Complementation of the embryo-lethal T-DNA insertion mutant of AUXIN-BINDING-PROTEIN 1 (ABP1) with abp1 point mutated versions reveals crosstalk of ABP1 and phytochromes

The function of the extracytoplasmic AUXIN-BINDING-PROTEIN1 (ABP1) is largely enigmatic. We complemented a homozygous T-DNA insertion null mutant of ABP1 in Arabidopsis thaliana Wassilewskia with three mutated and one wild-type (wt) ABP1 cDNA, all tagged C-terminally with a strepII-FLAG tag upstream the KDEL signal. Based on in silico modelling, the abp1 mutants were predicted to have altered geometries of the auxin binding pocket and calculated auxin binding energies lower than the wt. Phenotypes linked to auxin transport were compromised in these three complemented abp1 mutants. Red light effects, such as elongation of hypocotyls in constant red (R) and far-red (FR) light, in white light supplemented by FR light simulating shade, and inhibition of gravitropism by R or FR, were all compromised in the complemented lines. Using auxin-or light-induced expression of marker genes, we showed that auxininduced expression was delayed already after 10 min, and light-induced expression within 60 min, even though TIR1/AFB or phyB are thought to act as receptors relevant for gene expression regulation. The expression of marker genes in seedlings responding to both auxin and shade showed that for both stimuli regulation of marker gene expression was altered after 10-20 min in the wild type and phyB mutant. The rapidity of expression responses provides a framework for the mechanics of functional interaction of ABP1 and phyB to trigger interwoven signalling pathways.

Aprender a ser. Aprender a conviver e aprender a desenvolver a consciência cidadã: um estudo de caso no contexto de 1º CEB

Relatório de Estágio apresentado à Escola Superior de Educação de Paula Frassinetti para obtenção de grau de Mestre em Educação Pré-Escolar e Ensino do 1º Ciclo do Ensino Básico

Do Pré-escolar ao 1.º Ciclo: O percurso da escrita

Relatório de Estágio apresentado à Escola Superior de Educação de Paula Frassinetti para obtenção de grau de Mestre em Educação Pré-Escolar e Ensino do 1.º Ciclo do Ensino Básico

Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos

studies using UV as a source of DNA damage. However, even though unrepaired UV-induced DNA damages are related to mutagenesis, cell death and tumorigenesis, they do not explain phenotypes such as neurodegeneration and internal tumors observed in patients with syndromes like Xeroderma Pigmentosum (XP) and Cockayne Syndrome (CS) that are associated with NER deficiency. Recent evidences point to a role of NER in the repair of 8-oxodG, a typical substrate of Base Excision Repair (BER). Since deficiencies in BER result in genomic instability, neurodegenerative diseases and cancer, it was investigated in this research the impact of XPC deficiency on BER functions in human cells. It was analyzed both the expression and the cellular localization of APE1, OGG1 e PARP-1, the mainly BER enzymes, in different NER-deficient human fibroblasts. The endogenous levels of these enzymes are reduced in XPC deficient cells. Surprisingly, XP-C fibroblasts were more resistant to oxidative agents than the other NER deficient fibroblasts, despite presenting the highest of 8-oxodG. Furthermore, subtle changes in the nuclear and mitochondrial localization of APE1 were detected in XP-C fibroblasts. To confirm the impact of XPC deficiency in the regulation of APE1 and OGG1 expression and activity, we constructed a XPC-complemented cell line. Although the XPC complementation was only partial, we found that XPC-complemented cells presented increased levels of OGG1 than XPC-deficient cells. The extracts from XPC-complemented cells also presented an elevated OGG1 enzimatic activity. However, it was not observed changes in APE1 expression and activity in the XPCcomplemented cells. In addition, we found that full-length APE1 (37 kDa) and OGG1- α are in the mitochondria of XPC-deficient fibroblasts and XPC-complemented fibroblasts before and after induction of oxidative stress. On the other hand, the expression of APE1 and PARP-1 are not altered in brain and liver of XPC knockout mice. However, XPC deficiency changed the APE1 localization in hypoccampus and hypothalamus. We also observed a physical interaction between XPC and APE1 proteins in human cells. In conclusion, the data suggest that XPC protein has a role in the regulation of OGG1 expression and activity in human cells and is involved mainly in the regulation of APE1 localization in mice. Aditionally, the response of NER deficient cells under oxidative stress may not be only associated to the NER deficiency per se, but it may include the new functions of NER enzymes in regulation of expression and cell localization of BER proteins

Projeto Fénix : inovação na organização pedagógica no 1º ciclo : estudo de caso

A dissertação que aqui se apresenta resulta de um trabalho de investigação junto de um Agrupamento de Escolas que adotou, a partir do ano letivo 2011/2012, como medida de promoção de sucesso educativo, o Projeto Fénix. Teoricamente, este estudo pretendeu analisar a escola enquanto organização social em que a comunidade escolar e social são partes interessadas na inovação e na mudança; esclarecer o conceito de projeto nos quadros dos novos regimes de autonomia e de gestão das escolas; compreender a importância e o papel dos projetos educacionais como fatores promotores de mudança e inovação pedagógica nas organizações escolares. Assim, adotando uma metodologia de trabalho de caráter qualitativo, descritivo e interpretativo procurou-se, através de um estudo de caso, analisar as medidas organizacionais que a escola assumiu para dar um apoio mais personalizado aos alunos que evidenciavam dificuldades de aprendizagem, nomeadamente, na disciplina de português; de que forma se procedeu à sua implementação na prática diária e o impacto do projeto nos professores e alunos; e, ao mesmo tempo analisar a relevância e o sentido da avaliação no decorrer e no término de um projeto, como importante processo de apoio à transformação, à melhoria, à transparência, no desenvolvimento de projetos educativos. A concretização destes objetivos só foi possível graças à colaboração da direção do agrupamento que autorizou a consulta de vários documentos essenciais e de todos os outros intervenientes no processo (alunos, professores, diretora, coordenadores e encarregados de educação), que prontamente se disponibilizaram para responderem às entrevistas. Sucintamente pode dizer-se que o estudo permitiu perceber que a implementação do Projeto Fénix, no geral, foi bem aceite por toda a comunidade educativa, mesmo tratando-se de um desafio ambicioso que exigiu determinação, rigor e trabalho de equipa, no qual alunos, professores e pais se comprometeram a melhorar o sucesso educativo. Apesar dos entraves que surgiram em consequência das políticas economicistas e de contenção, do quadro político atual, e da falta de recursos humanos (bolsa de professores de apoio), a nova organização pedagógica fundamentada pelo Projeto Fénix conseguiu melhorar o clima de aprendizagem, recuperar os alunos com mais dificuldades de aprendizagem e aprimorar as excelências.

Le diagnostic anténatal modifie-t-il la prise en charge néonatale et le devenir à 1 an des enfants suivis pour atrésie de l’œsophage de type III ?

International audience

Rôle de la conformation des glycoprotéines de l’enveloppe du VIH-1 dans la réponse cytotoxique cellulaire dépendante des anticorps et impact des protéines virales Nef et Vpu

Alors que d’énormes efforts sont mis de l’avant pour mettre en place des stratégies thérapeutiques contre l’infection au VIH-1, il est nécessaire de mieux cerner les déterminants viraux qui aideront à l’efficacité de celles-ci. En ce sens, une volumineuse littérature scientifique suggère que les anticorps contre le VIH-1 possédant une capacité à induire une réponse effectrice dépendante de leur portion Fc puissent jouer un rôle important dans la prévention de l’infection et dans la progression de la maladie. Cependant, peu d’information est disponible concernant les déterminants reconnus par ces anticorps et comment le virus s’en protège. Le but des travaux présentés dans cette thèse est donc d’élucider les mécanismes viraux contrôlant la reconnaissance des cellules infectées par ces anticorps capables d’induire une réponse effectrice. De par les corrélats de protection identifiés au cours de l’essai vaccinal RV144, les travaux présentés ici se concentrent sur la réponse cytotoxique dépendante des anticorps (ADCC), puisqu’il s’agit d’une réponse effectrice suggérée pour avoir joué un rôle dans la protection observée dans le RV144, seul essai vaccinal anti-VIH à avoir démontré un certain degré de protection. De plus, plusieurs anticorps capables d’induire cette réponse contre le VIH sont connus pour reconnaître les glycoprotéines de surface du virus (Env) dans une conformation dite ouverte, c’est-à-dire la conformation adoptée lors de la liaison d’Env avec son récepteur CD4 (épitopes CD4i). Nous avons mis au point deux techniques in vitro permettant d’étudier ces changements de conformation ainsi que leur impact sur la réponse ADCC. Les techniques mises au point, un ÉLISA sur base cellulaire pour mesurer les changements de conformation d’Env ainsi que la mesure de la réponse ADCC par cytométrie en flux, nous ont permis de démontrer comment le virus empêche l’exposition des épitopes d’Env CD4i. L’activité simultanée des protéines accessoires virales Nef et Vpu sur le retrait du récepteur CD4 de la surface des cellules infectées et l’inhibition du facteur de restriction Tétherine / BST-2 par Vpu contrôlent à la fois les niveaux d’Env et de CD4 à la surface cellulaire et donc modulent l’interaction Env-CD4 et ultimement la susceptibilité à la réponse ADCC contre les épitopes CD4i reconnus par des anticorps hautement prévalents lors de l’infection au VIH. Également, nous démontrons comment de petits composés mimant la liaison de CD4 sur Env sont capables de forcer l’exposition des épitopes CD4i, même en présence des protéines Nef et Vpu, et donc d’augmenter la susceptibilité des cellules infectées à la réponse ADCC. Une autre découverte présentée ici est la démonstration que la portion soluble d’Env produite par les cellules infectées peut interagir avec le récepteur CD4 des cellules non-infectées avoisinantes et induire leur reconnaissance et élimination par la réponse ADCC contre Env. Somme toute, la modulation de la réponse ADCC par l’interaction Env–CD4 représente un important pilier de la relation hôte – pathogène du VIH-1 de la perspective des réponses Fc-dépendantes. Les travaux présentés dans cette thèse ont le potentiel d’être utilisés dans l’élaboration de nouvelles stratégies antivirales tout en élargissant les connaissances fondamentales de cette interaction hôte – pathogène.

Concepções dos alunos sobre a avaliação das aprendizagens na disciplina de matemática em anos terminais dos 1º e 2º ciclos do ensino básico

A presente investigação tem como objectivo o estudo das concepções dos alunos sobre a avaliação das aprendizagens na disciplina de Matemática em anos terminais dos 1º e 2° ciclos do Ensino Básico. Em particular, procurou-se estudar e comparar as concepções que alunos desses anos tinham sobre a avaliação e as práticas avaliativas dos seus professores, e compreender se existiam algumas relações de dependência entre essas concepções e a perspectiva face à Matemática e o desempenho escolar desses alunos. O enquadramento teórico está organizado em dois capítulos. O primeiro relacionado com a avaliação e o segundo referente às concepções. Este estudo segue uma metodologia de natureza interpretativa. A recolha de dados foi feita através da aplicação de um questionário a quatro turmas, uma do 4o e outra do 6° ano de escolaridade de dois agrupamentos distintos, um de Elvas e outro de Portalegre, e de entrevista semi-estruturada a dois alunos por turma. A análise de dados foi organizada em tomo de duas categorias: (i) perspectivas face à Matemática e (ii) perspectiva face à avaliação das aprendizagens. Os resultados do estudo indicam que as concepções sobre a avaliação das aprendizagens em Matemática dos alunos participantes incidem, preferencialmente, sobre sentimentos, consequências, funções e instrumentos de avaliação. Verifica-se uma tendência para a existência de relações de dependência entre a imagem negativa da Matemática escolar e a concepção de avaliação associada aos sentimentos. Alunos com classificações negativas a Matemática associa, igualmente, a avaliação a sentimentos. Já os alunos que têm uma imagem positiva da Matemática, assim com os que têm classificações mais elevadas, tendem a associar a avaliação às suas consequências. No que diz respeito às práticas avaliativas que lhes têm sido proporcionadas, os alunos do 1º e do 2°ciclos apresentam concepções quase semelhantes, reconhecendo as fichas de avaliação como os instrumentos com mais peso para o professor na atribuição de notas no final do período. Os alunos do 1º ciclo são os que mais revelam concordar que o professor está atento às suas dificuldades. Porém, quer os alunos do 1 o e do 2°ciclos não reconhecem poder combinar com o professor a forma como são avaliados. ABSTRACT; The main purpose of the present work is to study the students' beliefs on assessment learning related to the Mathematics subject in ending years of 1st and 2nd key stage of elementary school. ln particular, the research was aimed to study and compare the students’ beliefs that pupils of those particular years had on assessment and the assessment practices of their teachers and also if there were any kind of dependence relationships between those beliefs and the perspective towards Mathematics and those students' school performance. The theoretical framework is organized in two chapters. The first related with the assessment and the second regarding the beliefs. This study follows a methodology of interpretative nature. The data gathering was done through the application of a questionnaire to four classes, one of the 4th and another of the 6th year of two different school groups, one belonging to Elva’s and another one to Portalegre and also through a semi-structured interview done to two students per group. The data analysis was organized around two categories: (i) perspectives towards Mathematics and (ii) perspective towards assessment learning. The results of the study show that the beliefs of those students on assessment learning on Mathematics are preferably based on feelings, consequences, functions and assessment instruments. ln general terms, there seems to be dependence relationships between the Mathematics negative image and the assessment conception associated to feelings. Students with negative marks at Mathematics also associate assessment to feelings. Those who have a positive Mathematics image, as well as those with higher marks at the subject, seem to associate assessment to its consequences. Concerning the assessment practices that have been provided to students from 1 51 and 2nd key stage of elementary school, these same pupils show very similar beliefs, pointing the summative tests as having higher importance when the assessment term comes. The students of the 1st key stage of elementary school are those who most agree that the teacher is attentive to their difficulties. Even so, both groups of students say that they cannot negotiate with their teacher the way they are supposed to be assessed.

Don’t expect me to vote for you just because I like you, even if you do make me feel warm inside: a comparison of the validity of non-ipsative measures of party support

The inclusion of non-ipsative measures of party preference (in essence ratings for each of the parties of a political system) has become established practice in mass surveys conducted for election studies. They exist in different forms, known as thermometer ratings or feeling scores, likes and dislikes scores, or support propensities. Usually only one of these is included in a single survey, which makes it difficult to assess the relative merits of each. The questionnaire of the Irish National Election Study 2002 (INES2002) contained three different batteries of non-ipsative party preferences. This paper investigates some of the properties of these different indicators. We focus in particular on two phenomena. First, the relationship between non-ipsative preferences and the choices actually made on the ballot. In Ireland this relationship is more revealing than in most other countries owing to the electoral system (STV) which allows voters to cast multiple ordered votes for candidates from different parties. Second, we investigate the latent structure of each of the batteries of party preferences and the relationships between them. We conclude that the three instruments are not interchangeable, that they measure different orientations, and that one –the propensity to vote for a party– is by far preferable if the purpose of the study is the explanation of voters’ actual choice behaviour. This finding has important ramifications for the design of election study questionnaires.

Validação de uma escala de atitudes de docentes do 1.º ciclo do ensino básico, relativamente à utilização da tecnologia em ambiente educativo

Dissertação de mest. em Observação e Análise da Relação Educativa, Faculdade de Ciências Humanas e Sociais, Univ. do Algarve, Escola Superior de Educação, Instituto Politécnico de Beja, 2004

Sub-epidemics explain localized high prevalence of reduced susceptibility to Rilpivirine in treatment-naive HIV-1-infected patients: subtype and geographic compartmentalization of baseline resistance mutations

This Open Access article is distributed under the terms of the Creative Commons Attribution Noncommercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.

Synthesis and antibacterial study of some s-substituted aliphatic analogues of 2-mercapto-5-(1-(4-toluenesulfonyl) piperidin-4-yl)-1,3,4-oxadiazole

Purpose: To synthesize a series of analogues of 1,3,4-oxadiazole and to evaluate their antibacterial activity. Methods: Ethyl piperidin-4-carboxylate (1) was mixed with 4-toluenesulfonyl chloride (2) in benignant conditions to yield ethyl 1-(4-toluenesulfonyl)piperidin-4-carboxylate (3) and then 1-(4- toluenesulfonyl)piperidin-4-carbohydrazide (4). Intermolecular cyclization of 4 into 2-mercapto-5-(1-(4- toluenesulfonyl) piperidin-4-yl)-1,3,4-oxadiazole (5) was obtained on reflux with CS2 in the presence of KOH. Molecule 5 was stirred with alkyl halides, 6a-i, in DMF in the presence of LiH to synthesize the final compounds, 7a-i. The structures of these molecules were elucidated by Fourier transform infra-red (FTIR) spectroscopy, proton nuclear magnetic resonance (1H-NMR) and electron impact mass spectrometry (EI-MS). Antibacterial activity was evaluated against five bacterial strains, namely, Salmonella typhi, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis, with ciprofloxacin used as standard antibacterial agent. Results: Out of nine synthesized derivatives, compound 7a was the most active against three bacterial strains, S. typhi, E. coli and P. aeruginosa, with minimum inhibitory concentration (MIC) of 9.11 ± 0.40, 9.89 ± 0.45 and 9.14 ± 0.72 μM, respectively, compared with 7.45 ± 0.58, 7.16 ± 0.58 and 7.14 ± 0.18 μM, respectively, for the reference standard (ciprofloxacin). Similarly, compounds 7a - 7c showed relatively good antibacterial activity against B. subtilis strain while compound 7e - 7g revealed good results against S. typhi bacterial strain. Conclusion: The results indicate that S-substituted derivatives of the parent compound are more effective antibacterial agents than the parent compound, even with minor differences in substituents

Intelligent outcome measures in liaison psychiatry:essential even if not desirable. Commentary on... a multidimensional Framework for Routine Outcome Measurement in Liaison Psychiatry (FROM-LP)

Service development is guided by outcome measures that inform service commissioners and providers. Those in liaison psychiatry should be encouraged to develop a positive approach that integrates the collection of outcome measures into everyday clinical practice. The Framework for Routine Outcome Measurement in Liaison Psychiatry (FROM-LP) is a very useful tool to measure service quality and clinical effectiveness, using a combination of clinician-rated and patient-rated outcome measures and patient-rated experience measures. However, it does not include measures of cost-effectiveness or training activities. The FROM-LP is a significant step towards developing nationally unified outcome measures.

Changes In Chromatin Structure In Nih 3t3 Cells Induced By Valproic Acid And Trichostatin A.

Valproic acid (VPA) and trichostatin A (TSA) are known histone deacetylase inhibitors (HDACIs) with epigenetic activity that affect chromatin supra-organization, nuclear architecture, and cellular proliferation, particularly in tumor cells. In this study, chromatin remodeling with effects extending to heterochromatic areas was investigated by image analysis in non-transformed NIH 3T3 cells treated for different periods with different doses of VPA and TSA under conditions that indicated no loss of cell viability. Image analysis revealed chromatin decondensation that affected not only euchromatin but also heterochromatin, concomitant with a decreased activity of histone deacetylases and a general increase in histone H3 acetylation. Heterochromatin protein 1-α (HP1-α), identified immunocytochemically, was depleted from the pericentromeric heterochromatin following exposure to both HDACIs. Drastic changes affecting cell proliferation and micronucleation but not alteration in CCND2 expression and in ratios of Bcl-2/Bax expression and cell death occurred following a 48-h exposure of the NIH 3T3 cells particularly in response to higher doses of VPA. Our results demonstrated that even low doses of VPA (0.05 mM) and TSA (10 ng/ml) treatments for 1 h can affect chromatin structure, including that of the heterochromatin areas, in non-transformed cells. HP1-α depletion, probably related to histone demethylation at H3K9me3, in addition to the effect of VPA and TSA on histone H3 acetylation, is induced on NIH 3T3 cells. Despite these facts, alterations in cell proliferation and micronucleation, possibly depending on mitotic spindle defects, require a longer exposure to higher doses of VPA and TSA.

Hypertrophic Adenoid Is A Major Infection Site Of Human Bocavirus 1.

Human bocavirus 1 (HBoV1) is associated with respiratory infections worldwide, mainly in children. Similar to other parvoviruses, it is believed that HBoV1 can persist for long periods of time in humans, probably through maintaining concatemers of the virus single-stranded DNA genome in the nuclei of infected cells. Recently, HBoV-1 was detected in high rates in adenoid and palatine tonsils samples from patients with chronic adenotonsillar diseases, but nothing is known about the virus replication levels in those tissues. A 3-year prospective hospital-based study was conducted to detect and quantify HBoV1 DNA and mRNAs in samples of the adenoids (AD), palatine tonsils (PT), nasopharyngeal secretions (NPS), and peripheral blood (PB) from patients undergoing tonsillectomy for tonsillar hypertrophy or recurrent tonsillitis. HBoV1 was detected in 25.3% of the AD samples, while the rates of detection in the PT, NPS, and PB samples were 7.2%, 10.5%, and 1.7%, respectively. The viral loads were higher in AD samples, and 27.3% of the patients with HBoV had mRNA detectable in this tissue. High viral loads and detectable mRNA in the AD were associated with HBoV1 detection in the other sample sites. The adenoids are an important site of HBoV1 replication and persistence in children with tonsillar hypertrophy. The adenoids contain high HBoV1 loads and are frequently positive for HBoV mRNA, and this is associated with the detection of HBoV1 in secretions.