Identification of Schistosoma mansoni microRNAs

Background: MicroRNAs (miRNAs) constitute a class of single-stranded RNAs which play a crucial role in regulating development and controlling gene expression by targeting mRNAs and triggering either translation repression or messenger RNA (mRNA) degradation. miRNAs are widespread in eukaryotes and to date over 14,000 miRNAs have been identified by computational and experimental approaches. Several miRNAs are highly conserved across species. In Schistosoma, the full set of miRNAs and their expression patterns during development remain poorly understood. Here we report on the development and implementation of a homology-based detection strategy to search for miRNA genes in Schistosoma mansoni. In addition, we report results on the experimental detection of miRNAs by means of cDNA cloning and sequencing of size-fractionated RNA samples. Results: Homology search using the high-throughput pipeline was performed with all known miRNAs in miRBase. A total of 6,211 mature miRNAs were used as reference sequences and 110 unique S. mansoni sequences were returned by BLASTn analysis. The existing mature miRNAs that produced these hits are reported, as well as the locations of the homologous sequences in the S. mansoni genome. All BLAST hits aligned with at least 95% of the miRNA sequence, resulting in alignment lengths of 19-24 nt. Following several filtering steps, 15 potential miRNA candidates were identified using this approach. By sequencing small RNA cDNA libraries from adult worm pairs, we identified 211 novel miRNA candidates in the S. mansoni genome. Northern blot analysis was used to detect the expression of the 30 most frequent sequenced miRNAs and to compare the expression level of these miRNAs between the lung stage schistosomula and adult worm stages. Expression of 11 novel miRNAs was confirmed by northern blot analysis and some presented a stage-regulated expression pattern. Three miRNAs previously identified from S. japonicum were also present in S. mansoni. Conclusion: Evidence for the presence of miRNAs in S. mansoni is presented. The number of miRNAs detected by homology-based computational methods in S. mansoni is limited due to the lack of close relatives in the miRNA repository. In spite of this, the computational approach described here can likely be applied to the identification of pre-miRNA hairpins in other organisms. Construction and analysis of a small RNA library led to the experimental identification of 14 novel miRNAs from S. mansoni through a combination of molecular cloning, DNA sequencing and expression studies. Our results significantly expand the set of known miRNAs in multicellular parasites and provide a basis for understanding the structural and functional evolution of miRNAs in these metazoan parasites

Nearest-Neighbour Interaction from an Abelian Symmetry and Deviations from Hermiticity

We show that Nearest-Neighbour Interaction (NNI) textures for the quark mass matrices can be obtained through the introduction of an Abelian flavour symmetry. The minimal realisation requires a Z_4 symmetry in the context of a two Higgs doublet model. It is further shown that the NNI textures can be in agreement with all present experimental data on quark masses and mixings, provided one allows for deviations of Hermiticity in the quark mass matrices at the 20% level.Comment: 11 pages, 1 figure; references and minor corrections adde

Desempenho de índices quantitativos de seca na estimativa da produtividade de arroz de terras altas

O objetivo deste trabalho foi caracterizar a intensidade e a ocorrência de seca pelo uso de índices quantitativos, e avaliar a relação entre esses índices e os dados da série histórica da produtividade ajustada do arroz de terras altas da microrregião de Goiânia, GO. O ajuste da série histórica foi realizado para minimizar os efeitos da variabilidade climática da região e dos avanços tecnológicos sobre a produtividade. Foram avaliados os seguintes índices: severidade de seca de Palmer (PDSI); Z de Palmer (Z-index); o de anomalia de chuva (RAI); e o padronizado de precipitação (SPI). Os índices de seca foram analisados com uso da correlação de Pearson, número e frequência de ocorrência da seca e percentual de acerto dos índices em relação à produtividade ajustada. O RAI quantificou o maior número de eventos extremos de seca, enquanto o PDSI não estimou nenhum caso. O Z-index apresentou o maior percentual de acerto, em relação às variações ocorridas na produtividade ajustada. Em períodos com variações da produtividade ajustada maior que 300 kg ha-1, Z-index, RAI e SPI apresentaram 78, 78 e 67% de percentuais de acerto, respectivamente. O Z-index teve o melhor desempenho na estimação da produtividade ajustada de arroz de terras altas