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In the present study, the effect of 6-((4-fluorophenyl) selanyl)-9H-purine (FSP) was tested against memory impairment and sensitivity to nociception induced by intracerebroventricular injection of amyloid-beta peptide (Aβ) (25-35 fragment), 3 nmol/3 μl/per site in mice. Memory impairment was determined by the object recognition task (ORT) and nociception by the Von-Frey test (VFT). Aβ caused neuroinflammation with upregulation of glial fibrillary acidic protein (GFAP) (in hippocampus), nuclear factor-κB (NF-κB), and the proinflammatory cytokines interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in cerebral cortex and hippocampus. Additionally, Aβ increased oxidant levels and lipid peroxidation in cerebral cortex and hippocampus, but decreased heme oxygenase-1 (HO-1) and peroxiredoxin-1 (Prdx1) expression in the hippocampus. Anti-neuroinflammatory effects of FSP were demonstrated by a decrease in the expression of GFAP and NF-κB in the hippocampus, as well as a decrease in proinflammatory cytokines in both the hippocampus and cerebral cortex FSP protected against oxidative stress by decreasing oxidant levels and lipid peroxidation and by increasing HO-1 and Prdx1 expressions in the hippocampus of mice. Moreover, FSP prevented the activation of nuclear factor erythroid 2-related factor 2 (Nrf-2) in the hippocampus of mice induced by Aβ. In conclusion, treatment with FSP attenuated memory impairment, nociception sensitivity by decreasing oxidative stress, and neuroinflammation in a mouse model of Alzheimer's disease.

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.

Biodiversity is commonly referred to as the biological diversity and variety of life forms, but these organisms should not be observed individually, but rather as a complex and functional structure that interacts with each other. Educating citizens from an early age to know the biodiversity of their region and then helping to conserve, preserve and protect it is a very important process. Our objective was to analyze the knowledge of students in the last year of high school on the topic of regional biodiversity. It is a qualitative and quantitative research, carried out in three schools, in three different municipalities, with 65 students. Among the three levels of biodiversity, students know the diversity of species. Most of them conceptualize native and endemic species, but do not know how to conceptualize exotic species. In general, they understand that the great importance of biodiversity is ecological balance and that the loss of biodiversity will lead to ecological imbalance. Native species, cited more frequently, are species present in urban areas and students with more contact with nature cite, on average, a greater number of species, indicating the importance of rescuing contact with nature. La biodiversidad se conoce comúnmente como la diversidad biológica y la variedad de formas de vida, pero estos organismos no deben observarse individualmente, sino como una estructura compleja y funcional que interactúa entre sí. Educar a los ciudadanos desde temprana edad para que conozcan la biodiversidad de su región y luego ayudar a conservarla, preservarla y protegerla es un proceso muy importante. Nuestro objetivo fue analizar los conocimientos de los estudiantes del último año de secundaria sobre el tema de la biodiversidad regional. Es una investigación cualitativa y cuantitativa, realizada en tres escuelas, en tres municipios diferentes, con 65 alumnos. Entre los tres niveles de biodiversidad, los alumnos conocen la diversidad de especies. La mayoría conceptualiza especies nativas y endémicas, pero no sabe conceptualizar especies exóticas. En general, entienden que la gran importancia de la biodiversidad es el equilibrio ecológico y que la pérdida de biodiversidad conducirá al desequilibrio ecológico. Las especies nativas, citadas con mayor frecuencia, son especies presentes en áreas urbanas y los estudiantes con más contacto con la naturaleza citan, en promedio, un mayor número de especies, lo que indica la importancia de rescatar el contacto con la naturaleza. A biodiversidade comumente é referenciada como a diversidade biológica e variedade de formas de vida, porém esses organismos não devem ser observados de uma forma individual, mas sim como uma estrutura complexa e funcional que interage entre si. Educar o cidadão desde cedo para conhecer a biodiversidade da sua região e então ajudar a conservá-la, preservá-la e protegê-la é um processo importantíssimo. Nosso objetivo foi analisar o conhecimento dos estudantes do último ano do Ensino Médio sobre o tema biodiversidade regional. É uma pesquisa qualitativa e quantitativa, realizada em três escolas, de três municípios distintos, com 65 estudantes. Dentre os três níveis da biodiversidade, os estudantes conhecem a diversidade de espécies. A maioria deles conceitua espécies nativas e endêmicas, mas não sabe conceituar espécies exóticas. De forma geral eles entendem que a grande importância da biodiversidade é o equilíbrio ecológico e que a perda da biodiversidade levará ao desequilíbrio ecológico. As espécies nativas, citadas com mais frequência, são espécies presentes nas áreas urbanas e os estudantes com mais contato com a natureza citam, em média, um número maior de espécies, indicando a importância de resgatar o contato com a natureza.

O artigo em questão tabula e analisa o Transtorno do Espectro Autista (TEA) a partir de análise documental das teses e dissertações que foram produzidos dentro dos programas de pós-graduação stricto sensu, tendo TEA como objeto de pesquisa, entre os anos de 2011 e 2020, por meio da plataforma BDTD. Para tal, foi utilizado a fundamentação teórica amparada no Manual Diagnóstico e Estatístico de Transtornos Mentais (DSM-V), e contribuições de Oliveira (2009), dentre outras. Na contextualização teórica apresentamos a definição dos eixos orientadores. A metodologia competiu análise de documentos oficiais, que são as teses e dissertações, defendidas no Brasil devem compor obrigatoriamente a plataforma Ibict. E concluímos, que se faz urgente a necessidade de aprofundar e incentivar pesquisas a nível de pós-graduação stricto sensu para melhorar o diagnóstico e acompanhamento de sujeitos com quadro diagnóstico de TEA.

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; Brasil Fil: Silveira, Tony L. R.. Universidade Federal de Pelotas; Brasil. Universidade Federal do Rio Grande; Brasil Fil: Remião, Mariana H.. Universidade Federal de Pelotas; Brasil Fil: Domingues, William Borges. Universidade Federal de Pelotas; Brasil Fil: Dellagostin, Eduardo N.. Universidade Federal de Pelotas; Brasil Fil: Varela Junior, Antônio Sergio. Universidade Federal do Rio Grande; Brasil. Universidade Federal de Pelotas; Brasil Fil: Corcini, Carine D.. Universidade Federal de Pelotas; Brasil Fil: Costa, Patrícia G.. Universidade Federal do Rio Grande; Brasil Fil: Bianchini, Adalto. Universidade Federal do Rio Grande; Brasil Fil: 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); Argentina Fil: Robaldo, Ricardo B.. Universidade Federal de Pelotas; Brasil Fil: Campos, Vinicius Farias. Universidade Federal de Pelotas; Brasil

Anaplasma marginale is transmitted biologically by infected ticks or mechanically by biting flies and contaminated fomites. In tick-free areas, such southern Uruguay, horseflies could be the principal vectors of this pathogen to bovines, causing anaplasmosis occurrence. Therefore, the objective of this work was to detect the presence of A. marginale by MSP-5 PCR and Sanger sequencing in the most prevalent species of horseflies using different collection methods in the localities of Colonia, Taquarembó and Paysandú, Uruguay. Eight horsefly species were tested (Dasybasis missionum, Poeciloderas lindneri, Tabanus campestris, T. claripennis, T. fuscofasciatus, T. platensis, T. tacuaremboensis and T. triangulum); four species were found positive for A. marginale, with D. missionum and P. lindneri having the most frequent infections, while only one individual each of T. fuscofasciatus and T. tacuaremboensis was positive. Both D. missionum and P. lindneri were positive for A. marginale in tick-free areas, with implications that are discussed in this report.

Obstructive Sleep Apnea and Hypopnea Syndrome (OSAHS) is a progressive disorder of the respiratory system that seriously affects the general health and, consequently, the quality of life of an individual. The aim of the present study was to report a case of moderate OSAHS treated with the supervised use of the Gradual Mandibular Advancement Intraoral Appliance (AIO). The treatment was followed up through the patient's report, evaluation of the Epworth Sleepiness Scale, and its result was confirmed by the baseline polysomnography exam. After the treatment period, there was an improvement in the symptoms reported by the patient and in the polysomnographic parameters. The patient went from a moderate OSAHS condition, with an Apnea and Hypopnea Index (AHI) of 24.7 events per hour, to a mild condition with an AHI of 10.7 events per hour. Thus, it can be concluded based on the present case that the AIO was effective for the conservative treatment of moderate OSAHS. El síndrome de apnea e hipopnea obstructiva del sueño (SAHOS) es un trastorno progresivo del sistema respiratorio que afecta gravemente la salud general y, en consecuencia, la calidad de vida de un individuo. El objetivo del presente estudio fue reportar un caso de SAHOS moderado tratado con el uso supervisado del Aparato Intraoral de Avance Mandibular Gradual (AIO). El tratamiento fue seguido a través del informe del paciente, la evaluación de la Escala de Somnolencia de Epworth y su resultado fue confirmado por el examen de polisomnografía basal. Después del período de tratamiento, hubo una mejoría en los síntomas informados por el paciente y en los parámetros polisomnográficos. El paciente pasó de un estado de SAHOS moderado, con un índice de apnea e hipopnea (IAH) de 24.7 eventos por hora, a un estado leve con un IAH de 10.7 eventos por hora. Por lo tanto, se puede concluir con base en el presente caso que el AIO fue efectivo para el tratamiento conservador del SAHOS moderado. A Síndrome da Apneia e Hipopneia Obstrutiva do Sono (SAHOS) é uma desordem progressiva do sistema respiratório que afeta seriamente a saúde geral e, consequentemente, a qualidade de vida do indivíduo. O objetivo do presente estudo foi relatar um caso de SAHOS moderada tratado com o uso supervisionado do Aparelho Intraoral de Avanço Mandibular Gradual (AIO). O tratamento foi acompanhado através do relato do paciente, avaliação da Escala de Sonolência de Epworth, e seu resultado foi confirmado pelo exame de polissonografia de base. Após o período de tratamento, houve melhora nos sintomas relatados pelo paciente e nos parâmetros polissonográficos. O paciente passou de um quadro de SAHOS moderado, com Índice de Apnéia e Hipopnéia (IAH) de 24.7 eventos por hora, para um quadro leve com IAH de 10.7 eventos por hora. Assim, pode-se concluir com base no presente caso que o AIO foi eficaz para o tratamento conservador da SAHOS moderada.

Growing evidence demonstrates that Oxaliplatin (OXA) is commonly associated with neurotoxicity that leads to emotional and cognitive impairments. The aim of the present study was to evaluate the OXA and Na+, K+-ATPase interaction and to correlate anxious behavior and cognitive impairment induced by this chemotherapeutic in Swiss mice. Also, considering the pharmacological modulation of Na+, K+-ATPase as a potential target for OXA-induced neurotoxicity, the therapeutic potential of 7-chloro-4-(phenylselanyl) quinoline (4-PSQ) was evaluated. Mice received OXA (10 mg kg-1) or vehicle by intraperitoneal route (days 0 and 2). Oral administration of 4-PSQ (1 mg kg-1) or vehicle was performed from days 2-14. Behavioral tasks started from day 12 onwards. On day 15, the animals were sacrificed, and the tissues collected. The effects of OXA and 4-PSQ on activity and expression level of Na+, K+-ATPase in the hippocampus and cerebral cortex, and the plasmatic corticosterone levels were determined. The findings demonstrated a significant positive correlation between anxious behavior and cognitive impairment induced by OXA. OXA caused an increase on the plasmatic corticosterone levels and reduced activity and expression level of Na+, K+-ATPase. 4-PSQ reduced both anxious behavior and cognitive impairment induced by OXA. 4-PSQ effect seems to be due to the modulation of Na+, K+-ATPase and reduction of corticosterone levels. Our results helped to expand knowledge about the mechanisms involved in the physiopathology of the OXA-induced neurotoxicity and strongly indicated that 4-PSQ may be a good prototype for the treatment of anxious behavior and cognitive impairment induced by OXA exposure.

Sperm-mediated gene transfer (SMGT) has a potential use for zebrafish transgenesis. However, transfection into fish sperm cells still needs to be improved. The objective was to demonstrate the feasibility of tip type electroporation in zebrafish sperm, showing a protocol that provide high transfection efficiency, with minimal side-effects. Sperm was transfected with a Cy3-labelled DNA using tip type electroporation with voltages ranging from 500 to 1500 V. Sperm kinetics parameters were assessed using Computer Assisted Semen Analysis (CASA) and cell integrity, reactive oxygen species (ROS), mitochondrial functionality and transfection rate were evaluated by flow cytometry. The transfection rates were positively affected by tip type electroporation, reaching 64.9% ± 3.6 in the lowest voltage used (500 V) and 86.6% ± 1.9 in the highest (1500 V). The percentage of overall motile sperm in the electrotransfected samples was found to decrease with increasing field strength (P 0.05). Overall results indicate that tip type electroporation enhances the internalization of exogenous DNA into zebrafish sperm cells with minimal harmful effects to sperm cells. Electronic supplementary material The online version of this article (10.1007/s11033-020-05658-2) contains supplementary material, which is available to authorized users.

In this study, granulated activated charcoal (GAC) and bio charcoal (BC) is used as a filler in P3 biosand bag filter to study their filtration performance against a range of fluoride impurities from 1–1400 mg/L. A set of experiments are done to analyze the filtration efficiency of the sandbag filter against fluoride impurities after incorporating different amounts (e.g., 0.2, 2 kg) and a combination of GAC and BC. A combination of filler GAC and BC (1 kg each) have exhibited excellent results with 100% fluoride removal efficiency against 5 mg/L fluoride impurities for an entire experimental time of 165 min. It is because of the synergetic effect of adsorption caused by the high surface area (739 m2/g) of GAC and hydroxyapatite groups in BC. The data from remediation experiments using individual GAC and BC are fitted into the Langmuir and Freundlich Isotherm Models to check their adsorption mechanism and determine GAC and BC’s maximum adsorption capacity (Qm). The remediation data for both GAC and BC have shown the better fitting to the Langmuir Isotherm Model with a high R2 value of 0.994 and 0.970, respectively, showing the excellent conformity with monolayer adsorption. While the GAC and BC have presented negative Kf values of −1.08 and −0.72, respectively, for Freundlich Model, showing the non-conformity to multilayer adsorption. The Qm values obtained from Langmuir Model for GAC is 6.23 mg/g, and for BC, it is 9.13 mg/g. The pH study on adsorption efficiency of individual GAC and BC against 5 mg/L of fluoride impurities indicates the decrease in removal efficiency with an increase in pH from 3 to 9. For example, BC has shown removal efficiency of 99.8% at pH 3 and 99.5% at pH 9, while GAC has exhibited removal efficiency of 96.1% at pH 3 and 95.9% at pH 9. Importantly, this study presents the significance of the synergetic application of GAC and BC in the filters, where GAC and BC are different in their origin, functionalities, and surface characteristics.

Made available in DSpace on 2022-04-29T08:39:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-04-20 Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul The limited regenerative capacity in mammals has serious implications for cardiac tissue damage. Meanwhile, zebrafish has a high regenerative capacity, but the regulation of the heart healing process has yet to be elucidated. The dynamic nature of cardiac regeneration requires consideration of the inherent temporal dimension of this process. Here, we conducted a systematic review to find genes that define the regenerative cell state of the zebrafish heart. We then performed an in silico temporal gene regulatory network analysis using transcriptomic data from the zebrafish heart regenerative process obtained from databases. In this analysis, the genes found in the systematic review were used to represent the final cell state of the transition process from a non-regenerative cell state to a regenerative state. We found 135 transcription factors driving the cellular state transition process during zebrafish cardiac regeneration, including Hand2, Nkx2.5, Tbx20, Fosl1, Fosb, Junb, Vdr, Wt1, and Tcf21 previously reported for playing a key role in tissue regeneration. Furthermore, we demonstrate that most regulators are activated in the first days post-injury, indicating that the transition from a non-regenerative to a regenerative state occurs promptly. Laboratório de Genômica Estrutural Programa de Pós-Graduação em Biotecnologia Centro de Desenvolvimento Tecnológico Universidade Federal de Pelotas Laboratório de Bioinformática e Proteômica Programa de Pós-Graduação em Biotecnologia Centro de Desenvolvimento Tecnológico Universidade Federal de Pelotas Laboratório Genômica e Evolução Molecular Departamento de Genética Instituto de Biociências de Botucatu Universidade Estadual Paulista (UNESP) Laboratório Genômica e Evolução Molecular Departamento de Genética Instituto de Biociências de Botucatu Universidade Estadual Paulista (UNESP)

Made available in DSpace on 2022-04-29T08:33:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-09-08 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Overexpression of growth hormone (GH) in gh-transgenic zebrafish of a highly studied lineage F0104 has earlier been reported to cause increased muscle growth. In addition to this, GH affects a broad range of cellular processes in transgenic fish, such as morphology, physiology, and behavior. Reports show changes such as decreased sperm quality and reduced reproductive performance in transgenic males. It is hypothesized that microRNAs are directly involved in the regulation of fertility potential during spermatogenesis. The primary aim of our study was to verify whether gh overexpression disturbs the sperm miRNA profile and influences the sperm quality in transgenic zebrafish. We report a significant increase in body weight of gh-transgenic males along with associated reduced sperm motility and other kinetic parameters in comparison to the non-transgenic group. MicroRNA transcriptome sequencing of gh-transgenic zebrafish sperms revealed expressions of 186 miRNAs, among which six miRNA were up-regulated (miR-146b, miR-200a-5p, miR-146a, miR-726, miR-184, and miR-738) and sixteen were down-regulated (miR-19d-3p, miR-126a-5p, miR-126b-5p, miR-22a-5p, miR-16c-5p, miR-20a-5p, miR-126b-3p, miR-107a-3p, miR-93, miR-2189, miR-202–5p, miR-221–3p, miR-125a, miR-125b-5p, miR-126a-3p, and miR-30c-5p) in comparison to non-transgenic zebrafish. Some of the dysregulated miRNAs were previously reported to be related to abnormalities in sperm quality and reduced reproduction ability in other species. In this study, an average of 134 differentially expressed miRNAs-targeted genes were predicted using the in silico approach. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis demonstrated that the genes of affected pathways were primarily related to spermatogenesis, sperm motility, and cell apoptosis. Our results suggested that excess GH caused a detrimental effect on sperm microRNAome, consequently reducing the sperm quality and reproductive potential of zebrafish males. Laboratório de Genômica Estrutural Programa de Pós-Graduação em Biotecnologia Centro de Desenvolvimento Tecnológico Universidade Federal de Pelotas Laboratório de Biologia Molecular Instituto de Ciências Biológicas Universidade Federal do Rio Grande Faculdade de Nutrição Universidade Federal de Pelotas Programa de Pós-Graduação em Veterinária Faculdade de Veterinária Universidade Federal de Pelotas Unidad de Biología Molecular Institut Pasteur Laboratório Genômica e Evolução Molecular Departamento de Genética Instituto de Biociências de Botucatu Universidade Estadual Paulista (UNESP) Laboratório Genômica e Evolução Molecular Departamento de Genética Instituto de Biociências de Botucatu Universidade Estadual Paulista (UNESP) CAPES: AUXPE #2537/2018 FAPESP: FAPERGS FAPESP #19/2551-0000953-3

A biodiversidade comumente é referenciada como a diversidade biológica e variedade de formas de vida, porém esses organismos não devem ser observados de uma forma individual, mas sim como uma estrutura complexa e funcional que interage entre si. Educar o cidadão desde cedo para conhecer a biodiversidade da sua região e então ajudar a conservá-la, preservá-la e protegê-la é um processo importantíssimo. Nosso objetivo foi analisar o conhecimento dos estudantes do último ano do Ensino Médio sobre o tema biodiversidade regional. É uma pesquisa qualitativa e quantitativa, realizada em três escolas, de três municípios distintos, com 65 estudantes. Dentre os três níveis da biodiversidade, os estudantes conhecem a diversidade de espécies. A maioria deles conceitua espécies nativas e endêmicas, mas não sabe conceituar espécies exóticas. De forma geral eles entendem que a grande importância da biodiversidade é o equilíbrio ecológico e que a perda da biodiversidade levará ao desequilíbrio ecológico. As espécies nativas, citadas com mais frequência, são espécies presentes nas áreas urbanas e os estudantes com mais contato com a natureza citam, em média, um número maior de espécies, indicando a importância de resgatar o contato com a natureza.

Search engines have become important tools for obtaining information and assistance in choosing products and services. An example is the Google search engine, which uses the Google My Business tool. Through this, websites related to brands and locations can be evaluated and receive comments from users. The organic participation of Internet citizens generates an earned media, important for the digital presence of the brand. The present study aims to compare online reviews of private basic education institutions, in order to verify which aspects were highlighted and what information a potential client finds when searching for the name of the particular institution. The basic education institutes have had few negative comments, although some stand out and can make a strong negative impression. This study is relevant to reflect on the online image of educational units and the opportunity to interact with the public, promoting a satisfactory earned media., Mecanismos de busca online tornaram-se importantes ferramentas para a obtenção de informações e auxílio para escolhas de produtos e serviços. Um exemplo é o buscador da Google, que conta com a ferramenta Google Meu Negócio. Por meio desta, páginas da internet relacionadas a marcas e locais podem ser avaliadas e receber comentários de usuários. A participação orgânica de cidadãos da internet gera uma mídia ganhada, importante para a presença digital da marca. O presente estudo pretende comparar avaliações online de instituições de ensino básico privadas, a fim de verificar que aspectos foram destacados e que informações um potencial cliente encontra ao pesquisar o nome da determinada instituição. Os Colégios tiveram poucos comentários negativos, embora alguns se destaquem e possam causar uma forte impressão negativa. Este estudo é relevante para refletir sobre a imagem online de unidades educacionais e a oportunidade de interação com o público, promovendo uma mídia ganhada satisfatória.

Submitted by Aline Batista (alinehb.ufpel@gmail.com) on 2021-02-23T11:43:56Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_William_Borges_Aldrighi.pdf: 1158289 bytes, checksum: d1b0db72da344096115f7b49e3907674 (MD5) Made available in DSpace on 2021-02-23T11:46:33Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_William_Borges_Aldrighi.pdf: 1158289 bytes, checksum: d1b0db72da344096115f7b49e3907674 (MD5) Previous issue date: 2020-10-30 Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq Esta dissertação foi construída a partir de reflexões advindas da dificuldade enfrentada pelos agricultores familiares em realizarem o escoamento da produção. Os mercados para a agricultura familiar apresentam diversas faces e avanços significativos foram observados, nas últimas duas décadas, no que tange às políticas públicas no Brasil. No entanto, o alcance destas políticas, assim como dos mercados, é limitado. Nesse sentido, é possível observar grupos de agricultores familiares que criam os seus próprios mecanismos de comercialização. Este trabalho, tem como objeto um mecanismo de comercialização da Rede Bem da Terra, que iniciou em dezembro de 2014 com uma proposta de comercialização virtual e distribuição através de um Grupo de Consumidores Responsáveis, no município de Pelotas – RS. Os projetos iniciais, em relação ao formato que a feira virtual pretendia assumir não se concretizaram, assim como, os objetivos que tangem às dimensões de consumo, produção e distribuição. O objetivo central deste trabalho se fundou em compreender as características do processo que levaram os atores envolvidos às atitudes de consumo e produção que fizeram com que não se atingisse as metas de comercialização projetadas para a Feira Virtual Bem da Terra, mas ao mesmo tempo permitiram a sua permanência, ainda que sem autonomia financeira. Para atingir este objetivo, buscou-se: (a) compreender quais são os projetos individuais e coletivos dos atores das arenas de produtores, consumidores e Incubadoras; (b) avaliar qual a relação dos projetos individuais e coletivos das arenas de consumidores e produtores com a dinâmica de consumo encontrada; (c) entender quais e como foram construídas as estratégias a partir da dinâmica de consumo médio e do número de consumidores nas diferentes arenas a partir da agência dos atores. Para melhor compreender o que se propôs, utilizou-se o aporte teórico da Perspectiva Orientada pelo Ator (POA) como ferramenta central de análise e de forma transversalizada, a teoria econômica, os mercados e a compreensão sobre as orientações acerca do consumo como movimento. O estudo foi realizado com atores de três municípios do estado do Rio Grande do Sul-Brasil, dentre os quais estão Canguçu, Morro Redondo e Pelotas, num total de 11 entrevistados entre produtores, consumidores e membros de incubadoras. A coleta dos dados foi realizada mediante entrevista com roteiro semiestruturado e, posteriormente, efetuou-e a análise de discurso. A pesquisa apresenta como características: estudo de caso, exploratório-descritiva explicativa, e qualitativa. Os resultados demonstraram que os agricultores estão imersos no universo da produção e focados nas suas reproduções sociais, o que lhes dificultou realizarem uma participação mais efetiva. Os consumidores possuem diferentes níveis de sensibilidade socioambiental, o que fez com que, por um lado, muitos abandonassem ao longo do tempo a proposta de consumo, e, por outro lado, outros embarcassem de modo a garantirem junto às incubadoras até a desincubação, a continuidade da feira virtual, ainda que sem autonomia financeira. This dissertation was built from reflections arising from the difficulty faced by family farmers in carrying out the production. The markets for family farming have several faces and significant advances have been observed in the last two decades when it comes to public policies in Brazil. However, the scopes of these policies, as well as of the markets, are limited. In this sense, it is possible to observe groups of family farmers who create their own marketing mechanisms. This work has as object a market mechanism called Feira Virtual Bem da Terra, which was created in December 2014 and has proposal for virtual marketing and distribution through a Group of Responsible Consumers in the municipality of Pelotas - RS. The projects in relation to the format that the Feira Virtual intended to assume did not materialize, as well as the objectives related to the dimensions it reaches in terms of consumption, production and distribution. The main objective of the work was based on understanding the characteristics of the process that led the actors involved to consumption and production attitudes that made it impossible to reach the marketing goals projected for Feira Virtual Bem da Terra, but at the same time allowed the their permanence, even if without financial autonomy. To achieve this objective, we sought to: (a) understand the individual and collective projects of the actors in the arenas of producers, consumers and Incubators; (b) assessing the relationship between individual and collective projects in consumer and producer arenas with the consumption dynamics found; (c) understand which and how the strategies were built from the dynamics of average consumption and the number of consumers in the different arenas from the agency of the actors. In order to better understand what was proposed, the theoretical contribution of the Actor-Oriented Perspective (POA) was used as a central analysis tool and in a transversal way, the economic theory, the markets and the understanding of the orientations about consumption as a movement. The study was carried out with actors from three municipalities in the state of Rio Grande do Sul-Brazil, among which are Canguçu, Morro Redondo and Pelotas, in a total of 11 interviewees among producers, consumers and members of incubators. Data collection was carried out through an interview with a semi-structured script and, subsequently, the discourse analysis was performed. The research presents as characteristics: case study, exploratory-descriptive, explanatory, and qualitative. The results showed that farmers are immersed in the universe of production and focused on their social reproductions, which made it difficult for them to carry out more effective participation. Consumers have different levels of socioenvironmental sensitivity, which has meant that, on the one hand, many have abandoned the proposal over time, on the other hand others have embarked in order to guarantee the continuity of the Feira Virtual together with the incubators until the end of the incubation, although without financial autonomy.

Made available in DSpace on 2020-12-12T02:14:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-10-15 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) MicroRNAs have been hypothesized to be involved in the regulation of male fertility potential. The primary aim of our study was to demonstrate the effects of transfection with dendrimer nanostructure on the parameters of bovine sperm quality and to investigate whether the microRNA profile could be disturbed after cationic dendrimer-mediated exogenous DNA transfection of bovine spermatozoa. The binding of exogenous DNA was significantly increased when dendrimer-based transfection was implemented. However, cationic dendrimer transfection induced detrimental changes in the kinetics and sperm quality parameters, such as membrane integrity, acrosome reaction, and mitochondrial membrane potential, when compared to the control group. Sperm microRNA sequencing revealed 218 known and 106 novel microRNAs in the sperm samples, among which nine were dysregulated after transfection (one was upregulated and eight were downregulated), in comparison to the non-transfected sperm. All the dysregulated microRNAs were related to sperm quality and embryonic development. These results suggest that the transfection process using the dendrimer nanostructure has an impact on the quality and microRNA profile of bovine sperm. Laboratório de Genômica Estrutural Programa de Pós-Graduação em Biotecnologia Centro de Desenvolvimento Tecnológico Universidade Federal de Pelotas Laboratório de Reprodução Animal Programa de Pós-Graduação em Veterinária Universidade Federal de Pelotas Laboratório de Bioinformática e Proteômica Programa de Pós-Graduação em Biotecnologia Centro de Desenvolvimento Tecnológico Universidade Federal de Pelotas Laboratório de Biotecnologia do Câncer Programa de Pós-Graduação em Biotecnologia Centro de Desenvolvimento Tecnológico Universidade Federal de Pelotas Laboratório Genômica e Evolução Molecular Departamento de Genética Instituto de Biociências de Botucatu Universidade Estadual Paulista (UNESP) Unidad de Biología Molecular Institut Pasteur Montevideo Faculdade de Nutrição Universidade Federal de Pelotas Department of Biopharmaceutical Sciences Programs in Human Genetics and Biological Sciences University of California Laboratório Genômica e Evolução Molecular Departamento de Genética Instituto de Biociências de Botucatu Universidade Estadual Paulista (UNESP) Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul: 17/2551-0000953-3 CAPES: AUXPE 2537/2018 FAPESP: FAPERGS-FAPESP #19/2551-0002283-2

Reference genes (RGs) must have a stable expression in tissues in all experimental conditions to normalize real-time quantitative reverse transcription PCR (qRT-PCR) data. F0104 is a highly studied lineage of zebrafish developed to overexpress the growth hormone (GH). It is assumed that the transgenic process may influence the expression levels of commonly used RGs. The objective of the present study was to make a comprehensive analysis of stability of canditade RGs actb1, actb2, b2m, eif2s2, eef1a1, gapdh, rplp2, rpl7, rpl13α, tuba1, and rps18, in gh-transgenic and non-transgenic zebrafish. Liver, brain, intestine and muscle samples from both groups had qRT-PCR results analyzed by dCt, geNorm, NormFinder, BestKeeper, and RefFinder softwares. Consensus analyses among software concluded that rpl13α, rpl7, and eef1a1 are the most stable genes for zebrafish, considering the studied groups and tissues. Gapdh, rps18, and tuba1 suffered variations in stability among different tissues of both groups, and so, they were listed as the genes with lowest stability. Results from an average pairwise variations test indicated that the use of two RGs would generate reliable results for gene expression analysis in the studied tissues. We conclude that genes that are commonly used in mammals for qRT-PCR assays have low stability in both non-transgenic and gh-transgenic zebrafish reinforcing the importance of using species-specific RGs.

Sperm-mediated gene transfer (SMGT) has a potential application in the generation of transgenic animals. Capillary electroporation consists of the application of electrical pulses, resulting in an increased transfection rate. Little is known about the impacts of the transfection of exogenous DNA on sperm epigenetics. MicroRNAs are epigenetic factors that are related to sperm motility. MiRNA-122-5p regulates genes that influence motility, and consequently, the fertilizing potential of sperm. Therefore, we aimed at identifying whether epigenetic factors such as microRNAs could be altered after DNA transfection, using the capillary electroporation technique. In this study, bull sperm was electroporated using voltages of 600 V, 1500 V, and 0 V (control group), with or without exogenous DNA. Parameters of sperm quality were analyzed using CASA and flow cytometry, and expression of the miRNA-122-5p was analyzed using RT-qPCR. It was observed that electroporation increased the internalization of exogenous DNA (P 0.05), but did not impair the mitochondrial activity (P 0.05). It reduced sperm motility (P 0.05). The expression of miRNA-122-5p was upregulated in sperm electroporated at 1500 V, and the presence of exogenous DNA did not affect its expression. Thus, we can conclude that electroporation influences the expression of miRNA-122-5p from bull sperm cells.

Synthetic rubber produced from nonrenewable fossil fuel requires high energy costs and is dependent on the presumed unstable petroleum price. Natural rubber latex (NRL) is one of the major alternative sustainable rubber sources since it is derived from the plant &lsquo, Hevea brasiliensis&rsquo, Our study focuses on integrating sustainably processed carboxycellulose nanofibers from untreated jute biomass into NRL to enhance the mechanical strength of the material for various applications. The carboxycellulose nanofibers (NOCNF) having carboxyl content of 0.94 mmol/g was prepared and integrated into its nonionic form (&ndash, COONa) for its higher dispersion in water to increase the interfacial interaction between NRL and NOCNF. Transmission electron microscopy (TEM) and atomic force microscopy (AFM) analyses of NOCNF showed the average dimensions of nanofibers were length (L) = 524 &plusmn, 203 nm, diameter (D) 7 &plusmn, 2 nm and thickness 2.9 nm. Furthermore, fourier transform infra-red spectrometry (FTIR) analysis of NOCNF depicted the presence of carboxyl group. However, the dynamic light scattering (DLS) measurement of NRL demonstrated an effective diameter in the range of 643 nm with polydispersity of 0.005. Tensile mechanical strengths were tested to observe the enhancement effects at various concentrations of NOCNF in the NRL. Mechanical properties of NRL/NOCNF films were determined by tensile testing, where the results showed an increasing trend of enhancement. With the increasing NOCNF concentration, the film modulus was found to increase quite substantially, but the elongation-to-break ratio decreased drastically. The presence of NOCNF changed the NRL film from elastic to brittle. However, at the NOCNF overlap concentration (0.2 wt. %), the film modulus seemed to be the highest.

Este artigo tem como objetivo cartografar a experiência de um acadêmico de psicologia em um encontro na cidade de XXXX/XX, com a temática de Redução de Danos (RD) e Gestão Autônoma da Medicação (GAM). Na ocasião estavam presentes usuários, estudantes, residentes, professores e profissionais do campo da saúde mental, álcool e outras drogas. O gatilho para o pesquisar foi a afetação do estudante a partir da fala de uma profissional que disse preferir um usuário de crack internado e amarrado do que oferecendo perigo para a sociedade, pois alguém que faz o consumo de um benzodiazepínico não mata para sustentar seu vício. Um usuário, por sua vez, a respondeu dizendo que ela não via isso acontecer porque o medicamento está disponível em qualquer farmácia. Efetuamos um olhar para as reverberações desse acontecimento, constituindo nossa trajetória/método. Buscamos construir uma narrativa histórica das drogas por meio do campo discursivo da RD e GAM, além de mapear a produção de subjetividade na relação com as drogas nos movimentos desse encontro. Consideramos, por fim, que há algo de novo entre RD e GAM: o uso de medicamentos vem se tornando um problema tão importante quanto o consumo de drogas ilícitas. São investidos saberes e práticas para dar conta de inventar e disputar formas de entender e conduzir os modos de vida que incluem as substâncias psicoativas.

Suppressive effect of bis (3-amino-2-pyridine) diselenide (BAPD) on psychiatric disorders - atopic dermatitis (AD) comorbidity in mice was investigated. To sensitize the animals, 2,4-dinitrochlorobenzene (DNCB) was applied to their dorsal skin on days 1-3. Mice were challenged with DNCB on their ears and dorsal skin on days 14, 17, 20, 23, 26, and 29. BAPD and Dexamethasone were administered to the animals, from days 14-29, and skin severity scores and behavioral tests were determined. Oxidative stress and inflammatory parameters were evaluated on the dorsal skin of mice. Na+, K+-ATPase activity and corticosterone levels were determined in hippocampus/cerebral cortex and plasma of mice, respectively. BAPD improved cutaneous damage, scratching behavior, inflammatory and oxidative stress markers. BAPD showed anxiolytic- and antidepressant-like effects and restored Na+, K+-ATPase activity and corticosterone levels. The present study was performed using female mice due the susceptibility for this disease. But, the evaluation of AD model in male mice would help to verify whether the male gender has the same predisposition to present this pathology. Our data demonstrated the suppressive effect of BAPD on psychiatric disorders - AD comorbidity by regulating inflammatory and oxidative status in mice.

Made available in DSpace on 2018-11-26T17:45:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-02-06 Ministerio da Ciencia Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Silversides are fish that inhabit marine coastal waters, coastal lagoons, and estuarine regions in southern South America. The freshwater (FW) silversides have the ability to tolerate salinity variations. Odontesthes humensis have similar habitats and biological characteristics of congeneric O. bonariensis, the most studied silverside species and with great economic importance. Studies revealed that O. bonariensis is not fully adapted to FW, despite inhabiting hyposmotic environments in nature. However, there is little information about stressful environments for cultivation of silverside O. humensis. Thus, the aim of this study was to evaluate the stress and osmoregulation responses triggered by the osmotic transfers on silverside O. humensis. Silversides were acclimated to FW (0 ppt) and to brackish water (BW, 10 ppt) and then they were exposed to opposite salinity treatment. Silverside gills and blood were sampled on pre-transfer (D0) and 1, 7, and 15 days (D1, D7, and D15) after changes in environmental salinity, the expression levels of genes atp1a3a, slc12a2b, kcnh1, and hspa1a were determined by quantitative reverse transcription-PCR for evaluation of osmoregulatory and stress responses. Furthermore, glycemia, hematocrit, and osmolality were also evaluated. The expression of atp1a3a was up-and down-regulated at D1 after the FW-BW and BW-FW transfers, respectively. Slc12a2b was up-regulated after FW-BW transfer. Similarly, kcnh1 and hspa1a were up-regulated at D1 after the BW-FW transfer. O. humensis blood osmolality decreased after the exposure to FW. It remained stable after exposure to BW, indicating an efficient hyposmoregulation. The glycemia had a peak at D1 after BW-FW transfer. No changes were observed in hematocrit. The return to the pre-transfer levels at D7 after the significant increases in responses of almost all evaluated molecular and blood parameters indicated that this period is enough for acclimation to the experimental conditions. In conclusion, our results suggest that BW-FW transfer is more stressful to O. humensis than FW-BW transfer and the physiology of O. humensis is only partially adapted to FW. Univ Fed Pelotas, Technol Dev Ctr, Lab Struct Genom, Pelotas, Brazil Univ Fed Pelotas, Inst Biol, Lab Physiol, Pelotas, Brazil Univ Fed Pelotas, Technol Dev Ctr, Lab Canc Biotechnol, Pelotas, Brazil Sao Paulo State Univ, Inst Biosci Botucatu, Dept Genet, Genom & Mol Evolut Lab, Botucatu, SP, Brazil Univ Fed Pelotas, Technol Dev Ctr, Lab Vaccinol, Pelotas, Brazil Sao Paulo State Univ, Inst Biosci Botucatu, Dept Genet, Genom & Mol Evolut Lab, Botucatu, SP, Brazil Ministerio da Ciencia: 422292/2016-8 CNPq: 472210/2013-0 CAPES: AUXPE 2900/2014

Considerando as orientações curriculares para o Ensino Médio, desenvolveu-se uma atividade didática para o ensino de Física. A prática pedagógica buscou o lúdico no ensino de eletrostática por meio de jogos didáticos, sem a finalidade de estimular a competição ou ranking acadêmico. A atividade denominada “Corrida de latas eletrostaticamente móveis” buscou contribuir para o ensino de eletrostática previamente introduzido na amostra de alunos, reforçando a aprendizagem numa experiência divertida.

O tratamento de fraturas tem como objetivo a consolidacao ossea e o retorno precoce da funcao do membro. A escolha do metodo de fixacao envolve varios fatores relacionados ao individuo e ao proprietario. A associacao de duas placas colocadas em diferentes faces do osso foi recentemente descrita por alguns autores com intuito de conferir maior resistencia ao sistema. Foram realizadas osteossinteses em 5 casos onde foram aplicadas a tecnica de duas placas sobrepostas como metodo de fixacao de fraturas em ossos longos. Atraves de acompanhamentos radiograficos os animais apresentaram consolidacao entre 45 e 120 dias. Nos relatos em questao a associacao de placas sobrepostas foi um recurso viavel, nao apresentaram complicacoes e propiciaram retorno funcional precoce do membro afetado.

CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Resumo - O tecido ósseo é fundamental na locomoção, proteção dos órgãos internos e na produção de células do sistema hematopoiético e imunológico dos animais. As lesões neste tecido, principalmente nos animais de companhia, normalmente ocorrem devido a traumas. As fraturas têm sido bastante discutidas na ortopedia veterinária, principalmente quanto aos métodos de tratamento, tempo de recuperação e complicações. Uma opção no tratamento das lesões ósseas são os enxertos. Com o objetivo de se estudar a influência do enxerto omental livre autógeno na reparação de falha óssea utilizou-se 32 coelhos adultos, machos, não castrados da raça Nova Zelândia, com idade entre 6 e 7 meses, divididos em dois grupos (GI controle e GII tratados com enxerto omental) com 16 animais cada. Falhas ósseas segmentares de 1cm foram criadas cirurgicamente no terço médio da diáfise radio direito. No mesmo momento cirúrgico realizou-se também a celiotomia para obtenção do enxerto a partir do omento, inclusive nos animais do GI. Os animais do GII tiveram as falhas ósseas cobertas com o enxerto omental livre autógeno. Imediatamente após as cirurgias, aos 45 e 90 dias, os animais tiveram os respectivos antebraços radiografados para posterior avaliação do processo de regeneração óssea. Também foi realizada coleta de sangue para a dosagem da fosfatase alcalina óssea nos dias 0 (antes das cirurgias), 45 e 90 (após as cirurgias). Metade dos animais de cada grupo foram eutanásiados aos 45dias e aos 90 dias respectivamente para realização da coleta de amostra da área anteriormente lesionada visando a confecção de laminas histológicas. Não houve diferença nos níveis séricos da FAO entre GI e GII aos 45 e nem aos 90 dias (p=0,42), nem na avaliação radiográfica comparando os grupos (p=0,99). Histologicamente também não foi notada diferença no processo de regeneração óssea entre GI e GII, exceto pela presença de resíduos do enxerto omental em absorção, sugerindo uma reação tipo corpo estranho. O enxerto omental autógeno livre, no tratamento de falhas ósseas com 1cm de extensão no osso Rádio de coelhos Nova Zelândia, não influenciou o processo de regeneração óssea. Abstract - Bone tissue is essential in locomotion, internal organs protection and in the hematopoietic and immunological systems cells production of animals. Lesions in this tissue, especially in pet animals, usually occur due to trauma. Fractures have been widely discussed in veterinary orthopedics, mainly regarding treatment methods, recovery time and complications. An option in the treatment of bone lesions are the grafts. with aim to study the influence of the autogenous free omental graft on the repair of bone defect, 32 male, uncastrated New Zealand rabbits aged between 6 and 7 months were divided into two groups (GI control and GII treated with free omental graft) with 16 animals each. 1 cm Segmental bone defects were surgically criated in the middle third of the right Radio. At the same surgical procedure, the celiotomy was also performed to obtain the graft from the omentum, including the GI animals. The GII animals had bony defects covered with autogenous free omental graft. Immediately after the surgeries and at 45 and 90 days, the animals had their respective forearms radiographed for later images evaluation. Blood samples were also collected for the measurement of bone alkaline phosphatase on days 0 (before surgery), 45 and 90 (after surgery). Half of the animals in each group were euthanized at 45 days and 90 days, respectively, to collect samples from the previously injured area, aiming the preparation of histological laminae. There was no difference in bone alkaline phosphatase serum levels between GI and GII at 45 and 90 days (p = 0.42), nor at the radiographic evaluation comparing the groups (p = 0.99). Histologically, no difference was also observed in the regeneration process of bone defects between GI and GII, except for the presence of residues from the omental graft under absorption, suggesting a foreign body reaction. The autogenous free omental graft, on treatment of 1 cm extension bone defects in New Zealand rabbit Radio, did not influence the bone regeneration process. Dissertação (Mestrado)

Some mammalian reference genes, which are widely used to normalize the qRT-PCR, could not be used for this purpose due to its high expression variation. The normalization with false reference genes leads to misinterpretation of results. The silversides (Odontesthes spp.) has been used as models for evolutionary, osmoregulatory and environmental pollution studies but, up to now, there are no studies about reference genes in any Odontesthes species. Furthermore, many studies on silversides have used reference genes without previous validations. Thus, present study aimed to was to clone and sequence potential reference genes, thereby identifying the best ones in Odontesthes humensis considering different tissues, ages and conditions. For this purpose, animals belonging to three ages (adults, juveniles, and immature) were exposed to control, Roundup®, and seawater treatments for 24 h. Blood samples were subjected to flow-cytometry and other collected tissues to RNA extraction; cDNA synthesis; molecular cloning; DNA sequencing; and qRT-PCR. The candidate genes tested included 18s, actb, ef1a, eif3g, gapdh, h3a, atp1a, and tuba. Gene expression results were analyzed using five algorithms that ranked the candidate genes. The flow-cytometry data showed that the environmental challenges could trigger a systemic response in the treated fish. Even during this systemic physiological disorder, the consensus analysis of gene expression revealed h3a to be the most stable gene expression when only the treatments were considered. On the other hand, tuba was the least stable gene in the control and gapdh was the least stable in both Roundup® and seawater groups. In conclusion, the consensus analyses of different tissues, ages, and treatments groups revealed that h3a is the most stable gene whereas gapdh and tuba are the least stable genes, even being considered two constitutive genes.

In this work, we present the synthesis, characterization, and computational study of the supramolecular arrangement of a new cinnamic acid derivative: ethyl-(2E)-3-(4-hydroxy-3,5-dimethoxyphenyl)-prop-2-enoate (EHD). Single crystals of EHD were obtained using ethyl ether as solvent and a slow evaporation technique. Its crystallographic structure, derived from X-ray diffraction experiments, includes a disordered water molecule on the EHD supramolecular structure. This water molecule participates in four O-H···O hydrogen bonds, which are arranged as a centrosymmetric H-bond array with the water at the center. Electronic and structural properties of the isolated EHD molecule and of the EHD molecule in the presence of one water molecule were calculated at the B3LYP/6-311++G(2d,2p) level of theory. These calculations show that the HOMO-LUMO energy gap of EHD decreases upon the introduction of the water molecule, suggesting that EHD becomes a stronger electron acceptor. These results indicate that the water molecule helps to stabilize the crystal structure in this system containing unequal numbers of acceptor and donor atoms. The supramolecular synthon involving the disordered water molecule and the supramolecular features presented here provide new possibilities in the design of functional materials and should also help us to gain a deeper understanding of the processes by which molecules recognize biological targets.

A sustentabilidade está no rol de paradigmas capazes de dar resposta à crise socioambiental que surge no decorrer das últimas décadas. As dimensões social e econômica se inserem no arcabouço do desenvolvimento e se relacionam estreitamente com a sustentabilidade na concepção contemporânea. O objetivo deste artigo é discutir a experiência de identificação de indicadores que expressem a percepção da sustentabilidade na perspectiva dos atores da Rede de Comércio Justo e Solidário Bem Terra através da análise de uma ferramenta interna de acreditação agroecológica. Para a realização deste trabalhoutilizou-se como metodologia pesquisa bibliográfica, seguida de pesquisa documental, em concomitância com a observação participante. A ferramenta de acreditação apresenta grande potencial de contribuir com a transformação social, econômica e ambiental, servindo de estímulo para as experiências de transição agroecológica ora em curso na Rede Bem da Terra, assim como para a construção de indicadores de sustentabilidade, que poderão contribuir para um modelo de desenvolvimento mais sustentável.

A new terpenoid-like bischalcone (1E,4E)-1-(4-nitrophenyl)-5-(2,6,6-trimethylcyclohex-1-enyl)-penta-1,4-dien-3-one was synthesized from a β-ionone and 4-nitrobenzaldehyde by Claisen-Schmidt condensation reaction and its structure was determined by single crystal X-ray diffraction analysis. Additionally, this compound was featured by powder X-ray diffraction, 1H, 13C NMR and IR spectroscopy techniques. The molecule in the asymmetric unit showed disordered occupancy sites over two positions for the trimethylcyclohexene atoms. These two conformations were related by a rotation of about 180° around the axis of the C-C bond linking the six-membered ring and the olefin carbons. Single-molecule calculations using the DFT method have strengthened this structural finding, since our theoretical approaches also suggest two well-defined conformations of similar energies which resemble the molecular geometries determined by X-ray diffraction. Furthermore, the inspection of the crystal packing revealed that the hydrogen bonding patterns are different for each conformation of the compound reported here.

Melatonin has been used as a supplement in culture medium to improve the efficiency of in vitro produced mammalian embryos. Through its ability to scavenge toxic oxygen derivatives and regulate cellular mRNA levels for antioxidant enzymes, this molecule has been shown to play a protective role against damage by free radicals, to which in vitro cultured embryos are exposed during early development. In vivo and in vitro studies have been performed showing that the use of nanocapsules as active substances carriers increases stability, bioavailability and biodistribution of drugs, such as melatonin, to the cells and tissues, improving their antioxidant properties. These properties can be modulated through the manipulation of formula composition, especially in relation to the supramolecular structures of the nanocapsule core and the surface area that greatly influences drug release mechanisms in biological environments. This study aimed to evaluate the effects of two types of melatonin-loaded nanocapsules with distinct supramolecular structures, polymeric (NC) and lipid-core (LNC) nanocapsules, on in vitro cultured bovine embryos. Embryonic development, apoptosis, reactive oxygen species (ROS) production, and mRNA levels of genes involved in cell apoptosis, ROS and cell pluripotency were evaluated after supplementation of culture medium with non-encapsulated melatonin (Mel), melatonin-loaded polymeric nanocapsules (Mel-NC) and melatonin-loaded lipid-core nanocapsules (Mel-LNC) at 10−6, 10−9, and 10−12 M drug concentrations. The highest hatching rate was observed in embryos treated with 10−9 M Mel-LNC. When compared to Mel and Mel-NC treatments at the same concentration (10−9 M), Mel-LNC increased embryo cell number, decreased cell apoptosis and ROS levels, down-regulated mRNA levels of BAX, CASP3, and SHC1 genes, and up-regulated mRNA levels of CAT and SOD2 genes. These findings indicate that nanoencapsulation with LNC increases the protective effects of melatonin against oxidative stress and cell apoptosis during in vitro embryo culture in bovine species.

Com o intuito de se conhecer a estrutura molecular e cristalina dos compostos químicos para posteriores análises físico-químicas e estudos de aplicação, lança-se mão da cristalografia como metodologia científica, que permite conhecer a estrutura de qualquer composto químico cristalino sem nenhuma informação a priori. Esta metodologia está baseada em conceitos como difração de raios X por materiais cristalinos, simetria, lei de Bragg e transformada de Fourier. O presente trabalho tem como finalidade apresentar as principais etapas do trabalho cristalográfico, desde a coleta de dados em monocristais, solução e refinamento da estrutura até as etapas de análise e validação do modelo cristalográfico obtido e posterior depósito no banco de dados apropriados.

A cruzaína, principal cisteíno protease do Trypanosoma cruzi, é um alvo terapêutico validado para o tratamento da doença de Chagas. Grande parte dos inibidores desta enzima é constituída de peptídeo-miméticos do tipo covalente irreversível cujo desenvolvimento, entretanto, tem sido evitado devido ao potencial efeito off target e ao perfil farmacocinético indesejável. O grupo de química medicinal NEQUIMED/ IQSC/USP vêm utilizando métodos avançados em quimioinformática para a identificação de inibidores reversíveis da cruzaína e de outras cisteíno proteases. Contudo, para que estes inibidores se desenvolvam a compostos matrizes mais eficientes, informações computacionais e estruturais de seus Modos de Interação (MOB) com a proteína alvo tornam-se fundamentais. Neste trabalho, a cristalografia de raios X foi utilizada para descrever em detalhes o MOB de três importantes inibidores reversíveis da cruzaína identificados pelo NEQUIMED: a dipeptidil-nitrila Neq0409 e os dois fragmentos moleculares Neq0147 e Neq0176. De modo a evitar a auto-proteólise experimentada pela cruzaína nativa que torna desafiadora a cristalização com inibidores de baixa afinidade como os fragmentos reversíveis, duas novas construções mutantes e inativas da cruzaína (a C25A e a C25S) foram desenvolvidas. A C25S foi validada como modelo cristalográfico representativo dado a coerência do MOB elucidado nesta enzima mutante e na cruzaína nativa para o inibidor mais estudado da cruzaína, o K777. A descrição da presença, orientação, conformação e modo de ação no sítio, além do completo padrão de interações fornecidos por estas estruturas cristalinas, validaram ortogonalmente os MOB preditos e os métodos de planejamento in silico usados no NEQUIMED, permitindo a identificação de inibidores muito mais potentes análogos ao Neq0147 e ao Neq0409. O grupo 2-acetamidotiofeno-3-carboxamida do Neq0176 e o anel heterocíclico de cinco membros baseado no 1,3,4-oxadiazol do Neq0147 foram identificados como alternativos aos tradicionais esqueletos peptídeo-miméticos. O impacto do efeito proteolítico na qualidade e na resolução de estruturas cristalográficas na cruzaína foi melhor compreendido a partir de duas estruturas de alta resolução obtidas para a cruzaína nativa em complexo com o metanotiossulfonato de metila (MMTS) e a iodoacetamida. Estes resultados permitiram compreender as bases experimentais para a cristalização de inibidores reversíveis de baixa afinidade. Todos os resultados estruturais obtidos via cristalografia de raios X neste trabalho são úteis para mapear as bases estruturais essenciais para o planejamento de futuros inibidores mais potentes e seletivos da cruzaína. Cruzain, the major Trypanosoma cruzi cysteine protease, is a validated therapeutic target for the search of new medicines for the treatment of Chagas disease. A myriad of inhibitors of this enzyme consists of covalent irreversible peptidomimetics whose development has been impaired due to potential off target effect and undesirable pharmacokinetic profile. Modern cheminformatic methods employed by The Medicinal Chemistry Group (NEQUIMED/IQSC/USP) were used to identify cruzain reversible inhibitors. Their optimization for more efficient compounds can be accomplished by the use of data and information gathered from computational and structural modes of interaction (MOB). In this doctoral thesis, the X-ray crystallography was used to describe in detail the MOB of three important new cruzain reversible inhibitors: the dipeptidil-nitrile Neq0409 and the two molecular fragmentos Neq0147 and Neq0176. In order to avoid cruzain self-proteolysis during crystallization with low affinity reversible inhibitors such as the identified fragments, two new mutant and inactive constructs of cruzain (the C25S and C25A) were designed to upholding the same properties of the wild type catalytic site. The C25S was validated as representative crystallographic model given the coherence of the MOB elucidated for the best-known and studied cruzain inhibitor, the K777. The description of the presence, orientation, conformation and mode of action at the site, besides the complete pattern of bimolecular interactions, provided by these crystalline structures, orthogonally validated the predicted in silico MOB and allowed the identification of other potent inhibitors analogous to the Neq0147 and the Neq0409. The 2-acetamidothiophene-3-carboxamide moiety (Neq0176) and heterocyclic five-membered ring based on the 1,3,4-oxadiazole (Neq0147) were thereby identified as attractive alternatives to traditional peptidomimetics. The impact of proteolytic effect on the quality and resolution of crystallographic structures in cruzain was best understood from two high-resolution structures obtained for the native cruzain in complex with methyl methanethiolsulfonate (MMTS) and iodoacetamide. These results allow the understanding of experimental basis for the crystallization with reversible inhibitors of low affinity such as fragments. All structural results obtained by X-ray crystallography together with the catalytic site depiction using GRID and SuperStar methods are useful for mapping the essential structural basis for the design of future more potent and selective cruzain inhibitors.

A series of compounds based on the dipeptidyl nitrile scaffold were synthesized and assayed for their inhibitory activity against the T. cruzi cysteine protease cruzain. Structure activity relationships (SARs) were established using three, eleven and twelve variations respectively at the P1, P2 and P3 positions. A K i value of 16 nM was observed for the most potent of these inhibitors which reflects a degree of non-additivity in the SAR. An X-ray crystal structure was determined for the ligand-protein complex for the structural prototype for the series. Twenty three inhibitors were also evaluated for their anti-trypanosomal effects and an EC50 value of 28 μM was observed for the most potent of these. Although there remains scope for further optimization, the knowledge gained from this study is also transferable to the design of cruzain inhibitors based on warheads other than nitrile as well as alternative scaffolds., Author Summary Chagas disease is a parasitic infection with high morbidity and mortality that is endemic in much of Latin America where it remains a serious public health problem. With increased migration, Chagas disease represents an emerging worldwide challenge and there is an urgent, unmet need for safe and effective medication. The available drugs to treat Chagas disease may be effective in the acute phase of the disease, but efficacy in the chronic phase remains controversial. They can cause serious side effects that lead sufferers to abandon treatment. Using a hypothesis-driven approach to molecular design and drawing on cysteine protease cruzain structural information, we have mapped structure-activity relationships for a dipeptidyl nitrile scaffold and demonstrated that compounds are competitive inhibitors, bind reversibly and bear trypanocidal activity. The binding mode revealed by the crystal structure of the protein-ligand complex for one of the inhibitors shows that binding involves the formation of a covalent bond between the catalytic cysteine and the nitrile carbon. As such, we believe that our study represents a valuable step in the search for new drugs for the treatment of a neglected disease that continues to affect the lives of millions of people.