Loom: Unifying Client-Side Web Technologies in a Single Programming Language

Modern client-centred web applications typically depend on a set of complementary languages to control different layers of abstraction in their interfaces: the behaviour, structure, and presentation layers (in order, traditionally: JavaScript, HTML, and CSS). Applications with dynamic interfaces whose structure and presentation depend on the data and state of the application require tight links between such layers; however, communicating between them is often non-trivial or simply cumbersome, mainly because they are effectively distinct languages—each with a specific way of being interacted with. Numerous technologies have been introduced in an attempt to simplify the interaction between the multiple layers; their main focus so far, however, regards the communication between structure and behaviour—leaving room for improvement in the field of presentation. This dissertation presents Loom: a novel reactive programming language that unifies the enunciated abstraction layers of a client-side web application. Loom allows the specification of an interface’s structure and presentation in a declarative, data-dependent, and reactive manner by means of signals—values that change over time—inspired by the field of functional reactive programming: reactive meaning that when the structure and presentation of an interface depend on application-data, changes to said data cause an automatic update of the application’s interface. We provide an implementation of the language’s compiler that allows the creation of interfaces with performance comparable to that of most existent frameworks

Lactobacillus crispatus represses vaginolysin expression by BV associated Gardnerella vaginalis and reduces cell cytotoxicity

Using a chemically-defined medium simulating genital tract secretions, we have shown that pre-adhering Lactobacillus crispatus to Hela epithelial cells reduced cytotoxicity caused by Gardnerella vaginalis. This effect was associated to the expression of vaginolysin and was specific to L. crispatus interference, as other vaginal facultative anaerobes had no protective effect.This work was supported by Portuguese National Funds (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684). JC, and MER acknowledge the financial support of individual Grants SFRH/BD/93963/2013, and SFRH/BPD/95401/2013 respectively. NC is an Investigador FCT.info:eu-repo/semantics/publishedVersio

Development of an in vitro vaginal exudate adhesion model for Bacterial Vaginosis

Microbiotec'17 - Congress of Microbiology and Biotechnology 2017Background: Bacterial vaginosis (BV) is the worldwide leading vaginal disorder commonly recognized between menarche and menopause in women of all ethnicities. It is associated with serious health problems relating to both fertility and pregnancy. This dysbiosis is characterized by a reduction in lactic acid-producing bacteria, mainly Lactobacillus spp., accompanied by an overgrowth of strict or facultative anaerobic bacteria, predominantly Gardnerella vaginalis. However, G. vaginalis is also present in healthy women and its vaginal colonization does not always lead to BV. To better understand the complex interactions that occur between host and microorganisms, and as well as between microorganisms in the vaginal microenvironment, development of in vitro models that can simulate the in vivo conditions is required, since no adequate animal model exists.This work was supported by national funds (FCT) by the strategic project of UID/BIO/04469/2013 and by BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte2020. AR had an individual FCT fellowship (PD/BD/128037/2016). NC is an Investigator FCT.info:eu-repo/semantics/publishedVersio

L. crispatus protects Hela cells against G. vaginalis cytotoxicity

Eurobiofilms 2017 - 5th European Congress on Microbial BiofilmsBacterial vaginosis (BV) is the most common lower genital tract disorder among women of reproductive age and is characterized by a shift in the vaginal flora from the dominant Lactobacillus to a polymicrobial flora, being Gardnerella vaginalis the predominant species of the biofilm mass. However, G. vaginalis vaginal colonization does not always result in BV. Accurate in vitro model systems mimicking in vivo conditions are required to better understand the complex host-microbe and microbe-microbe interactions. In effort to analyse the adaptation and interaction of the commensal vaginal microbiota and pathogens in the vaginal environment, we used a genital tract simulant medium (mGTS) to evaluate the growth of resident vaginal Lactobacillus crispatus and G. vaginalis in the adopted growth medium. Also, to understand the differences between G. vaginalis strains isolated from women with and without BV, we performed in vitro assays to compare the virulence properties of G. vaginalis strains. In this sense, G. vaginalis strains were characterised for their cytotoxicity activity after adhesion on a monolayer of epithelial cells pre-adhered with L. crispatus, mimicking the healthy vagina environment. Furthermore, transcript levels of vaginolysin and sialidase genes were also evaluated. These assays revealed that a BV isolate of G. vaginalis was significantly more cytotoxic than a non-BV isolate after 3 hours in the contact with a monolayer of HeLa cells. However, when L. crispatus was pre-adhered on a monolayer of HeLa cells, the cytotoxicity effect of both strains observed was drastically reduced. Significant differences in the transcript levels of both genes were also observed in the presence of L. crispatus. Thus, this work highlights not only the discrepant virulence potential of two distinct variants of G. vaginalis but also the beneficial role of vaginal lactobacilli in protecting the vaginal epithelium from G. vaginalis infection.info:eu-repo/semantics/publishedVersio

Compressed earth blocks stabilized with glass waste and fly ash activated with a recycled alkaline cleaning solution

Sustainable alternatives are increasingly demanded as a sound response, from the construction industry, to the worldwide growing concerns with the environment. Such effort is justifiable by the degree of the contribution of this human activity to the problem, and it has thus propelled the development of a major trend in terms of funded research. The study reported in this paper focused on the physical-mechanical properties of compacted earth blocks formed by a common Portuguese silty clay (as the mineral skeleton), stabilized with a sustainable alkali activated cement exclusively produced from wastes and residues, including coal fly ash and glass waste, in a 50/50 wt ratio combination, and activated with an alkaline solution from the aluminium industry, using activator/precursor weight ratios of 0.50, 0.57 and 0.75. After optimising the alkaline activated cement (AAC), the AAC/Soil blocks were fabricated, using the response surface method to define their composition based on curing periods of 28 and 180 days at controlled ambient temperature. Uniaxial compressive strength (UCS) and several durability tests were performed, and the material was characterised using FTIR and SEM. The results evidenced the effectiveness of the alkaline cementing agent in forming a binding matrix for the soil particles. An average compressive strength of 17.23 MPa, in unsaturated conditions, was obtained for the blocks. The newly formed soil-binder structure was very capable to withstand wetting and drying cycles, ice-thaw cycles and erosion. The microstructure of the material was further analysed, using scanning electron microscopy and energy dispersive spectroscopy. The results demonstrated the real possibility of using this type of cement as a viable alternative to traditional soil stabilisation binders used in earth construction.This work was funded by the R&D Project JUSTREST-Development of Alkali Binders for Geotechnical Applications Made Exclusively from Industrial Waste, with reference PTDC/ECM-GEO/0637/2014, financed by the Foundation for Science and Technology - FCT/MCTES (PIDDAC).The research was supported by the GEO-DESIGN project, no17501, co-financed by the European Regional Development Fund (ERDF) through NORTE 2020 (North Regional Operational Program, 2014/2020)

Bacterial vaginosis multi-species biofilms: can standard quantification methods accurately quantify in vitro biofilms?

Background. While it is well established that Bacterial Vaginosis (BV), the most common cause of vaginal discharge, involves the presence of a multi-species biofilm adhered to vaginal epithelial cells, in-depth study has been limited due to the complexity of the bacterial community comprising the biofilm. Assessing bacterial interactions between bacterial species that inhabit the BV biofilm can provide key information regarding synergism or antagonism between these species and provide insights into the pathogenesis of BV. Thus, proper biofilm quantification approaches are essential to further this body of research. Objectives. To evaluate BV biofilm formation by several key individual BV-associated bacteria (Gardnerella vaginalis, Fannyhessea vaginae, and Prevotella bivia) and compare with a multispecies biofilm formed simultaneously by all three bacterial species. Methods. Single- or multi-species biofilms were quantified by the crystal violet (CV) staining method, total cell counts by epifluorescence microscopy, and the plate counting technique (CFU); individual traits were assessed by bacterial species. Results. Each individual species had a unique signature assessed by the distinct relationship between the total number of cells, CFUs, and total biofilm biomass. Conclusions & Significance: The assessment of multi-species BV biofilm quantification results in significant bias, mainly since individual species quantification signatures cant be related to the multi-species consortia. To minimize this bias, a multiple-technical approach should be considered when quantifying multi-species BV biofilms, to circumvent the caveats of individual techniques alone, tailoring a more complete picture of the biofilm-forming capacity of key bacterial species and furthering the field of BV pathogenesis research.This work was partially supported by the Portuguese Foundation for Science and Technology (FCT) by the research project [PTDC/BIA-MIC/28271/2017] under the scope of COMPETE 2020 [POCI-01-0145-FEDER 028271] and by the strategic funding of unit [UID/BIO/04469/2020]. It was also partially funded by the National Institute of Allergy and Infectious Diseases (R01AI146065-01A1)info:eu-repo/semantics/publishedVersio

projecto interdisciplinar/metodologias integradas

Depois de descrever e fundamentar a concepção do PIMI (Projecto Interdisciplinar/Metodologias integradas), apresenta-se um conjunto de intervenções elaboradas em equipes de 2 a 3 professores da ESELx referentes a projectos realizados nos jardins de infância cooperantes da prática pedagógica ao longo do ano lectivo de 2008/09. Com estas breves apresentações pretende-se cruzar a visão inter/multidisciplinar inerente ao trabalho de projecto com crianças (Katz e Chard, 2009) com os saberes e reflexões específicos das diferentes metodologias relativas às áreas de conteúdo tal como estão expressas nas Orientações Curriculares para a Educação Pré-Escolar (1997/2002). Apresentam-se reflexões a duas ou três mãos sobre projectos específicos que conduzem a uma visão crítica sobre como se constrói a transversalidade e a especificidade no currículo, isto é, como se constrói o conhecimento (Roldão, 2004). Finalmente, apontam-se alguns dados avaliativos do processo e tecem-se desafios finais e implicações para a supervisão da prática profissional do 2º ciclo de formação (Bolonha), à luz do Decreto-Lei nº 47 /2007 de 22 de Fevereiro. O Projecto Interdisciplinar/Metodologias Integradas (PIMI) dos cursos de Formação Inicial de Educadores de Infância da ESELx, na sua concepção original, terminará no ano lectivo de 2009/2010

New integrative computational approaches unveil the Saccharomyces cerevisiae pheno-metabolomic fermentative profile and allow strain selection for winemaking

During must fermentation by Saccharomyces cerevisiae strains thousands of volatile aroma compounds are formed. The objective of the present work was to adapt computational approaches to analyze pheno-metabolomic diversity of a S. cerevisiae strain collection with different origins. Phenotypic and genetic characterization together with individual must fermentations were performed, and metabolites relevant to aromatic profiles were determined. Experimental results were projected onto a common coordinates system, revealing 17 statistical-relevant multi-dimensional modules, combining sets of most-correlated features of noteworthy biological importance. The present method allowed, as a breakthrough, to combine genetic, phenotypic and metabolomic data, which has not been possible so far due to difficulties in comparing different types of data. Therefore, the proposed computational approach revealed as successful to shed light into the holistic characterization of S. cerevisiae pheno-metabolome in must fermentative conditions. This will allow the identification of combined relevant features with application in selection of good winemaking strains.Ines Mendes was recipient of a fellowship from the Portuguese Science Foundation, FCT (SFRH/BD/74798/2010). This work was supported by FCT I.P. through the strategic funding UID/BIA/04050/2013, and the project PTDC/AGR-ALI/121062/2010

Intracellular autofluorescence as a new biomarker for cancer stem cells in glioblastoma

The identification of cancer stem cells (CSCs), which are implicated in tumor initiation, progression, therapy resistance, and relapse, is of great biological and clinical relevance. In glioblastoma (GBM), this is still a challenge, as no single marker is able to universally identify populations of GBM cancer stem cells (GSCs). Indeed, there is still controversy on whether biomarker-expressing cells fulfill the functional criteria of bona fide GSCs, despite being widely used. Here, we describe a novel subpopulation of autofluorescent (Fluo+) cells in GBM that bear all the functional characteristics of GSCs, including higher capacity to grow as neurospheres, long-term self-renewal ability, increased expression of stem cell markers, and enhanced in vivo tumorigenicity. Mechanistically, the autofluorescent phenotype is largely due to the intracellular accumulation of riboflavin, mediated by the ABC transporter ABCG2. In summary, our work identifies an intrinsic cellular autofluorescent phenotype enriched in GBM cells with functional stem cells features that can be used as a novel, simple and reliable biomarker to target these highly malignant tumors, with implications for GBM biological and clinical research.This research was funded by FEDER funds through the Operational Programme Competitiveness Factors–COMPETE and National Funds through FCT under the projects UIDB/50026/2020, UIDP/50026/2020, and POCI-01-0145-FEDER-007038; by the project NORTE-01-0145-FEDER-000013, NORTE-01-0246-FEDER-000012, and NORTE-01-0145-FEDER-000023, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). J.V.d.C., C.S.G., E.P.M., and B.M.C. was funded by FCT-Foundation for Science and Technology (SFRH/BD/88121/2012 to J.V.d.C.; SFRH/BD/92786/2013 to C.S.G.; PD/BDE/143154/2019 to E.P.M.; and PTDC/SAUGMG/113795/2009, IF/00601/2012 and CEECIND/00072/2018 to B.M.C.). B.M.C. was also funded by Fundação Calouste Gulbenkian and Liga Portuguesa Contra o Cancro

WNT6 is a novel oncogenic prognostic biomarker in human glioblastoma

Glioblastoma (GBM) is a universally fatal brain cancer, for which novel therapies targeting specific underlying oncogenic events are urgently needed. While the WNT pathway has been shown to be frequently activated in GBM, constituting a potential therapeutic target, the relevance of WNT6, an activator of this pathway, remains unknown. Methods: WNT6 protein and mRNA levels were evaluated in GBM. WNT6 levels were silenced or overexpressed in GBM cells to assess functional effects in vitro and in vivo. Phospho-kinase arrays and TCF/LEF reporter assays were used to identify WNT6-signaling pathways, and significant associations with stem cell features and cancer-related pathways were validated in patients. Survival analyses were performed with Cox regression and Log-rank tests. Meta-analyses were used to calculate the estimated pooled effect. Results: We show that WNT6 is significantly overexpressed in GBMs, as compared to lower-grade gliomas and normal brain, at mRNA and protein levels. Functionally, WNT6 increases typical oncogenic activities in GBM cells, including viability, proliferation, glioma stem cell capacity, invasion, migration, and resistance to temozolomide chemotherapy. Concordantly, in in vivo orthotopic GBM mice models, using both overexpressing and silencing models, WNT6 expression was associated with shorter overall survival, and increased features of tumor aggressiveness. Mechanistically, WNT6 contributes to activate typical oncogenic pathways, including Src and STAT, which intertwined with the WNT pathway may be critical effectors of WNT6-associated aggressiveness in GBM. Clinically, we establish WNT6 as an independent prognostic biomarker of shorter survival in GBM patients from several independent cohorts. Conclusion: Our findings establish WNT6 as a novel oncogene in GBM, opening opportunities to develop more rational therapies to treat this highly aggressive tumor.FCT - Foundation for Science and Technology (PTDC/SAU-GMG/113795/2009 and IF/00601/2012 to B.M.C.; SFRH/BD/92786/2013 to C.S.G.; SFRH/BD/88121/2012 to J.V.C.; SFRH/BD/81042/2011 to M.P.; SFRH/BD/93443/2013 to S.Q.) and Fundação Calouste Gulbenkian (B.M.C.), by FEDER funds through the Operational Programme Competitiveness Factors - COMPETE and National Funds through FCT under the project POCI-01-0145-FEDER-007038; by the project NORTE-01-0145-FEDER-000013 and NORTE-01-0246-FEDER-000012, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF); and by the project NORTE-01-0145-FEDER-000023, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER)info:eu-repo/semantics/publishedVersio