Estimativa do parentesco numa população de melhoramento de Eucalyptus globulus através de microsatélites nucleares

É situação comum desconhecer-se o grau de parentesco entre a população na origem da maioria dos programas de melhoramento genético de espécies florestais. Para resolver este problema, desenvolvemos um protocolo de avaliação do parentesco utilizando 125 indivíduos e 16 microssatélites, da população base ou de referência (PR) de Eucalyptus globulus do RAIZ. Através da recombinação gamética in silico foram simulados 105 indivíduos com diferentes graus de parentesco: descendentes de autopolinização, meiosirmãos, irmãos completos e indivíduos não aparentados. Por simulação Monte-Carlo foram calculados o valor médio e a variância associada à média dos diferentes grupos de parentesco, com quatro coeficientes de similaridade genética. Compararam-se as funções densidade dos diferentes grupos de parentesco, obtidas com quatro coeficientes de parentesco, utilizando o valor crítico correspondente à intercepção das funções densidade dos indivíduos não aparentados e dos meios-irmãos. O estimador escolhido foi aplicado à PR. Detectaram-se 4,4% de pares de indivíduos potencialmente aparentados, com um erro de tipo II de 8%. Inferimos também, o parentesco de um conjunto de 24 clones elite e encontrámos 4 pares que são potencialmente aparentados. Futuros cruzamentos entre estes indivíduos deverão ser evitados

Optimizing the choice of microsatellite markers for fingerprinting eucalyptus

In this study we have analyzed the information provided by 17 publicly available Eucalyptus microsatellite (SSR) markers (Brondani et al. 1998, 2002; Jones et al. 2002; Steane et al. 2001) in the context of genetic identification within a sample of 140 individuals from an elite collection (denoted hereafter base) of RAIZ genetic improvement population

Impact of inbreeding on growth and development of young open-pollinated progeny of Eucalyptus globulus

The use of open-pollinated seeds from seed orchards is a common strategy for the deployment of genetically improved eucalypts, including Eucalyptus globulus, an important pulpwood tree in many temperate climate areas. However, seed quality can be affected by the rate of selfing and to a lesser extent by contamination from pollen outside the orchard. Inbreeding between related parents and especially from self-crosses is known to cause diminished growth and developmental abnormalities in the resulting progeny. This study looks at the magnitude and variation in selfing and the impact in inbreeding depression across several E. globulus families collected over the years in a seed orchard. The effects on growth and development of outcrossed and selfed progeny were studied across five progeny trials, after pedigree reconstruction of the open pollinated progeny based on SSR genotyping. An additive genetic mixed linear model was fitted to the data to evaluate the impact of inbreeding on height growth. The results showed a significant inbreeding depression, with a height growth reduction of 15% in selfed progeny, when compared with crosses from unrelated parents. These inbreeding depression values varied among families, ranging between 7% and 24%, evidencing the importance of genetic background. Contamination rates were on average 10% suggesting long distance pollen dispersal was present. A small number of abnormal phenotypes (less than 10%) was observed in the field. This was associated with specific, unrelated, crosses and not to high inbreeding rates such as found among selfed progeny. The relevance of these results for orchard management and parent selection is discussed.info:eu-repo/semantics/publishedVersio

Molecular estimates of similarity in Eucalyptus globulus

The base populations used in most forest tree genetic improvement programs usually lack detailed pedigree information. Molecular markers, such as microsatellites (SSR), can be used to estimate individuals’ pairwise relatedness, which is based on the probabilities’ ratios of the identity in state between the individuals compared and the reference unrelated population These estimates can be very useful to infer the level of relationship among sub-populations of elite material and/or for the design of controlled crosses between putatively unrelated parents. Using 113 putatively unrelated individuals - genotyped with 18 SSR - self, full-sib, half-sib and unrelated were simulated, and four pairwise similarity coefficients were tested: Queller & Goodnight 1989; Li et al. 1993; Ritland 1996, and Lynch & Ritland 1999. The Lynch & Ritland (1999) coefficient was selected (Figure 1), for it displayed a better adjustment with the expected level of relatedness and narrower standard errors (SE). SE were calculated through Monte- Carlo techniques, to avoid unequal sample size bias, by using 105 simulations for each relatedness group. To illustrate the usefulness of molecular estimates of similarity in genetic improvement programs, a clustering (UPGMA) based on the pairwise Lynch & Ritland (1999) coefficient (LR) values was performed to infer about the putative relationship among individuals of the subgroups of E. globulus elite individuals. From that analysis at least two pairs might be related and a PCA analysis confirmed the clustering results

Assessment of a Large-Scale Unbiased Malignant Pleural Effusion Proteomics Study of a Real-Life Cohort

Background: Pleural effusion (PE) is common in advanced-stage lung cancer patients and is related to poor prognosis. Identification of cancer cells is the standard method for the diagnosis of a malignant PE (MPE). However, it only has moderate sensitivity. Thus, more sensitive diagnostic tools are urgently needed. Methods: The present study aimed to discover potential protein targets to distinguish malignant pleural effusion (MPE) from other non-malignant pathologies. We have collected PE from 97 patients to explore PE proteomes by applying state-of-the-art liquid chromatography-mass spectrometry (LC-MS) to identify potential biomarkers that correlate with immunohistochemistry assessment of tumor biopsy or with survival data. Functional analyses were performed to elucidate functional differences in PE proteins in malignant and benign samples. Results were integrated into a clinical risk prediction model to identify likely malignant cases. Sensitivity, specificity, and negative predictive value were calculated. Results: In total, 1689 individual proteins were identified by MS-based proteomics analysis of the 97 PE samples, of which 35 were diagnosed as malignant. A comparison between MPE and benign PE (BPE) identified 58 differential regulated proteins after correction of the p-values for multiple testing. Furthermore, functional analysis revealed an up-regulation of matrix intermediate filaments and cellular movement-related proteins. Additionally, gene ontology analysis identified the involvement of metabolic pathways such as glycolysis/gluconeogenesis, pyruvate metabolism and cysteine and methionine metabolism. Conclusion: This study demonstrated a partial least squares regression model with an area under the curve of 98 and an accuracy of 0.92 when evaluated on the holdout test data set. Furthermore, highly significant survival markers were identified (e.g., PSME1 with a log-rank of 1.68 × 10−6 ).info:eu-repo/semantics/publishedVersio

Produtores de vinho do Algarve no Facebook: oportunidades e desafios

No marketing de relacionamento o Facebook é uma das redes sociais mais difundidas e que, por isso, pode ser mais relevante para consolidar a notoriedade das marcas de vinho algarvias e atrair e fidelizar públicos locais e dos principais mercados turísticos, configurando-se como elemento de elevado potencial para consolidar a atividade vinícola algarvia. A presença dos produtores de vinho do algarve no Facebook vem dar visibilidade ao vinho, produto que é parte integrante do património e da cultura algarvia, no ambiente digital baseado na internet, onde os restantes atores do sistema turístico algarvio desenvolvem já uma forte atividade. Esta investigação pretende descrever e analisar a presença dos produtores de vinho do Algarve na rede social Facebook, descrever e comparar a presença na referida rede bem como avaliar as principais diferenças nos perfis de utilização. Partindo da identificação dos produtores que atualmente produzem e comercializam vinho do Algarve, procedeu-se à identificação das respetivas páginas na rede social, bem como a recolha de dados com recurso à aplicação Likealyzer. Os dados que caracterizam a presença na rede social foram triangulados com dados secundários do desempenho económico e financeiro das empresas. Os resultados obtidos permitiram identificar e posicionar os produtores de vinho do Algarve de acordo com o seu nível de maturidade no uso da rede social, bem como aferir diferenças entre grupos.info:eu-repo/semantics/publishedVersio

Aberrant MEK5/ERK5 signalling contributes to human colon cancer progression via NF-κB activation

© 2015 Macmillan Publishers Limited All rights reserved. Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0This study was designed to evaluate MEK5 and ERK5 expression in colon cancer progression and to ascertain the relevance of MEK5/ERK5 signalling in colon cancer. Expression of MEK5 and ERK5 was evaluated in 323 human colon cancer samples. To evaluate the role of MEK5/ERK5 signalling in colon cancer, we developed a stable cell line model with differential MEK5/ERK5 activation. Impact of differential MEK5/ERK5 signalling was evaluated on cell cycle progression by flow cytometry and cell migration was evaluated by wound healing and transwell migration assays. Finally, we used an orthotopic xenograft mouse model of colon cancer to assess tumour growth and progression. Our results demonstrated that MEK5 and ERK5 are overexpressed in human adenomas (P<0.01) and adenocarcinomas (P<0.05), where increased ERK5 expression correlated with the acquisition of more invasive and metastatic potential (P<0.05). Interestingly, we observed a significant correlation between ERK5 expression and NF-κB activation in human adenocarcinomas (P<0.001). We also showed that ERK5 overactivation significantly accelerated cell cycle progression (P<0.05) and increased cell migration (P<0.01). Furthermore, cells with overactivated ERK5 displayed increased NF-κB nuclear translocation and transcriptional activity (P<0.05), together with increased expression of the mesenchymal marker vimentin (P<0.05). We further demonstrated that increased NF-κB activation was associated with increased IκB phosphorylation and degradation (P<0.05). Finally, in the mouse model, lymph node metastasis was exclusively seen in orthotopically implanted tumours with overactivated MEK5/ERK5, and not in tumours with inhibited MEK5/ERK5. Our results suggested that MEK5/ERK5/NF-κB signalling pathway is important for tumour onset, progression and metastasis, possibly representing a novel relevant therapeutic target in colon cancer treatment.This study was supported by Fundação para a Ciência e Tecnologia (HMSP-ICT/0018/2011, SFRH/BD/96517/2013, SFRH/BD/88619/2012 and SFRH/BD/79356/2011).info:eu-repo/semantics/publishedVersio

Genetic control of Eucalyptus urophylla and E. grandis resistance to canker caused by Chrysoporthe cubensis

Chrysophorte cubensis induced canker occurs in nearly all tropical and subtropical regions where eucalypts are planted, causing losses in both wood quality and volume productivity, especially so in the warmer and more humid regions of Brazil. The wide inter and intra-specific genetic variability of resistance to canker among Eucalyptus species facilitates the selection of resistant plants. In this study, we evaluated resistance to this pathogen in five Eucalyptus grandis (G) and 15 E. urophylla (U) trees, as well as in 495 individuals from 27 progenies derived from crosses between the trees. In the field, six-months-old test seedlings were inoculated with C. cubensis. Lesion length in the xylem and bark was measured eight months later. The results demonstrated that xylem lesions could preferentially be used for the selection of resistant clones. Eight trees (7 U and 1 G) were susceptible, and the remainder (8 U and 4 G) resistant. Individual narrow and broad sense heritability estimates were 17 and 81%, respectively, thereby suggesting that canker resistance is quantitative and highly dependent on dominance and epistasis