Circulation of Different Lineages of Dengue Virus 2, Genotype American/Asian in Brazil: Dynamics and Molecular and Phylogenetic Characterization

The American/Asian genotype of Dengue virus type 2 (DENV-2) was introduced into the Americas in the 80′s. Although there is no data showing when this genotype was first introduced into Brazil, it was first detected in Brazil in 1990. After which the virus spread throughout the country and major epidemics occurred in 1998, 2007/08 and 2010. In this study we sequenced 12 DENV-2 genomes obtained from serum samples of patients with dengue fever residing in São José do Rio Preto, São Paulo (SJRP/SP), Brazil, in 2008. The whole open reading frame or envelope sequences were used to perform phylogenetic, phylogeographic and evolutionary analyses. Isolates from SJRP/SP were grouped within one lineage (BR3) close to isolates from Rio de Janeiro, Brazil. Isolates from SJRP were probably introduced there at least in 2007, prior to its detection in the 2008 outbreak. DENV-2 circulation in Brazil is characterized by the introduction, displacement and circulation of three well-defined lineages in different times, most probably from the Caribbean. Thirty-seven unique amino acid substitutions were observed among the lineages, including seven amino acid differences in domains I to III of the envelope protein. Moreover, we dated here, for the first time, the introduction of American/Asian genotype into Brazil (lineage BR1) to 1988/89, followed by the introduction of lineages BR2 (1998-2000) and BR3 (2003-05). Our results show a delay between the introduction and detection of DENV-2 lineages in Brazil, reinforcing the importance and need for surveillance programs to detect and trace the evolution of these viruses. Additionally, Brazilian DENV-2 differed in genetic diversity, date of introduction and geographic origin and distribution in Brazil, and these are important factors for the evolution, dynamics and control of dengue. © 2013 Drumond et al.

Histomorphometrical and molecular evaluation of endosseous dental implants sites in humans: Correlation with clinical and radiographic aspects

Objective: To evaluate the correlations between clinical-radiographical aspects and histomorphometric-molecular parameters of endosseous dental implant sites in humans. Material and methods: The study sample consisted of bone implant sites from the jawbones of 32 volunteers, which were classified according to two different systems: (1) based only on periapical and panoramic images (PP); (2) as proposed by Lekholm & Zarb (L&Z). Bone biopsies were removed using trephine during the first drilling for implant placement. Samples were stained with haematoxylin-eosin (HE), and histomorphometric analysis was performed to obtain the following parameters: trabecular thickness (Tb.Th), trabecular number, bone volume density (BV/TV), bone specific surface (BS/BV), bone surface density and trabecular separation (Tb.Sp). In addition, immunohistochemistry analysis was performed on bone tissue samples for the proteins, Receptor activator of nuclear factor kappa-B (RANK), RANK ligand (RANKL), osteoprotegerin (OPG) and Osteocalcin (OC). Also, the determination of the relative levels of gene expression was performed using Reverse transcription-real-time Polymerase Chain Reaction (RT-PCR). Results: PP and L&Z classification systems revealed a moderate correlation with BV/TV, BS/BV, Tb.Th and Tb.Sp. L&Z's system identified differences among bone types when BV/TV, BS/BV, Tb.Th and Tb.Sp were compared. A weak correlation between PP/L&Z classifications and the expression of bone metabolism regulators (RANK, RANKL, OPG e OC) was found. The analysis of mRNA expression showed no difference between the bone types evaluated. Conclusions: Our results suggest that PP and L&Z subjective bone-type classification systems are related to histomorphometric aspects. These data may contribute to the validation of these classifications. Bone remodelling regulatory molecules do not seem to influence morphological aspects of the jawbone © 2011 John Wiley & Sons A/S.

Detection of post-F1 fish hybrids in broodstock using molecular markers: Approaches for genetic management in aquaculture

Herein, we have developed molecular markers for nuclear genes to use in multiplex-PCR and PCR-RFLP, with the goal of characterising hybrid lines derived from crosses between pintado Pseudoplatystoma corruscans and cachara P. reticulatum. These markers, together with others described previously, were used to perform molecular identification analyses as genetic subsidies for Brazilian aquaculture. These analyses were performed due to the problems of high mortality in the offspring reported by the aquaculturist. From a total of 16 broodstock samples, 13 were genetically identified as hybrids; surprisingly, nine of these hybrids were found to be post-F1 lineages. These data show that the fertility of these animals can seriously affect the cultivated stocks, thus causing financial damage in this aquaculture system. The establishment of PCR-RFLP and multiplex-PCR as molecular techniques allows for both the correct management of these animals and the routine monitoring of production and trade of fish hybrids in aquaculture. Consequently, such tools will enable a sustainable development in the aquaculture industry. © 2012 Blackwell Publishing Ltd.

Elucidation of carbohydrate molecular interaction mechanism of recombinant and native ArtinM

The quartz crystal microbalance (QCM) technique has been applied for monitoring the biorecognition of ArtinM lectins at low horseradish peroxidase glycoprotein (HRP) concentrations, using a simple kinetic model based on Langmuir isotherm in previous work.18 The latter approach was consistent with the data at dilute conditions but it fails to explain the small differences existing in the jArtinM and rArtinM due to ligand binding concentration limit. Here we extend this analysis to differentiate sugar-binding event of recombinant (rArtinM) and native (jArtinM) ArtinM lectins beyond dilute conditions. Equivalently, functionalized quartz crystal microbalance with dissipation monitoring (QCM-D) was used as real-time label-free technique but structural-dependent kinetic features of the interaction were detailed by using combined analysis of mass and dissipation factor variation. The stated kinetic model not only was able to predict the diluted conditions but also allowed to differentiate ArtinM avidities. For instance, it was found that rArtinM avidity is higher than jArtinM avidity whereas their conformational flexibility is lower. Additionally, it was possible to monitor the hydration shell of the binding complex with ArtinM lectins under dynamic conditions. Such information is key in understanding and differentiating protein binding avidity, biological functionality, and kinetics. © 2013 American Chemical Society.

Expression, purification and molecular analysis of the human ZNF706 protein

Background: The ZNF706 gene encodes a protein that belongs to the zinc finger family of proteins and was found to be highly expressed in laryngeal cancer, making the structure and function of ZNF706 worthy of investigation. In this study, we expressed and purified recombinant human ZNF706 that was suitable for structural analysis in Escherichia coli BL21(DH3). Findings. ZNF706 mRNA was extracted from a larynx tissue sample, and cDNA was ligated into a cloning vector using the TOPO method. ZNF706 protein was expressed according to the E. coli expression system procedures and was purified using a nickel-affinity column. The structural qualities of recombinant ZNF706 and quantification alpha, beta sheet, and other structures were obtained by spectroscopy of circular dichroism. ZNF706's structural modeling showed that it is composed of α-helices (28.3%), β-strands (19.4%), and turns (20.9%), in agreement with the spectral data from the dichroism analysis. Conclusions: We used circular dichroism and molecular modeling to examine the structure of ZNF706. The results suggest that human recombinant ZNF706 keeps its secondary structures and is appropriate for functional and structural studies. The method of expressing ZNF706 protein used in this study can be used to direct various functional and structural studies that will contribute to the understanding of its function as well as its relationship with other biological molecules and its putative role in carcinogenesis. © 2013 Colombo et al.; licensee BioMed Central Ltd.

Description of three new species of Hepatozoon (Apicomplexa, Hepatozoidae) from Rattlesnakes (Crotalus durissus terrificus) based on molecular, morphometric and morphologic characters

Hepatozoon spp. are commonly found infecting snakes. Since the latter are parasitized by diverse forms and data in the literature show divergence, we studied Hepatozoon spp. diversity on Crotalus durissus terrificus snakes using both molecular and morphological approaches. Naturally infected animals were employed. Blood was collected, blood smears were prepared and an aliquot was stored at -20. °C for DNA extraction. Five specimens of C. durissus terrificus were selected, each of them infected with one gamont type. Morphological and morphometric analyses of the found gamonts led to their grouping into three populations. For molecular characterization, seven oligonucleotide pairs that amplify distinct regions of rDNA gene were tested by adopting the PCR technique. Only the oligonucleotide pairs HepF300/Hep900 and HEMO1/HEMO2 were efficient in amplifying and distinguishing different isolates of Hepatozoon spp. from snakes. The better results were obtained when both oligonucleotide pairs were used in association. Based on the molecular and morphologic differences, three new species were proposed: Hepatozoon cuestensis sp. nov.; Hepatozoon cevapii sp. nov. and Hepatozoon massardii sp. nov. This is the first description of new Hepatozoon species from snakes, based on molecular characterization and morphological data, in South America. © 2013 Elsevier Inc.

Estruturação do Eumenine Mastroparano por dinâmica molecular em misturas de TFE-água

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Interação do peptídeo MP-I com micela de SDS em solução aquosa: um estudo por dinâmica molecular

Pós-graduação em Biofísica Molecular - IBILCE

Análise molecular em morcegos do gênero Artibeus Leach, 1821 (Chiroptera, Phyllostomidae) a partir de espécimes depositados em coleção científica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Marcadores citológicos no cariótipo de espécies de Leptodactylus (Amphibia, Anura, Leptodactylidae) analisado com técnicas de citogenética clássica e molecular

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Carreamento nasal de Staphylococcus aureus na população de Botucatu, São Paulo: prevalência, fatores de risco, resistência a antimicrobianos e epidemiologia molecular

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo da usinabilidade do polietileno de ultra alto peso molecular pela análise da força de corte

Pós-graduação em Engenharia Mecânica - FEB

Avanços recentes em biologia celular e molecular, questões éticas implicadas e sua abordagem em aulas de biologia no ensino médio: um estudo de caso

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização molecular e bioquímica de uma esterase macho-específica em Zaprionus indianus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ocorrência e caracterização molecular de Cryptosporidium spp. em aves selvagens brasileiras

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