Análise estrutural “in-sílico” de uma proteína glutationa-stransferase de arroz, com potencial aplicação na resistência de plantas e herbicidas

O arroz é uma cultura de grande interesse e importância mundial, tanto na esfera agroindustrial quanto nutricional, sendo inclusive intitulada fonte primária do layout nutricional de países asiáticos em desenvolvimento, mas estudos para minimizar os problemas intrínsecos ao cultivo desta cerealífera ainda são escassos. Sabe-se que a superfamília de enzimas glutationa s-transferases (GSTs) confere em arroz (Oryza sativa) proteção a estresse biótico e abiótico e que sua estrutura tridimensional, que em estudos de natureza proteômica servem para identificar a funcionalidade da proteína, ainda não foi elucidada. Nesse contexto, o presente trabalho busca contribuir para a solução desta problemática, cujo desenvolvimento se deu numa sequência de três etapas gerais, complementares entre si, quais sejam: construção de um modelo teórico válido para a proteína glutationa-S-transferas classe Tau4 de Oryza sativa (OsGSTU4) em plataforma computacional, por meio de modelagem por homologia. Para tato, usou-se como template (molde) uma sequência de uma GSTs classe Tau 4 de trigo (Aegilops tauschii) (AtGSTU4) por ser uma sequência ortóloga a OsGSTU4; estudo da natureza funcional da proteína modelada, em que foi simulado o processo de ligação por meio do programa AUTODOCK 1.5.6, entre a proteína e ligantes (herbicidas Bentazon e Metsulfuron), no qual foi verificado, como resultado, processo de conjugação entre a proteína e o herbicida Metuslfuron, evidenciando o processo de desintoxicação do herbicida por meio da rota metabólica da proteína OsGSTU4; análise da dinâmica (atomística) molecular por meio de simulação computacional, de forma que possibilitou observar, numa fração de tempo, os movimentos de grande amplitude por meio de Análise de Modos Normais (NMA) pelo servidor ÉlNemo e movimentos de grande e média amplitude por meio do pacote de programa GROMACS. O conjunto de técnicas desta última etapa permitiu avaliar a cinética estrutural ao longo de toda a cadeia da proteína e evidenciar a estabilidade da estrutura, revelando que o modelo obtido pela primeira etapa da metodologia apresenta-se em seu estado nativo. Conclui-se que o modelo obtido apresenta alta confiabilidade estrutural, estabilidade físico-química e revela potencial de ação catalítica para os herbicidas estudados. Além disto, os resultados forneceram dados que podem auxiliar posteriores pesquisas visando o desenvolvimento de tecnologias voltadas para outras culturas agronomicamente importantes, como o arroz.Rice is one of the most important crops in the world, and this includes the fact that’s being called the primary source of nutritional layout in developing countries in Asian developing countries, however there are few studies of agro-industrial and technological bias for rice. It is known that the superfamily of s-transferase enzymes glutathione (GST) confer rice (oryzae Sativa) protection to biotic and abiotic stress and its three dimensional structure, which in proteomics studies serve to identify the protein functionality, that have not been elucidated. In this context, this work aims to contribute to solution of this problem and has been developed in a sequence of three general complementary steps, which are: construction of a valid theoretical model for glutathione-S-transferas protein Tau4 class oriza Sativa (OsGSTU4) in computational platform through homology modeling, for which was used a wheat template sequence of one GSTs Tau 4 Aegilops tauschii (AtGSTU4) ortholog sequence to the OsGSTU4; Study of the functional nature of the patterned protein, which the binding process between the protein and ligands (herbicides, Bentazon and Metsulfuron), that was simulated in AutoDock 1.5.6 program, which was found as a result of the conjugation process between protein and Metuslfuron herbicide, showing the herbicide detoxification process by metabolic route OsGSTU4 protein; dynamic (atomistic) analysis through computer simulation enabling to observe, in a fraction of time, the movements of large amplitude by Normal Mode Analysis (NMA) by ÉlNemo server and large movements and average amplitude through GROMACS program package. The set of techniques of this latter step allows to evaluate the structural kinetic throughout the protein chain and show the structure stability, revealing that the model obtained by the first step of the methodology presented itself in its native state. We conclude that the model obtained in the first methodology step shows high structural reliability, physical and chemical stability and reveals catalytic action potential for the studied herbicides. In addition, the results provide data that can aid further research aimed at developing technologies for agronomically important crops such as rice

Molecular Docking for Detoxifying Enzyme Studies

In this chapter, we pointed some relevant results obtained by protein-ligand docking simulations in the context of insecticide and herbicide resistance performed by glutathione S-transferases (GSTs), a detoxifying superfamily enzyme. We present here some in silico evidences of GST binding against chemical insecticides in the malaria and dengue vectors (Anopheles gambiae and Aedes aegypti mosquitoes) and against chemical herbicides used on rice (Oryza sativa) culture. Our findings suggest that some members from epsilon class (GSTE2, GSTE5) can metabolize some insecticide compounds and that a tau class member (GSTU4) can metabolize some herbicides. The results reinforce the importance of docking studies for enzyme activity comprehension. These information can allow in the future the implementation of new strategies for mosquito control and herbicide management on rice culture through biotechnological improvements designed to specific GST targets. Induced mutations on catalytic binding sites of GSTU4 could improve rice herbicide resistance and minimize produce damage, while rational compounds can be designed to inhibit GSTE members to decline insecticide resistance on mosquito control. In both cases, biotechnological tools could be developed focusing on GSTs that would reduce environmental impact by the use of insecticide and herbicide

Análise comparativa entre variações no custo de uma estrutura de concreto armado dimensionada com alterações na locação dos pilares

In the field of civil construction each step requires the elaboration of a specific project, whichare fundamental for the quality of service. In this sense, the present work is focused on the area of structures, since the needs of the Brazilian scenario in cost reduction, this study aims to perform a comparative analysis of a reinforced concrete structure, meeting the recommendations of ABNT NBR 6118 (2014). The building consists of four residential floors, located in the region of Teófilo Otoni, with an area of about 130 m² on each floor. Variations are made in the locations of the structural elements, verifying and analyzing the financial impacts, in order to identify the percentage of increase and its relevance in the final value. In the design the TQS software was used, for the quantification of costs it was adopted as base the data of the National Construction Costs and Indexes Research System (SINAPI) for the region of Minas Gerais. From the budget spreadsheets with the final costs, they obtained considerable differences, presenting a savings percentage of approximately 10%, between the worst case and the cheapest. Thus, it is concluded that the final price changes affect and show that there is the possibility of generating a cheaper structure, following the requirements of the standards and the safety of users.En el campo de la construcción civil, cada paso requiere la elaboración de un proyecto específico, que es fundamental para la calidad del servicio, en este sentido, el presente trabajo se centra en el área de las estructuras, ya que, en vista de las necesidades del escenario brasileño en la reducción de costos, Este estudio tiene como objetivo realizar un análisis comparativo de una estructura de hormigón armado, cumpliendo con las recomendaciones de ABNT NBR 6118 (2014). La construcción y el compostaje de cuatro aceras residenciales, ubicadas en la región de Teófilo Otoni, con un área de aproximadamente 130 m² en cada acera, se realizan variaciones en las ubicaciones de los elementos estructurales , verificando y analizando los impactos financieros para identificar o porcentaje de aumento y relevancia, no el valor final. En el diseño se utilizó el software TQS, para la cuantificación de costos se adoptó como base los datos del Sistema Nacional de Encuesta de Costos e Índices de Construcción (SINAPI) para la región de Minas Gerais. De acuerdo con los planes de la organización con las contribuciones finales, considerando un porcentaje de ahorro de aproximadamente 10% entre el más bajo y el más barato. Por lo tanto, se concluye que las variaciones finales en el precio han impactado y muestran que existe la posibilidad de generar una estructura más barata, siguiendo los requisitos de las normas y respetando la seguridad de los usuarios.No campo da construção civil cada etapa exige a elaboração de um projeto específico, que são fundamentais para a qualidade do serviço, nesse sentido o presente trabalho é voltado para a área de estruturas, visto que diante as necessidades do cenário brasileiro na redução dos custos, este estudo tem como objetivo realizar uma análise comparativa de uma estrutura de concreto armado, atendendo as recomendações da ABNT NBR 6118 (2014). A edificação é composta por quatro pavimentos residenciais, situado na região de Teófilo Otoni, com área de cerca de 130 m² em cada pavimento, são feitas variações nas locações dos elementos estruturais, constatando e analisando os impactos financeiros, a fim de identificar o percentual de aumento e sua relevância no valor final. No dimensionamento utilizou-se o software TQS, para a quantificação dos custos adotou-se como base os dados do Sistema Nacional de Pesquisa de Custos e Índices da Construção (SINAPI) para região de Minas Gerais. A partir das planilhas de orçamento com os custos finais, obtiveram diferenças consideráveis, apresentando um percentual de economia em aproximadamente 10%, entre o pior caso e o mais barato. Com isso conclui-se que as variações finais no preço impactaram e mostram que existe a possibilidade de gerar uma estrutura mais barata, seguindo os requisitos das normas e presando pela segurança dos usuários

Análise comparativa entre variações no custo de uma estrutura de concreto armado dimensionada com alterações na locação dos pilares

In the field of civil construction each step requires the elaboration of a specific project, whichare fundamental for the quality of service. In this sense, the present work is focused on the area of structures, since the needs of the Brazilian scenario in cost reduction, this study aims to perform a comparative analysis of a reinforced concrete structure, meeting the recommendations of ABNT NBR 6118 (2014). The building consists of four residential floors, located in the region of Teófilo Otoni, with an area of about 130 m² on each floor. Variations are made in the locations of the structural elements, verifying and analyzing the financial impacts, in order to identify the percentage of increase and its relevance in the final value. In the design the TQS software was used, for the quantification of costs it was adopted as base the data of the National Construction Costs and Indexes Research System (SINAPI) for the region of Minas Gerais. From the budget spreadsheets with the final costs, they obtained considerable differences, presenting a savings percentage of approximately 10%, between the worst case and the cheapest. Thus, it is concluded that the final price changes affect and show that there is the possibility of generating a cheaper structure, following the requirements of the standards and the safety of users.No campo da construção civil cada etapa exige a elaboração de um projeto específico, que são fundamentais para a qualidade do serviço, nesse sentido o presente trabalho é voltado para a área de estruturas, visto que diante as necessidades do cenário brasileiro na redução dos custos, este estudo tem como objetivo realizar uma análise comparativa de uma estrutura de concreto armado, atendendo as recomendações da ABNT NBR 6118 (2014). A edificação é composta por quatro pavimentos residenciais, situado na região de Teófilo Otoni, com área de cerca de 130 m² em cada pavimento, são feitas variações nas locações dos elementos estruturais, constatando e analisando os impactos financeiros, a fim de identificar o percentual de aumento e sua relevância no valor final. No dimensionamento utilizou-se o software TQS, para a quantificação dos custos adotou-se como base os dados do Sistema Nacional de Pesquisa de Custos e Índices da Construção (SINAPI) para região de Minas Gerais. A partir das planilhas de orçamento com os custos finais, obtiveram diferenças consideráveis, apresentando um percentual de economia em aproximadamente 10%, entre o pior caso e o mais barato. Com isso conclui-se que as variações finais no preço impactaram e mostram que existe a possibilidade de gerar uma estrutura mais barata, seguindo os requisitos das normas e presando pela segurança dos usuários.En el campo de la construcción civil, cada paso requiere la elaboración de un proyecto específico, que es fundamental para la calidad del servicio, en este sentido, el presente trabajo se centra en el área de las estructuras, ya que, en vista de las necesidades del escenario brasileño en la reducción de costos, Este estudio tiene como objetivo realizar un análisis comparativo de una estructura de hormigón armado, cumpliendo con las recomendaciones de ABNT NBR 6118 (2014). La construcción y el compostaje de cuatro aceras residenciales, ubicadas en la región de Teófilo Otoni, con un área de aproximadamente 130 m² en cada acera, se realizan variaciones en las ubicaciones de los elementos estructurales , verificando y analizando los impactos financieros para identificar o porcentaje de aumento y relevancia, no el valor final. En el diseño se utilizó el software TQS, para la cuantificación de costos se adoptó como base los datos del Sistema Nacional de Encuesta de Costos e Índices de Construcción (SINAPI) para la región de Minas Gerais. De acuerdo con los planes de la organización con las contribuciones finales, considerando un porcentaje de ahorro de aproximadamente 10% entre el más bajo y el más barato. Por lo tanto, se concluye que las variaciones finales en el precio han impactado y muestran que existe la posibilidad de generar una estructura más barata, siguiendo los requisitos de las normas y respetando la seguridad de los usuarios

Análise da distribuição de focos de calor no município de Novo Progresso, Pará

Fires are a major cause of damage to forest ecosystems. There are several ways of detecting hotspots. However, for countries with a vast territory as Brazil, the monitoring by satellite images is more efficient and cheaper. The aim of this study was to analyze the distribution of hotspots in the county of Novo Progresso, Pará, from 2010 to 2015 through data monitoring performed by reference satellites of the National Institute for Space Research. After organizing the information obtained through the Hotspots Database, analyses of socioeconomic and environmental factors that enhance the appearance of fire outbreaks were made with the assistance of scientific literature. It was observed that the outbreaks are concentrated in the northeastern area of the county along the BR-163, presenting higher occurrence in August and September, which may be related to the drought period of the city.Os incêndios são um dos principais causadores de danos aos ecossistemas florestais. Existem diversas formas de detecção dos focos de calor. Entretanto, para países com grande extensão territorial como o Brasil, é mais eficiente e barato o monitoramento por imagens de satélite. A partir disso, objetivou-se analisar a distribuição de focos de calor no município de Novo Progresso, Pará no período de 2010 a 2015, por meio de dados do monitoramento realizado por satélites de referência pertencentes ao Instituto Nacional de Pesquisas Espaciais. Após a organização das informações obtidas através do Banco de Dados de Queimadas, foram feitas análises dos fatores socioeconômicos e ambientais que intensificam o surgimento de focos de incêndios com o auxílio da literatura científica. Observou-se que os focos se concentraram na região nordeste do munícipio, ao longo da BR-163, apresentando maior ocorrência nos meses de agosto e setembro, o que pode estar relacionado com o período de estiagem do município

Plant Antimicrobial Peptides as Potential Tool for Topic Treatment of Hidradenitis Suppurativa

Among chronic skin autoinflammatory diseases, Hidradenitis Suppurativa (HS) stands out for its chronicity, highly variable condition, and profound impact on the patients’ quality of life. HS is characterized by suppurative skin lesions in diverse body areas, including deep-seated painful nodules, abscesses, draining sinus, and bridged scars, among others, with typical topography. To date, HS is considered a refractory disease and medical treatments aim to reduce the incidence, the infection, and the pain of the lesions. For this purpose, different classes of drugs, including anti-inflammatory molecules, antibiotics and biological drugs are being used. Antimicrobial peptides (AMPs), also called defense peptides, emerge as a new class of therapeutic compounds, with broad-spectrum antimicrobial action, in addition to reports on their anti-inflammatory, healing, and immunomodulating activity. Such peptides are present in prokaryotes and eukaryotes, as part of the innate eukaryotic immune system. It has been proposed that a deregulation in the expression of AMPs in human epithelial tissues of HS patients may be associated with the etiology of this skin disease. In this scenario, plant AMPs stand out for their richness, diversity of types, and broad antimicrobial effects, with potential application for topical systemic use in patients affected by HS.This work was supported by a Biomolecular Analyses for Tailored Medicine in AcneiNversa (BATMAN) project, funded by ERA PerMed (JTC_2018), by a grant from the Institute for Maternal and Child Health IRCCS ‘Burlo Garofolo’/Italian Ministry of Health (RC16/2018), and by a Starting Grant (SG-2019-12369421) funded by Italian Ministry of Health

Lipid transfer proteins (Ltps)- structure, diversity and roles beyond antimicrobial activity

Lipid transfer proteins (LTPs) are among the most promising plant-exclusive antimicrobial peptides (AMPs). They figure among the most challenging AMPs from the point of view of their structural diversity, functions and biotechnological applications. This review presents a current picture of the LTP research, addressing not only their structural, evolutionary and further predicted functional aspects. Traditionally, LTPs have been identified by their direct isolation by biochemical techniques, whereas omics data and bioinformatics deserve special attention for their potential to bring new insights. In this context, new possible functions have been identified revealing that LTPs are actually multipurpose, with many additional predicted roles. Despite some challenges due to the toxicity and allergenicity of LTPs, a systematic review and search in patent databases, indicate promising perspectives for the biotechnological use of LTPs in human health and also plant defense.The research received financial support of FACEPE (Fundação de Amparo à Pesquisa do Estado de Pernambuco, Brazil) CNPq (Conselho Nacional de Desenvolvimento Científico Tecnológico, no. 313581/2020-7, 442019/2019-0, and 433931/2018-3) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, no. 88881.507105/2020-01) throught research funding and fellowships grants

Prospective observational cohort study on grading the severity of postoperative complications in global surgery research

Background The Clavien–Dindo classification is perhaps the most widely used approach for reporting postoperative complications in clinical trials. This system classifies complication severity by the treatment provided. However, it is unclear whether the Clavien–Dindo system can be used internationally in studies across differing healthcare systems in high- (HICs) and low- and middle-income countries (LMICs). Methods This was a secondary analysis of the International Surgical Outcomes Study (ISOS), a prospective observational cohort study of elective surgery in adults. Data collection occurred over a 7-day period. Severity of complications was graded using Clavien–Dindo and the simpler ISOS grading (mild, moderate or severe, based on guided investigator judgement). Severity grading was compared using the intraclass correlation coefficient (ICC). Data are presented as frequencies and ICC values (with 95 per cent c.i.). The analysis was stratified by income status of the country, comparing HICs with LMICs. Results A total of 44 814 patients were recruited from 474 hospitals in 27 countries (19 HICs and 8 LMICs). Some 7508 patients (16·8 per cent) experienced at least one postoperative complication, equivalent to 11 664 complications in total. Using the ISOS classification, 5504 of 11 664 complications (47·2 per cent) were graded as mild, 4244 (36·4 per cent) as moderate and 1916 (16·4 per cent) as severe. Using Clavien–Dindo, 6781 of 11 664 complications (58·1 per cent) were graded as I or II, 1740 (14·9 per cent) as III, 2408 (20·6 per cent) as IV and 735 (6·3 per cent) as V. Agreement between classification systems was poor overall (ICC 0·41, 95 per cent c.i. 0·20 to 0·55), and in LMICs (ICC 0·23, 0·05 to 0·38) and HICs (ICC 0·46, 0·25 to 0·59). Conclusion Caution is recommended when using a treatment approach to grade complications in global surgery studies, as this may introduce bias unintentionally

The surgical safety checklist and patient outcomes after surgery: a prospective observational cohort study, systematic review and meta-analysis

© 2017 British Journal of Anaesthesia Background: The surgical safety checklist is widely used to improve the quality of perioperative care. However, clinicians continue to debate the clinical effectiveness of this tool. Methods: Prospective analysis of data from the International Surgical Outcomes Study (ISOS), an international observational study of elective in-patient surgery, accompanied by a systematic review and meta-analysis of published literature. The exposure was surgical safety checklist use. The primary outcome was in-hospital mortality and the secondary outcome was postoperative complications. In the ISOS cohort, a multivariable multi-level generalized linear model was used to test associations. To further contextualise these findings, we included the results from the ISOS cohort in a meta-analysis. Results are reported as odds ratios (OR) with 95% confidence intervals. Results: We included 44 814 patients from 497 hospitals in 27 countries in the ISOS analysis. There were 40 245 (89.8%) patients exposed to the checklist, whilst 7508 (16.8%) sustained ≥1 postoperative complications and 207 (0.5%) died before hospital discharge. Checklist exposure was associated with reduced mortality [odds ratio (OR) 0.49 (0.32–0.77); P\u3c0.01], but no difference in complication rates [OR 1.02 (0.88–1.19); P=0.75]. In a systematic review, we screened 3732 records and identified 11 eligible studies of 453 292 patients including the ISOS cohort. Checklist exposure was associated with both reduced postoperative mortality [OR 0.75 (0.62–0.92); P\u3c0.01; I2=87%] and reduced complication rates [OR 0.73 (0.61–0.88); P\u3c0.01; I2=89%). Conclusions: Patients exposed to a surgical safety checklist experience better postoperative outcomes, but this could simply reflect wider quality of care in hospitals where checklist use is routine