On Hepatitis C Virus Evolution: The Interaction between Virus and Host towards Treatment Outcome

Background:Hepatitis C is a disease spread throughout the world. Hepatitis C virus (HCV), the etiological agent of this disease, is a single-stranded positive RNA virus. Its genome encodes a single precursor protein that yields ten proteins after processing. NS5A, one of the non-structural viral proteins, is most associated with interferon-based therapy response, the approved treatment for hepatitis C in Brazil. HCV has a high mutation rate and therefore high variability, which may be important for evading the immune system and response to therapy. The aim of this study was to analyze the evolution of NS5A quasispecies before, during, and after treatment in patients infected with HCV genotype 3a who presented different therapy responses.Methods:Viral RNA was extracted, cDNA was synthesized, the NS5A region was amplified and cloned, and 15 clones from each time-point were sequenced. The sequences were analyzed for evolutionary history, genetic diversity and selection.Results:This analysis shows that the viral population that persists after treatment for most non-responder patients is present in before-treatment samples, suggesting it is adapted to evade treatment. In contrast, the population found in before treatment samples from most end-of-treatment responder patients either are selected out or appears in low frequency after relapse, therefore changing the population structure. The exceptions illustrate the uniqueness of the evolutionary process, and therefore the treatment resistance process, in each patient.Conclusion:Although evolutionary behavior throughout treatment showed that each patient presented different population dynamics unrelated to therapy outcome, it seems that the viral population from non-responders that resists the treatment already had strains that could evade therapy before it started. © 2013 Bittar et al.

Genetic structure of Aedes aegypti populations determined using pairwise comparisons

The biological characteristics of Aedes aegypti (Diptera, Culicidae), which is a vector of dengue and yellow fever, make this organism a good model for studying population structure and the events that may influence it under the effect of human activity. We assessed the genetic variability of five A. aegypti populations using RAPD-PCR technique and six primers. Four populations were from Brazil and one was from the USA. A total of 165 polymorphic DNA loci were generated. Considering the six primers and the five populations, the mean value of inter-population genetic diversity (Gst) was 0.277, which is considered high according to the Wright classification. However, pairwise comparisons of the populations gave variable Gst values ranging from 0.044 to 0.289. This variation followed the population's geographic distance to some extent but was also influenced by human activity. The lowest Gst values were obtained in the comparison of populations from cities with intensive commercial and medical contacts. These mosquito populations were previously classified as insecticide resistant, susceptible, or with decreased susceptibility; this parameter apparently had an effect on the Gst values obtained in the pairwise comparisons. ©FUNPEC-RP.

Estudos de variabilidade genética em populações de Chrysoperla externa Hagen, 1861 (Neuroptera: Chrysopidae) do estado de São Paulo, Brasil

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Prospecção de gênes de celulase presentes em biblioteca metagenômica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização molecular de isolados de Paracoccidioides brasiliensis obtidos de Dasypus novemcinctus (tatu) e sua correlação com virulência e origem geográfica

Pós-graduação em Patologia - FMB

Enxertia e caracterização de rambutanzeiro por aspectos foliares

Pós-graduação em Agronomia (Produção Vegetal) - FCAV

Caracterização molecular e biológica do Lettuce big-vein associated vírus e Mirafiori lettuce big-vein vírus e estudo da ocorrência em relação à época e sintoma em plantas de alface no Estado de São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Simbioses de rizóbios com leguminosas arbóreas na pré-amazônia maranhense

The tree leguminous gliricídia (Gliricidia sepium), acácia (Acacia mangium), leucena (Leucaena leucocephala) and sombreiro (Clitoria fairchildiana) are indicated for agroforestry systems, reclamation of degraded lands, reforestation and other purposes in the wet tropic. Despite the importance of legumes the preamazon region it is lacking in information about the symbiotic capacity and diversity of indigenous rhizobia of these legumes. The aim of this work was to evaluate the phenotypic and genetic diversity of rhizobia species nodulating gliricidia sombreiro, leucena and acacia in the Maranhão pre-Amazon region and authenticate isolates of these species in siratro (Macroptilium atropurpureum). For this they were installed two experiments. Sampling was carried out on a alley cropping system, was sampled 20 plants of each species by collecting 10 nodules per plant. It was made isolation, cultural characterization, partial 16S rRNA gene sequencing and analysis of the symbiotic ability of bacterial strains with siratro. The authentication experiment was done in two steps for each legume (gliricidia, acácia, sombreiro and leucena), in the greenhouse and in a completely randomized design with three replications with sterile Hoagland nutrient solution as substrate. Gliricídia, Sombreiro, leucena and acacia are colonized by distinct groups of rhizobia. Gliricidia nodulate preferably with Rhizobium, sombreiro and acacia nodulate preferably with Bradyrhizobium and leucena has Mesorhizobium main symbiote. Endophytic strains of ten genera were found colonizing the gliricidia, sombrero, leucena and acacia nodules and a strain of Arthrobacter sp. had a positive nodulation with siratro. This is the first report on isolation of Methylobacterium sp. in gliricidia nodules and endophytic ability of Terriglobus sp. strains. Indigenous strains of pre-Amazon region of Bradyrhizobium, Mesorhizobium and Rhizobium genus nodulate with siratro, but are ineficiente or had low efficiency to promote their growth.

Distância e padrões de dispersão contemporânea de pólen e sistema de reprodução em pequeno fragmento isolado de Copaifera Langsdorfii Desf. (Leguminosae – Caesalpinoideae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diversidade genética em populações de Myracrodruon urundeuva Fr. All., sob diferentes tipos de perturbação antrópica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análises genéticas em progênies de Pinus caribaea Morelet var. caribaea por caracteres quantitativos e marcadores moleculares

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da organização da variabilidade genética em populações de anfíbios de hábitats antropizados por meio marcadores microssatélites

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evolução das quasiespécies da proteína NS5A do vírus da hepatite C genótipo 3a

Pós-graduação em Genética - IBILCE

Genetic variability and efficiency of DNA microsatellite markers for paternity testing in horse breeds from the Brazilian Marajó archipelago

In this study, 15 microsatellite DNA loci used in comparative tests by the International Society for Animal Genetics were applied to the evaluation of genetic diversity and management, and the efficiency of paternity testing in Marajoara horses and Puruca ponies from the Marajó Archipelago. Based on the genotyping of 93 animals, mean allelic diversity was estimated as 9.14 and 7.00 for the Marajoara and Puruca breeds, respectively. While these values are similar to those recorded in most European breeds, mean levels of heterozygosity were much lower (Marajoara 49%, Puruca 40%), probably as a result of high levels of inbreeding in the Marajó populations. The mean informative polymorphic content of this 15-marker system was over 50% in both breeds, and was slightly higher in the Marajoara horses. The discriminative power and exclusion probabilities derived from this system were over 99% for both populations, emphasizing the efficacy of these markers for paternity testing and genetic management in the two breeds.

Genetic characterisation of populations of the critically endangered Goliath grouper (Epinephelus itajara, Serranidae) from the Northern Brazilian coast through analyses of mtDNA

The Goliath grouper (Epinephelus itajara) is one of the most endangered species of fish of the subfamily Epinephelinae. Slow to develop and mature, and dependent on mangrove habitats for breeding, the species also suffers intense harvesting, which has reduced drastically in numbers in many areas. To contribute to the understanding of the characteristics of E. itajara populations, we conducted a molecular genetics study of the species, focusing on populations from the Northern Brazilian coast. The mtDNA control region (D-loop) of 116 individuals from five localities (Bragança, Ajuruteua, Parnaíba, Fortaleza and Natal) was analysed, and a sequence of 499 base pairs identified. Analyses of the sequences indicated that genetic variability was generally lower in E. itajara than in other endangered species of the genus. AMOVA found no significant grouping structure among the populations. Nested Clade Analysis revealed a significant association between genetic variability and geographic distribution among only three populations (Ajuruteua, Parnaíba and Natal). Genetic diversity was higher in populations from the Amazon region, which may be related to the better conservation of mangrove habitats in this area. Therefore, the present study could be used for the implementation of conservation and management measures in order to protect and consolidate these populations.