Local fetal lung renin-angiotensin system as a target to treat congenital diaphragmatic hernia

Antenatal stimulation of lung growth is a reasonable approach to treat congenital diaphragmatic hernia (CDH), a disease characterized by pulmonary hypoplasia and hypertension. Several evidences from the literature demonstrated a possible involvement of renin-angiotensin system (RAS) during fetal lung development. Thus, the expression pattern of renin, angiotensin-converting enzyme, angiotensinogen, type 1 (AT₁) and type 2 (AT₂) receptors of angiotensin II (ANGII) was assessed by immunohisto-chemistry throughout gestation, whereas the function of RAS in the fetal lung was evaluated using fetal rat lung explants. These were morphometrically analyzed and intracellular pathway alterations assessed by Western blot. In nitrofen-induced CDH model, pregnant rats were treated with saline or PD-123319. In pups, lung growth, protein/DNA ratio, radial saccular count, epithelial differentiation and lung maturation, vascular morphometry, right ventricular hypertrophy and overload molecular markers, gasometry and survival time were evaluated. Results demonstrated that all RAS components were constitutively expressed in the lung during gestation and that ANGII had a stimulatory effect on lung branching, mediated by AT₁ receptor, through p44/42 and Akt phosphorylation. This stimulatory effect on lung growth was mimicked by AT₂-antagonist (PD-123319) treatment. In vivo antenatal PD-123319 treatment increased lung growth, ameliorated indirect parameters of pulmonary hypertension, improved lung function and survival time in nonventilated CDH pups, without maternal or fetal deleterious effects. Therefore, this study demonstrated a local and physiologically active RAS during lung morphogenesis. Moreover, selective inhibition of AT₂ receptor is presented as a putative antenatal therapy for CDH

Target score—a proteomics data selection tool applied to esophageal cancer identifies glut1-sialyl tn glycoforms as biomarkers of cancer aggressiveness

Esophageal cancer (EC) is a life-threatening disease, demanding the discovery of new biomarkers and molecular targets for precision oncology. Aberrantly glycosylated proteins hold tremendous potential towards this objective. In the current study, a series of esophageal squamous cell carcinomas (ESCC) and EC-derived circulating tumor cells (CTCs) were screened by immunoassays for the sialyl-Tn (STn) antigen, a glycan rarely expressed in healthy tissues and widely observed in aggressive gastrointestinal cancers. An ESCC cell model was glycoengineered to express STn and characterized in relation to cell proliferation and invasion in vitro. STn was found to be widely present in ESCC (70% of tumors) and in CTCs in 20% of patients, being associated with general recurrence and reduced survival. Furthermore, STn expression in ESCC cells increased invasion in vitro, while reducing cancer cells proliferation. In parallel, an ESCC mass spectrometry-based proteomics dataset, obtained from the PRIDE database, was comprehensively interrogated for abnormally glycosylated proteins. Data integration with the Target Score, an algorithm developed in-house, pinpointed the glucose transporter type 1 (GLUT1) as a biomarker of poor prognosis. GLUT1-STn glycoproteoforms were latter identified in tumor tissues in patients facing worst prognosis. Furthermore, healthy human tissues analysis suggested that STn glycosylation provided cancer specificity to GLUT1. In conclusion, STn is a biomarker of worst prognosis in EC and GLUT1-STn glycoforms may be used to increase its specificity on the stratification and targeting of aggressive ESCC forms.The authors wish to acknowledge the Portuguese Foundation for Science and Technology (FCT) for the human resources grants: PhD grant SFRH/BD/111242/2015 (AP), SFRH/BD/146500/2019 (MRS), SFRH/BD/142479/2018 (JS), SFRH/BD/105355/2014 (RA) and FCT assistant researcher grant CEECIND/03186/2017 (JAF). FCT is co-financed by European Social Fund (ESF) under Human Potential Operation Programme (POPH) from National Strategic Reference Framework (NSRF). The authors also acknowledge FCT the funding for CI-IPOP research unit (PEst-OE/SAU/UI0776/201) and LAQV-REQUIMTE research unit (UIDB/50006/2020), the Portuguese Oncology Institute of Porto Research Centre (CI-IPOP-29-2016-2020; CI-IPOP-58-2016-2020; CI-IPOP-Proj.70-bolsa2019-GPTE) and PhD Program in Biomedical Sciences of ICBAS-University of Porto. The author also thanks “Early stage cancer treatment, driven by context of molecular imaging (ESTIMA)” framework (NORTE-01-0145-FEDER-000027) and IPO-Score (DSAIPA/DS/0042/2018) for financial support. This work was financed by FEDER—Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020—Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020 and by Portuguese funds through FCT/MCTES—Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior. The authors also acknowledge support from the Portuguese League against Cancer grant LPCC-NRN-2020 (DF)

Leukemia Inhibitory Factor in Rat Fetal Lung Development: Expression and Functional Studies

Background: Leukemia inhibitory factor (LIF) and interleukin-6 (IL-6) are members of the family of the glycoprotein 130 (gp130)-type cytokines. These cytokines share gp130 as a common signal transducer, which explains why they show some functional redundancy. Recently, it was demonstrated that IL-6 promotes fetal lung branching. Additionally, LIF has been implicated in developmental processes of some branching organs. Thus, in this study LIF expression pattern and its effects on fetal rat lung morphogenesis were assessed. Methodology/Principal Findings: LIF and its subunit receptor LIFRa expression levels were evaluated by immunohistochemistry and western blot in fetal rat lungs of different gestational ages, ranging from 13.5 to 21.5 days post-conception. Throughout all gestational ages studied, LIF was constitutively expressed in pulmonary epithelium, whereas LIFRa was first mainly expressed in the mesenchyme, but after pseudoglandular stage it was also observed in epithelial cells. These results point to a LIF epithelium-mesenchyme cross-talk, which is known to be important for lung branching process. Regarding functional studies, fetal lung explants were cultured with increasing doses of LIF or LIF neutralizing antibodies during 4 days. MAPK, AKT, and STAT3 phosphorylation in the treated lung explants was analyzed. LIF supplementation significantly inhibited lung growth in spite of an increase in p44/42 phosphorylation. On the other hand, LIF inhibition significantly stimulated lung growth via p38 and Akt pathways

Retinoic acid regulates avian lung branching through a molecular network

Retinoic acid (RA) is of major importance during vertebrate embryonic development and its levels need to be strictly regulated otherwise congenital malformations will develop. Through the action of specific nuclear receptors, named RAR/RXR, RA regulates the expression of genes that eventually influence proliferation and tissue patterning. RA has been described as crucial for different stages of mammalian lung morphogenesis, and as part of a complex molecular network that contributes to precise organogenesis; nonetheless, nothing is known about its role in avian lung development. The current report characterizes, for the first time, the expression pattern of RA signaling members (stra6, raldh2, raldh3, cyp26a1, rar alpha, and rar beta) and potential RA downstream targets (sox2, sox9, meis1, meis2, tgf beta 2, and id2) by in situ hybridization. In the attempt of unveiling the role of RA in chick lung branching, in vitro lung explants were performed. Supplementation studies revealed that RA stimulates lung branching in a dose-dependent manner. Moreover, the expression levels of cyp26a1, sox2, sox9, rar beta, meis2, hoxb5, tgf beta 2, id2, fgf10, fgfr2, and shh were evaluated after RA treatment to disclose a putative molecular network underlying RA effect. In situ hybridization analysis showed that RA is able to alter cyp26a1, sox9, tgf beta 2, and id2 spatial distribution; to increase rar beta, meis2, and hoxb5 expression levels; and has a very modest effect on sox2, fgf10, fgfr2, and shh expression levels. Overall, these findings support a role for RA in the proximal-distal patterning and branching morphogenesis of the avian lung and reveal intricate molecular interactions that ultimately orchestrate branching morphogenesis.The authors would like to thank Ana Lima for slide sectioning and Rita Lopes for contributing to the initiation of this project. This work has been funded by FEDER funds, through the Competitiveness Factors Operational Programme (COMPETE), and by National funds, through the Foundation for Science and Technology (FCT), under the scope of the Project POCI-01-0145-FEDER-007038; and by 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). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.info:eu-repo/semantics/publishedVersio

A user-centered interface for scheduling problem definition

In this paper we present a user-centered interface for a scheduling system. The purpose of this interface is to provide graphical and interactive ways of defining a scheduling problem. To create such user interface an evaluation-centered user interaction development method was adopted: the star life cycle. The created prototype comprises the Task Module and the Scheduling Problem Module. The first one allows users to define a sequence of operations, i.e., a task. The second one enables a scheduling problem definition, which consists in a set of tasks. Both modules are equipped with a set of real time validations to assure the correct definition of the necessary data input for the scheduling module of the system. The usability evaluation allowed us to measure the ease of interaction and observe the different forms of interaction provided by each participant, namely the reactions to the real time validation mechanism

Spatial-Temporal modelization of the NO2 concentration data in mainland Portugal

In this work, we propose characterizing the evolution of the NO2 levels in Portugal, by using geostatistical approaches that deal with both the space and time coordinates and by establishing different factors affecting the concentrations detected. We also discuss appropriate tools for modelling the presence of a trend or a seasonal effect, together with specific approaches for estimation of the space-time variability. The correlation structure of the NO2 levels can be approximated, enabling to make use of the kriging techniques for prediction, without requiring data from a dense monitoring network. Proceeding in this way, the spatio-temporal patterns of the NO2 data can be derived, as well as the corresponding deviation errors. Among the potentialities of the information provided, compliance with the regulations about the NO2 concentrations can be checked

Sage drinking improves plasma lipid profile, erythrocyte antioxidant defences and increases lymphocyte HSP70

Salvia officinalis (common sage) is a medicinal plant to which antioxidant, anti-inflammatory and antimutagenic properties have been attributed. Recent results from our laboratory showed cellular and in vivo antioxidant effects of sage as well as metformin-like effects at the rat liver level, suggesting an antidiabetic potential for sage. In order to test these effects in humans, we performed a pilot trial with six healthy female volunteers. The trial was carried out in three phases, which includes two weeks of baseline, four weeks of sage treatment (drinking of a sage infusion twice a day) and two weeks of wash-out. Sage treatment positively affected the erythrocyte antioxidant status as shown by increased SOD and CAT activities. Cholesterol and LDL levels significantly decreased and HDL levels significantly increased after treatment, indicating benefits also in lipid metabolism. However, no changes in glucose clearance were observed in the oral glucose tolerance tests at the end of treatment period. In addition, a reduction of in vitro lymphocyte DNA damage induced by H2O2 was observed during the treatment period, which was maintained through the wash-out period. During the S. officinalis drinking period, lymphocyte Hsp70 protein expression was significantly increased (about 2.25 times) and decreased to baseline following the wash-out period. Overall these results confirm the health improving potential of sage infusion drinking

Improved lymphocyte response to H2O2 after regular intake of Sage tea (Salvia officinalis) involves induction of HSP70

Sage (Salvia officinalis) has recently been shown to have plasma glucose lowering potential. This suggests an antidiabetic potential for this plant alongside with other well-known properties that include antioxidant, anti-inflammatory and anti-mutagenic activities. Diabetes, a disease caused by loss of control of glucose homeostasis, also involves imbalances in the internal metabolic environment that can lead to irreversible oxidative damage in some cell populations. In the present study, we investigated the protective effect of the regular intake of Sage tea on the antioxidant response of peripheral blood lymphocytes (PBLs) challenged with H2O2. Lymphocytes were isolated from six healthy human volunteers at several times during a pilot trial of four weeks of sage tea treatment (twice a day). We also collected blood samples in the pre- and post-treatment periods referred to as baseline and wash-out, respectively. Damage to DNA was evaluated by Comet Assay. Significant reduction of damage induced by 200 μM H2O2 after two weeks of treatment were observed, indicative of improved cell defences. To further explore the mechanisms of cellular protection conferred by tea drinking to lymphocytes, we assessed effects on Hsp70 expression levels in PBLs by use of Western Blotting analysis. Besides being associated with improved stress resistance, the increased expression of Hsp is regarded as one of the most powerful means of cytoprotection against protein misfolding and aggregation. Our results show Hsp70 levels were significantly elevated in treatment compared with the baseline, which suggests that an induction of Hsp70 may be, at least in part, responsible for the improved cellular response to H2O2 induced damage.Fundação para a Ciência e a Tecnologia (FCT) - grant SFR/BD/12527/200, POCTI/AGR/62040/2004

Effects of the regular intake of sage tea (Salvia Officinalis) in humans : a pilot trial

Cells are constantly subjected to oxidative stress and oxidative damage is frequently involved in the development of several pathologies, such as cancer, hypertension and diabetes. Salvia officinalis (common sage) extracts and constituents are known for their antioxidant, antiinflammatory and anti-mutagenic proprieties. Recent experimental studies have shown that a water extract of Salvia officinalis reduces liver glucose production and fasting plasma glucose levels in normal rats, suggesting an antidiabetic potential. Therefore, we believe that common sage may be beneficial to diabetic patients where it may improve antioxidant defences and have a role in glycaemia control. The present work was undertaken to assess the effects of a sage water extract (tea) in human volunteers. To evaluate the effects of water extract of Salvia officinalis, a medicinal plant, on blood parameters and on erythrocyte antioxidant enzymes activities, we developed a pilot trial with six healthy volunteers. This trial was carried out in three phases which include two weeks of baseline, four weeks of sage tea treatment (sage tea was taken twice a day) and two weeks of wash out. During the study, blood samples were collected and analysed for haemoglobin as well as ALT and AST activities, fasting and post-prandial glucose, LDL, HDL, and cholesterol. Erythrocyte antioxidant enzymes activities (SOD and CAT) were measured from haemolysed samples. Effects on weight, blood pressure, heart rate at rest, perceived negative events (and concomitant medication) were recorded at the end of every week. The results indicate that sage tea had no toxic effects to the liver enzyme activities (ALT, AST) and increased SOD and CAT activities, which indicate that two weeks of sage tea treatment positively affect the antioxidant status. Cholesterol and LDL levels significantly decreased and HDL levels significantly increased after four weeks of treatment, which in turn suggests a beneficial effect also in lipid metabolism. We also found that two weeks of sage treatment increased blood haemoglobin levels and did not change significantly plasma glucose following oral glucose tolerance tests. Despite the fact that there were no effects on blood glucose, improvements on lipid and antioxidant balance suggest health improving effects to sage tea drinking.Fundação para a Ciência e a Tecnologia (FCT)FCT Bolsa SFRH/BD/6942/2001, POCTI/AGR/62040/200

Spatial modelling of factor analysis scores : the case of heavy metal biomonitoring in mainland Portugal

The use of mosses as biomonitors operates as an indicator of their concentration in the environment, becoming a methodology which provides a significant interpretation in terms of environmental quality. The different types of pollution are variables that can not be measured directly in the environment - latent variables. Therefore, we propose the use of factor analysis to estimate these variables in order to use them for spatial modelling. On the contrary, the main aim of the commonly used principal components analysis method is to explain the variability of observed variables and it does not permit to explicitly identify the different types of environmental contamination. We propose to model the concentration of each heavy metal as a linear combination of its main sources of pollution, similar to the case of multiple regression where these latent variables are identified as covariates, though these not being observed. Moreover, through the use of geostatistical methodologies, we suggest to obtain maps of predicted values for the different sources of pollution. With this, we summarize the information acquired from the concentration measurements of the various heavy metals, and make possible to easily determine the locations that suffer from a particular source of pollution.PTDC/MAT/112338/200