Patterning of the vertebrate ventral spinal cord

We review investigations that have lead to a model of how the ventral spinal cord of higher vertebrate embryos is patterned during development. Central to this model is the secreted morphogen protein, Sonic hedgehog. There is now considerable evidence that this molecule acts in a concentration-dependent manner to direct the development of the spinal cord. Recent studies have suggested that two classes of homeodomain proteins are induced by threshold concentrations of Sonic hedgehog. Reciprocal inhibition between the two classes acts to convert the continuous gradient of Sonic hedgehog into defined domains of transcription factor expression. However, a number of aspects of ventral spinal cord patterning remain to be elucidated. Some issues currently under investigation involve temporal aspects of Shh-signalling, the role of other signals in ventral patterning and the characterisation of ventral interneurons. In this review, we discuss the current state of knowledge of these issues and present some preliminary studies aimed at furthering understanding of these processes in spinal cord patterning.

Macropodid herpesvirus 1 encodes genes for both thymidylate synthase and ICP34.5

Macropodid herpesvirus 1 (MaHV-1) is an unclassified alphaherpesvirus linked with the fatal infections of kangaroos and other marsupials. During the characterisation of the internal repeat region of MaHV-1, an open reading frame (ORF) encoding for thymidylate synthase (TS) gene was identified and completely sequenced. Southern blot analysis confirmed the presence of two copies of the TS gene in the MaHV-1 genome as expected. Computer analysis of the TS ORF showed it was 948 nucleotides in length. A putative polyadenylation signal was identified 17-22 bp inside the ORF implying a minimal or absent 3' untranslated region. The predicted polypeptide was 316 amino acid residues in length and contained the highly conserved motifs for folate binding and F-dUMP binding, typical of all TS enzymes. Interestingly, MaHV-1 TS polypeptide had highest similarity to the human TS polypeptide (81%) compared to the TS polypeptides of other herpesviruses (72-75%). Immediately upstream of the TS gene, a second ORF of 510 bp, encoding a polypeptide with 170 amino acid residues, was identified. The carboxyl domain of this MaHV-1 polypeptide shared 68% similarity to a 59 amino acid motif of human herpesvirus 1 ICP34.5, identifying it as the MaHV-1 ICP34.5 homologue. This is the first report of a herpesvirus that encodes for both TS and ICP34.5.

Study of n-fold Weibull competing risk model

This paper deals with an n-fold Weibull competing risk model. A characterisation of the WPP plot is given along with estimation of model parameters when modelling a given data set. These are illustrated through two examples. A study of the different possible shapes for the density and failure rate functions is also presented. (C) 2003 Elsevier Ltd. All rights reserved.

A molecular mechanism for mRNA trafficking in neuronal dendrites

Specific neuronal mRNAs are localized in dendrites, often concentrated in dendritic spines and spine synapses, where they are translated. The molecular mechanism of localization is mostly unknown. Here we have explored the roles of A2 response element (A2RE), a cis-acting signal for oligodendrocyte RNA trafficking, and its cognate trans-acting factor, heterogeneous nuclear ribonucleoprotein ( hnRNP) A2, in neurons. Fluorescently labeled chimeric RNAs containing A2RE were microinjected into hippocampal neurons, and RNA transport followed using confocal laser scanning microscopy. These RNA molecules, but not RNA lacking the A2RE sequence, were transported in granules to the distal neurites. hnRNP A2 protein was implicated as the cognate trans-acting factor: it was colocalized with RNA in cytoplasmic granules, and RNA trafficking in neurites was compromised by A2RE mutations that abrogate hnRNP A2 binding. Coinjection of antibodies to hnRNP A2 halved the number of trafficking cells, and treatment of neurons with antisense oligonucleotides also disrupted A2RE - RNA transport. Colchicine inhibited trafficking, whereas cells treated with cytochalasin were unaffected, implicating involvement of microtubules rather than microfilaments. A2RE-like sequences are found in a subset of dendritically localized mRNAs, which, together with these results, suggests that a molecular mechanism based on this cis-acting sequence may contribute to dendritic RNA localization.

Surface plasmon resonance as a tool in the functional analisys of an immunodominant site in foot-and-mouth disease virus

A fast and direct surface plasmon resonance (SPR) method for the kinetic analysis of the interactions between peptide antigens and immobilised monoclonal antibodies (mAb) has been established. Protocols have been developed to overcome the problems posed by the small size of the analytes (< 1600 Da). The interactions were well described by a simple 1:1 bimolecular interaction and the rate constants were self-consistent and reproducible. The key features for the accuracy of the kinetic constants measured were high buffer flow rates, medium antibody surface densities and high peptide concentrations. The method was applied to an extensive analysis of over 40 peptide analogues towards two distinct anti-FMDV antibodies, providing data in total agreement with previous competition ELISA experiments. Eleven linear 15-residue synthetic peptides, reproducing all possible combinations of the four replacements found in foot-and-mouth disease virus (FMDV) field isolate C-S30, were evaluated. The direct kinetic SPR analysis of the interactions between these peptides and three anti-site A mAbs suggested additivity in all combinations of the four relevant mutations, which was confirmed by parallel ELISA analysis. The four-point mutant peptide (A15S30) reproducing site A from the C-S30 strain was the least antigenic of the set, in disagreement with previously reported studies with the virus isolate. Increasing peptide size from 15 to 21 residues did not significantly improve antigenicity. Overnight incubation of A15S30 with mAb 4C4 in solution showed a marked increase in peptide antigenicity not observed for other peptide analogues, suggesting that conformational rearrangement could lead to a stable peptide-antibody complex. In fact, peptide cyclization clearly improved antigenicity, confirming an antigenic reversion in a multiply substituted peptide. Solution NMR studies of both linear and cyclic versions of the antigenic loop of FMDV C-S30 showed that structural features previously correlated with antigenicity were more pronounced in the cyclic peptide. Twenty-six synthetic peptides, corresponding to all possible combinations of five single-point antigenicity-enhancing replacements in the GH loop of FMDV C-S8c1, were also studied. SPR kinetic screening of these peptides was not possible due to problems mainly related to the high mAb affinities displayed by these synthetic antigens. Solution affinity SPR analysis was employed and affinities displayed were generally comparable to or even higher than those corresponding to the C-S8c1 reference peptide A15. The NMR characterisation of one of these multiple mutants in solution showed that it had a conformational behaviour quite similar to that of the native sequence A15 and the X-ray diffraction crystallographic analysis of the peptide ? mAb 4C4 complex showed paratope ? epitope interactions identical to all FMDV peptide ? mAb complexes studied so far. Key residues for these interactions are those directly involved in epitope ? paratope contacts (141Arg, 143Asp, 146His) as well as residues able to stabilise a particular peptide global folding. A quasi-cyclic conformation is held up by a hydrophobic cavity defined by residues 138, 144 and 147 and by other key intrapeptide hydrogen bonds, delineating an open turn at positions 141, 142 and 143 (corresponding to the Arg-Gly-Asp motif).

Essential oils of Origanum vulgare L. subsp glandulosum (Desf.) letswaart from Tunisia: chemical composition and antioxidant activity

BACKGROUND: Characterisation of the essential oils from O. glandulosum collected in three locations of Tunisia, chemical composition and the evaluation of their antioxidant activities were carried out. RESULTS: The essential oils from Origanum vulgare L. subsp. glandulosum (Desf.) letswaart collected from three localities of north Tunisia - Krib, Bargou and Nefza - were obtained in yields of 2.5, 3.0 and 4.6% (v/w), respectively. The essential oils were analysed by GC and GC/MS and assayed for their total phenolics content, by the Folin-Ciocalteu method, and antioxidant effectiveness, using the 2,2-diphenyl-1-picrylhydrazil (DPPH) radical scavenging assay. The main components of these essential oils, from Nefza, Bargou and Krib, were p-cymene (36%, 40% and 46%), thymol (32%, 39% and 18%), gamma-terpinene (24%, 12% and 16%) and carvacrol (2%, 2% and 15%), respectively). The ability to scavenge the DPPH radicals, expressed by IC50, ranged from 59 to 80 mg L-1. The total phenolic content, expressed in gallic acid equivalent (GAE) g kg(-1) dry weight, varied from 9.37 to 17.70 g kg(-1) dw. CONCLUSIONS: A correlation was identified between the total phenolic content of the essential oils and DPPH radical scavenger capacity. The occurrence of a p-cymene chemotype of O. glandulosum in the northern region of Tunisia is demonstrated.

Semiconductor Device as Optical Demultiplexer for Short Range Optical Communications

In this paper we present results on the use of a multilayered a-SiC:H heterostructure as a wavelength-division demultiplexing device for the visible light spectrum. The proposed device is composed of two stacked p-i-n photodiodes with intrinsic absorber regions adjusted to short and long wavelength absorption and carrier collection. An optoelectronic characterisation of the device was performed in the visible spectrum. Demonstration of the device functionality for WDM applications was done with three different input channels covering the long, the medium and the short wavelengths in the visible range. The recovery of the input channels is explained using the photocurrent spectral dependence on the applied voltage. An electrical model of the WDM device is proposed and supported by the solution of the respective circuit equations. Short range optical communications constitute the major application field, however other applications are also foreseen.

Investimento pessoal e independência funcional em idosos

Enquadramento: Se o envelhecimento populacional é encarado como um triunfo da sociedade atual, constitui ao mesmo tempo um desafio a toda a sociedade no sentido de se conseguir que este envelhecimento seja ativo e com qualidade de vida. O investimento na vida pessoal realizado pelos idosos de forma a obter a melhor qualidade de vida bem como a manutenção da sua independência funcional são indicadores importantíssimos para um envelhecimento bem-sucedido. Objetivos: Identificar a perceção dos idosos sobre o seu investimento na vida pessoal, avaliar níveis de independência funcional e analisar associações entre varáveis sociodemográficas clínicas e psicossociais e o investimento pessoal e a independência funcional. Métodos: Realizou-se um estudo do tipo transversal, analítico-correlacional de natureza quantitativa e de cariz descritivo, com uma amostra não probabilística constituída por 103 idosos da Santa Casa da Misericórdia de Mogadouro. Para a mensuração das variáveis aplicou-se um questionário que integra uma secção de caraterização socio demográfica e clínica, a Escala de Apgar Familiar, o índice de Barthel e a Escala de Investimento Pessoal. Resultados: As evidências encontradas neste estudo demonstram que a nossa amostra apresenta níveis elevados de independência funcional (40,8%) e de investimento na vida pessoal (89,3%). As variáveis que se associam de forma significativa com a independência funcional foram o género, local de institucionalização e o exercício físico. Já as que se associaram ao investimento pessoal foram o estado civil, local de residência, a prática de exercício físico e a funcionalidade familiar. Conclusão: Apesar da elevada média de idades dos nossos idosos estes apresentam bastante funcionalidade e moderado investimento na vida pessoal, contudo as variáveis associadas de forma significativa a estes constructos são: ser do género masculino, ser casado ou viver em união de facto, o residir no próprio domicílio, praticar regularmente exercício físico e percecionar famílias funcionais.

Estudo geológico e geotécnico de taludes rochosos em S. Pedro da Afurada (Vila Nova de Gaia): contributos para a avaliação da estabilidade estrutural

Mestrado em Engenharia Geotécnica e Geoambiente

Efeitos tridimensionais no escoamento em torno de cilindros confinados

Mestrado em Engenharia Química

Optical demultiplexer based on an a-SiC:H voltage controlled device

In this paper we present results on the use of a semiconductor heterostructure based on a-SiC:H as a wavelength-division demultiplexer for the visible light spectrum. The proposed device is composed of two stacked p-i-n photodiodes with intrinsic absorber regions adjusted to short and long wavelength absorption and carrier collection. An optoelectronic characterisation of the device was performed in the visible spectrum. Demonstration of the device functionality for WDM applications was done with three different input channels covering the long, the medium and the short wavelengths in the visible range. The recovery of the input channels is explained using the photocurrent spectral dependence on the applied voltage. An electrical model of the WDM device is proposed and supported by the solution of the respective circuit equations. Short range optical communications constitute the major application field however other applications are foreseen. (C) 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Cartografia geotécnica subterrânea do maciço granítico de Arca d’Água (Paranhos): implicações para o modelo geomecânico comportamental

Mestrado em Engenharia Geotécnica e Geoambiente

Cartografia geotécnica subterrânea do maciço granítico de Arca D'Água (sector de Carvalhido–Burgães): implicações para o modelo geomecânico comportamental

Este trabalho pretende enfatizar a importância da elaboração de uma base de dados extensa e robusta para o melhoramento da cartografia geotécnica subterrânea do maciço granítico do sector de Carvalhido–Burgães (Porto). Este estudo envolveu a análise e o refinamento das principais características geomecânicas ao longo de um traçado com cerca de 1220 m, para a caracterização do maciço rochoso da galeria subterrânea Carvalhido–Burgães. Para esta caracterização foram coligidos e uniformizados dados geológicos, geotécnicos e geomecânicos relativos a várias campanhas de campo, realizadas entre 2005 e 2011, tendo estado a técnica da amostragem linear aplicada ao grau de compartimentação do maciço rochoso na base do seu estudo. Além disso, procedeu‐se a um tratamento estatístico das descontinuidades, bem como dos parâmetros geológico‐geotécnicos e geomecânicos a estas associados. O zonamento geotécnico do maciço granítico foi realizado sempre em estreita ligação com o conhecimento das características do maciço in situ. Pretende‐se que esta metodologia contribua para um melhor conhecimento da compartimentação dos maciços rochosos em geral e, em particular, do modelo geomecânico comportamental do maciço rochoso do Porto.

Prospecção de recursos hidrominerais e geotérmicos na Orla Ocidental Portuguesa: os casos das Termas dos Cucos e de Monte Real

A cobertura sedimentar da região Oeste portuguesa é constituída por uma série possante de sedimentos com uma variedade de fácies com idades compreendidas entre o Triásico Superior e o actual. Estes sedimentos foram depositados numa bacia alongada com direcção NNE‐SSW. A tectónica desta cobertura sedimentar é condicionada pelas falhas tardi‐Variscas que afectaram o substrato e pelo complexo evaporítico depositado na base das séries sedimentares. Séries evaporíticas espessas de idade Hetangiana formaram numerosas estruturas diapíricas. Na região Oeste de Portugal existem diversas nascentes minerais e termais usadas para hidroterapia. Esta tese tem por objectivo estudar as concessões hidrominerais das Termas dos Cucos e das Termas de Monte Real (Portugal Central), bem como as suas áreas envolventes. Estas actividades hidroterapêuticas são muito relevantes em termos sócio‐económicos para os concelhos de Torres Vedras e Leiria. Os estudos contemplados nesta tese (que incluíram trabalho de campo no domínio da geotectónica, geomorfologia e hidrogeologia) permitiram delinear modelos hidrogeológicos conceptuais, apoiados, ainda, pela re‐interpretação de estudos geofísicos e hidrogeológicos prévios. A caracterização destas áreas foi suportada por inventários hidrogeológicos, tendo sido determinante para o projecto dos furos de captação, incluindo localização e profundidade. Todos os dados compilados foram representados cartograficamente numa base de Sistemas de Informação Geográfica (SIG).

Avaliação geológica e geotécnica do maciço da Pedreira de S. Domingos n.º 2 (Armamar)

Neste trabalho, apresentam-se e discutem-se os resultados da aplicação da técnica de amostragem linear de descontinuidades em faces expostas do maciço rochoso da pedreira granítica de S. Domingos Nº 2 (Fontelo, Armamar; N de Portugal). É, igualmente, utilizada informação sobre a rede de fracturação regional, obtida através da análise morfoestrutural de mapas topográficos e mapas geológicos. São ainda referidos os métodos utilizados no tratamento dos dados de terreno com o objectivo de definir as famílias de descontinuidades e de caracterizar estatísticamente a sua atitude, espaçamento e extensão. Os resultados obtidos são comparados, à mega escala e macro-escala, no sentido de averiguar a presença de um padrão de fracturação com dimensão multiescala. Esta abordagem foi refinada através da aplicação de Sistemas de Informação Geográfica. A aplicação desta técnica para a caracterização da compartimentação do maciço poderá contribuir para aperfeiçoar a gestão sustentável do georrecurso da pedreira de S. Domingos Nº 2 (Fontelo). O controlo geomecânico do desmonte do maciço rochoso é salientado com o intuito de uma abordagem de geo-engenharia integrada dos maciços rochosos.