RCAAP Repository

RNA-sequencing (RNA-seq) is rapidly emerging as the technology of choice for whole-transcriptome studies. However, RNA-seq is not a bias free technique. It requires large amounts of RNA and library preparation can introduce multiple artifacts, compounded by problems from later stages in the process. Nevertheless, RNA-seq is increasingly used in multiple studies, including the characterization of tissue-specific transcriptomes from invertebrate models of human disease. The generation of samples in this context is complex, involving the establishment of mutant strains and the delicate contamination prone process of dissecting the target tissue. Moreover, in order to achieve the required amount of RNA, multiple samples need to be pooled. Such datasets pose extra challenges due to the large variability that may occur between similar pools, mostly due to the presence of cells from surrounding tissues. Therefore, in addition to standard quality control of RNA-seq data, analytical procedures for control of "biological quality" are critical for successful comparison of gene expression profiles. In this study, the transcriptome of the central nervous system (CNS) of a Drosophila transgenic strain with neuronal-specific RNAi of an ubiquitous gene was profiled using RNA-seq. After observing the existence of an unusual variance in our dataset, we showed that the expression profile of a small panel of marker genes, including GAL4 under control of a tissue specific driver, can identify libraries with low levels of contamination from neighboring tissues, enabling the selection of a robust dataset for differential expression analysis. We further analyzed the potential of profiling a complex tissue to identify cell-type specific changes in response to target gene down-regulation. Finally, we showed that trimming 5' ends of reads decreases nucleotide frequency biases, increasing the coverage of protein coding genes with a potential positive impact in the incurrence of systematic technical errors.

Although, the precise molecular mechanisms underlying Parkinson's disease (PD) are still elusive, it is now known that spreading of alpha-synuclein (aSyn) pathology and neuroinflammation are important players in disease progression. Here, we developed a novel microfluidic cell-culture platform for studying the communication between two different cell populations, a process of critical importance not only in PD but also in many biological processes. The integration of micro-valves in the device enabled us to control fluid routing, cellular microenvironments, and to simulate paracrine signaling. As proof of concept, two sets of experiments were designed to show how this platform can be used to investigate specific molecular mechanisms associated with PD. In one experiment, naïve H4 neuroglioma cells were co-cultured with cells expressing aSyn tagged with GFP (aSyn-GFP), to study the release and spreading of the protein. In our experimental set up, we induced the release of the contents of aSyn-GFP producing cells to the medium and monitored the protein's diffusion. In another experiment, H4 cells were co-cultured with N9 microglial cells to assess the interplay between two cell lines in response to environmental stimuli. Here, we observed an increase in the levels of reactive oxygen species in H4 cells cultured in the presence of activated N9 cells, confirming the cross talk between different cell populations. In summary, the platform developed in this study affords novel opportunities for the study of the molecular mechanisms involved in PD and other neurodegenerative diseases.

Aim: To describe the prevalence of periapical lesions, root canal treatments and coronal restorations on teeth adjacent to either implant- or natural tooth-supported crowns using cone-beam computed tomography (CBCT) assessment compared to nonadjacent teeth. Methodology: A total of 1249 CBCT volumes were screened by five pre-calibrated observers in 11 health centres. A step-by-step screening protocol was implemented and reviewed every 3 months, and mandatory setting characteristics were established for all CBCT scans using dedicated visualization software. Intra- and inter-rater reliability tests were conducted. The prevalence of periapical lesions, root canal treatments and coronalrestorations wasrecorded on both teeth adjacent and nonadjacent to implant- (predictor 1) or natural tooth-supported crowns (predictor 2). A binary logistic model (Generalizing Estimating Equations test) was used to verify whether the prevalence of periapical lesion, root canal filling and the tooth restorative status are altered when the assessed tooth is adjacent or not to an implant-supported crown (predictor 1); or to a natural tooth-supported crown (predictor 2). Odds ratio and confidence intervals for the dependent variables at both predictors were obtained. The significance level was set at .05. Results: A global sample of 22 899 teeth was included. Compared to nonadjacent teeth, the prevalence of periapical lesion, root canals treatments and restorative procedures when adjacent to implant-supported crowns was 10.7%, 19.6% and 22.9% higher, respectively, and when adjacent to tooth-supported crowns was 19.3%, 35.6% and 37.4% higher respectively. These results were significant only for variables root canal filling and coronal restoration (p < .0001). Odds ratio to present root canal treatment is 2.57 times higher (CI 1.95–3.39, p = .0001) when the tooth is adjacent to implant-supported crown and 4.39 times (CI 3.49–5.53, p = .0001) when adjacent to tooth-supported crown, whilst for restorative procedure, the odds are, respectively, 1.63 (CI 1.29–2.06, p = .0001) and 2.30 (CI 1.92–2.76, p = .0001).

Heart failure (HF) is an increasingly prevalent syndrome and a leading cause of both first hospitalization and readmissions. Strikingly, up to 25% of the patients are readmitted within 30 to 60-days, accounting for HF as the primary cause for readmission in the adult population. Given its poor prognosis, one could describe it as a "malignant condition". Acute decompensation is intrinsically related to increased right heart tele-diastolic pressures and often related to congestive symptoms. In-hospital strategies to adequately compensate and timely discharge patients are limited. Conversely, the fragile early postdischarge phase is a vulnerable period when one could potentially intervene cost-effectively to improve survival and to reduce morbidity. Promising transitional hospital-to-home programs may have a broader role in the near future, namely for selected higher risk patients. However, identifying patients at risk for hospital readmission has been challenging. Novel approaches, such as ferric carboxymaltose and valsartan/sacubitril, and reemerging drugs, particularly digoxin, may reduce hospitalizations. Despite this, optimizing the use of "older" therapies is still warranted. Right heart pressures monitoring may provide novel insights into promptly outpatient management. Unfortunately, randomized trials in the specific ADHF population are scarce. A novel paradigmatic approach is needed in order to suitably improve the currently poor prognosis of ADHF. Both improving survival and reducing hospitalizations are, therefore, primordial therapy goals. Lastly, no single drug has consistently proved to improve survival in HF with preserved ejection fraction (HFpEF); yet, some approaches may efficiently reduce hospitalizations. Awareness on HFpEF management beyond the failing heart is imperative.

The unstrained S-allyl cysteine amino acid was site-specifically installed on apoptosis protein biomarkers and was further used as a chemical handle and ligation partner for 1,2,4,5-tetrazines by means of an inverse-electron-demand Diels-Alder reaction. We demonstrate the utility of this minimal handle for the efficient labeling of apoptotic cells using a fluorogenic tetrazine dye in a pre-targeting approach. The small size, easy chemical installation, and selective reactivity of the S-allyl handle towards tetrazines should be readily extendable to other proteins and biomolecules, which could facilitate their labeling within live cells.

The human pathogenic parasite Trypanosoma brucei possess both de novo and salvage routes for the biosynthesis of pyrimidine nucleotides. Consequently, they do not require salvageable pyrimidines for growth. Thymidine kinase (TK) catalyzes the formation of dTMP and dUMP and is one of several salvage enzymes that appear redundant to the de novo pathway. Surprisingly, we show through analysis of TK conditional null and RNAi cells that TK is essential for growth and for infectivity in a mouse model, and that a catalytically active enzyme is required for its function. Unlike humans, T. brucei and all other kinetoplastids lack dCMP deaminase (DCTD), which provides an alternative route to dUMP formation. Ectopic expression of human DCTD resulted in full rescue of the RNAi growth phenotype and allowed for selection of viable TK null cells. Metabolite profiling by LC-MS/MS revealed a buildup of deoxypyrimidine nucleosides in TK depleted cells. Knockout of cytidine deaminase (CDA), which converts deoxycytidine to deoxyuridine led to thymidine/deoxyuridine auxotrophy. These unexpected results suggested that T. brucei encodes an unidentified 5'-nucleotidase that converts deoxypyrimidine nucleotides to their corresponding nucleosides, leading to their dead-end buildup in TK depleted cells at the expense of dTTP pools. Bioinformatics analysis identified several potential candidate genes that could encode 5'-nucleotidase activity including an HD-domain protein that we show catalyzes dephosphorylation of deoxyribonucleotide 5'-monophosphates. We conclude that TK is essential for synthesis of thymine nucleotides regardless of whether the nucleoside precursors originate from the de novo pathway or through salvage. Reliance on TK in the absence of DCTD may be a shared vulnerability among trypanosomatids and may provide a unique opportunity to selectively target a diverse group of pathogenic single-celled eukaryotes with a single drug.

O presente estudo exploratório pretende caracterizar as crenças de carreira e as crenças de auto-eficácia dos estudantes universitários finalistas e a relação entre ambos os constructos, fundamentados pela Teoria da Aprendizagem de Krumboltz (1996) e pela Teoria do Desenvolvimento da Carreira (Lent, Hackett & Brown, 1994). À amostra, constituída por 170 jovens adultos de diferentes Faculdades da região de Lisboa, foi aplicado o Inventário de Crenças de Carreira (ICC; versão adaptada por Rafael, 2001) e a Escala de Auto-eficácia na Transição para o Trabalho (AETT; Vieira & Coimbra, 2005). Os resultados indicam que as variáveis representam diferentes dimensões psicológicas e apresentam, em geral, associações positivas baixas entre si. Sugere-se que estes estudantes percepcionam com alguma confiança esta etapa da carreira e que, de acordo com a literatura, algumas das suas crenças parecem corresponder a factores facilitadores para uma transição mais adaptativa; outras aparentam estar desajustadas face às exigências do mercado de trabalho. Este padrão foi semelhante para ambos os géneros. Os dados obtidos evidenciam a importância em explorar as crenças de carreira e em promover as crenças de auto-eficácia na transição do ensino superior para o trabalho.

This article is about the equivoques of anthropology’s colonial encounter as well as the story of intellectual artefacts. [1] It addresses an old debate on the genealogy of anthropological knowledge, at the core of which is a shared assumption: anthropologists and historians today perceive the history of anthropology as intertwined with colonial history. Few, if any, anthropologists or historians would disagree that the anthropology of non-Western peoples is genealogically embedded in imperial expansion or colonial contexts, and that we can hardly imagine colonial power without some sort of anthropological knowledge as bedside company. It is also true that this assumption has not been at all devoid of strong moral convictions about the evil nature of colonialism and its friendly anthropologies, a moral impetus particularly evident in earlier approaches to the subject in the 1960s and ’70s. Since that time, though, with the exception of Talal Asad’s seminal assessment in 1973, historians have given it little systematic attention as a subject in its own right. [2] As George Stocking noted in 1991, the ‘assumption that anthropology was linked to Western colonialism’ seems to have survived in the professional culture of anthropologists more in the form of a ‘commonplace of disciplinary discourse’ than as a ‘serious interest in the history of anthropology in colonial context’. [3] Although the Portuguese case has been entirely left out of this picture by English literature, Stocking’s remarks apply to the Portuguese anthropological discourse since the 1970s. Having its professional identity closely associated with political opposition to the authoritarian Estado Novo regime, Portuguese social anthropology, born into institutional autonomy out of the April Revolution in 1974, has been eager to accuse earlier generations of (physical) anthropologists of racial prejudice and complicity with the imperial state. Very few have undertaken a serious and critical historical approach to such a pervasive disciplinary assumption.

Grazing exclusion may be used to promote the recovery of disturbed ecosystems. A promising way for the evaluation of its effectiveness is through the monitoring of key biological groups, particularly those more responsive to disturbance and playing key roles in ecosystem functioning. Ants have been used as ecological indicators as they are abundant, diverse and sensitive to environmental changes. Here, we aimed to evaluate changes in ant taxonomic and functional structure and diversity, using functional groups, along a post-grazing succession in a Mediterranean oak woodland and to understand which environmental variables drive them. The post-grazing succession comprised a chronosequence of grazing excluded sites for 8, 12 and 18 years and a grazed control site. We found that ant species richness, functional structure and diversity increased with years since grazing exclusion: Generalist/Opportunist and the Hot Climate specialists increased in the 18 years grazing excluded site, while the Cryptic Species group increased in the 12 years grazing excluded site. Yet, their responses were not linear over time. Time since grazing exclusion and vegetation structure explained differences in ant taxonomic and functional structure and diversity. The Invasive/Exotic group dominated in all sites, except in the longest excluded site, where it occurred in the lowest proportion. The invasive Argentine ant dominated the grazed site, where it may have led to ant taxonomic and functional homogenization. Our results suggest that the time and changes in habitat structure may favour the recovery of ant biodiversity, although the presence of the invasive Argentine ant species may have slowed it down.

Durante o seu ciclo olímpico (2009-2016), num momento histórico em que governos e agentes privados de todo o mundo buscavam superar os efeitos de uma grave crise económica evidenciada justamente na relação entre capitais financeiros e a urbanização, a cidade do Rio de Janeiro recebeu avultados investimentos para políticas urbanas. O bairro Madureira, representado pelas suas tradições afro-brasileiras, foi um lugar privilegiado para esses investimentos e fundamental para a compreensão da eficácia de curto prazo desse projeto de cidade e dos seus conflitos e reviravoltas políticas decorrentes. O objetivo deste artigo é apresentar razões, ações e omissões para a implementação desse projeto de cidade, com as suas potencialidades, contradições e os seus efeitos imediatos.

Primary examples in vaccine design have shown good levels of carbohydrate-specific antibody generation when raised using extracted or fully synthetic capsular polysaccharide glycans covalently coupled to a protein carrier. Herein, we cover recent clinical developments of carbohydrate-based vaccines and describe how novel cutting-edge methodology for the total synthesis of oligosaccharides and for the precise placement of carbohydrates at pre-determined sites within a protein may be used to further improve the safety and efficacy of glycovaccines.

The present paper is limited to a discussion of the principles of an iodometric and a colorimetric method, which are both more reliable than the usual titration, and to showing that their results agree closely with those obtained by the Clark & Collip (1921) method, when applied to 2 ml. samples of serum.

As matérias-primas têm as marcas da sua origem, da sua localidade. Mas também é verdade que o centro de um processo educativo é não raramente desequilibrado pelas tensões hegemónicas e culturais. A reprodução do saber e da habilitação simbólica é uma das agências da hegemonia cultural. Perante os discursos hegemónicos, pós colonizadores, há propostas de resistência: a contextualização, o valor cultural local, a identidade. Entre as expressões dos vários países o tema é tratado em alguns dos 17 artigos que se apresentam neste 15º número da revista Matéria-Prima.

A Educação Artística joga-se em muito mais locais que no ensino formal. As oportunidades formativas têm sido aproveitadas por museus, bibliotecas, centros culturais, exposições, festivais, associações e plataformas culturais, eventos, plataformas de disseminação artística, edições. Os artistas individuais têm vindo a integrar as dinâmicas relacionais e de criação de públicos nas suas obras, ao convocarem as audiências e implicarem o espectador. O terreno é limitado apenas pela imaginação, e as oportunidades de convocação alargam-se aos novos conteúdos e plataformas digitais, a par com a valorização do que é local e identitário: a revolução pode fazer-se pela cidadania.

This retrospective study analyzed implant survival of immediate implant-supported fixed complete denture (IFCD) treatment options (TOs) based on the level of alveolar atrophy (CC). Records of 882 patients receiving a total of 6042 implants at one private referral clinic between 2004 and 2020 were considered. The mean follow-up period was 3.8 ± 2.7 years. Cumulative implant survival rates (CSRs) were analyzed as a function of CCs and TOs according to Mantel-Haenszel and Mantel-Cox. Hazard risk ratios for implant loss were compared using Cox regression. Confounding factors were identified using mixed Cox regression models. The 2- and 5-year CSRs were 98.2% and 97.9%, respectively. Maxillary 2- and 5-year CSRs were lower (97.7% and 97.3%) compared to mandibular CSRs (99.8% and 98.6%) (p = 0.030 and 0.0020, respectively). The CC did not influence CSRs of IFCDs in the mandible (p = 0.1483 and 0.3014, respectively) but only in the maxilla (p = 0.0147 and 0.0111), where CSRs decreased with increasing atrophy. TOs did not statistically differ in terms of survival rate for a given level of alveolar atrophy. The adaption of IFCD treatments to the level of atrophy and patient-specific risk factors can result in high CSRs, even at different levels of bone atrophy.

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Risco: educação artística: A educação artística enfrenta o risco de desaparecimento gradual do seu habitat da educação formal ao mesmo tempo que a sociedade civil cada vez mais a reclama, no contexto das instituições que dependem do sucesso dos públicos e dos visitantes. Assiste-se a uma contradição de termos que parece ser originária de uma liberalização cognitiva: este parece ser um paradigma para ficar. O neo-liberalismo vigente contribui de modo contraditório. Por um lado, diminui as cargas horárias e extingue disciplinas, expulsa os profissionais artísticos da sala de aula: cada vez há menos horas de contacto de educação artística, do ensino do desenho. Por outro lado, a dependência das lógicas de sponsorização obriga os gestores culturais a urdirem estratégias educativas dirigidas a massas cada vez maiores, a convocarem e a reivindicarem posicionamentos formativos, a procurarem implicar e criar mais audiências. O sistema precisa dos alunos que quer reduzir, numa contradição de termos evidente. O professor de artes, o mediador entre o mundo da arte e a premência educativa vê-se com uma injunção que dele exige uma operatividade quer como pedagogo, quer como artista, integrando várias dimensões numa só, numa incarnação exigente. A educação artística parece depender da criatividade dos seus agentes, da sua mobilização, do seu entusiasmo, da sua competência mobilizadora: os tempos estão adversos ao conformismo e assim se anotam muitas ações desassossegadas e, cremos, implicadas.

Os objetivos do estudo recaem sobre a investigação, em adolescentes em meio escolar, do contributo do perfecionismo e das suas dimensões autorientada e socialmente prescrita, bem como da sintomatologia depressiva e/ou ansiosa, para a sintomatologia de perturbação alimentar. Participaram no estudo 531 adolescentes, de idades compreendidas entre os 12 e os 18 anos de ambos os sexos. Os participantes responderam a um Questionário de Dados Pessoais e Sociodemográficos, à subescala de perfecionismo do Eating Disorder Inventory 2 (Garner, Olmstead & Polivy, 1983), ao Eating Attitudes Test 26 (Garner, Olmstead, Bohr & Garfinkel, 1982) e ao Cuestionario Educativo-Clínico: Ansiedad y Depresión (González, Cueto & Fernández, 2007). A subescala do EDI2 foi, neste estudo, repartida em perfecionismo autoorientado e perfecionismo socialmente prescrito. Os participantes do sexo masculino apresentaram menor número de casos de dieta e níveis mais baixos de sintomatologia depressiva-ansiosa, depressão, ansiedade e de sintomatologia de perturbação alimentar em comparação com as raparigas, não tendo sido observadas diferenças entre sexos no que se refere ao perfecionismo global, perfecionismo socialmente prescrito e perfecionismo auto-orientado. As correlações encontradas entre as variáveis em estudo foram mais elevadas e significativas no grupo das raparigas. Também entre estas foi possível explicar consideravelmente a sintomatologia de perturbação alimentar através da análise de regressões múltiplas hierárquicas, que revelou que a idade, a dieta, o perfecionismo auto-orientado e a depressão contribuíam para explicar 42% da variância da sintomatologia de perturbação alimentar. Tal não foi observado em rapazes pois relativamente a estes só foi possível explicar 10% da variância de sintomatologia de perturbação alimentar para a qual apenas contribuíam perfecionismo global e sintomatologia depressiva-ansiosa. O significado dos resultados é discutido em termos das implicações para o conhecimento acerca dos fatores de risco para o desenvolvimento de perturbações alimentares em ambos os sexos.

In recent years, journalism in Portugal has faced significant structural changes, particularly in terms of employment and working conditions. The aim of this chapter is to analyse the relationship between job loss and re-employment in Portuguese journalism. Job terminations may result from forced redundancies, through the non-renewal of temporary work contracts or the removal of permanent staff. Alternatively, they may be instigated by journalists, whether through resignations, including company buyout programs, or by way of voluntary non-renewals, in the case of precarious workers. The main argument presented in this chapter is that the form of job termination, along with age difference, plays a significant role in influencing the coping process in dealing with the aftermath of job loss and, ultimately, the job prospects on offer. The research follows a biographical approach, centred on professional trajectories, developed through semi-structured interviews with 28 ex-journalists.

Arte invisível: A relação entre arte e sociedade é mediada pelos artistas e pelos educadores. Entre uns e outros há um segredo que nenhum deles partilha. Os artistas conseguem ensinar as gerações que ainda não nasceram, os educadores conseguem dar sentido aos novos públicos que já nasceram, aqui e agora, e formar os novos artistas. Não devia ser assim tão difícil falar de arte, ou melhor, de educação artística. Poderia ser mais transparente a sua necessidade, devia ser óbvia a sua utilidade, devia ser evidente o seu benefício. Porque é tão elusiva? Porque foge do aprisionamento das didáticas? A chave encontra-se do lado dos professores, pela razão de serem eles a formar os públicos e a formarem os próprios artistas de entre estes públicos. A exigência torna-se clara, o investimento na formação exige novidade, introspeção, inovação, crítica, criaçã. A educação artística olha-se de modo renovado, espantado, interventivo, inovador: está tudo por fazer, suspeitam os seus agentes, ao mesmo tempo que os decisores dela desinvestem continuadamente, visando indicadores contáveis. As cargas horárias são reduzidas, confunde-se criatividade com empreendedorismo, cultura com capital. Tem-se assim um contexto de crise na Educação Artística, que é também o desafio. Para isso pede-se um professor que possa ser ao mesmo tempo artista e criador, uma formação mais profissionalizada nos domínios artísticos, uma maior intervenção dos artistas junto das escolas, um apoio às redes colaborativas e às intervenções de disseminação.