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The assertive online strategies of Portuguese luxury fashion brands/designers (LFBDs) are essential for managers to succeed in the digital medium and establish long-lasting relationships with clients. This paper explores the challenges in digitizing all the distinctive elements of a luxury fashion brand: the product characteristics, the luxury atmosphere, the personalization of the service, and the services provided. Fourteen Portuguese brands/designers were selected for content analysis. According to Hansen’s framework, 43% of Portuguese LFBDs fail to share information about their companies on their websites, and only 43% include promotions. Only 36% have a community outside the websites, compared to 93% of the international luxury fashion brands. Moreover, Portuguese websites lack 360° technology and augmented reality. Considering the Digital Visual Merchandising – MVD Grid, Portuguese LFBDs do not allow multiple zooms and 2D F/T cursor change, while international brands feature these tools. Regarding the Social Media Performance Analysis Grid, the global average engagement rate of Portuguese LFBDs on Facebook is 0.19% and on Instagram – 0.89%. According to Social Media Content Analysis Grid, Portuguese brands bet a lot on sharing content in story format, mainly on Instagram; however, Facebook has a higher network value. Therefore, the findings show that Portuguese LFBDs should be present and market their products online. This online presence must be supported by a multi-channel strategy while maintaining luxury product characteristics and elements of differentiation. AcknowledgmentThis study is partially financed by the Portuguese national funds through FCT – Fundação para a Ciência e Tecnologia, under the project UIDB/05422/2020.

Background Jet fires and boiling liquid expanding vapour explosions (BLEVEs) are potential events when a vessel containing liquefied petroleum gas (LPG) is exposed to fire. Events involving domestic LPG tanks have occurred at wildland–urban interface areas in Portugal, the USA, Spain and Greece. Aims Evaluation of the pressure relief device (PRD), the type of cylinder and the effects of jet fires and BLEVEs. Methods LPG cylinders manufactured with steel and composite materials were exposed to fire. Hydrostatic tests were carried out to compare the burst pressure without fire influence. Fourteen accidents that occurred during wildfires are described. Key results The presence of a PRD and the right choice of type of cylinder as well as cylinder location may avert major accidents. The jet fires and fireball radiation are also described, as well as the maximum distance reached by cylinder fragments. Conclusions The tests showed that the burst pressure in the test of a cylinder without PRD under fire was significantly lower when compared with the hydrostatic tests. Implications Lastly, recommendations to avoid accidents are made.

Background In Pedrógão Grande on 17 June 2017, two fire fronts merged and the propagation of the fire was influenced by the interaction of these non-symmetric fire fronts. Aims This wildfire motivated us to study a junction fire with two non-symmetrical fire fronts. The analysis of the movement of the intersection point and the angle (γ) between the bisector of the fire lines and the maximum rate of spread (ROS) direction is of particular relevance. Methods The study was carried out at Forest Fire Laboratory of the University of Coimbra in Lousã (Portugal) with laboratory experiments. Key results We found that, for small rotation angles (δ), the non-dimensional ROS of the intersection point depends on the slope angle (α) and the initial angle between fire fronts. Conclusions For high α, the non-dimensional ROS was highly influenced by the convection process and γ where the maximum ROS occurred, increased when δ increased. However, the radiation process was more relevant for lower α and influenced the non-dimensional ROS. For these cases, the maximum spread direction was close to that of the fire line bisector. Implications The present work aimed to explain fire behaviour during the Pedrógão Grande wildfire.

We control phase separation/gelation inside of emulsion droplets to reproducibly achieve SiO2–TiO2 based microspheres with tailored interconnected coral-like macroporosity. These microspheres (MICROSCAFS®) are key enablers of emergent applications.

Cellular structures are formed by struts and edges, which makes them difficult to fabricate by conventional technologies. The emergence of 3D printing has enabled the production of such complex structures. In the present work, the Fused Filament Fabrication method was used to obtain cellular structures formed by the repetition of triply periodic minimal surfaces unit cells. Due to the complexity of the cells, several defects occurred until the procedure was fine-tuned. Failure analysis of samples was performed after bending and compression tests. Failure occurs at the same locations of the samples independently of their relative density, for the three densities tested. These fracture initiation locations correspond to the zones where the von Mises stress is highest.

Background Chagas disease (CD), caused by the parasite Trypanosoma cruzi, is a serious public health concern in Latin America. Nifurtimox and benznidazole (BZ), the only two drugs currently approved for the treatment of CD, have very low efficacies in the chronic phase of the disease and several toxic side effects. Trypanosoma cruzi strains that are naturally resistant to both drugs have been reported. We performed a comparative transcriptomic analysis of wild-type and BZ-resistant T. cruzi populations using high-throughput RNA sequencing to elucidate the metabolic pathways related to clinical drug resistance and identify promising molecular targets for the development of new drugs for treating CD. Methods All complementary DNA (cDNA) libraries were constructed from the epimastigote forms of each line, sequenced and analysed using the Prinseq and Trimmomatic tools for the quality analysis, STAR as the aligner for mapping the reads against the reference genome (T. cruzi Dm28c—2018), the Bioconductor package EdgeR for statistical analysis of differential expression and the Python-based library GOATools for the functional enrichment analysis. Results The analytical pipeline with an adjusted P-value of < 0.05 and fold-change > 1.5 identified 1819 transcripts that were differentially expressed (DE) between wild-type and BZ-resistant T. cruzi populations. Of these, 1522 (83.7%) presented functional annotations and 297 (16.2%) were assigned as hypothetical proteins. In total, 1067 transcripts were upregulated and 752 were downregulated in the BZ-resistant T. cruzi population. Functional enrichment analysis of the DE transcripts identified 10 and 111 functional categories enriched for the up- and downregulated transcripts, respectively. Through functional analysis we identified several biological processes potentially associated with the BZ-resistant phenotype: cellular amino acid metabolic processes, translation, proteolysis, protein phosphorylation, RNA modification, DNA repair, generation of precursor metabolites and energy, oxidation–reduction processes, protein folding, purine nucleotide metabolic processes and lipid biosynthetic processes. Conclusions The transcriptomic profile of T. cruzi revealed a robust set of genes from different metabolic pathways associated with the BZ-resistant phenotype, proving that T. cruzi resistance mechanisms are multifactorial and complex. Biological processes associated with parasite drug resistance include antioxidant defenses and RNA processing. The identified transcripts, such as ascorbate peroxidase (APX) and iron superoxide dismutase (Fe-SOD), provide important information on the resistant phenotype. These DE transcripts can be further evaluated as molecular targets for new drugs against CD. Graphical Abstract

This work aims to produce a cartographic model that identifies areas with a greater probability of finding karstic caves - the endokarst potential - in the northern sector of Santo António Plateau (Estremadura Limestone Massif, Central Portugal). Geological, topographic, hydrogeological, and vegetation cover data were collected, processed, and integrated into a spatial database using a geographic information system (GIS). The location of known cave entrances in the study area was also identified from the records of local public institutions and speleological teams. Four conditioning factors were extracted from the collected data, including lithostratigraphic units, fracture density, relief energy, and land cover. In a multi-criteria decision-making analysis framework, each previously chosen conditioning factor and respective classes/categories were weighted using the analytic hierarchy process (AHP). With an agreement of 81.9%, the results of the cartographic model constructed in GIS seem to be promising, considering that the entrances of the known caves are mainly located in areas classified as having high to very high endokarst potential. This prototype of the endokarst potential map for the study area can be used in strategic and operational environmental planning (at least on a local scale), as it can assist decision-makers, competent authorities, and local speleological teams in a more accurate and thoughtful definition of the areas that should be investigated, providing a substantial reduction in times and costs field prospecting.

Avian metapneumovirus (aMPV) causes a highly contagious upper respiratory and reproductive disease in chickens, turkeys, and ducks. Here, complete genome sequences of aMPV-B vaccine strains BR/1890/E1/19 (PL21, Nemovac; Boehringer Ingelheim Animal Health, Brazil) and BR/1891/E2/19 (1062; Hipraviar, France) were sequenced and compared with the pathogenic field strain VCO3/60616.

Leishmania are kinetoplastid pathogens that cause leishmaniasis, a debilitating and potentially life-threatening infection if untreated. Unusually, Leishmania regulate their gene expression largely post-transcriptionally due to the arrangement of their coding genes into polycistronic transcription units that may contain 100s of functionally unrelated genes. Yet, Leishmania are capable of rapid and responsive changes in gene expression to challenging environments, often instead correlating with dynamic changes in their genome composition, ranging from chromosome and gene copy number variations to the generation of extrachromosomal DNA and the accumulation of point mutations. Typically, such events indicate genome instability in other eukaryotes, coinciding with genetic abnormalities, but for Leishmania, exploiting these products of genome instability can provide selectable substrates to catalyse necessary gene expression changes by modifying gene copy number. Unorthodox DNA replication, DNA repair, replication stress factors and DNA repeats are recognised in Leishmania as contributors to this intrinsic instability, but how Leishmania regulate genome plasticity to enhance fitness whilst limiting toxic under- or over-expression of co-amplified and co-transcribed genes is unclear. Herein, we focus on fresh, and detailed insights that improve our understanding of genome plasticity in Leishmania. Furthermore, we discuss emerging models and factors that potentially circumvent regulatory issues arising from polycistronic transcription. Lastly, we highlight key gaps in our understanding of Leishmania genome plasticity and discuss future studies to define, in higher resolution, these complex regulatory interactions.

Additive manufacturing (AM) of polymer large parts is a technological research area with great growth potential if the main barriers to its implementation are successfully addressed. In this research, a review of large-format AM (LFAM) processes for polymers is presented, followed by market research concerning the identification of large-format polymer commercial printers. An overview was performed covering the current LFAM systems configurations and their control aspects. The design and modelling approaches related with the fabrication of polymer large parts by AM, and the materials currently being applied and under development, were described. Finally, a summary of LFAM applications with a focus in the Transportation, Academic, Construction and Energy sectors, was presented. The current main advances in the LFAM of polymers are linked with the possibility of producing large parts in a faster, cheaper, and reliable way. The market research analysis concerning results for all AM families involving polymer materials reveals that, currently, the material extrusion AM process family is potentially the most suitable to produce large parts, with a significant number of applications attesting its capability to produce such large-format components.

Repetitive elements cause assembly fragmentation in complex eukaryotic genomes, limiting the study of their variability. The genome of Trypanosoma cruzi, the parasite that causes Chagas disease, has a high repetitive content, including multigene families. Although many T. cruzi multigene families encode surface proteins that play pivotal roles in host-parasite interactions, their variability is currently underestimated, as their high repetitive content results in collapsed gene variants. To estimate sequence variability and copy number variation of multigene families, we developed a read-based approach that is independent of gene-specific read mapping and

/SummaryBackgroundVisceral leishmaniasis (VL) is a potentially fatal disease mainly caused by Leishmania infantum in South America and L. donovani in Asia and Africa. Disease outcomes have been associated with patient genotype, nutrition, age, sex, comorbidities, and co-infections. In this study, we examine the effects of parasite genetic variation on VL disease severity in Brazil.MethodsWe collected and sequenced the genomes of 109 L. infantum isolates from patients in northeast Brazil and retrieved matching patient clinical data from medical records, including mortality, sex, HIV co-infection and laboratory data (creatinine, haemoglobin, leukocyte and platelet counts). We identified genetic differences between parasite isolates, including single nucleotide polymorphisms (SNPs), small insertions/deletions (indels), and variations in genic, intergenic, and chromosome copy numbers (copy number variants, CNVs). To describe associations between the parasite genotypes and clinical outcomes, we applied quantitative genetics methods of heritability and genome-wide association studies (GWAS), treating clinical outcomes as traits that may be influenced by parasite genotype.FindingsMultiple aspects of the genetic analysis indicate that parasite genotype affects clinical outcomes. We estimate that parasite genotype explains 83% chance of mortality (narrow sense heritability, h2 = 0·83±0·17), and has a significant relationship with patient sex (h2 = 0·60±0·27). Impacts of parasite genotype on other clinical traits are lower (h2 ≤0·34). GWAS analysis identified multiple parasite genetic loci that were significantly associated with clinical outcomes; 17 CNVs that were significantly associated with mortality, two with creatinine and one with bacterial co-infection, jaundice and HIV co-infection; and two SNPs/indels and six CNVs that associate with age, jaundice, HIV and bacterial co-infections, creatinine, and/or bleeding sites.InterpretationParasite genotype is an important factor in VL disease severity in Brazil. Our analysis indicates that specific genetic differences between parasites act as virulence factors, enhancing risks of severe disease and mortality. More detailed understanding of these virulence factors could be exploited for novel therapies.Author SummaryMultiple factors contribute to the risk of mortality from visceral leishmaniasis (VL), including, patient genotype, comorbidities, and nutrition. Many of these factors will be influenced by socio-economic biases 1. Our work suggests that the virulence of the infecting parasite is an important risk factor for mortality. We pinpoint some specific genomic markers that are associated with mortality, which can lead to a greater understanding of the molecular mechanisms that cause severe VL disease, to genetic markers for virulent parasites and to the development of drug and vaccine therapies.

This paper is focused on the formability of wire-arc additively manufactured AISI 316L stainless steel sheets with the purpose of analysing the feasibility of including this material in hybrid additive manufacturing chains involving sheet metal forming operations. Conventional tensile tests performed in specimens obtained from different orientations to the building direction were carried out to characterise the mechanical properties, the strain loading paths and the limiting strains at fracture. Microstructure observations combined with fractography analysis add insight to the results by establishing a link between the forming limits by fracture and the crack opening mechanisms. Results obtained for wrought commercial AISI 316L stainless steel sheets are included for comparison purposes and reveal that the additively manufactured sheets have a much stronger anisotropic behaviour and poorer formability due to their dendritic-based microstructure. Still, the forming limits obtained from the experiments allow concluding that the additively manufactured sheets can withstand large plastic deformations and, therefore, can be used in hybrid additive manufacturing routes.

Additive manufacturing technologies are becoming increasingly popular due to their advantages over traditional subtracting manufacturing technologies. Despite advances in this field, fixed and maintenance costs for additive manufacturing with metals remain high. The introduction of low-cost metal machines in the additive manufacturing market considerably reduces the cost of acquiring and maintaining this type of equipment. This work aims to establish the process requirements for a low-cost selective powder deposition process, and validate it through the production of specimens in the laboratory and evaluate their mechanical properties. Tin bronze specimens were produced under different manufacturing conditions, namely powder dimensions, type of crucible and coke, firing segments and casting strategy. The morphology and chemical composition of the specimens were carried out combining the scanning electron microscopy and energy dispersive X-Ray spectroscopy techniques, respectively. It was observed that crucibles and coke with impurities that react with the metal powders and infill in a reducing atmosphere have influence in the final quality of parts. Tested samples displayed high variability of results which can be correlated with different manufacturing conditions. The selection of the appropriate print parameters led to the manufacture of tin bronze specimens with mechanical properties comparable to those reported in the literature. Overall, low-cost selective powder deposition is a promising technology, if identified manufacturing issues are addressed.

Não tenho do meu curso de História, na época de 1960 a 1966, ano em que defendi a tese de licenciatura, uma imagem essencialmente positiva, conforme já o disse num livro, História… Que História?, publicado pela editora Temas e Debates em 2015. A nossa ciência era então vista de uma forma “narrativista” e “acontecimental”. As cadeiras que estimulavam a minha reflexão crítica —aprendida no liceu com Alberto Martins de Carvalho — fui buscá-las particularmente à Filosofia e à História da Cultura, em que ensinavam professores, de diversas concepções, oriundos de outras áreas: Sílvio Lima (em 1935 demitido por Salazar, mas que pudera regressar nos anos 40), o qual reflectia, nas suas lições inesquecíveis, sobre os princípios da teoria da História

Intracellular parasites of the Leishmania donovani species complex cause visceral leishmaniasis (VL). For parasitic diseases, VL has a mortality rate second only to malaria, and is associated with poverty-stricken areas of the world: primarily Brazil, East Africa and the Indian subcontinent (ISC). Miltefosine (MIL) and the antimonal sodium stibogluconate (SSG) are drugs used in the treatment of leishmaniasis. However, treatment efficacy is variable, and the numbers of reports of parasite resistance to both drugs have risen since their introductions, particularly in the ISC. To assess the level of parasite genotype contribution to drug resistance, we utilised the sequencing and associated drug susceptibility data from Imamura et al. (2016) to estimate heritability and GWAS using LDAK. We obtained strong heritability results, with mainly SNP/indel variations associated with SSG and copy number variants associated with MIL resistance, respectively. However, GWAS results were inconclusive, suggesting that, although the parasite genotype directly influences drug resistance, the effect might be multifactorial.

A new generation of biodegradable metal alloys with a porous structure has been receiving growing attention as temporary bone scaffolds for tissue regeneration. The mechanical response of the scaffolds depends upon several factors including the properties of the metal itself, the amount of porosity, the geometrical topology and the immersion conditions. The purpose of this study is to evaluate the degradation behaviour and the mechanical properties of porous iron samples with porosities in the range of 20–30%. Besides the amount of porosity, the effect of topology was evaluated with the study of different arrangement of pores, as well as pore shapes. The specimens were subjected to chemical degradation by immersion of the iron samples in body fluid simulation conditions. The mechanical properties of the samples prior and after the degradation process were assessed by three-point bending tests. Numerical simulations were carried out and the results were compared with the experimental results. The degradation operated by body fluids tends to reduce the mechanical properties. In comparison with the compact structures, porous structures exhibit lower mechanical strength, but still with reasonable values for the use in temporary implants, which also allows reducing the stress shielding effect, keeping the biodegradable advantages. The present work also confirms that the topological design has a strong influence on the mechanical properties of the specimens and on the biodegradation behaviour.

An estimated 400 million people are infected by parasites of the genus Ascaris and the existing control measures are inefficient. Vaccine development using B cell antigens is a promising strategy for increased protection against this parasite. The present study aimed at developing a chimeric protein capable of conferring protection against infection by Ascaris sp. For this purpose, we performed B-cell epitope predictions on previously described vaccine candidate proteins from Ascaris suum and the corresponding peptides were used to construct a chimeric protein. Female BALB / c mice were immunized subcutaneously in three doses at 10 day intervals with a vaccine formulation comprised of the chimeric protein together with monophosphoryl lipid A (MPLA). Control groups included protein alone, MPLA, or PBS. After challenge infection, animals vaccinated with chimeric protein plus MPLA showed a reduction of 73.54% of larval load in the lung compared to control group animals. Animals immunized with chimeric protein plus MPLA also display higher IgG response and a reduction in lung inflammation. Our study highlights how chimeric proteins containing more than one B cell epitope can enhance immune protection against helminthic infection and offer new approaches to the development of Ascaris vaccines.

Introdução: Um dos princípios da Atenção Primária à Saúde (APS) é a abordagem do indivíduo dentro do seu contexto familiar. Com isso, a ESF lança mão de ferramentas provenientes de áreas como a sociologia e a psicologia, tais como o genograma, ecomapa, ciclos de vida, F.I.R.O, P.R.A.C.T.I.C.E e a conferência familiar, para intervir em demandas familiares que são direcionadas ao serviço de saúde. Objetivo: Relatar a experiência de uma equipe saúde da família (eSF) com a aplicação de ferramentas de abordagem familiar no cuidado de um indivíduo idoso. Métodos: Trata-se de um trabalho de abordagem descritiva e qualitativa, do tipo relato de experiência, em que as ferramentas de abordagem familiar foram utilizadas como método de diagnóstico e intervenção para avaliação da dinâmica familiar do paciente. Relato de experiência: A.B., 81 anos, polifarmacêutico, procurou a Unidade Básica de Saúde (UBS) com queixa de pressão na cabeça. Depois de escuta por parte dos profissionais, foi percebido que os sintomas se iniciaram após a morte da companheira, há seis anos. Foi decidido utilizar as ferramentas de abordagem familiar para intervenção nos problemas identificados. Logo após análise das ferramentas, foi percebido como ações de intervenção realizar orientações quanto ao autocuidado e principalmente reiterar o sujeito quanto ao convívio social, lidando com as questões do luto e relacionamento familiar com seus filhos. Conclusão: Por meio das ferramentas de abordagem familiar, foram propostas intervenções eficazes para melhora do paciente com a utilização de recursos que incluíam não somente o cuidado clínico- médico, mas também o envolver em espaços e atividades que favoreçam a qualidade de vida.

Sandwich structures are frequently used in automotive, aerospace and marine industries, as they provide adequate functional properties. The two-dimensional regular hexagonal cell shape, i.e. honeycomb is the most used core structure in sandwich panels. Recently, a new type of cellular structures composed of lattice struts has been proposed, as they combine high stiffness, strength and energy absorption with low weight. The main purpose of this research is to investigate the effect of the lattice topology on the flexural behaviour of sandwich panels. Five lattice geometries inspired in crystalline structures were designed, namely, body-centred parallelepiped, body-centred parallelepiped with struts in z-axis, body- and face-centred parallelepiped with struts in z-axis, face-centred parallelepiped with struts in z-axis and parallelepiped simple. The relative density of all the lattices was kept constant as 0.3. Both numerical and experimental approaches were used to evaluate the flexural properties and failure behaviour of the sandwich structures under three-point bending tests. The numerical analysis was undertaken with the finite element software NX Nastran. Taking advantage of additive manufacturing technologies, material extrusion was used to produce polylactic acid samples with the configurations aforementioned. The sandwich panels are composed by a single layer formed by the lattice core and two thin plates, at the bottom and top. The three parts of the panel were manufactured all together. The simulation results indicate that, among the lattices studied, topologies body-centred parallelepiped with struts in z-axis and body- and face-centred parallelepiped with struts in z-axis exhibit higher strength, while body- and face-centred parallelepiped with struts in z-axis shows higher stiffness and higher energy absorption, attaining values that do not differ much from the ones obtained with a two-dimensional hexagonal cellular structure, with the same relative density. As a consequence, some of the geometries studied may have the potential to be considered as alternatives to conventional structures in the design of sandwich structures.

O artigo teve como objetivo traçar o perfil profissional ideal e real dos professores de Educação Física do Ensino Médio que atuam em escolas da cidade de Porto Velho/RO. Foi realizada uma pesquisa de campo, descritiva, de cunho quantitativo e qualitativo, método dedutivo. A amostra foi de 10 professores que atuam especificamente no ensino médio. Foi aplicado um questionário misto com 11 perguntas. A pesquisa foi realizada nas escolas: Centro de Ensino Classe A, Colégio e Cursos Sapiens – Unidade Jardim América, Instituição adventista de Educação Noroeste Brasileira, Colégio Tiradentes da Polícia Militar de Rondônia, E.E.E.M. Major Guapindaia, E.E.E.F.M. Prof. Daniel Neri da Silva, da cidade de Porto Velho, Rondônia. O perfil do professor de Educação Física do ensino médio é de um indivíduo que se preocupa com sua formação, tem prazer em atuar na docência de adolescentes, se propõe a resolver pequenas questões relacionadas ao convívio com os alunos, Concluiu-se que os professores pesquisados estão satisfeitos com suas condições atuais, sendo cientes de suas obrigações e responsabilidades como professores, todos possuem especialização, e a atuação em uma única escola é opção dos próprios professores que possuem atividades complementares que se divergem do âmbito escolar. Palavras-chave:Perfil profissional. Educação Física. Ensino médio. Professor.

As the global demand for energy rises, renewable sources are increasingly becoming more accessible, and oceans show an excellent possibility for the expansion of wind energy. In this work, the stru...

Chagas disease, caused by the protozoan Trypanosoma cruzi, is a serious chronic parasitic disease, currently treated with Nifurtimox (NFX) and Benznidazole (BZ). In addition to high toxicity, these drugs have low healing efficacy, especially in the chronic phase of the disease. The existence of drug-resistant T. cruzi strains and the occurrence of cross-resistance between BZ and NFX have also been described. In this context, it is urgent to study the metabolism of these drugs in T. cruzi, to better understand the mechanisms of resistance. Prostaglandin F2α synthase (PGFS) is an enzyme that has been correlated with parasite resistance to BZ, but the mechanism by which resistance occurs is still unclear. Our results show that the genome of the CL Brener clone of T. cruzi, contains five PGFS sequences and three potential pseudogenes. Using CRISPR/Cas9 we generated knockout cell lines in which all PGFS sequences were disrupted, as shown by PCR and western blotting analyses. The PGFS deletion did not alter the growth of the parasites or their susceptibility to BZ and NFX when compared to wild-type (WT) parasites. Interestingly, NTR-1 transcripts were shown to be upregulated in ΔPGFS mutants. Furthermore, the ΔPGFS parasites were 1.6 to 1.7-fold less tolerant to oxidative stress generated by menadione, presented lower levels of lipid bodies than the control parasites during the stationary phase, and were less infective than control parasites.

Ultrasonic fatigue testing is a recent fatigue methodology that applies resonant principles to induce cyclic stresses in a modally designed specimen enabling the application of fatigue damage at very high frequencies. Its primary purpose is to study fatigue life in the Very High Cycle Fatigue regime between 10E07 and 10E09 cycles with a higher performance of time and energy wise compared to conventional servo-hydraulic fatigue testing machines. In this study, an ultrasonic fatigue testing machine was used to carry out multiaxial tension/torsion fatigue tests at a frequency of 20 kHz. The objective was to reach a reliable multiaxial fatigue testing method by modifying only the specimen, with the ability to induce different shear to axial stress ratios. The improved design/methodology of the testing ultrasonic multiaxial method focused first on the innovative design specimens and setup modal behaviour by conducting both numerical and experimental analyses. Laser displacement measurements, thermographic imaging, and rosette strain gauges applied to the specimen’s fatigue stress region were carried out to validate the new design concept and compared with the computed results obtained by finite element. A good numerical to experimental results agreement was achieved. A series of fatigue tests were carried out in tension/torsion fatigue to three different combining dimension specimens allowing three different shear/axial stress ratios. The resulting crack initiation angles and fracture surfaces were analysed and compared not only between each other but also with tension-compression and pure torsion ultrasonic tested specimens.

In this work, the magnesium alloy AZ31BF subjected to proportional and non-proportional loads has been studied. For this purpose, a series of experimental multiaxial fatigue tests were carried out according to the ASTM E466 protocol. The main objective was to determine the relationship between the multiaxial fatigue strength of this alloy under these two different types of loading. The results showed that the AZ31BF magnesium alloy has different fatigue strengths depending on the loading type. Based on these results, it was found that the ratio between proportional and non-proportional damage in AZ31BF magnesium alloy varies depending on the number of loading cycles. To represent this variation, parameter Y was used to modulate the non-proportional damage of AZ31BF. In this way, two Y functions were considered, one for the normal stress component and the other for the shear stress component. The results obtained for the non-proportional parameter Y are of particular interest since the multiaxial fatigue models do not distinguish between these two types of loading when evaluating fatigue life. In this sense, the results of this study can be used in these models to overcome this limitation.

Trypanosoma cruzi, the etiological agent of Chagas disease, exhibits extensive inter- and intrastrain genetic diversity. As we have previously described, there are some genetic differences between the parental G strain and its clone D11, which was isolated by the limiting dilution method and infection of cultured mammalian cells. Electrophoretic karyotyping and Southern blot hybridization of chromosomal bands with specific markers revealed chromosome length polymorphisms of small size with additional chromosomal bands in clone D11 and the maintenance of large syntenic groups. Both G strain and clone D11 belong to the T. cruzi lineage TcI. Here, we designed intraspecific array-based comparative genomic hybridization (aCGH) to identify chromosomal regions harboring copy-number variations between clone D11 and the G strain. DNA losses were more extensive than DNA gains in clone D11. Most alterations were flanked by repeated sequences from multigene families that could be involved in the duplication and deletion events. Several rearrangements were detected by chromoblot hybridization and confirmed by aCGH. We have integrated the information of genomic sequence data obtained by aCGH to the electrophoretic karyotype, allowing the reconstruction of possible recombination events that could have generated the karyotype of clone D11. These rearrangements may be explained by unequal crossing over between sister or homologous chromatids mediated by flanking repeated sequences and unequal homologous recombination via break-induced replication. The genomic changes detected by aCGH suggest the presence of a dynamic genome that responds to environmental stress by varying the number of gene copies and generating segmental aneuploidy.

Trans-sialidases (TS) are unusual enzymes present on the surface of Trypanosoma cruzi, the causative agent of Chagas disease. Encoded by the largest gene family in the T. cruzi genome, only few members of the TS family have catalytic activity. Active trans-sialidases (aTS) are responsible for transferring sialic acid from host glycoconjugates to mucins, also present on the parasite surface. The existence of several copies of TS genes has impaired the use of reverse genetics to study this highly polymorphic gene family. Using CRISPR-Cas9, we generated aTS knockout cell lines displaying undetectable levels of TS activity, as shown by sialylation assays and labeling with antibodies that recognize sialic acid-containing mucins. In vitro infection assays showed that disruption of aTS genes does not affect the parasite’s capacity to invade cells or to escape from the parasitophorous vacuole but resulted in impaired differentiation of amastigotes into trypomastigotes and parasite egress from the cell. When inoculated into mice, aTS mutants were unable to establish infection even in the highly susceptible gamma interferon (IFN-γ) knockout mice. Mice immunized with aTS mutants were fully protected against a challenge infection with the virulent T. cruzi Y strain. Altogether, our results confirmed the role of aTS as a T. cruzi virulence factor and indicated that aTS play a major role during the late stages of intracellular development and parasite egress. Notably, mutants lacking TS activity are completely avirulent in animal models of infection and may be used as a live attenuated vaccine against Chagas disease.

Leishmania braziliensis is the main causative agent of Tegumentary Leishmaniasis in the Americas. However, difficulties related to genome manipulation, experimental infection, and parasite growth have so far limited studies with this species. CRISPR-Cas9-based technology has made genome editing more accessible, and here we have successfully employed the LeishGEdit approach to attenuate L. braziliensis. We generated a transgenic cell line expressing Cas9 and T7 RNA polymerase, which was employed for the targeted deletion of centrin, a calcium-binding cytoskeletal protein involved in the centrosome duplication in eukaryotes. Centrin-deficient Leishmania exhibit growth arrest at the amastigote stage. Whole-genome sequencing of centrin-deficient L. braziliensis (LbCen−/−) did not indicate the presence of off-target mutations. In vitro, the growth rates of LbCen−/− and wild-type promastigotes were similar, but axenic and intracellular LbCen−/− amastigotes showed a multinucleated phenotype with impaired survival following macrophage infection. Upon inoculation into BALB/c mice, LbCen−/− were detected at an early time point but failed to induce lesion formation, contrary to control animals, infected with wild-type L. braziliensis. A significantly lower parasite burden was also observed in mice inoculated with LbCen−/−, differently from control mice. Given that centrin-deficient Leishmania sp. have become candidates for vaccine development, we propose that LbCen−/− can be further explored for the purposes of immunoprophylaxis against American Tegumentary Leishmaniasis.

The practice of feeding dogs raw meat-based diets (RMBDs) is growing in several countries, and the risks associated with the ingestion of pathogenic and antimicrobial-resistant

Fuel moisture content is one of the fundamental parameters in forest fire research and management given its implications for many aspects of fire danger systems. In Portugal, such as in many other countries, to classify the days with more favourable conditions for wildfires it is common to use the Canadian Forest Fire Weather Index System (CFFWIS) which is based on the estimation of moisture content of several fuel components. The mathematical structure of CFFWIS requires as input the daily meteorological parameters which are used to estimate the moisture content of the soil for different layers that are the primary outputs of the system – the fire moisture codes. The final output parameter of the system is the Fire Weather Index (FWI) which represents a measure of the fire danger due to meteorological conditions. The temporal scale of the study is from 2018 and 2021 and the study area is in Lousã, a central region of Portugal. In this work, in addition to the meteorological data, we will use as input the direct measurements of dead fuels in the CFFWIS and analyse its influence on the FWI. The fuel moisture content (mf) is determined through the sample collection of dead pine needles in Lousã. For this temporal scale, mf by sampling is significantly lower than the modelled mf using meteorological parameters. An advantage of using the mf measurements is to increase the range of FWI variation, giving a higher sensitivity to the index to more easily discriminate the days with high fire danger and large burned areas. Two methods are addressed: “FWI a” which represents the traditionally FWI determined only by meteorological parameters, and the “FWI b” which is determined with fuel moisture content measurements and with meteorological data for the days that we did not have measurements. The original FWI, based only meteorological parameters, is compared with the FWI determined using mf measurements. The different methods will be related with the number of fires and burned area to analyse their performance. The results show a good fit between mf and FWI for days with extreme weather conditions (mf

The merging of two linear forest fire fronts that intersect at a small angle creates an accelerating fire in a relatively short time. In the majority of cases during the interaction, the fire fronts are non-symmetric. In Pedrogão Grande on the 17th June 2017, two-fire front merged and the propagation of the fire was influenced by the interaction of these two non-symmetric fire fronts. This forest fire motivates us to study the Junction Fire with two non-symmetrical fire fronts. The analysis of the interaction of point A and the angle between the bisector of the fire lines and the maximum ROS () is of particular relevance. It is found that with these preliminary laboratory tests, the non-ROS of point A for small rotation angles () depends on the slope angle () and the initial angle between fire fronts. For the highest slope angles, the non-ROS is the highest influence by the convection process, and the angle  where the maximum ROS occurred, increases when  increases. However, for the lowest slope angles, the radiation process is dominant and influences the non-ROS. For these cases, the angle ï§ï€ is near to the bisector of the fire lines.

O presente artigo foi realizado com o objetivo de aferir como as ferramentas de Business Intelligence apoiam as tomadas de decisão dos gestores na logística e nos negócios eletrónicos. Para esse efeito, inicialmente foi abordada a definição de Business Intelligence e os seus sistemas, bem como a compreensão da sua implementação numa organização e as ferramentas utilizadas durante esse processo, com o objetivo de conseguir perceber quais as suas vantagens e os seus benefícios a longo prazo para as empresas. Além disso, foi abordado o tema do Business Intelligence na área da logística, de forma a verificar onde se insere e quais as suas vantagens nessa área. Por fim, é abordado o tema de avaliação de maturidade de Business Intelligence com o intuito de verificar como o BI está a ser implementado e aceite numa empresa.

The influence of Pinus Pinaster needles packing ratio on fire spread at laboratory scale is investigated experimentally and compared to other wildland fuels. The packing ratio was varied from up to 10% by changing the fuel bed load and thickness. An optimum ratio of 5.5% is obtained corresponding to a maximum rate of spread. This optimum ratio is close to that found by Rothermel for crib fuels. However, the rate of spread increases again for higher porosity, which suggests the existence of a second optimum ratio. Two optimum ratios were also observed by Rothermel for excelsior fuels and recently by He et al for experiments using laser-cut cardboard. Recently, two optimum ratios corresponding to a minimum ignition time were observed using Pinus Halepensis needles. The minimum ignition time for spreading fuels correspond to a maximum rate of spread. These ratios have been attributed to a maximum heat flux absorption by the fuel. The heat flux absorption effect on the rate of spread is discussed by using the fuel optical length. The obtained optimum ratio corresponds to a small optical length (12 mm), which means that the heat flux is absorbed by the surface of the fuel bed. The second optimum ratio may correspond to an optical length much larger than the fuel bed thickness (around 50 mm). This corresponds to a bulk absorption of the heat flux. Further analyses of the mass-loss rate are provided to discuss thermochemical effects.

Forest fires are one of the main disasters that devastate many countries every year. The destruction caused by these phenomena generates social impacts putting at risk the population lives, environmental impacts due to the extreme deforestation and the high number of pollutants that are released to the atmosphere but also extremely important economic impacts caused by the destruction of a wide range of infrastructures and essential goods. Therefore, as it is impossible to remove all the infrastructures from the forest and from the wildland-urban interface, the development and installation of protection systems is essential. The main objective of this work is the development of a low cost protection system, with rigid panels, requiring a simple and easy installation, to protect outdoor infrastructures such as telecommunications stations, shelters, roadside enclosures, power cabinets, and other structures. A study was carried out on panels that could be used for the protection in order to determine whether the protective material would be more appropriate. Taking into account the fire resistance behaviour, thermal and structural properties and cost, the panels selected were the magnesium oxide fibreglass reinforced. The protection was constructed, installed on a telecommunication cabinet and experimentally laboratory tested in a wind combustion tunnel. To collect the data InfraRed and video cameras, heat flux sensors and sheathed thermocouples were used to determine the fire propagation, heat flux and temperatures, respectively. The data obtained in the experimental tests show clearly that the simple low cost protection is effective for the protection of telecommunication cabinets and other similar infrastructures against forest fires.