A baculovirus-mediated strategy for full-length plant virus coat protein expression and purification

Background: Garlic production is severely affected by virus infection, causing a decrease in productivity and quality. There are no virus-free cultivars and garlic-infecting viruses are difficult to purify, which make specific antibody production very laborious. Since high quality antisera against plant viruses are important tools for serological detection, we have developed a method to express and purify full-length plant virus coat proteins using baculovirus expression system and insects as bioreactors. Results: In this work, we have fused the full-length coat protein (cp) gene from the Garlic Mite-borne Filamentous Virus (GarMbFV) to the 3′-end of the Polyhedrin (polh) gene of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). The recombinant baculovirus was amplified in insect cell culture and the virus was used to infect Spodoptera frugiperda larvae. Thus, the recombinant fused protein was easily purified from insect cadavers using sucrose gradient centrifugation and analyzed by Western Blotting. Interestingly, amorphous crystals were produced in the cytoplasm of cells infected with the recombinant virus containing the chimeric-protein gene but not in cells infected with the wild type and recombinant virus containing the hexa histidine tagged Polh. Moreover, the chimeric protein was used to immunize rats and generate antibodies against the target protein. The antiserum produced was able to detect plants infected with GarMbFV, which had been initially confirmed by RT-PCR. Conclusions: The expression of a plant virus full-length coat protein fused to the baculovirus Polyhedrin in recombinant baculovirus-infected insects was shown to produce high amounts of the recombinant protein which was easily purified and efficiently used to generate specific antibodies. Therefore, this strategy can potentially be used for the development of plant virus diagnostic kits for those viruses that are difficult to purify, are present in low titers or are present in mix infection in their plant hosts

Curso de atualização em informática para escritório para os discentes da Associação de Pais e Amidos dos Surdos (APAS) do Município de Concórdia - SC

Anais do 35º Seminário de Extensão Universitária da Região Sul - Área temática: EducaçãoNos últimos anos, a inserção de pessoas com deficiência (PCDs) no mercado de trabalho formal tem se expandido de forma notável. Especificamente no caso das pessoas surdas ou perda auditiva, essa estatística deve-se, entre outros fatores, a falta de qualificação profissional que capacite intelectualmente e que seja na sua língua, ou seja, na Língua Brasileira de Sinais - Libras. Muitas dessas pessoas surdas atuam no mercado de trabalho realizando tarefas que não exigem qualificação, constituindo uma mão de obra para atividades que desfavorece o conhecimento. Neste contexto, se objetiva um projeto de qualificação profissional e inserção no mercado de trabalho, porém a construção do mesmo, está divido em processos de ações, sendo: qualificação profissional e ações afirmativas junto as empresas. Construindo a primeira ação, o presente projeto propõe um curso de qualificação profissional de curta duração para capacitar essas pessoas surdas na utilização de softwares para escritório. O curso introduz estes discentes à um conjunto de sinais específicos na área de Informática. A programação do curso é composta pelas seguintes linhas: 1) Introdução ao conjunto de sinais específicos da área de Informática; 2) Operação básica de computadores e conceitos introdutórios à internet e serviços em nuvem; 3) Operação de processadores de texto: Microsoft Word, OpenOffice Writer e Documentos do Google; 4) Operação de planilhas eletrônicas: Microsoft Excel, OpenOffice Calc e Planilhas do Google; e 5) Operação de editores de apresentação de slides: Microsoft PowerPoint, OpenOffice Draw e Apresentações do Google. O curso está em andamento e até o momento foi completado a etapa 2. O público-alvo demonstrou compreensão no desenvolvimento dos assuntos e atividades, obtendo interação entre os docentes e discentes no decorrer de cada aula, principalmente com a troca de saberes com a Língua de Sinais à Língua Ora

Breast reconstruction with implant: creating a pocket with a reverse serratus anterior muscle flap

Introduction: Immediate breast reconstruction plays an important role in the treatment of breast cancer and relatively promotes patients' emotional and physical recovery. It may be difficult to cover the entire prosthesis with a muscle flap in single-stage breast reconstructions based on a permanent implant. This study aimed to present a muscle pocket for the implant using a reverse anterior serratus muscle flap associated with submuscular dissection of the pectoralis major muscle. Methods: This was a prospective study comprising 61 patients undergoing mastectomy followed by immediate reconstruction (74 reconstructions) with implant and anterior serratus muscle reverse flap associated with submuscular pectoralis dissection between January 2017 and July 2018. In this study, age, adjuvant and neoadjuvant therapies, implant volume, length of hospital stay, follow-up, and complications, including functional deficit and reconstruction failure, were analyzed. Results: The technique described was used to operate 74 patients with a mean age of 49.2 years. The volume of the implant varied from 200 to 500 cc, and the mean follow-up time was 14.9 months. Complications such as hematoma, suture dehiscence, skin flap necrosis, and implant extrusion were observed in 14 patients (18.9%). Conclusion: In most cases, breast reconstruction with an anterior serratus muscle reverse flap associated with submuscular dissection of the pectoralis major muscle allows the complete muscle coverage of the implant, reduces the occurrence of major surgical complications, and has a good aesthetic result

Molecular characterization of Hovenia dulcis-associated virus 1 (HDaV1) and 2 (HDaV2) : new tentative species within the order picornavirales

In a systematic field survey for plant-infecting viruses, leaf tissues were collected from trees showing virus-like symptoms in Brazil. After viral enrichment, total RNA was extracted and sequenced using the MiSeq platform (Illumina). Two nearly full-length picorna-like genomes of 9534 and 8158 nucleotides were found associated with Hovenia dulcis (Rhamnaceae family). Based upon their genomic information, specific primers were synthetized and used in RT-PCR assays to identify plants hosting the viral sequences. The larger contig was tentatively named as Hovenia dulcis-associated virus 1 (HDaV1), and it exhibited low nucleotide and amino acid identities with Picornavirales species. The smaller contig was related to insect-associated members of the Dicistroviridae family but exhibited a distinct genome organization with three non-overlapping open reading frames (ORFs), and it was tentatively named as Hovenia dulcis-associated virus 2 (HDaV2). Phylogenetic analysis using the amino acid sequence of RNA-dependent RNA polymerase (RdRp) revealed that HDaV1 and HDaV2 clustered in distinct groups, and both viruses were tentatively assigned as new members of the order Picornavirales. HDaV2 was assigned as a novel species in the Dicistroviridae family. The 50 ends of both viruses are incomplete. In addition, a nucleotide composition analysis (NCA) revealed that HDaV1 and HDaV2 have similarities with invertebrate-infecting viruses, suggesting that the primary host(s) of these novel virus species remains to be discovered

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost

COVID-19 symptoms at hospital admission vary with age and sex: results from the ISARIC prospective multinational observational study

Background: The ISARIC prospective multinational observational study is the largest cohort of hospitalized patients with COVID-19. We present relationships of age, sex, and nationality to presenting symptoms. Methods: International, prospective observational study of 60 109 hospitalized symptomatic patients with laboratory-confirmed COVID-19 recruited from 43 countries between 30 January and 3 August 2020. Logistic regression was performed to evaluate relationships of age and sex to published COVID-19 case definitions and the most commonly reported symptoms. Results: ‘Typical’ symptoms of fever (69%), cough (68%) and shortness of breath (66%) were the most commonly reported. 92% of patients experienced at least one of these. Prevalence of typical symptoms was greatest in 30- to 60-year-olds (respectively 80, 79, 69%; at least one 95%). They were reported less frequently in children (≤ 18 years: 69, 48, 23; 85%), older adults (≥ 70 years: 61, 62, 65; 90%), and women (66, 66, 64; 90%; vs. men 71, 70, 67; 93%, each P < 0.001). The most common atypical presentations under 60 years of age were nausea and vomiting and abdominal pain, and over 60 years was confusion. Regression models showed significant differences in symptoms with sex, age and country. Interpretation: This international collaboration has allowed us to report reliable symptom data from the largest cohort of patients admitted to hospital with COVID-19. Adults over 60 and children admitted to hospital with COVID-19 are less likely to present with typical symptoms. Nausea and vomiting are common atypical presentations under 30 years. Confusion is a frequent atypical presentation of COVID-19 in adults over 60 years. Women are less likely to experience typical symptoms than men

Senescência de inflorescências de copo-de-leite: influência de diferentes armazenamentos e procdimentos pós-colheita

O copo-de-leite é bastante apreciado para corte devido às suas características favoráveis para composição de arranjos florais. No entanto, a durabilidade de suas inflorescências é curta e informações sobre a fisiologia pós-colheita dessa espécie são restritas. Dessa forma, objetivou-se neste trabalho analisar as mudanças verificadas durante o processo de senescência de inflorescências de copo-de-leite em diferentes condições de armazenamento e procedimentos pós-colheita. Foram realizados experimentos para avaliar a necessidade ou não de água para imersão das bases das hastes durante o transporte em curtas distâncias (25 minutos), os melhores tipos de armazenamento e a influência de diferentes valores de pH da água utilizada para conservação das inflorescências em câmara fria. Não foi observada a necessidade da imersão das hastes de copo-de-leite em água para transporte, mas essas devem ser armazenadas em câmara fria (4°C) após a colheita a fim de retardar o processo de senescência. A utilização de diferentes níveis de pH não influenciou na qualidade pós-colheita das inflorescências. Independentemente dos tratamentos, o tempo de armazenamento de 10 dias foi excessivo, pois não houve durabilidade considerável após a retirada das inflorescências da câmara fria