A novel microfluidic cell co-culture platform for the study of the molecular mechanisms of Parkinson's Disease and other synucleinopathies

Copyright © 2016 Fernandes, Chutna, Chu, Conde and Outeiro. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Although, the precise molecular mechanisms underlying Parkinson's disease (PD) are still elusive, it is now known that spreading of alpha-synuclein (aSyn) pathology and neuroinflammation are important players in disease progression. Here, we developed a novel microfluidic cell-culture platform for studying the communication between two different cell populations, a process of critical importance not only in PD but also in many biological processes. The integration of micro-valves in the device enabled us to control fluid routing, cellular microenvironments, and to simulate paracrine signaling. As proof of concept, two sets of experiments were designed to show how this platform can be used to investigate specific molecular mechanisms associated with PD. In one experiment, naïve H4 neuroglioma cells were co-cultured with cells expressing aSyn tagged with GFP (aSyn-GFP), to study the release and spreading of the protein. In our experimental set up, we induced the release of the contents of aSyn-GFP producing cells to the medium and monitored the protein's diffusion. In another experiment, H4 cells were co-cultured with N9 microglial cells to assess the interplay between two cell lines in response to environmental stimuli. Here, we observed an increase in the levels of reactive oxygen species in H4 cells cultured in the presence of activated N9 cells, confirming the cross talk between different cell populations. In summary, the platform developed in this study affords novel opportunities for the study of the molecular mechanisms involved in PD and other neurodegenerative diseases.JF was supported by FCT (SFRH/BD/73908/2010). TO is supported by the DFG Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB). The work was also supported by FCT through the Associated Laboratory IN—Institute of Nanoscience and Nanotechnology and the research project EXCL/CTM-NAN/0441/2012.info:eu-repo/semantics/publishedVersio

Novel biosensing device for point-of-care applications with plastic antibodies grown on Au-Screen Printed Electrodes

A gold screen printed electrode (Au-SPE) was modified by merging Molecular Imprinting and Self-Assembly Monolayer techniques for fast screening cardiac biomarkers in point-of-care (POC). For this purpose, Myoglobin (Myo) was selected as target analyte and its plastic antibody imprinted over a glutaraldehyde (Glu)/cysteamine (Cys) layer on the gold-surface. The imprinting effect was produced by growing a reticulated polymer of acrylamide (AAM) and N,N′-methylenebisacrylamide (NNMBA) around the Myo template, covalently attached to the biosensing surface. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) studies were carried out in all chemical modification steps to confirm the surface changes in the Au-SPE. The analytical features of the resulting biosensor were studied by different electrochemical techniques, including EIS, square wave voltammetry (SWV) and potentiometry. The limits of detection ranged from 0.13 to 8 μg/mL. Only potentiometry assays showed limits of detection including the cut-off Myo levels. Quantitative information was also produced for Myo concentrations ≥0.2 μg/mL. The linear response of the biosensing device showed an anionic slope of ~70 mV per decade molar concentration up to 0.3 μg/mL. The interference of coexisting species was tested and good selectivity was observed. The biosensor was successfully applied to biological fluids

Metaheuristic Approaches for Hydropower System Scheduling

This paper deals with the short-term scheduling problem of hydropower systems. The objective is to meet the daily energy demand in an economic and safe way. The individuality of the generating units and the nonlinearity of their efficiency curves are taken into account. The mathematical model is formulated as a dynamic, mixed integer, nonlinear, nonconvex, combinatorial, and multiobjective optimization problem. We propose two solution methods using metaheuristic approaches. They combine Genetic Algorithm with Strength Pareto Evolutionary Algorithm and Ant Colony Optimization. Both approaches are divided into two phases. In the first one, to maximize the plant’s net generation, the problem is solved for each hour of the day (static dispatch). In the second phase, to minimize the units’ switching on-off, the day is considered as a whole (dynamic dispatch). The proposed methodology is applied to two Brazilian hydroelectric plants, in cascade, that belong to the national interconnected system. The nondominated solutions from both approaches are presented. All of them meet demand respecting the physical, electrical, and hydraulic constraints

Evaluation of antifungal activity and potential application as fluorescent probes of indolenine and benzo[e]indole-based squarylium dyes

The antifungal performance and the possible use as fluorescent probes of a series of squarylium dyes derived from indolenine and benzo[e]indole previously synthesized was evaluated. Some photophysical properties were performed in ethanol and phosphate buffer, and the type of aggregates form in phosphate buffer was analyzed. Using the 1,3-diphenylisobenzofuran assay, a qualitative assessment of the capacity of dyes to produce singlet oxygen after irradiation was performed. Regarding the antifungal activity, this was studied through a broth microdilution assay using Saccharomyces cerevisiae PYCC 4072 as a biological model. The effect of irradiation of the dyes, with an appropriate light emitting diode system, on the antifungal activity was also evaluated, and it was verified that some of the dyes improve their activity after irradiation. Using fluorescence microscopy techniques, the colocalization of dyes in S. cerevisae cells was investigated and it was possible to verify that some of the squarylium dyes with a barbituric moiety in the four-membered central ring stained and accumulated preferentially in the mitochondrial web and perinuclear membrane of the cells. The possible use as a fluorescent probe for the detection of HSA was also evaluated for one of the dyes of the series, demonstrating a linear variation of the fluorescence intensity accompanied by the increase of the protein concentration.We thank to Fundação para a Ciência e Tecnologia (FCT), Comissão de Coordenação e Desenvolvimento Regional do Norte (CCDR-N) and FEDER (European Fund for Regional Development)-COMPETEQREN-EU for financial support to the research centers CQ/UM (UIDB/00686/2020), CBMA (UID/BIA/04050/2020), CQ/VR (UID/QUI/UI0616/2019) and CICSUBI (POCI-01-0145-FEDER-007491), as well as PhD grants to V.S.D.G. (UMINHO/BD/43/2016) and J.C.C.F. (SFRH/BD/133207/2017)

Phytosterols and novel triterpenes recovered from industrial fermentation coproducts exert in vitro anti‐inflammatory activity in macrophages

The unstoppable growth of human population that occurs in parallel with all manufacturing activities leads to a relentless increase in the demand for resources, cultivation land, and energy. In response, currently, there is significant interest in developing strategies to optimize any available resources and their biowaste. While solutions initially focused on recovering biomolecules with applications in food, energy, or materials, the feasibility of synthetic biology in this field has been demonstrated in recent years. For instance, it is possible to genetically modify Saccharomyces cerevisiae to produce terpenes for commercial applications (i.e., against malaria or as biodiesel). But the production process, similar to any industrial activity, generates biowastes containing promising biomolecules (from fermentation) that if recovered may have applications in different areas. To test this hypothesis, in the present study, the lipid composition of by‐products from the industrial production of β‐farnesene by genetically modified Saccharomyces cerevisiae are studied to identify potentially bioactive compounds, their recovery, and finally, their stability and in vitro bioactivity. The assayed biowaste showed the presence of triterpenes, phytosterols, and 1‐ octacosanol which were recovered through molecular distillation into a single fraction. During the assayed stability test, compositional modifications were observed, mainly for the phytosterols and 1‐octacosanol, probably due to oxidative reactions. However, such changes did not affect the in vitro bioactivity in macrophages, where it was found that the obtained fraction decreased the production of TNF‐α and IL‐6 in lipopolysaccharide (LPS)‐induced inflammation.info:eu-repo/semantics/publishedVersio

Promotion of mental health literacy and mental well-being in a Portuguese unemployed population sample : effectiveness assessment of a capacity building community-based intersectoral intervention

Copyright © 2017 Published by Elsevier Masson SASIntroduction: Economic crises have consequences on labor market, with impacts on mental health (MH) and psychological well-being (PWB). We describe the effectiveness of an intervention among unemployed, performed within EEA Grants Healthy Employment project.Objectives Evaluate the effectiveness of an intervention for MH literacy, PWB and resilience among unemployed. Aims MH and PWB promotion, common mental disorders prevention and inequalities reduction linked to unemployment.Methods A five modules intervention (life-work balance; impact of unemployment on PWB and MH; stigma; depression and anxiety; health promotion) distributed by 20 hours was developed based on literature reviews and a Delphi panel. It was delivered to unemployees from two public employment centers (PECs).Inclusion criteria: 18–65 years old; registration in PEC for less than a year; minimum of nine years of formal education. Control groups from the same PECs received the care-as-usual. Measures of psychological WB, MH self-reported symptoms, life satisfaction,resilience and mental health literacy were collected through an online survey before and one week after intervention.Results Overall, 87 unemployed participated,48% allocated to the intervention group (IG); 56% women (21–64 years old), average education was 15 years. Mixed measures ANOVA showed that the interaction between time and group was significant for PWB and MH literacy measures. The IG showed better self-reported PWB and improved MH literacy after intervention, compared to controls. No significant interactions were found for MH symptoms, life satisfaction and resilience.Conclusions This study shows the contribution of short-term community-based interventions in increasing MH literacy and PWB among unemployed.info:eu-repo/semantics/publishedVersio

Human naïve regulatory T-cells feature high steady-state turnover and are maintained by IL-7

Naïve FoxP3-expressing regulatory T-cells (Tregs) are essential to control immune responses via continuous replenishment of the activated-Treg pool with thymus-committed suppressor cells. The mechanisms underlying naïve-Treg maintenance throughout life in face of the age-associated thymic involution remain unclear. We found that in adults thymectomized early in infancy the naïve-Treg pool is remarkably well preserved, in contrast to conventional naïve CD4 T-cells. Naïve-Tregs featured high levels of cycling and pro-survival markers, even in healthy individuals, and contrasted with other circulating naïve/memory CD4 T-cell subsets in terms of their strong γc-cytokine-dependent signaling, particularly in response to IL-7. Accordingly, ex-vivo stimulation of naïve-Tregs with IL-7 induced robust cytokine-dependent signaling, Bcl-2 expression, and phosphatidylinositol 3-kinase (PI3K)-dependent proliferation, whilst preserving naïve phenotype and suppressive capacity. Altogether, our data strongly implicate IL-7 in the thymus-independent long-term survival of functional naïve-Tregs, and highlight the potential of targeting the IL-7 pathway to modulate Tregs in different clinical settings.This work was supported by Fundação para a Ciência e Tecnologia (FCT; POCI2010/IC/83068/2007 to RMMV; PTDC/SAU-MIC/109786/2009 to AES), and Gulbenkian Foundation (96526/2009 to JF; P132532/2013 to AES). SLS, ASA, RBF, ARP, PM and SMF received FCT scholarships

May Measurement Month 2017: an analysis of blood pressure screening in Angola-Sub-Saharan Africa

Elevated blood pressure (BP) is a growing burden worldwide, leading to over 10 million deaths each year. May Measurement Month (MMM) is a global initiative aimed at raising awareness of high BP and to act as a temporary solution to the lack of screening programmes worldwide. There is increasing prevalence of hypertension in developing countries leading to increased risk of adverse outcomes. An opportunistic cross-sectional survey of volunteers aged ≥18 was carried out in May 2017. Blood pressure measurement, the definition of hypertension and statistical analysis followed the standard MMM protocol. In Angola, 17 481 individuals were screened in six provinces with an average BP of 126/78 mmHg. After multiple imputation, 6022 (34.5%) had hypertension. Of individuals not receiving antihypertensive medication, 4080 (26.3%) were hypertensive. Of those receiving antihypertensive medication, 1159 (59.7%) had uncontrolled BP. MMM17 was the largest BP screening campaign undertaken in Angola. These results suggest that there is still a significant percentage of patients who are not medicated and possibly require medication, and more than half of the hypertensive patients do not have controlled BP demonstrating the need for a more targeted and comprehensive screening of BP and more effective treatment

Risk response strategies for collaborative university-industry R&D funded programs

Universities are centers of knowledge in our societies and their role when it comes to innovation has become more important over the years. Companies have several reasons to engage in research collaborations with universities, namely to gain access to innovative technologies. University-Industry R&D collaborations are expected to play an important role in regional economies, and to fulfill the industry’s demand for innovative products, technologies and processes. However, the knowledge on what are the potential risks resulting from these collaborations and the risk response strategies to reduce the negative risk impacts and to enhance positive risk impacts is still limited. Thus, this paper aims to fill the gap in literature when it comes to risk identification and risk responses’ planning, by identifying, based on a case study analysis, 19 potential risks and 53 potential risk response strategies.INCT-EN - Instituto Nacional de Ciência e Tecnologia para Excitotoxicidade e Neuroproteção(SFRH/BPD/111033/2015

Leaf heteroblasty in Passiflora edulis as revealed by metabolic profiling and expression analyses of the microRNAs miR156 and miR172

Juvenile-to-adult phase transition is marked by changes in leaf morphology, mostly due to the temporal development of the shoot apical meristem, a phenomenon known as heteroblasty. Sugars and microRNA-controlled modules are components of the heteroblastic process in Arabidopsis thaliana leaves. However, our understanding about their roles during phase-changing in other species, such as Passiflora edulis, remains limited. Unlike Arabidopsis, P. edulis (a semi-woody perennial climbing vine) undergoes remarkable changes in leaf morphology throughout juvenile-to-adult transition. Nonetheless, the underlying molecular mechanisms are unknown.Here we evaluated the molecular mechanisms underlying the heteroblastic process by analysing the temporal expression of microRNAs and targets in leaves as well as the leaf metabolome during P. edulis development.Metabolic profiling revealed a unique composition of metabolites associated with leaf heteroblasty. Increasing levels of glucose and α-trehalose were observed during juvenile-to-adult phase transition. Accumulation of microRNA156 (miR156) correlated with juvenile leaf traits, whilst miR172 transcript accumulation was associated with leaf adult traits. Importantly, glucose may mediate adult leaf characteristics during de novo shoot organogenesis by modulating miR156-targeted PeSPL9 expression levels at early stages of shoot development.Altogether, our results suggest that specific sugars may act as co-regulators, along with two microRNAs, leading to leaf morphological modifications throughout juvenile-to-adult phase transition in P. edulis