Lisete Sousa

The apoplast is the first hub of plant−pathogen communication where pathogen effectors are recognized by plant defensive proteins and cell receptors, thus activating signal transduction pathways. As a result of this first contact, the host triggers a defense response that involves the modulation of extra- and intracellular proteins. In grapevine−pathogen interactions, little is known about the trafficking between extra- and intracellular spaces. Grapevine is an economically important crop that relies on heavy fungicide use to control several diseases, and a deeper knowledge on the activation of its immune response is crucial to define new control strategies. In this study, we focused on the first 6 h postinoculation with Plasmopara viticola to evaluate grapevine proteome modulation in the apoplast. The in planta P. viticola proteome was also assessed to enable a deeper understanding of plant−pathogen communication. Our results showed that several plant mechanisms are triggered in th...

Plant apoplast is the first hub of plant-pathogen communication where pathogen effectors are recognized by plant defensive proteins and cell receptors and several signal transduction pathways are activated. As a result of this first contact, the host triggers a defence response that involves the modulation of several extra and intracellular proteins. In grapevine-pathogen interactions, little is known about the communication between cells and apoplast. Also, the role of apoplastic proteins in response to pathogens still remains a blackbox. In this study we focused on the first 6 hours after Plasmopara viticola inoculation to evaluate grapevine proteome modulation in the apoplastic fluid (APF) and whole leaf tissue. Plasmopara viticola proteome was also assessed enabling a deeper understanding of plant and pathogen communication. Our results showed that oomycete recognition, plant cell wall modifications, ROS signalling and disruption of oomycete structures are triggered in Regent af...

Nacional de Saúde Doutor Ricardo Jorge, IP, (Documento VIH/SIDA; 148). 2017. 3. PEPFAR. President’s Emergency Plan for AIDS Relief. U.S. Department of State Office of the U.S. Global AIDS Coordinator and Health Diplomacy 2017. https://www.pepfar.gov/documents/organization/267809.pdf 4. Charmaz K. Constructing Grounded Theory. 2nd edition. London: Sage Publications Limited ; 2014. 5. Meleis AI, Sawyer LM, Im EO, Hilfinger Messias DK, Schumacher K. Experiencing Transitions: An Emerging Middle-Rang Theory. ANS Adv Nurs Sci. 2000;23(1):12-28.

Autism Spectrum Disorder (ASD): Common neurodevelopmental disorder, global prevalence ~1 %; Persistent deficits in social communication and social interaction; restricted and repetitive behavior, interests, or activities – highly heterogeneous clinical presentation; Male to female ratio ~4:1

MA is supported by the Fundacao para a Ciencia e Tecnologia, Portugal (SFRH/BD/52485/2014). Patients and parents were genotyped in the context of the Autism Genome Project (AGP), funded by NIMH, HRB, MRC, Autism Speaks, Hilibrand Foundation, Genome Canada, OGI, CIHR.

Microarray allow to monitoring simultaneously thousands of genes, where the abundance of the transcripts under a same experimental condition at the same time can be quantified. Among various available array technologies, double channel cDNA microarray experiments have arisen in numerous technical protocols associated to genomic studies, which is the focus of this work. Microarray experiments involve many steps and each one can affect the quality of raw data. Background correction and normalization are preprocessing techniques to clean and correct the raw data when undesirable fluctuations arise from technical factors. Several recent studies showed that there is no preprocessing strategy that outperforms others in all circumstances and thus it seems difficult to provide general recommendations. In this work, it is proposed to use exploratory techniques to visualize the effects of preprocessing methods on statistical analysis of cancer two-channel microarray data sets, where the cancer types (classes) are known. For selecting differential expressed genes the arrow plot was used and the graph of profiles resultant from the correspondence analysis for visualizing the results. It was used 6 background methods and 6 normalization methods, performing 36 pre-processing methods and it was analyzed in a published cDNA microarray database (Liver) available at http://genome-www5.stanford.edu/ which microarrays were already classified by cancer type. All statistical analyses were performed using the R statistical software

RESUMO Objetivo: identificar os indicadores clínicos do diagnóstico de enfermagem disfunção sexual em mulheres grávidas. Método: revisão integrativa da literatura, com pesquisa em bases de dados, utilizando os descritores "sexual*", "pregnan*" e"function*". Foram incluídos estudos com resumo disponível para análise, referentes a grávidas com idade igual ou superior a 18 anos, escritos em português, francês, espanhol e inglês, com data de publicação entre 2010 e 2014. Foram excluídos estudos que reportassem grávidas com patologia associada. Resultados: a disfunção sexual na grávida é consistente na literatura. Foram identificadas nove características definidoras e 16 fatores relacionados, alguns não classificados na NANDA Internacional. Conclusão: indicadores clínicos podem ser adicionados ao diagnóstico de enfermagem de modo a favorecer um diagnóstico acurado e intervenções efetivas na vigilância da gravidez como um período de vivência sexual saudável.

Among various available array technologies, double-channel cDNA microarray experiments provide numerous technical protocols associated with functional genomic studies. The chapter begins by detailing the arrow plot, which is a recent graphical-based methodology to detect differentially expressed (DE) genes, and briefly mentions the significance analysis of microarrays (SAM) procedure, which is, in contrast, quite well known. Next, it introduces the correspondence analysis (CA) and explains how the resultant graphic can be interpreted. Then, CA in both class comparison and class prediction applications and over the data sets lymphoma (lym), lung (lun), and liver (liv) is executed. The CA is applied to all three databases in order to obtain graphical representations of background correction (BC) and normalization (NM) profiles in a two-dimensional reduced space. Whenever possible, more than one preprocessing strategy on microarray data could be applied and results from preprocessed data should be compared before any conclusion and subsequent analysis.info:eu-repo/semantics/publishedVersio

Grapes are economically the most important fruit worldwide. However, the complexity of biological events that lead to ripening of nonclimacteric fruits is not fully understood, particularly the role of polyamines' catabolism. The transcriptional and metabolic profilings complemented with biochemical data were studied during ripening of Trincadeira grapes submitted to guazatine treatment, a potent inhibitor of polyamine oxidase activity. The mRNA expression profiles of one time point (EL 38) corresponding to harvest stage was compared between mock and guazatine treatments using Affymetrix GrapeGen(®) genome array. A total of 2113 probesets (1880 unigenes) were differentially expressed between these samples. Quantitative RT-PCR validated microarrays results being carried out for EL 35 (véraison berries), EL 36 (ripe berries) and EL 38 (harvest stage berries). Metabolic profiling using HPLC and (1)H NMR spectroscopy showed increase of putrescine, proline, threonine and 1-O-ethyl-β-glucoside in guazatine treated samples. Genes involved in amino acid, carbohydrate and water transport were down-regulated in guazatine treated samples suggesting that the strong dehydrated phenotype obtained in guazatine treated samples may be due to impaired transport mechanisms. Genes involved in terpenes' metabolism were differentially expressed between guazatine and mock treated samples. Altogether, results support an important role of polyamine catabolism in grape ripening namely in cell expansion and aroma development.

The oomycete pathogen Plasmopara viticola (Berk. et Curt.) Berl. et de Toni is the causing agent of the destructive downy mildew disease in grapevine. Despite the advances towards elucidation of grapevine resistance mechanisms to downy mildew, increased knowledge of the biological and genetic components of the pathosystem is important to design suitable breeding strategies. Previously, a cDNA microarray approach was used to compare two Vitis vinifera genotypes Regent and Trincadeira (resistant and susceptible to downy mildew, respectively) in field conditions. The same cDNA microarray chip was used to confirm field-based results and to compare both genotypes under greenhouse conditions at 0, 6, and 12 h post-inoculation with P. viticola. Results show that when comparing both cultivars after pathogen inoculation, there is a preferential modulation of several defense, signaling, and metabolism associated transcripts in Regent. Early transcriptional changes are discussed in terms of genetic background and resistance mechanism. This study is the first to directly compare resistant and susceptible cultivars responses as early as 6 hpi with P. viticola, providing several candidate genes potentially related to the expression of resistance traits.

Objective to determine the psychometric properties of the international 7-item Falls Efficacy Scale. Method a psychometric study. Convenience sample consisting of 170 older adults living in the Madeira Autonomous Region, Portugal. A two-part instrument was used (sociodemographic characterization and the Falls Efficacy Scale-International-Portugal). The starting point was the translation and transcultural adaptation already carried out for the Falls Efficacy Scale - International (16 items). Construct validity (factorial analysis and discriminant validity) and the reliability (Cronbach's α) of the 7-item scale were evaluated. Previous authorization was obtained from the Ethics Commission and from the people involved. Results in the exploratory factorial analysis, the International 7-item Falls Efficacy Scale presents an explained variance of 65.8%. The Spearman's correlation between the score obtained based on the 7 items and the score obtained based on the 16 items is signif...

Introdução: As incapacidades resultantes do envelhecimento ganham destaque com as alterações demográficas. A queda é um dos preditores principais em idosos. O medo de cair provoca falta de confiança nas tarefas do quotidiano, restrição nas atividades sociais e aumento da dependência. A investigação demonstra que o medo de cair é causador de institucionalização, mesmo quando os idosos não caíram. A Falls Efficacy Scale International (FES-I) é constituída por 16 itens, utilizada internacionalmente em diversos contextos. Objetivos: Identificar as propriedades psicométricas das adaptações culturais da FES-I na população idosa residente na comunidade. Metodologia: Revisão sistemática da literatura, com a questão: Quais as propriedades psicométricas da FES-I em idosos residentes na comunidade? Pesquisa realizada em 2015, na plataforma EBSCOHost®, com a fórmula de pesquisa: (Validation) AND (Reliability) AND (Accidental Falls) AND (FES-I OR Falls Efficacy Scale International). Utilizou-se ...

These report presents two methods for the identi cation of signal peptides and their cleavage sites. The rst method is based on based neural networks and the second on hidden Markov models. The transmembrane protein topology can also be identi ed by a method based on hidden Markov models, which is described here in detail. The methodologies are then applied to identify a signal peptide in fOg44 lysin and to determine the topology of the transmembrane protein holin, also in fOg44 virus. Finally an outlook for possible improvements in these methodologies combining, somehow, hidden Markov models and neural networks.

Abstract Just behind wheat and rice, maize is the third most important grain crop in the world, grown in many agricultural areas. In Mozambique, rainfall is declining, posing serious threats to national food sovereignty and security, particularly in southern regions of the country. Despite this fact, few research studies have been conducted on the performance of Mozambique´s maize germplasm under drought stress conditions. A randomized design combining eight maize genotypes (six traditional Mozambican genotypes: LVA34, TCDE, G234, D456C, MABC, C123; and two commercial cultivars from DuPont Pioneer: P0023 and P9838) and three water regimes (control (irrigated), stress and recovery) was used to characterize the maize resilience to water deficit. The photochemical performance was accessed by rapid chlorophyll fluorescence induction curves (JIP-test). The use of Performance Index based dendrogram analysis allowed to explain genotypes behavior on the different experimental sets (control, drought stress and recovery). G234 and C123 presented the best performance with the lowest mortality rates, showing a better ability to cope with drought stress when compared to other genotypes from Mozambique and to the commercial cultivars from DuPont Pioneer.

Biological sequences, such as proteins, have been provided with annotations that assign functional information. These functional annotations are associations of proteins (or other biological sequences) with descriptors characterizing their biological roles. However, not all proteins are fully (or even at all) annotated. This annotation incompleteness limits our ability to make sound assertions about the functional coherence within sets of proteins. Annotation incompleteness is a problematic issue when measuring semantic functional similarity of biological sequences since they can only capture a limited amount of all the semantic aspects the sequences may encompass. Instead of relying uniquely on single (reductive) metrics, this work proposes a comprehensive approach for assessing functional coherence within protein sets. The approach entails using visualization and term enrichment techniques anchored in specific domain knowledge, such as a protein family. For that purpose we evaluat...

ABSTRACT Microarray allow to monitoring simultaneously thousands of genes, where the abundance of the transcripts under a same experimental condition at the same time can be quantified. Among various available array technologies, double channel cDNA microarray experiments have arisen in numerous technical protocols associated to genomic studies, which is the focus of this work. Microarray experiments involve many steps and each one can affect the quality of raw data. Background correction and normalization are preprocessing techniques to clean and correct the raw data when undesir-able fluctuations arise from technical factors. Several recent studies showed that there is no preprocessing strategy that outperforms others in all circumstances and thus it seems difficult to provide general recommendations. In this work, it is proposed to use exploratory techniques to visualize the effects of preprocessing methods on statistical analysis of cancer two-channel microarray data sets, where the cancer types (classes) are known. For selecting differential expressed genes the arrow plot was used and the graph of profiles resultant from the correspondence analysis for visualizing the results. It was used 6 background methods and 6 normalization methods, performing 36 pre-processing methods and it was analyzed in a published cDNA microarray database (Liver) available at http://genome-www5.stanford.edu/ which microar-rays were already classified by cancer type. All statistical analyses were performed using the R statistical software.

One of the most remarkable pollination strategy in orchids biology is pollination by sexual deception, in which the modified petal labellum lures pollinators by mimicking the chemical (e.g. sex pheromones), visual (e.g. colour and shape/size) and tactile (e.g. labellum trichomes) cues of the receptive female insect species. The present study aimed to characterize the transcriptional changes occurring after pollination in the labellum of a sexually deceptive orchid (Ophrys fusca Link) in order to identify genes involved on signals responsible for pollinator attraction, the major goal of floral tissues. Novel information on alterations in the orchid petal labellum gene expression occurring after pollination demonstrates a reduction in the expression of alkene biosynthetic genes using O. fusca Link as the species under study. Petal labellum transcriptional analysis revealed downregulation of transcripts involved in both pigment machinery and scent compounds, acting as visual and olfactory cues, respectively, important in sexual mimicry. Regulation of petal labellum senescence was revealed by transcripts related to macromolecules breakdown, protein synthesis and remobilization of nutrients.

Transcriptional changes in Pisolithus tinctorius leading to ectomycorrhizal formation in P. tinctorius- Castanea sativa were investigated using a 12-h fungal interaction in vitro system. Using a 3107-cDNA clone microarray, 34 unique expressed sequence tags (ESTs) were found to be differentially expressed. These ESTs represent 14 known genes, 5 upregulated and 9 downregulated, and 20 orphan sequences. Some transcripts of upregulated genes (with unknown function) were previously identified in other mycorrhizal Pisolithus spp. associations. ESTs for S-adenosyl-L-homocysteine hydrolase and several orphan sequences were identified in our system. The identified transcript of downregulated genes involved hydrophobins, 5S, 18S, and 28S ribosomal RNA genes, large subunits of ribosomal RNA (mitochondrial gene), and two types of heat shock proteins. This study demonstrates the high complexity of molecular events involved in the preinfection steps and suggests the utilization of different fungal gene repertories before ectomycorrhizal formation. These data constitute a first contribution for the molecular understanding of early signaling events between P. tinctorius and C. sativa roots during ectomycorrhizal formation.

Two multivariate statistical methods, factor analysis (FA) and hierarchical cluster analysis (HCA), were applied to experimental data set to evaluate their usefulness in selecting the adequate expression system and optimal growth parameters for recombinant cyprosin B production. Using FA, the large data set was reduced to two factors representing 73.4% of variability. Factor 1, with 53.5% of variability, corresponds to recombinant cyprosin B expression and efficient secretion, while factor 2, accounting for 19.9% of variability, represents cell growth and physiological characteristics. FA and HCA allowed the establishment of correlations among different variables and the clusters obtained providing clear identification of the experimental parameters related to cyprosin B production, which results on more accurate scientific output and time saving when selection of an adequate expression system is concerned.

Pre-mRNA splicing is an essential step in the post-transcriptional gene expression control involving protein-splicing factors like U2AF, which is exported to the cytoplasm and implicated in additional cellular functions. Identification of U2AF-associated mRNAs under native conditions was performed by immunoprecipitation and hybridization to Affymetrix GeneChip. Normalization and gene selection methods were performed, but the results were not reliable as they were different for different procedures, mainly because more than 20% of the mRNAs detected are differently enriched and the common normalization methods are based on small differences between them. We implemented a background correction method inspired in a non-specific hybridization method used for pre-processing data from ChIP-Chip technology. In this work, linear regression models are used to model in each array the non-specific hybridization, accounting for interactions between each three consecutive nucleotides into the probe sequence. Every probe intensity on the array was standardized using its predicted intensity and the probes' variance for similar predicted intensities. The standardized probe intensity values showed no need for further normalization and could be directly compared. We propose a probe set score, and a probe set enrichment value (ENRval) and its respective p-value for gene enrichment selection.