Characterization and expression analysis of the cytochrome bd oxidase operon from Desulfovibrio gigas

Abstract: Although classified as anaerobic, Desulfovibrio gigas contains a functional canonical membrane respiratory chain, including a cytochrome bd quinol oxidase as its terminal element. In the present study, we report the identification of the operon cydAB encoding the two subunits of cytochrome bd from this bacterium. Two hypothetical promoter regions and sequences resembling transcriptional regulators-binding sites have been identified. Amino acid sequence analysis revealed a high similarity to cytochrome bd from other organisms, presenting the conserved residues typical from these proteins. Reverse transcription polymerase chain reaction (RT-PCR) and Northern blot analysis confirmed the operon transcription. Gene expression was assessed by real-time RT-PCR in cells grown in different media and under exposure to oxygen and nitric oxide. mRNA levels were slightly enhanced in the presence of 150 mu M NO. However, in the presence of 10 mu M NO, a decrease was observed of the steady-state population of cydAB mRNA. No considerable effect was observed in the presence of fumarate/sulfate medium, 60 mu M O-2 or 10 mu M NO

A chemotaxis operon in the bacterium Desulfovibrio gigas is induced under several growth conditions

Abstract: The chemosensory system of bacteria controls their motility and behaviour in different environments. In the present study, we report the identification of the first chemotaxis operon in Desulfovibrio gigas. Amino acid sequence analysis revealed seven coding regions for polypeptides with a high similarity to chemotaxis proteins from other organisms. D. gigas chemotaxis operon has a similar genetic organisation to chemotaxis operons found in the sequenced genomes of Desulfovibrio desulfuricans and Desulfovibrio vulgaris. Control of gene expression was assessed by real-time reverse transcription-PCR in cells grown under different conditions. mRNA levels were enhanced in the presence of thiosulfate and sulfite and decreased upon exposure to NO. No effect was observed in the presence of O-2, NaNO2, pyruvate or fumarate. These results show that the expression of the chemotaxis operon is enhanced in the presence of thiosulfate and sulfite indicating that under these compounds a chemotactic response seems to be triggered in D. gigas

O LUGAR DOS POBRES E A VIOLÊNCIA NA CIDADE: UM ESTUDO PARA O MUNICÍPIO DE SÃO PAULO

This paper discusses the relationship between poverty and urban violence, taking São Paulo Municipality as example. It is argued that the local of residence, and not the income, is the most adequate variable to evaluate the association between poverty and violence in the city. The spatial cut allows to observe the inter-relations among social, economic and law dimensions of the violence dynamic, which could not be understood through the observation of the income dimension only. The local of residence serves to identify where the law enforcement agencies fail and also to identify the environment that, due to its socio-economic characteristics are favorable to violence diffusion. The paper focuses on the life conditions of poor communities, highlighting slums areas. It stress that is precisely in these localities where the law enforcement agencies action is more problematical. Besides that, it reasons that precarious housing conditions are fundamental to explain the context of low socio-economic mobility, since the precarious space constrain the access to the resources which could help to overcome the community's disadvantages with respect to other social groups within the city. Finally, it shows the results of an exercise checking the correlation between, from on side, bad housing conditions and other socio-economical variables and, from the other side, the levels of violence in the city districts.

Powdery mildew and grapevine: which genes, enzymes and metabolites in resistant and susceptible cultivars?

Tese de mestrado, Biologia dos Recursos Vegetais, Universidade de Lisboa, Faculdade de Ciências, 2020The Eurasian Vitis vinifera, one of the most cultivated horticultural crops, has its economic impact derived from the fruits which are utilized to produce mainly wine, table grapes, raisins. The process of domestication originated a superior species with desirable agronomic traits and highly adaptable to external environment but prevented a coevolution with natural biotic stresses. In result, V. vinifera is highly susceptible to several pathogens. Erysiphe necator is one of the most threatening pathogens, being the etiologic agent of powdery mildew (PM). As an obligate biotrophic fungus, E. necator differentiates specialized infection (appressorium) and feeding (haustorium) structures to invade grapevine cells and acquired resources to complete his life cycle. At these stages of infection, a complex pathogen-host interaction occurs, and the outcome is the establishment of an incompatible (resistance) or compatible (disease) interaction. In grapevine, constitutive defences will avoid adhesion, germination and penetration. Induced defences organized in a two-layered system will block the endophytic growth and infection propagation. In these, in an intricate network, hormones act as secondary messengers to trigger downstream structural and biochemical changes. Based on additional undergoing studies and previous reports transcriptome and proteome reprogramming related with signalling and hormonal metabolism occurs; our study was performed to give some clues about the phytohormone network upon E. necator infection. For that, through LC-MS/MS, was quantified the content of some hormones in non-infected and E. necator-infected leaves of the resistant hybrid Vitis rupestris × riparia cv. 101-14 Millardet et de Grasset and the susceptible V. vinifera cv. Aragonez. Quantitative real time-PCR (qPCR) was performed to analyse the expression profiling of genes putatively involved in the chosen phytohormones pathways. At constitutive levels, a higher content of salicylic acid (SA) and auxins (mainly indole acetic acid; IAA) and a specific selection of jasmonates (JAs) and abscisic acid (ABA) were present in resistant and susceptible species, respectively. Upon infection, an accumulation of these hormones in each species was noticed. Also, a putative phytohormonal network based on a blend between SA/IAA and JAs/ABA was present in resistant and susceptible species, respectively. Altogether, these results suggest a role of hormones, and particularly the balance among them, in modulating the type of interaction and a possible involvement in response to E. necator. Nevertheless, the gene expression analysis revealed a more complex regulation of hormonal pathways than in others reported plant-PM interactions. Research on grapes to understand the defences responses upon E. necator infection have been neglected. In parallel with a broader transcriptomic and metabolomic-based project, and as for leaves, it was quantified the hormonal content and analysed the gene expression of susceptible grapes of V. vinifera cv. Carignan. In these, even delayed, a putative involvement of SA and JA pathways seems to respond to E. necator infection. Together with the gene expression analysis, the study pointed a different network of phytohormones and activation of different gene isoforms among leaves and grapes in response to E. necator infection. Reports also indicate a reprogramming of metabolism upon E. necator infection. Regarding the secondary metabolism, the phenylpropanoid pathway produce a plethora of phenolic compounds and phytoalexins important in defence responses against E. necator. Through spectrophotometry, was performed a quantification of total phenolic content in leaves and grapes. Additionally, in grapes, the phenylalanine ammonia lyase (PAL) biochemical activity was determined. In both organs, no change was noticed in total phenolic content upon E. necator infection nor in PAL activity in grapes. This suggest a reprogramming of phenylpropanoid pathway in order to accumulate specific phenolic compounds. The new obtained data in leaves and grapes of grapevine upon E. necator infection, once fully dissected and validated, will be useful for breeding programmes in order to improve tolerance against this fungus

The marble façade of the Ducal Palace of Vila Viçosa in the context of the 1940 Centenary Commemorations: cleaning and consolidation

Espera realização de congresso "Consolidation 2021. Stone Consolidation in Cultural Heritage" (20-22 outubro 2021). [Apoio FCT (ref.UIDB/04189/2020)]N/

Fatores determinantes da opção entre o Regime Simplificado e o Regime Geral de Tributação na perspetiva dos Contabilistas Certificados: O caso do Distrito de Portalegre

Com este projeto pretende-se compreender, na perceção dos Contabilistas Certificados (CC), quais são os aspetos considerados pelos contribuintes e por si aquando da opção entre o regime geral/contabilidade e os regimes simplificados (RST), conhecer ainda se os fatores são diferentes em Imposto sobre o Rendimento das Pessoas Coletivas (IRC) e Imposto sobre o Rendimento das Pessoas Singulares (IRS), bem como se variam em função de quem faz a opção. Assim, foi lançado um questionário dirigido aos Contabilistas Certificados do distrito de Portalegre, obtendo-se 54 questionários válidos, abrangendo informação sobre 1.279 empresas. Depois de analisados os resultados conclui-se que, de acordo com a perceção dos CC, quem opta pelo regime de tributação são, maioritariamente, os contribuintes aconselhados pelos CC. Verifica-se, ainda, que, independentemente de quem faz a opção pelo regime de tributação (clientes/empregador ou CC), bem como independentemente de se tratar da opção em IRC ou em IRS, o objetivo final é sempre o de maximização da poupança fiscal; fator decisivo ao qual acresce, nos sujeitos passivos de IRS, a preocupação com os custos de cumprimento. Quanto ao RST concluímos que tem maior aderência em sede de IRS quando comparado com o IRC, bem como que apesar das alterações ao RST em IRC em 2014, os dados não apontam para que tal facto tenha aumentado a aderência ao mesmo.This project intends to understand the perception of the certified accountants on the aspects considered by the taxpayers and by themselves when choosing between tax regimes in the corporate and income taxation, we also intend to know if the factors are the same in corporate taxation and income individual taxation, as well as if there are differences between taxpayers and accountants decisions. Thus, a questionnaire was applied to the certified accountants from the district of Portalegre, obtaining 54 valid questionnaires, covering information on 1,279 companies. Then we can concluded that, according to the accountants’ perception, the option for tax regime are, mostly, do by the taxpayers advised by their accountants. It is also verified that, regardless of who makes the option (taxpayers or accountants), and regardless of whether it is the option in IRC or IRS, the goal is always the maximum tax saving; moreover, in IRS context, there is also a concerning with the compliance costs. Regarding the simplified regime, we conclude that it has a higher adherence in IRS then in IRC context, and despite the changes to simplified regime in IRC, in 2014, the data suggest that those changes do not increased the regime’s adherenc

Procriação medicamente assistida: presente e futuro

Neste trabalho são apresentadas algumas considerações sobre as técnicas de Procriação Medicamente Assistida (PMA). Numa idade em que a ideia de planeamento familiar é unicamente direcionada no sentido de não ter filhos, fomos desafiados a desenvolver um projeto que tinha como principal objetivo a construção de um jogo Playdecide que permitisse a divulgação dos progressos científicos nesta área e que, ao mesmo tempo, suscitasse a discussão das questões éticas, morais e sociais ligadas à PMA. Para a construção do jogo revelou-se fundamental a pesquisa de informação, a participação num fórum de discussão com cientistas e médicos integrantes do Conselho Nacional de Procriação Medicamente Assistida (CNPMA) e a presença no colóquio realizado na Fundação Calouste Gulbenkian, “PMA: Presente e Futuro – questões emergentes nos contextos científico, ético, social e legal”, em Lisboa, no dia 9 de Janeiro. A realização deste trabalho permitiu-nos concluir que Portugal se encontra num patamar técnico-científico semelhante aos dos países mais desenvolvidos da Europa

Human-canine dyads : identifying dysfunctional relationships, a portuguese case

Tese de Doutoramento em Ciências Veterinárias na Especialidade de ClínicaThe human-dog dyad is thought to be the oldest existing domestic partnership and is generally mutually beneficial for both members of the partnership. Dysfunction in the human-dog dyad, however, produces serious consequences for each member of the partnership and also for society at large. Research into these relationships has addressed only the consequences of dysfunction, making prevention difficult. This project set out to evaluate the possibility of pre-emptively identifying dysfunction in such dyads by using dog health histories easily available in clinical contexts. To that end, the researcher developed a simple, one-page questionnaire that was disseminated in the greater metropolitan areas of Lisbon, Portugal, and was made available online. By identifying a dog’s biting history, trauma, or involvement in a vehicular accident, the researcher was able to suggest the possibility of the dog’s involvement in a dysfunctional dyad. To classify the canine behaviour traits essential for establishing the general characteristics of dysfunctional dyads, the researcher developed the European Portuguese Canine Behaviour Assessment and Research Questionnaire (C-BARQ). The psychometric properties were evaluated, and the instrument showed excellent to respectable consistency. The result was a canine behavioural questionnaire that established 13 different personality traits. A more extensive questionnaire was then administered to the same population in Lisbon, Portugal, aimed at identifying husbandry and noting dog and human characteristics within dysfunctional dyads. The results suggest that dogs housed on verandas or on plots of land, dogs that were fed diets purchased at agricultural cooperatives, dogs with C-BARQ scores showing high owner-directed aggression (ODA), dog-directed agressoion/fear (DAF) and dog rivalry (DR) were more likely to be part of dysfunctional dyads. Similarly, owners with high neuroticism scores and low lie/social desirability scores on the Eysenck Personality Questionnaire (EPQ-R) were also more likely to be part of these partnerships. These characteristics were then used to develop two predicative models – the Predicted Dysfunction with Dog and Owner Characteristics (PDDOC) and the Predicted Dysfunction with Dog Characteristics (PDDC) – that successfully predicted dysfunction in 79.7% and 80.1% of cases respectively. These findings reveal the feasibility of pre-emptively identifying dysfunctional human-dog dyads. As a result, this pre-emptive identification can be used to take preventative action – specifically the development of educational programs, the improvement of human-dog pairings, and the equipping of veterinarians to better prevent and/or correct dysfunction.RESUMO - A díade homem-cão é considerada a mais antiga parceria doméstica, sendo tida como mutualmente benéfica para ambos os membros. Quando estas díades se tornam disfuncionais pode haver sérias consequências, não apenas para os membros da díade, mas para a sociedade no seu todo. A disfuncionalidade de díades tem sido abordada em diversos estudos, contudo somente após se terem sentido as suas consequências nefastas, o que dificulta o processo de implementação de medidas preventivas. Este projecto teve como objetivo a sua identificação precoce, usando para isso, o historial de saúde do animal disponibilizado em contexto clínico. Foi desenvolvido um questionário sucinto de uma página, o qual foi distribuído a proprietários em Centros de Atendimento Médico-Veterinário (CAMV) na Área Metropolitana de Lisboa e também em formato online. A identificação de ocorrência de mordedura, trauma ou atropelamento foi associado a díade disfuncional. Foi desenvolvido o European Portuguese Canine Behaviour Assessment and Research Questionnaire – C-BARQ (Questionário de Investigação e Avaliação de Comportamento Canino) com o intuito de estabelecer bases gerais que permitissem classificar alguns aspetos do comportamento canino. Avaliaram-se as propriedades psicométricas e o instrumento mostrou um intervalo de consistência do respeitável ao excelente. O resultado final foi um questionário de comportamento canino que estabeleceu 13 traços de personalidade diferentes. Administrou-se seguidamente um questionário mais extenso à mesma população, mas agora com a finalidade de identificar características tanto do homem como do cão nestas díades disfuncionais. Observou-se que cães alojados em varandas ou em terrenos, alimentados com rações compradas em cooperativas agrícolas ou que apresentaram valores elevados nos scores de ODA, DAF e DR no C-BARQ, têm uma maior tendência de fazer parte de uma díade disfuncional. Proprietários que no questionário de personalidade humana, EPQ-R apresentaram um valor elevado em neuroticismo e baixo em mentira/desejabilidade social também partilham esta tendência. Estas características foram então usadas no desenvolvimento de dois modelos preditivos (PDDOC e PDDC), cujos resultados previram disfunções em 79,7% e 80,1% dos casos, respetivamente. Estes resultados, possibilitarão o desenvolvimento de programas educacionais, escolha mais informada na adoção de animais em abrigos, bem como dar aos médicos veterinários ferramentas para identificar e eventualmente prevenir e/ou corrigir algumas destas disfunções.N/