Efeitos do tratamento com inibidor da rock durante o estágio de pós-aquecimento de oócitos bovinos vitrificados sobre desenvolvimento embrionário / Effects of treatment with rock inhibitor during the post-warming stage of vitrified bovine oocytes on embryo development

Diversas biotécnicas vem sendo utilizadas para a preservação de material genético e manutenção a medio e longo prazo de gametas e embrioes de elevado valor genético, contribuindo para o avanço das estratégias para produção pecuária. O processo de vitrificação de oocitos bovinos é uma delas, entretanto com aplicação ainda restrita, pois durante o processo  podem ocorrer varias crio injurias. Afim de reduzir os danos causados pelo congelamento de oócitos e melhorar as taxas de  clivagem e desenvolvimento embrionário obtidas no cultivo in vitro, utilizou-se a exposição a Y-27632, um inibidor seletivo de quinases de proteínas específicas associadas com Rho durante o estágio de pós-aquecimento.  Nosso estudo demonstrou que a utilização de Y-27632 em oocitos bovinos maturados e submetidos a vitrificação promoveu  taxas mais elevadas de sobrevivência bem como maior número de embriões atingindo o etágio de blastocisto.

Acute exposition to Roundup Transorb® induces systemic oxidative stress and alterations in the expression of newly sequenced genes in silverside fish (Odontesthes humensis)

Roundup Transorb® (RDT) is a glyphosate-based herbicide commonly used in agricultural practices worldwide. This herbicide exerts negative effects on the aquatic ecosystem and affects bioenergetic and detoxification pathways, oxidative stress, and cell damage in marine organisms. These effects might also occur at the transcriptional level; however, the expression of genes associated with oxidative stress has not been studied well. Odontesthes humensis is a native Brazilian aquatic species naturally distributed in the habitats affected by pesticides, including Roundup Transorb® (RDT). This study evaluated the toxic effects of short-term exposure to RDT on O. humensis. Moreover, the genes related to oxidative stress were sequenced and characterized, and their expressions in the gills, hepatopancreas, kidneys, and brain of the fish were quantified by quantitative reverse transcription-polymerase chain reaction. The animals were exposed to two environmentally relevant concentrations of RDT (2.07 and 3.68 mg L−1) for 24 h. Lipid peroxidation, reactive oxygen species (ROS), DNA damage, and apoptosis in erythrocytes were quantified by flow cytometry. The expression of the target genes was modulated in most tissues in the presence of the highest tested concentration of RDT. In erythrocytes, the levels of lipid peroxidation, ROS, and DNA damage were increased in the presence of both the concentrations of RDT, whereas cell apoptosis was increased in the group exposed to 3.68 mg L−1 RDT. In conclusion, acute exposure to RDT caused oxidative stress in the fish, induced negative effects on cells, and modulated the expression of genes related to the enzymatic antioxidant system in O. humensis.Fil: Martins, Amanda Weege S.. Universidade Federal de Pelotas; BrasilFil: Silveira, Tony L. R.. Universidade Federal de Pelotas; Brasil. Universidade Federal do Rio Grande; BrasilFil: Remião, Mariana H.. Universidade Federal de Pelotas; BrasilFil: Domingues, William Borges. Universidade Federal de Pelotas; BrasilFil: Dellagostin, Eduardo N.. Universidade Federal de Pelotas; BrasilFil: Varela Junior, Antônio Sergio. Universidade Federal do Rio Grande; Brasil. Universidade Federal de Pelotas; BrasilFil: Corcini, Carine D.. Universidade Federal de Pelotas; BrasilFil: Costa, Patrícia G.. Universidade Federal do Rio Grande; BrasilFil: Bianchini, Adalto. Universidade Federal do Rio Grande; BrasilFil: Somoza, Gustavo Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); ArgentinaFil: Robaldo, Ricardo B.. Universidade Federal de Pelotas; BrasilFil: Campos, Vinicius Farias. Universidade Federal de Pelotas; Brasi

Validity and reliability of a health care service evaluation instrument for tuberculosis

OBJECTIVE To evaluate the validity and reliability of an instrument that evaluates the structure of primary health care units for the treatment of tuberculosis.METHODS This cross-sectional study used simple random sampling and evaluated 1,037 health care professionals from five Brazilian municipalities (Natal, state of Rio Grande do Norte; Cabedelo, state of Paraíba; Foz do Iguaçu, state of Parana; Sao José do Rio Preto, state of Sao Paulo, and Uberaba, state of Minas Gerais) in 2011. Structural indicators were identified and validated, considering different methods of organization of the health care system in the municipalities of different population sizes. Each structure represented the organization of health care services and contained the resources available for the execution of health care services: physical resources (equipment, consumables, and facilities); human resources (number and qualification); and resources for maintenance of the existing infrastructure and technology (deemed as the organization of health care services). The statistical analyses used in the validation process included reliability analysis, exploratory factor analysis, and confirmatory factor analysis.RESULTS The validation process indicated the retention of five factors, with 85.9% of the total variance explained, internal consistency between 0.6460 and 0.7802, and quality of fit of the confirmatory factor analysis of 0.995 using the goodness-of-fit index. The retained factors comprised five structural indicators: professionals involved in the care of tuberculosis patients, training, access to recording instruments, availability of supplies, and coordination of health care services with other levels of care. Availability of supplies had the best performance and the lowest coefficient of variation among the services evaluated. The indicators of assessment of human resources and coordination with other levels of care had satisfactory performance, but the latter showed the highest coefficient of variation. The performance of the indicators “training” and “access to recording instruments” was inferior to that of other indicators.CONCLUSIONS The instrument showed feasibility of application and potential to assess the structure of primary health care units for the treatment of tuberculosis.OBJETIVO Analisar a validade e confiabilidade de instrumento de avaliação da estrutura dos serviços de atenção básica para o tratamento da tuberculose.MÉTODOS Estudo transversal com amostragem aleatória simples para 1.037 profissionais de saúde de cinco municípios (Natal, RN; Cabedelo, PB; Foz do Iguaçu, PR; São José do Rio Preto, SP e Uberaba, MG) em 2011. Foram identificados e validados indicadores estruturais, considerando as diferentes modalidades de organização do sistema de saúde de municípios de diferentes portes populacionais. A “estrutura” correspondeu aos recursos existentes para a execução dos serviços: físicos (equipamentos, materiais de consumo e instalações); humanos (número e qualificação); e fontes para manutenção da infraestrutura e da tecnologia existente, entendida como organização do serviço de saúde. Os procedimentos estatísticos utilizados no processo de validação foram análise de confiabilidade, análise fatorial exploratória e confirmatória.RESULTADOS O processo de validação determinou a retenção de cinco fatores com 85,9% de variância total explicada, consistência interna entre 0,6460 e 0,7802 e qualidade de ajustamento da análise confirmatória de 0,995 para ogoodness-of-fit index. Os fatores retidos compuseram os cinco indicadores estruturais: profissionais envolvidos com o atendimento de tuberculose; capacitação; acesso a instrumentos de registro; disponibilidade de insumos; e articulação do serviço de saúde com outros níveis de atenção. O indicador com melhor desempenho foi disponibilidade de insumos, com menor coeficiente de variação nos serviços avaliados. Os indicadores de avaliação dos recursos humanos e articulação com outros níveis de serviços apresentaram percentuais satisfatórios; entretanto, este último apresentou o maior coeficiente de variação. Os indicadores de capacitação e acesso a instrumentos de registro apresentaram percentuais de avaliação inferiores aos demais.CONCLUSÕES O instrumento apresentou viabilidade de aplicação e potencial de avaliação da estrutura dos serviços de saúde para o tratamento da tuberculose na atenção básica

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

Large expert-curated database for benchmarking document similarity detection in biomedical literature search

Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe

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

Sperm microRNAs related to fertility

Spermatozoa is a cell type that carry much more than just male genomic DNA during the fertilization process. In recent years, epigenetic mechanisms related to sperm quality have been extensively studied. Among the mechanisms, microRNAs deserve to be highlighted because they act as powerful post-transcriptional regulation of their target genes, being correlated with spermatogenesis, sperm functionality and reproductive success in different species. It is hypothesized that changes in sperm kinetic and morphofunctional parameters, as well as in testicular physiology, may disturb sperm microRNA levels. Thus, the main objective was to elucidate whether the levels of microRNAs related to fertility is affected exogenous DNA transfected-sperm, and in sperm samples from transgenic animals overexpressing growth hormone. In the first study, through a systematic review of scientific literature, it was possible to identify the microRNAs let-7c, let-7d, let-7g, miR-1, miR-100, miR-148b-3p, miR-151-3p, miR-20a, miR-26a-5p, miR-30b-5p, miR-30c-5p, miR-34c-5p, and miR-378, with different levels according to the contrasting sperm motility profile in livestock species, such as cattle and pigs. In a second study, through sperm-mediated gene transfer technique, it was observed that transfection of exogenous DNA associated with cationic nanostructure affects not only kinetic and morphofunctional parameters, but also the microRNA levels related to sperm functionality in bovine, such as, bta-miR-30b-5p and bta-miR-34c. Finally, in a third study, using a transgenic zebrafish model overexpressing growth hormone, it was observed a reduction in sperm quality parameters and a modulation of twenty two sperm microRNAs levels, among them, dre-miR-20a-5p, dre-miR-202-5p, dre-miR-146b, and dre-miR-200a-5p. Based on the results achieved using bovine and zebrafish model organisms, it was possible to infer that sperm exogenous DNA uptake by sperm, and physiological changes in testicular tissue affects some microRNAs. Finally, it is important to note that a better understanding of relationship of sperm microRNAs with fertility becomes a stimulus for validation and using these small RNA molecules as a biotechnological tool.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESEspermatozoides são células que carregam muito mais que somente o DNA genômico masculino durante o processo de fertilização. Nos últimos anos, mecanismos epigenéticos relacionados a qualidade das células espermáticas têm sido extensivamente estudados. Dentre os mecanismos, os microRNAs merecem destaque por terem um alto poder de regulação pós-transcricional de inúmeros genes, fazendo com que os mesmos estejam relacionados aos processo de espermatogênese, funcionalidade espermática e sucesso reprodutivo em diferentes espécies. Hipotetiza-se que alterações nos parâmetros cinéticos e morfofuncionais dos espermatozoides, bem como na fisiologia testicular, podem perturbar os níveis de microRNAs. Sendo assim, a presente tese teve como objetivo principal elucidar se os níveis de microRNAs relacionados com a fertilidade podem ser afetados em espermatozoides transfectados com DNA exógeno e em amostras seminais de animais transgênicos superexpressando o hormônio do crescimento. No primeiro estudo, através de uma revisão sistemática da literatura científica, foi possível identificar os microRNAs let-7c, let-7d, let-7g, miR-1, miR-100, miR-148b-3p, miR-151-3p, miR-20a, miR-26a-5p, miR-30b-5p, miR-30c-5p, miR-34c-5p e miR-378 com níveis alterados em distintos padrões de alta e baixa motilidade espermática em animais domésticos voltados à produção, como bovinos e suínos. No segundo estudo, através da transferência gênica mediada por espermatozoides, foi observado que a transfecção de DNA exógeno associado a nanoestrutura catiônica afeta não somente os parâmetros cinéticos e morfofuncionais espermáticos em bovinos, mas também os níveis de microRNAs ligados à funcionalidade destas células, como por exemplo, btamiR-30b-5p e bta-miR-34c. Por fim, em um terceiro estudo, utilizando um modelo transgênico de zebrafish para superexpressão do hormônio do crescimento, foi possível observar a redução de parâmetros de qualidade espermática e a alteração dos níveis de vinte e dois microRNAs espermáticos, dentre eles, os dre-miR-20a-5p, dre-miR-202-5p, dre-miR-146b e dre-miR-200a-5p. Com base nos resultados alcançados nos modelos bovino e zebrafish, foi possível inferir que a transfecção de DNA exógeno em espermatozoides e mudanças fisiológicas no tecido testicular, causam alterações em alguns microRNAs. Por fim, é importante ressaltar que uma maior compreensão da relação dos microRNAs espermáticos com a fertilidade, acaba por ser um estímulo para validação e utilização destas pequenas moléculas de RNA como ferramentas biotecnológicas

Efeito de extrato de mirtilo na expressão de citocinas inflamatórias em córtex cerebral de camundongos submetidos a um modelo experimental de comportamento tipo-depressivo

O objetivo do presente trabalho foi avaliar o efeito do tratamento com extrato de mirtilo na expressão gênica de TNF-α, IL-1β e IL-10 em córtex cerebral de camundongos submetidos ao modelo experimental de comportamento tipo-depressivo induzido por lipopolissacarídeo (LPS)