Analysis of HCV quasispecies dynamic under selective pressure of combined therapy

Background: The quasispecies composition of Hepatitis C virus (HCV) could have important implications with regard to viral persistence and response to interferon-based therapy. The complete NS5A was analyzed to evaluate whether the composition of NS5A quasispecies of HCV 1a/1b is related to responsiveness to combined interferon pegylated (PEG-IFN) and ribavirin therapy.Methods: Viral RNA was isolated from serum samples collected before, during and after treatment from virological sustained responder (SVR), non-responder (NR) and the end-of-treatment responder patients (ETR). NS5A region was amplified, cloned and sequenced. Six hundred and ninety full-length NS5A sequences were analyzed.Results: This study provides evidence that lower nucleotide diversity of the NS5A region pre-therapy is associated with viral clearance. Analysis of samples of NRs and the ETRs time points showed that genetic diversity of populations tend to decrease over time. Post-therapy population of ETRs presented higher genetic distance from baseline probably due to the bottleneck phenomenon observed for those patients in the end of treatment. The viral effective population of those patients also showed a strong decrease after therapy. Otherwise, NRs demonstrated a continuous variation or stability of effective populations and genetic diversity over time that did not seem to be related to therapy. Phylogenetic relationships concerning complete NS5A sequences obtained from patients did not demonstrate clustering associated with specific response patterns. However, distinctive clustering of pre/post-therapy sequences was observed. In addition, the evolution of quasispecies over time was subjected to purifying or relaxed purifying selection. Codons 157 (P03), 182 and 440 (P42), 62 and 404 (P44) were found to be under positive selective pressure but it failed to be related to the therapy.Conclusion: These results confirm the hypothesis that a relationship exists between NS5A heterogeneity and response to therapy in patients infected with chronic hepatitis C. © 2013 Jardim et al.; licensee BioMed Central Ltd.

O modelo 'hydrotime' pode descrever a resposta de sementes de feijão comum (phaseolus vulgaris l.) à temperatura e potenciais de água reduzidos

Germination parameters of the response to temperature and water potential from four common bean (Phaseolus vulgaris) lines based on thermal-time and hydrotime concepts were estimated to verify to what extent they can predict germination under different thermal and water conditions. The cultivars IPR Uirapuru and IAPAR 81 (drought-tolerant), and Grauna and Carioca (not tolerant) were used. The isothermal assays were performed in a temperature gradient block, and the assays with different osmotic potentials (PEG 6000) were performed in germination chambers. Seeds from drought-tolerant cultivars spent less time to germinate at supra-optimum temperatures than non-tolerant ones, and the cultivar Uirapuru (drought-tolerant) germinated faster in response to reduced Ψ and low temperatures. The parameter Ψb(50) did not discriminate between drought-tolerant and non-tolerant lines at the infraoptimum temperature range, but it can be used to identify drought-tolerant lines at high temperatures. In general, the hydrotime model reproduced the actual germination data relatively well, chiefly at higher temperatures. This study evidenced that the hydrotime model can be used to describe the germination of common bean seeds under reduced water potentials, and as a screening tool for drought-tolerant bean genotypes.

PEGylation of SPIONs by polycondensation reactions: A new strategy to improve colloidal stability in biological media

In this study, we report on a new route of PEGylation of superparamagnetic iron oxide nanoparticles (SPIONs) by polycondensation reaction with carboxylate groups. Structural and magnetic characterizations were performed by X-ray diffractometry (XRD), transmission electron microscopy (TEM), thermogravimetric analysis (TGA), and vibrating sample magnetometry (VSM). The XRD confirmed the spinel structure with a crystallite average diameter in the range of 3.5-4.1 nm in good agreement with the average diameter obtained by TEM (4.60-4.97 nm). The TGA data indicate the presence of PEG attached onto the SPIONs' surface. The SPIONs were superparamagnetic at room temperature with saturation magnetization (M S) from 36.7 to 54.1 emu/g. The colloidal stability of citrate- and PEG-coated SPIONs was evaluated by means of dynamic light scattering measurements as a function of pH, ionic strength, and nature of dispersion media (phosphate buffer and cell culture media). Our findings demonstrated that the PEG polymer chain length plays a key role in the coagulation behavior of the Mag-PEG suspensions. The excellent colloidal stability under the extreme conditions we evaluated, such as high ionic strength, pH near the isoelectric point, and cell culture media, revealed that suspensions comprising PEG-coated SPION, with PEG of molecular weight 600 and above, present steric stabilization attributed to the polymer chains attached onto the surface of SPIONs. © 2013 Springer Science+Business Media Dordrecht.

Identification of duplicated and stress-inducible Aox2b gene co-expressed with Aox1 in species of the Medicago genus reveals a regulation linked to gene rearrangement in leguminous genomes

In flowering plants, alternative oxidase (Aox) is encoded by 3-5 genes distributed in 2 subfamilies (Aox1 and Aox2). In several species only Aox1 is reported as a stress-responsive gene, but in the leguminous Vigna unguiculata Aox2b is also induced by stress. In this work we investigated the Aox genes from two leguminous species of the Medicago genus (Medicago sativa and Medicago truncatula) which present one Aox1, one Aox2a and an Aox2b duplication (named here Aox2b1 and Aox2b2). Expression analyses by semi-quantitative RT-PCR in M. sativa revealed that Aox1, Aox2b1 and Aox2b2 transcripts increased during seed germination. Similar analyses in leaves and roots under different treatments (SA, PEG, H2O2 and cysteine) revealed that these genes are also induced by stress, but with peculiar spatio-temporal differences. Aox1 and Aox2b1 showed basal levels of expression under control conditions and were induced by stress in leaves and roots. Aox2b2 presented a dual behavior, i.e., it was expressed only under stress conditions in leaves, and showed basal expression levels in roots that were induced by stress. Moreover, Aox2a was expressed at higher levels in leaves and during seed germination than in roots and appeared to be not responsive to stress. The Aox expression profiles obtained from a M. truncatula microarray dataset also revealed a stress-induced co-expression of Aox1, Aox2b1 and Aox2b2 in leaves and roots. These results reinforce the stress-inducible co-expression of Aox1/Aox2b in some leguminous plants. Comparative genomic analysis indicates that this regulation is linked to Aox1/Aox2b proximity in the genome as a result of the gene rearrangement that occurred in some leguminous plants during evolution. The differential expression of Aox2b1/2b2 suggests that a second gene has been originated by recent gene duplication with neofunctionalization. © 2013 Elsevier GmbH. All rights reserved.

The tonoplast intrinsic aquaporin (TIP) subfamily of Eucalyptus grandis: Characterization of EgTIP2, a root-specific and osmotic stress-responsive gene

Aquaporins have important roles in various physiological processes in plants, including growth, development and adaptation to stress. In this study, a gene encoding a root-specific tonoplast intrinsic aquaporin (TIP) from Eucalyptus grandis (named EgTIP2) was investigated. The root-specific expression of EgTIP2 was validated over a panel of five eucalyptus organ/tissues. In eucalyptus roots, EgTIP2 expression was significantly induced by osmotic stress imposed by PEG treatment. Histochemical analysis of transgenic tobacco lines (Nicotiana tabacum SR1) harboring an EgTIP2 promoter:GUS reporter cassette revealed major GUS staining in the vasculature and in root tips. Consistent with its osmotic-stress inducible expression in eucalyptus, EgTIP2 promoter activity was up-regulated by mannitol treatment, but was down-regulated by abscisic acid. Taken together, these results suggest that EgTIP2 might be involved in eucalyptus response to drought. Additional searches in the eucalyptus genome revealed the presence of four additional putative TIP coding genes, which could be individually assigned to the classical TIP1-5 groups. © 2013 Elsevier B.V.

Consulta de enfermagem como instrumento de avaliação do protocolo assistencial de gastrostomia endóscópica percutânea

Pós-graduação em Bases Gerais da Cirurgia - FMB

Expressão de genes em sementes de lobeira (Solanum Lycocarpum St Hill) submetidas ao condicionamento fisiológico

Pós-graduação em Agronomia (Agricultura) - FCA

Ciclo reprodutivo de machos de Dendropsophus minutus (Anura, Hylidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Temperatura e osmocondicionamento na germinação de sementes de Urochloa brizantha (STAPF) Webster cv. Basilisk

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Dosagem de etanol utilizando alcool desidrogenase de levedura de panificação

Pós-graduação em Alimentos e Nutrição - FCFAR

Honduras: evolución económica durante 2008 y perspectivas para 2009

Incluye Bibliografía

Análise comparativa da variação entre quasiespecies do Vírus da Hepatite C genótipo 1 em amostras prétratamento de pacientes tratados com Peginterferon

Pós-graduação em Genética - IBILCE

Desempenho e caracterização de sementes de diferentes procedências de Cedrela fissilis Velozzo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expressão diferencial da aquaporina ScPIP2;1 em genótipos de cana-de-açúcar contrastantes para a tolerância à seca

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Eficiência alimentar e desempenho de bovinos nelore selecionados para peso pós-desmame

Pós-graduação em Zootecnia - FMVZ