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    A betabaculovirus encoding a gp64 homolog

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    Abstract\ud \ud Background\ud A betabaculovirus (DisaGV) was isolated from Diatraea saccharalis (Lepidoptera: Crambidae), one of the most important insect pests of the sugarcane and other monocot cultures in Brazil.\ud \ud \ud Results\ud The complete genome sequence of DisaGV was determined using the 454-pyrosequencing method. The genome was 98,392 bp long, which makes it the smallest lepidopteran-infecting baculovirus sequenced to date. It had a G + C content of 29.7 % encoding 125 putative open reading frames (ORF). All the 37 baculovirus core genes and a set of 19 betabaculovirus-specific genes were found. A group of 13 putative genes was not found in any other baculovirus genome sequenced so far. A phylogenetic analysis indicated that DisaGV is a member of Betabaculovirus genus and that it is a sister group to a cluster formed by ChocGV, ErelGV, PiraGV isolates, ClanGV, CaLGV, CpGV, CrleGV, AdorGV, PhopGV and EpapGV. Surprisingly, we found in the DisaGV genome a G protein-coupled receptor related to lepidopteran and other insect virus genes and a gp64 homolog, which is likely a product of horizontal gene transfer from Group 1 alphabaculoviruses.\ud \ud \ud Conclusion\ud DisaGV represents a distinct lineage of the genus Betabaculovirus. It is closely related to the CpGV-related group and presents the smallest genome in size so far. Remarkably, we found a homolog of gp64, which was reported solely in group 1 alphabaculovirus genomes so far.BTP received a scholarship from CAPES through the Graduate Program in\ud Biotechnology from the University of São Paulo and Butantan Institute. This project was partially funded by CNPq grant 481741/2008-9. We thank Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF)

    The Amazonia, a shared sea

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    Territoire en apparence vierge et immaîtrisable, l’Amazonie était aux yeux des premiers scientifiques un espace quasi déserté par l’homme. L’archéologie propose désormais une vision diamétralement opposée: villes, agriculture, cultures florissantes, etc. Les populations anciennes habitant la plus grande forêt du monde maîtrisaient parfaitement leur environnement, sans aucun doute mieux que leurs successeurs venus d’Europe.Território aparentemente virgem e desconhecido, a Amazônia era aos olhos dos primeiros cientistas um espaço quase desabitado pelo homem. A Arqueologia propõe agora uma visão diametralmente oposta: cidades, agricultura, culturas florescentes. As populações antigas que habitavam a maior floresta do mundo conheciam e dominavam seu ambiente, certamente melhor do que seus sucessores advindos da Europa.Territory apparently virgin and unknown, the Amazon was the sight of the first scientists an almost uninhabited space by man. Archaeology is now proposing a diametrically opposite view: cities, agriculture, flourishing cultures. The ancient people who inhabited the world's forest knew and dominated their environment, certainly better than his successors coming from Europe

    Fatal factitious Cushing syndrome (Münchhausen’s syndrome) in a patient with macroprolactinoma and silent corticotrophinoma: case report and literature review

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    Abstract Münchhausen’s syndrome (MS) is a chronic factitious disorder characterized by the intentional production of clinical symptoms without external incentive. One type of MS is factitious Cushing syndrome, an extremely rare clinical situation in which the diagnosis is challenging mainly due to interference of the exogenous medication in cortisol immunoassays. We described a 26-year-old woman who was originally diagnosed with a macroprolactinoma and during follow-up developed clinical and laboratorial hypercortisolism. A transsphenoidal surgery was performed and immunohistochemistry revealed positive and diffuse staining for both hormones. Four years later, her hypercortisolism recurred and the confirmation of factitious Cushing syndrome was delayed due to conflicting laboratorial results. There are few cases in the literature of factitious Cushing syndrome, and only one had a fatal outcome. The diagnosis of this condition is complex and includes cyclic Cushing syndrome in the differential diagnosis. These patients have high morbidity and increased mortality risk and are likely to have other psychiatric disorders. Prednisone was identified as the culprit in the majority of the cases

    Modularity and evolutionary constraints in a baculovirus gene regulatory network

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    Abstract Background The structure of regulatory networks remains an open question in our understanding of complex biological systems. Interactions during complete viral life cycles present unique opportunities to understand how host-parasite network take shape and behave. The Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) is a large double-stranded DNA virus, whose genome may encode for 152 open reading frames (ORFs). Here we present the analysis of the ordered cascade of the AgMNPV gene expression. Results We observed an earlier onset of the expression than previously reported for other baculoviruses, especially for genes involved in DNA replication. Most ORFs were expressed at higher levels in a more permissive host cell line. Genes with more than one copy in the genome had distinct expression profiles, which could indicate the acquisition of new functionalities. The transcription gene regulatory network (GRN) for 149 ORFs had a modular topology comprising five communities of highly interconnected nodes that separated key genes that are functionally related on different communities, possibly maximizing redundancy and GRN robustness by compartmentalization of important functions. Core conserved functions showed expression synchronicity, distinct GRN features and significantly less genetic diversity, consistent with evolutionary constraints imposed in key elements of biological systems. This reduced genetic diversity also had a positive correlation with the importance of the gene in our estimated GRN, supporting a relationship between phylogenetic data of baculovirus genes and network features inferred from expression data. We also observed that gene arrangement in overlapping transcripts was conserved among related baculoviruses, suggesting a principle of genome organization. Conclusions Albeit with a reduced number of nodes (149), the AgMNPV GRN had a topology and key characteristics similar to those observed in complex cellular organisms, which indicates that modularity may be a general feature of biological gene regulatory networks

    Behavior of adhesion forces of the aqueous-based\ud polychloroprene adhesive magnetically\ud conditioned

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    This research is an innovation proposal for magnetic conditioning of the aqueous-based polychloroprene contact adhesive with the purpose of increasing the adherence capacity between the adhesive and the substrate. Today, the aqueous-based polychloroprene contact adhesive presents an adhesion capacity variation between 1.15 and up to 2.75 kgf/cm2. On the other hand, the solvent-based polychloroprene adhesives support average tensions of shearing of 3.8 kgf/cm2. To promote an increase in adhesion to the aqueous-based polychloroprene contact adhesive, we formulated one adhesive using a nano-additive, carbon dioxide as catalyst, and a magnetic conditioning process before the phase of application on the substrates. The results obtained show an average increase of 292% in the shearing tension of the adhesive magnetically conditioned when compared with an adhesive of same formulation without the magnetic conditioning and when compared to the commercial aqueous-based polichloroprene adhesive, the increase reaches 122%

    Atraumatic Restorative Treatment compared to the Hall Technique for occluso-proximal cavities in primary molars: study protocol for a randomized controlled trial

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    Abstract\ud \ud Background\ud In many parts of the world, school-age children have high dental treatment needs; however, there is often low, or no, dental care provision. Although Atraumatic Restorative Treatment (ART) was developed to address this, its survival rate in occluso-proximal lesions is low. An alternative, the Hall Technique (HT) has shown better relative outcomes for occluso-proximal lesions, but has not been directly compared to ART or tested in field settings. This trial will compare ART and the HT for the most clinically- and cost-effective strategy for managing occluso-proximal lesions in primary molars, in a school setting, using low-technology and child-friendly dental techniques.\ud \ud \ud Methods/Design\ud This two-arm, parallel group, patient-randomized controlled, superiority trial will have treatment provided in schools. Schoolchildren (n = 124, age 6–8) with at least one occluso-proximal carious primary molar lesion will have random allocation to treatment with ART or HT. Baseline measures and outcome data will be assessed through participant report, clinical examination and parent report/questionnaires. The primary outcome is survival rate, a composite measure of absence of Minor Failures (a defect in the restoration/crown, but not interfering with tooth health) and Major Failures (signs or symptoms of irreversible pulp damage, such as dental fistula/abscess, tooth fracture or failures that cannot be repaired). Secondary outcomes are: (1) child-reported discomfort, (2) childrens’ and (3) parents’ concerns around dental appearance and (4) acceptability of treatments, (5) occlusal-vertical dimensions (OVD) changes, (6) plaque index, (7) gingival health, (8) decayed, missing, filled teeth in permanent teeth (DMFT)/decayed, missing, filled teeth in primary teeth (dmft), (9) oral health-related-quality of life, reported by children and parents/caregivers, (10) the incremental cost-effectiveness, and (11) operator effect. A trained and calibrated examiner will evaluate the treated teeth after 1 week, then 1, 6, 12, 24 and 36 months post treatment. Kaplan-Meier and Cox regression tests will be used to investigate the primary outcome. The Mann-Whitney or t test, Friedman test, paired t test or Wilcoxon test and Ordinal Logistic Regression Analysis will be used to analyze the secondary outcomes.\ud \ud \ud Discussion\ud The results of this trial will support decision-making by clinicians and policy-makers for managing occluso-proximal lesions in settings with constrained resources and limited dental access.\ud \ud \ud Trial registration\ud www.clinicaltrials.gov, \ud NCT02569047\ud \ud , registered 5 October 2015.The authors would like to declare our gratitude for the Brazilians Funding\ud agencies (CNPq, FAPESP and CAPES) for providing scholarships for some of\ud the staff involved in this study. Also, GC Europe and 3 M-ESPE donated some\ud of the materials (glass ionomer cement, crowns and orthodontic separators).\ud All other materials and instruments were not from donations. Staff time and\ud resources for the trial has been supported through the University of São\ud Paulo (USP), the Academic Centre for Dentistry Amsterdam (ACTA) and the\ud University of Dundee

    Synthesis of Ca-doped spinel by ultrasonic spray pyrolysis

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    MgAl2O4 is a stable catalyst support with potential for replacing gamma-alumina in several applications. However, synthesis of magnesium spinel requires elevated temperatures to avoid phase separation (in MgO and Al2O3) at low temperatures, leading to coarsening and reduction of active surface area. In this work, nano CaO-doped and undoped magnesium aluminate were successfully prepared by Ultrasonic Spray Pyrolysis (USP), using a simple adapted experimental set-up operating at 1100 °C. During the process, the particles stay at high temperatures for a short period of time, allowing phase stability and limited coarsening. The influence of calcium oxide on the particles morphology and structure was investigated via X-ray diffraction, N2 adsorption, X-ray fluorescence, scanning electron microscopy and transmission electron microscopy. The spinel nanopowders were obtained as spherical porous agglomerates of ~1 µm. The resulting powder showed low crystallite sizes in the 5–10 nm range and high specific surface area from 110.0 to 76.6 m2 g−1.CNP

    Spatial genetic structure, genetic diversity and pollen dispersal in a harvested population of Astrocaryum aculeatum in the Brazilian Amazon

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    Abstract\ud \ud Background\ud \ud Astrocaryum aculeatum is a palm tree species native to the tropical regions of South America, exploited commercially by local farmers for the pulp extracted from its fruits. The objective of this research was to compare the genetic diversity between adult plants and seedlings from open-pollinated seeds, quantify the pollen flow and dispersal, the spatial genetic structure, and the effective size of a population that has been continuously harvested for its fruits. The study was carried out in a natural population of A. aculeatum distributed over approximately 8 ha in the State of Amazonas (Brazil), separated by 400 m from the closest neighboring population. In total, 112 potential pollen donors, 12 mother plants and 120 offspring were mapped and genotyped.\ud \ud \ud Results\ud Genetic diversity was high for parents and the offspring. The fixation indexes for adults (F = -0.035) and offspring (F = -0.060) were negative and not significant. A significant spatial genetic structure was detected for the adult plants (up to the distance of 45 m) indicating short-distance seed dispersal. Paternity analysis detected 9.2 % of pollen immigration and the average distance of pollination within the population was 81 m. The average effective pollination neighborhood area between plants was 1.51 ha.\ud \ud \ud Conclusions\ud Our results indicate that substantial introduction of new alleles has occurred in the population through pollen immigration, contributing to the maintenance of genetic diversity. Conservation efforts aimed at maintaining the gene pool of the current population or establishing new populations should utilize offspring from mother plants selected to be spaced by at least 50 m to prevent collecting seeds from relatives.The authors thank the Fundação de Amparo à Pesquisa do Estado do Amazonas\ud (FAPEAM) for a scholarship to SLFR and supporting the project "Publicação de\ud Artigos Científicos em Genética e Melhoramento de Plantas" - PAPAC (process\ud 062.02685 / 2014); the Conselho Nacional de Desenvolvimento Científico e\ud Tecnológico (CNPq) for scholarships to AMS, MTGL, EAV; the Embrapa Amazônia\ud Ocidental through the project “Pesquisa, desenvolvimento e inovação em\ud palmáceas para a produção de óleo e aproveitamento econômico de coprodutos\ud e resíduos – PROPALMA” (process 01.10.0343.00); and the Coordenação\ud de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) - Programa de\ud Excelência Acadêmica (Proex) of the Genetics Department of ESALQ-USP for\ud supporting this research; and the Laboratório Temático de Biologia Molecular -\ud Instituto Nacional de Pesquisas da Amazônia (LTBM – INPA) for its facilities. The\ud authors also thank Mrs. Angela Mary Wray for the great contribution in the English\ud language and two anonymous reviewers for the excellent suggestions in the\ud manuscript

    Genome-wide metabolic (re-) annotation of Kluyveromyces lactis

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    Abstract Background Even before having its genome sequence published in 2004, Kluyveromyces lactis had long been considered a model organism for studies in genetics and physiology. Research on Kluyveromyces lactis is quite advanced and this yeast species is one of the few with which it is possible to perform formal genetic analysis. Nevertheless, until now, no complete metabolic functional annotation has been performed to the proteins encoded in the Kluyveromyces lactis genome. Results In this work, a new metabolic genome-wide functional re-annotation of the proteins encoded in the Kluyveromyces lactis genome was performed, resulting in the annotation of 1759 genes with metabolic functions, and the development of a methodology supported by merlin (software developed in-house). The new annotation includes novelties, such as the assignment of transporter superfamily numbers to genes identified as transporter proteins. Thus, the genes annotated with metabolic functions could be exclusively enzymatic (1410 genes), transporter proteins encoding genes (301 genes) or have both metabolic activities (48 genes). The new annotation produced by this work largely surpassed the Kluyveromyces lactis currently available annotations. A comparison with KEGG’s annotation revealed a match with 844 (~90%) of the genes annotated by KEGG, while adding 850 new gene annotations. Moreover, there are 32 genes with annotations different from KEGG. Conclusions The methodology developed throughout this work can be used to re-annotate any yeast or, with a little tweak of the reference organism, the proteins encoded in any sequenced genome. The new annotation provided by this study offers basic knowledge which might be useful for the scientific community working on this model yeast, because new functions have been identified for the so-called metabolic genes. Furthermore, it served as the basis for the reconstruction of a compartmentalized, genome-scale metabolic model of Kluyveromyces lactis, which is currently being finished

    Evaluation of real-time PCR assay to detect Schistosoma mansoni infections in a low endemic setting

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    Abstract Background Schistosomiasis constitutes a major public health problem, and 200 million people are estimated to be infected with schistosomiasis worldwide. In Brazil, schistosomiasis has been reported in 19 states, showing areas of high and medium endemicity and a wide range of areas of low endemicity (ALE). Barra Mansa in Rio de Janeiro state has an estimated prevalence of 1%. ALE represent a new challenge for the helminth control because about 75% of infected individuals are asymptomatic and infections occur with a low parasite load (<100 eggs per gram of feces), causing a decrease in sensitivity of stool parasitological techniques, which are a reference for the laboratory diagnosis of this helminth. The objective of this study was to evaluate the performance of a TaqMan quantitative polymerase chain reaction (qPCR) technique in serum and feces DNA samples using the techniques of Kato-Katz (KK), Hoffman, Pons and Janer (HH) as references, during an epidemiological survey using fecal samples and sera from randomized residents from an ALE. Methods A cross-sectional study conducted from April to December 2011 using a probabilistic sampling that collected 572 fecal and serum samples. The laboratory diagnostic techniques used were: KK, HH and qPCR (feces and serum). Results We obtained the following results using the different diagnostic techniques: KK and HH, 0.9% (n =5); qPCR-feces, 9.6% (n =55); and qPCR-serum, 1.4% (n =8). The qPCR-feces presented the highest positivity, whereas the techniques of HH and KK were the least sensitive to detect infections (0.8%). Compared to HH and KK, qPCR-feces showed a statistically significant difference in positivity (p <0.05), although with poor agreement. Conclusion The positivity rate presented by the qPCR approach was far higher than that obtained by parasitological techniques. The lack of adequate surveillance in ALE of schistosomiasis indicates a high possibility of these areas being actually of medium and high endemicity. This study presents a control perspective, pointing to the possibility of using combined laboratory tools in the diagnosis of schistosomiasis in ALE

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