUCPVCs). In the present work our aim is to make a comprehensive review on MSC populations of the UC and how these cell populations may be used for future applications in CNS regenerative medicine. Following a brief insight on the general characteristics of MSC like cells, we will discuss the possible sources of stem cells within the WJ and the cord itself (apart UC blood), as well as their phenotypic character. As it has already been shown that these cells hold a strong trophic support to neighbouring cell populations, we will then focus on their secretome, namely which molecules have already been identified within it and their role in phenomena such as immunomodulation. The possible applications of these cell populations to CNS regenerative medicine will be addressed by critically reviewing the work that has been performed so far in this field. Finally, a brief insight will be made on what in the authors’ opinion are the major challenges in the field for the future application of these cell populations in CNS regenerative medicine.funds attributed by Fundação Calouste de Gulbenkian to A.J. Salgado under the scope of the The Gulbenkian Programme to Support research in Life Sciences; Portuguese Foundation for Science and Technology (FCT) (PhD scholarship to M.M. Carvalho - SFRH / BD / 51061 / 2010)

Behavioral characterization of the 6-hydroxidopamine model of Parkinson's disease and pharmacological rescuing of non-motor deficits

BACKGROUND: Parkinson's disease (PD) is a chronic neurodegenerative condition that is characterized by motor symptoms as a result of dopaminergic degeneration, particularly in the mesostriatal pathway. However, in recent years, a greater number of clinical studies have focused on the emergence of non-motor symptoms in PD patients, as a consequence of damage on the mesolimbic and mesocortical dopaminergic networks, and on their significant impact on the quality of life of PD patients. Herein, we performed a thorough behavioral analysis including motor, emotional and cognitive dimensions, of the unilateral medial forebrain bundle (MFB) 6-hydroxidopamine (6-OHDA)-lesioned model of PD, and further addressed the impact of pharmacological interventions with levodopa and antidepressants on mood dimensions. RESULTS: Based on apomorphine-induced turning behaviour and degree of dopaminergic degeneration, animals submitted to MFB lesions were subdivided in complete and incomplete lesion groups. Importantly, this division also translated into a different severity of motor and exploratory impairments and depressive-like symptoms; in contrast, no deficits in anxiety-like and cognitive behaviors were found in MFB-lesioned animals. Subsequently, we found that the exploratory and the anhedonic behavioural alterations of MFB-lesioned rats can be partially improved with the administration of both levodopa or the antidepressant bupropion, but not paroxetine. CONCLUSIONS: Our results suggest that this model is a relevant tool to study the pathophysiology of motor and non-motor symptoms of PD. In addition, the present data shows that pharmacological interventions modulating dopaminergic transmission are also relevant to revert the non-motor behavioral deficits found in the disease.We would like to acknowledge the funds attributed by Fundacao Calouste de Gulbenkian to A.J. Salgado under the scope of the The Gulbenkian Program to Support Research in the Life Sciences, and Portuguese Foundation for Science and Technology: Ciencia 2007 Program to A.J. Salgado; the PhD scholarships to M. M. Carvalho (SFRH/BD/51061/2010) and F. L. Campos (SFRH/BD/47311/2008), and the Post-Doctoral Fellowship to A.J. Rodrigues (SFRH/BPD/33611/2009) We want to further acknowledge Carina Cunha, Fabio Teixeira, Joao Bessa and Joao Cerqueira for their contribution to this work

Conformational dynamics of soy protein isolate under ohmic heating

Ohmic heating (OH) brings a promise of sustainable thermal processing by assuring high levels of heating efficiency and reduction of water consumption. It is important to gather fundamental knowledge about the way how this emerging heating method technology can affect the biological, functional, and technological properties of important molecules such as proteins during food processing. Vegetable proteins are attracting scientific interest in the food arena, mostly due to the need of a protein transition but also the most recent consumer trends. This work describes the effects of OH on structural aspects of soy protein isolate (SPI). OH was applied at different electrical frequencies (i.e. from 50 Hz to 20 kHz) and moderate electric fields (i.e. from 0 V/cm to 20 V/cm) at the onset denaturation temperature of SPI (95ºC). Different SPI fractions, such as legumins and vicilins, were identified through non-denaturing SDS-PAGE. Through fluorescence spectroscopy it was observed that frequencies below 500 Hz quenches tryptophan emission, suggesting structural rearrangements of the protein folding behavior; this effect is more pronounced with increasing electric field intensity. OH at 50 Hz resulted in a decrease of maximum intrinsic fluorescence intensity (p <0.05) of 27 % and 21 % when compared with the non-heated SPI and heated sample without the presence of an electric field, respectively. At the same time, treatments at 20 kHz presented a higher intrinsic fluorescence intensity than the other treatments (p < 0.05). The exposition of hydrophobic pockets appears to be less dependent on the electrical treatments applied. By reducing the thermal load, OH induces fewer changes on the protein´s structure, but the inherent electrochemical reactions and release of metals from the electrodes surface (confirmed by inductively coupled plasma mass spectrometry) can promote complexation reactions, thus changing protein folding dynamics. OH processing parameters when properly controlled result on differentiated protein binding properties which may be of essential importance regarding their functional properties but also for different applications in bioscience fields.info:eu-repo/semantics/publishedVersio

Tunning pectinase activity under the effects of electric fields in the enhanced clarification of wine must

Introduction: Pectinolytic enzymes are of great importance for the clarification process of “Vinho Verde” wine must, contributing to the reduction of haze development. During the last decade, a growing body of knowledge has been established about the effects of electric fields on the activation of important food enzymes. However, the influence of electrical parameters on catalytic activity is enzyme-dependent and should be evaluated on a case-by-case basis. This work describes, for the first time, the effects of electric fields and electrical frequency on the activity of pectinase (PEC) in the accelerated clarification of “Vinho Verde” must. Method: Moderate electric fields (MEF) with intensities below 20 V/cm and at electrical frequencies ranging from 50 to 20 kHz were applied at temperatures between 15 and 35°C. Enzymatic activity was measured for 25 min, and the initial rate of reaction was determined by the coefficient of the linear plot of galacturonic acid (GAL) production as a function of time. Results: The results show that electrical frequency can increase enzymatic activity depending on temperature conditions; at 20°C and with electrical frequencies of 2 and 20 kHz, enzymatic activity increased by up to 40 and 20%, respectively, when compared with the control sample (without the application of MEF). Temperature dependence was evaluated through the Arrhenius equation, showing that energy of activation (Ea) can be reduced from 9.2 to 6.6 kJ/mol at sub-optimal temperatures for PEC activity when MEF is applied. Discussion: Electrical parameters, when combined with temperatures below 20°C, reduced pectin concentration in “Vinho Verde” wine must by up to 42% of its initial content. This emergent treatment can be integrated in relevant environmental conditions, presenting an opportunity to increase enzyme efficiency even in low-temperature conditions, which favors the winemaking process.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit, LABBELSAssociate Laboratory in Biotechnology, Bioengineering and Microelectromechanical Systems, LA/P/0029/2020, and funded by BIOECONORTE (ref. NORTE-01-0145-FEDER-000070) with a funding body from the Norte Portugal Regional Coordination and Development Commission (CCDR-N), NORTE2020, Portugal 2020 and ESIF - European Structural and Investment Funds.info:eu-repo/semantics/publishedVersio

Manual técnico da macieira

info:eu-repo/semantics/publishedVersio

Development of Inhalable Superparamagnetic Iron Oxide Nanoparticles (SPIONs) in microparticulate system for antituberculosis drug delivery

Tuberculosis (TB) is an infectious disease which affects millions of people worldwide. Inhalable polymeric dry powders are promising alternatives as anti-TB drug carriers to the alveoli milieu and infected macrophages, with potential to significantly improve the therapeutics efficiency. Here, the development of a magnetically responsive microparticulate system for pulmonary delivery of an anti-TB drug candidate (P3) is reported. Microparticles (MPs) are developed based on a cast method using calcium carbonate sacrificial templates and incorporate superparamagnetic iron oxide nanoparticles to concentrate MPs in alveoli and enable drug on demand release upon actuation of an external alternate magnetic field (AMF). The MPs are shown to be suitable for P3 delivery to the lower airways and for alveolar macrophage phagocytosis. The developed MPs reveal unique and promising features to be used as an inhalable dry powder allowing the AMF control over dosage and frequency of drug delivery anticipating improved TB treatments.The authors wish to acknowledge the financial support from the Portuguese Foundation for Science and Technology (FCT) for the postdoctoral grant of M.S.M. (SFRH/BPD/110868/2015) and R.M.A.D (SFRH/BPD/112459/2015), FCT grant of E.T. (IF/01390/2014) and Recognize project (UTAP-ICDT/CTM-BIO/0023/2014). This article is also a result of the project “Accelerating tissue engineering and personalized medicine discoveries by the integration of key enabling nanotechnologies, marine-derived biomaterials and stem cells,” supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). The authors acknowledge the financial support from the European Union Framework Programme for Research and Innovation HORIZON 2020, under the TEAMING Grant Agreement No. 739572 – The Discoveries CTR.info:eu-repo/semantics/publishedVersio

Arbustus unedo essence: morphological and genetic characterization of the strawberry tree of Castelo de Paiva

O medronheiro é um arbusto da região mediterrânica que pode ser encontrada por todo o país. Ao contrário do que verifica na região sul do país, no concelho de Castelo de Paiva é atribuída uma reduzida importância económica a esta espécie. Com o intuito de preservar e potenciar a produção desta espécie e contribuir para a dinamização da economia do concelho, procedeu-se à caracterização morfológica e genética de uma amostra da população de medronheiros de Castelo de Paiva. A caracterização morfológica e genética foi realizada para um total de 10 genótipos. Para tal recolheram-se 70 folhas aleatoriamente em cada árvore. Em 40 folhas mediu-se o comprimento, largura, comprimento do pedúnculo, peso fresco, peso seco e determinou-se a área foliar. Dos caracteres morfológicos analisados, aqueles que se revelaram mais úteis na distinção dos vários genótipos foram: comprimento do pedúnculo, peso fresco e peso seco. As restantes 30 folhas foram utilizadas para a caracterização genética. Esta caracterização foi realizada recorrendo a um marcador de DNA, ISSR. Os 5 primeiros exemplaresutilizados na técnica de ISSR demonstraram-se polimórficos. Os resultados da caracterização genética sugerem que a variabilidade genética na população é média a alta.The strawberry tree is a shrub native in the Mediterranean region and it can be found throughout Portugal. Unlike the case in the southern region of the country, in Castelo de Paiva a minor economic importance is given to this species. In order to preserve, to enhance the production of this species and to contribute to the boosting of the economy of the region, we proceeded to the characterization of a small sample population of this fruit tree of Castelo de Paiva in what concerns to its morphology and genetics. The morphological and genetic characterization was performed for a total of 10 genotypes. For this, 70 leaves were randomly collected from each tree. For 40 leaves, it was measured the length, the width, the peduncle length, the wet weight, the dry weight and determined the leaf area. Of the morphological characteristics analyzed, the ones that proved most useful in distinguishing the various genotypes were: the length peduncle, the wet weight and the dry weight. The remaining 30 leaves were used in the genetic characterization. This characterization was performed using a DNA marker, the ISSR. The 5 primers used in the ISSR technique proved to be polymorphic. The results from the genetic characterization suggest that variability in population genetics is medium to high

Capacitação de trabalhadores em suporte básico de vida

Introdução: A capacitação dos cidadãos no auxílio a vítimas em risco de vida é fundamental em qualquer sociedade. Objetivo: avaliar os conhecimentos sobre Suporte Básico de Vida dos trabalhadores de uma Câmara Municipal do norte de Portugal, antes e após a implementação do programa de intervenção “Capacitar para Salvar” e avaliar as competências demonstradas, após a participação no programa de intervenção. Materiais e Métodos: Trata-se de um estudo do tipo quase-experimental e longitudinal, onde foi utilizado um questionário, constituído por 2 partes (caraterização sociodemográfica e avaliação de conhecimentos) e uma grelha de avaliação de competências. No 1º momento foram avaliados os conhecimentos sobre Suporte Básico de Vida e implementado um programa de intervenção, sendo que no 2º momento, foram avaliados os conhecimentos e competências sobre Suporte Básico de Vida. Resultados: Participaram no estudo 97 trabalhadores. Os resultados mostraram que os trabalhadores possuem conhecimentos e competências insuficientes em Suporte Básico de Vida. Discussão e Conclusões: Verificaram-se diferenças estatisticamente significativas nos conhecimentos e competências em Suporte Básico de Vida. Implicações para a prática: implementação de estratégias que promovam a formação em Suporte Básico de Vida a todos cidadãos, quer ao nível da transmissão de conhecimentos, quer da formação prática.Como citar este artigo: Monteiro MJFSP, Pereira MCARS, Carvalho RMBC, Carril ÉSB, Carril MFB, Rodrigues VMCP. Capacitação de trabalhadores em suporte básico de vida. Rev Cuid. 2018; 9(2): 2117-26. http://dx.doi.org/10.15649/cuidarte.v9i2.50

A prediction rule to stratify mortality risk of patients with pulmonary tuberculosis

Tuberculosis imposes high human and economic tolls, including in Europe. This study was conducted to develop a severity assessment tool for stratifying mortality risk in pulmonary tuberculosis (PTB) patients. A derivation cohort of 681 PTB cases was retrospectively reviewed to generate a model based on multiple logistic regression analysis of prognostic variables with 6-month mortality as the outcome measure. A clinical scoring system was developed and tested against a validation cohort of 103 patients. Five risk features were selected for the prediction model: hypoxemic respiratory failure (OR 4.7, 95% CI 2.8-7.9), age >= 50 years (OR 2.9, 95% CI 1.7-4.8), bilateral lung involvement (OR 2.5, 95% CI 1.44.4), >= 1 significant comorbidity-HIV infection, diabetes mellitus, liver failure or cirrhosis, congestive heart failure and chronic respiratory disease-(OR 2.3, 95% CI 1.3-3.8), and hemoglobin = 6) mortality risk. The mortality associated with each group was 2.9%, 22.9% and 53.9%, respectively. The model performed equally well in the validation cohort. We provide a new, easy-to-use clinical scoring system to identify PTB patients with high-mortality risk in settings with good healthcare access, helping clinicians to decide which patients are in need of closer medical care during treatment.This work was supported by Fundacao Amelia de Mello/Jose de Mello Saude and Sociedade Portuguesa de Pneumologia (SPP). This work was developed under the scope of the project NORTE-01-0145-FEDER-000013, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER). NSO is a FCT (Fundacao para a Ciencia e Tecnologia) investigator. MS is an Associate FCT Investigator. The fundershad no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript