Development of a Low Bias Method for Characterizing Viral Populations Using Next Generation Sequencing Technology

Background: With an estimated 38 million people worldwide currently infected with human immunodeficiency virus (HIV), and an additional 4.1 million people becoming infected each year, it is important to understand how this virus mutates and develops resistance in order to design successful therapies. Methodology/Principal Findings: We report a novel experimental method for amplifying full-length HIV genomes without the use of sequence-specific primers for high throughput DNA sequencing, followed by assembly of full length viral genome sequences from the resulting large dataset. Illumina was chosen for sequencing due to its ability to provide greater coverage of the HIV genome compared to prior methods, allowing for more comprehensive characterization of the heterogeneity present in the HIV samples analyzed. Our novel amplification method in combination with Illumina sequencing was used to analyze two HIV populations: a homogenous HIV population based on the canonical NL4-3 strain and a heterogeneous viral population obtained from a HIV patient's infected T cells. In addition, the resulting sequence was analyzed using a new computational approach to obtain a consensus sequence and several metrics of diversity. Significance: This study demonstrates how a lower bias amplification method in combination with next generation DNA sequencing provides in-depth, complete coverage of the HIV genome, enabling a stronger characterization of the quasispecies present in a clinically relevant HIV population as well as future study of how HIV mutates in response to a selective pressure

Data standardization of plant-pollinator interactions

Background: Animal pollination is an important ecosystem function and service, ensuring both the integrity of natural systems and human well-being. Although many knowledge shortfalls remain, some high-quality data sets on biological interactions are now available. The development and adoption of standards for biodiversity data and metadata has promoted great advances in biological data sharing and aggregation, supporting large-scale studies and science-based public policies. However, these standards are currently not suitable to fully support interaction data sharing. Results: Here we present a vocabulary of terms and a data model for sharing plant–pollinator interactions data based on the Darwin Core standard. The vocabulary introduces 48 new terms targeting several aspects of plant–pollinator interactions and can be used to capture information from different approaches and scales. Additionally, we provide solutions for data serialization using RDF, XML, and DwC-Archives and recommendations of existing controlled vocabularies for some of the terms. Our contribution supports open access to standardized data on plant–pollinator interactions. Conclusions: The adoption of the vocabulary would facilitate data sharing to support studies ranging from the spatial and temporal distribution of interactions to the taxonomic, phenological, functional, and phylogenetic aspects of plant–pollinator interactions. We expect to fill data and knowledge gaps, thus further enabling scientific research on the ecology and evolution of plant–pollinator communities, biodiversity conservation, ecosystem services, and the development of public policies. The proposed data model is flexible and can be adapted for sharing other types of interactions data by developing discipline-specific vocabularies of terms.Fil: Salim, José A. Universidade de Sao Paulo; BrasilFil: Saraiva, Antonio M.. Universidade de Sao Paulo; BrasilFil: Zermoglio, Paula Florencia. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Patagonia Norte. Instituto de Investigaciones En Recursos Naturales, Agroecologia y Desarrollo Rural. - Universidad Nacional de Rio Negro. Instituto de Investigaciones En Recursos Naturales, Agroecologia y Desarrollo Rural.; ArgentinaFil: Agostini, Kayna. Universidade Federal do São Carlos; BrasilFil: Wolowski, Marina. Universidade Federal de Alfenas; BrasilFil: Drucker, Debora P.. Empresa Brasileira de Pesquisa Agropecuaria (embrapa);Fil: Soares, Filipi M.. Universidade de Sao Paulo; BrasilFil: Bergamo, Pedro J.. Jardim Botânico do Rio de Janeiro; BrasilFil: Varassin, Isabela G.. Universidade Federal do Paraná; BrasilFil: Freitas, Leandro. Jardim Botânico do Rio de Janeiro; BrasilFil: Maués, Márcia M.. Empresa Brasileira de Pesquisa Agropecuaria (embrapa);Fil: Rech, Andre R.. Universidade Federal dos Vales do Jequitinhonha e Mucuri; BrasilFil: Veiga, Allan K.. Universidade de Sao Paulo; BrasilFil: Acosta, Andre L.. Instituto Tecnológico Vale; BrasilFil: Araujo, Andréa C. Universidade Federal do Mato Grosso do Sul; BrasilFil: Nogueira, Anselmo. Universidad Federal do Abc; BrasilFil: Blochtein, Betina. Pontificia Universidade Católica do Rio Grande do Sul; BrasilFil: Freitas, Breno M.. Universidade Estadual do Ceará; BrasilFil: Albertini, Bruno C.. Universidade de Sao Paulo; BrasilFil: Maia Silva, Camila. Universidade Federal Rural Do Semi Arido; BrasilFil: Nunes, Carlos E. P.. University of Stirling; BrasilFil: Pires, Carmen S. S.. Empresa Brasileira de Pesquisa Agropecuaria (embrapa);Fil: Dos Santos, Charles F.. Pontificia Universidade Católica do Rio Grande do Sul; BrasilFil: Queiroz, Elisa P.. Universidade de Sao Paulo; BrasilFil: Cartolano, Etienne A.. Universidade de Sao Paulo; BrasilFil: de Oliveira, Favízia F. Universidade Federal da Bahia; BrasilFil: Amorim, Felipe W.. Universidade Estadual Paulista Julio de Mesquita Filho; BrasilFil: Fontúrbel, Francisco E.. Pontificia Universidad Católica de Valparaíso; ChileFil: da Silva, Gleycon V.. Ministério da Ciência, Tecnologia, Inovações. Instituto Nacional de Pesquisas da Amazônia; BrasilFil: Consolaro, Hélder. Universidade Federal de Catalão; Brasi

The importance of Portuguese Continental Shelf Waters to Balearic Shearwaters revealed by aerial census

The Balearic shearwater Puffinus mauretanicus is one of the most threatened seabirds in the world. To evaluate the abundance and distribution of Balearic Shearwaters in Portuguese Continental Shelf Waters, during the post-breeding period when migrating birds are outside the Mediterranean Sea, we conducted 5 aerial surveys between 2010 and 2014 (21 survey days covering 62,716 km2). Following a line transect method, observers recorded a total of 181 Balearic Shearwaters sightings. Using Distance sampling software, we estimated an overall species abundance (2010–2014) of 10,182, ranging between 2338 in 2010 and 23,221 individuals in 2012. During the 2012 post-breeding period, the Portuguese Continental Shelf Waters were used by up to 96.8% of the latest migratory population assessment. Considering Balearic Shearwater estimates per sampling block, there was a preference for the North and Center sectors of the Portuguese coast (respectively, 7058 and 1366 individuals) where several SPAs were already designated. We computed the annual and overall habitat predictive models for Balearic Shearwaters using a maximum entropy algorithm on MaxEnt software. In all models, the Balearic shearwater distribution was best predicted by mean chlorophyll concentration. Balearic Shearwaters are mostly present in shallow shelf and coastal waters particularly in the widest portions of the continental shelf. These areas are strongly influenced by upwelling, which concurs with the chlorophyll concentration being the most important predicting variable. Portuguese Continental Shelf Waters are one of the most important post-breeding grounds to the Balearic ShearwaterPortuguese Wildlife Society and projects SafeSea EEA-Grants, FAME (Proj. 2009-1/089) and European Commission’s Life Programme (MarPro NAT/PT/00038). This study was also partly supported by the Portuguese Foundation for Science and Technology (FCT) with Grants SFRH/ BD/30240/2006 to M. Ferreira and SFRH/BD/32841/2006 to P. C. Rodrigues. C. Eira is supported by FCT through CESAM UID/AMB/50017/2013 co-funded by FCT/MEC and FEDER, within PT2020 and Compete 2020 and S. Monteiro is financed by a Grant (BPD/0043/AMB/50017) from UID/AMB/50017/2013. This work was also partially supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by FCT and by ERDF (COMPETE2020). The authors thank observers and airplane pilots who contributed to this workinfo:eu-repo/semantics/publishedVersio

Data standardization of plant–pollinator interactions

Background: Animal pollination is an important ecosystem function and service, ensuring both the integrity of natural systems and human well-being. Although many knowledge shortfalls remain, some high-quality data sets on biological interactions are now available. The development and adoption of standards for biodiversity data and metadata has promoted great advances in biological data sharing and aggregation, supporting large-scale studies and science-based public policies. However, these standards are currently not suitable to fully support interaction data sharing. Results: Here we present a vocabulary of terms and a data model for sharing plant–pollinator interactions data based on the Darwin Core standard. The vocabulary introduces 48 new terms targeting several aspects of plant–pollinator interactions and can be used to capture information from different approaches and scales. Additionally, we provide solutions for data serialization using RDF, XML, and DwC-Archives and recommendations of existing controlled vocabularies for some of the terms. Our contribution supports open access to standardized data on plant–pollinator interactions. Conclusions: The adoption of the vocabulary would facilitate data sharing to support studies ranging from the spatial and temporal distribution of interactions to the taxonomic, phenological, functional, and phylogenetic aspects of plant–pollinator interactions. We expect to fill data and knowledge gaps, thus further enabling scientific research on the ecology and evolution of plant–pollinator communities, biodiversity conservation, ecosystem services, and the development of public policies. The proposed data model is flexible and can be adapted for sharing other types of interactions data by developing discipline-specific vocabularies of termsinfo:eu-repo/semantics/publishedVersio

Small bowel enteroscopy - A joint clinical guideline from the spanish and portuguese small bowel study groups

The present evidence-based guidelines are focused on the use of device-assisted enteroscopy in the management of small-bowel diseases. A panel of experts selected by the Spanish and Portuguese small bowel study groups reviewed the available evidence focusing on the main indications of this technique, its role in the management algorithm of each indication and on its diagnostic and therapeutic yields. A set of recommendations were issued accordingly.Estas recomendações baseadas na evidência detalham o uso da enteroscopia assistida por dispositivo no manejo clínico das doenças do intestino delgado. Um conjunto de Gastrenterologistas diferenciados em patologia do intestino delgado foi selecionado pelos grupos de estudos Espanhol e Português de intestino delgado para rever a evidência disponível sobre as principais indicações desta técnica, o seu papel nos algoritmos de manejo de cada indicação e sobre o seu rendimento diagnóstico e terapêutico. Foi gerado um conjunto de recomendações pelos autores

Convection patterns gradients of non-living and living micro-entities in hydrogels

"Available online 28 October 2020"Inducing thermal gradients in two injected fluid systems results in the temporal formation of mixing conductive streams. If preserved through sol-gel transition, this mechanism can be used to drive and pattern non-living and living entities in mixed hydrogels. Interfaces are vital in nature, where gradients of non-living and living entities build distinct yet continuous integrated living tissues. However, the common tissue fabrication methodologies often result in dissimilar interfaces, lacking continuity through the interfaced engineered tissues. Thus, there is an urgent need for the fabrication of heterotypic but continuous engineered tissues with spatial control over biomimetic features. Here, we demonstrate the influence of gel injection temperature on the patterning of gradients of non-living and living entities. The experimental part was confirmed by numerical modelling, showing the formation of convective lines which spatially drive microscale microparticle and cells when different temperatures are applied in the sequential injection of two gels. Based on this finding, pure gellan gum (GG) and blended GG with methacrylated gelatin (GelMA) systems were used to program the formation of gradient features in hydrogels, such as microparticle and cells distribution patterns, polymeric bioactivity, degradation, controlled release, and stiffness. The correlation between gel injection temperature and gradients formation can be applied to tissue interface modelling, regeneration, drug release systems, and broader materials engineering fields.The authors are grateful for the Portuguese Foundation for Science and Technology (FCT) distinctions attributed to R. F. Canadas (SFRH/BD/92565/2013), D. Caballero (CEECIND/00352/2017), and to J. M. Oliveira (IF/00423/2012, IF/01285/2015). R. F. Canadas and J. M. Oliveira are also thankful to FCT, Fundo Europeu de Desenvolvimento Regional (FEDER), and Programa Operacional Competitividade e Internacionalização (POCI) for funding the B-Liver Project (PTDC/EMD-EMD/29139/2017). D. Caballero and S.C. Kundu also acknowledge the FCT for the financial support under the scope of the 2MATCH Project (02/SAICT/2017 – n° 028070). V. Brancato and S.C. Kundu acknowledge FCT project BREAST-IT (grant agreement: PTDC/BTM-ORG/28168/2017). The authors are also thankful to FCT for supporting the project Hierarchitech (M-ERA-NET/0001/2014). Pedro Patrício and Nuno A. M. Araújo acknowledge financial support from the FCT under Contracts no. PTDC/FIS-MAC/28146/2017 (LISBOA-01-0145-FEDER-028146) and UID/FIS/00618/2019. The authors acknowledge that this material and collaboration is based in part upon work supported by FLAD (2016/CON15/CAN6). This work was in part supported by European Research Council Grant agreement ERC-2012-ADG 20120216-321266 for project ComplexiTE

Small bowel enteroscopy - A joint clinical guideline from the spanish and portuguese small bowel study groups

The present evidence-based guidelines are focused on the use of device-assisted enteroscopy in the management of small-bowel diseases. A panel of experts selected by the Spanish and Portuguese small bowel study groups reviewed the available evidence focusing on the main indications of this technique, its role in the management algorithm of each indication and on its diagnostic and therapeutic yields. A set of recommendations were issued accordingly.Estas recomendações baseadas na evidência detalham o uso da enteroscopia assistida por dispositivo no manejo clínico das doenças do intestino delgado. Um conjunto de Gastrenterologistas diferenciados em patologia do intestino delgado foi selecionado pelos grupos de estudos Espanhol e Português de intestino delgado para rever a evidência disponível sobre as principais indicações desta técnica, o seu papel nos algoritmos de manejo de cada indicação e sobre o seu rendimento diagnóstico e terapêutico. Foi gerado um conjunto de recomendações pelos autores