Desenvolvimento de metodologias de controle do Aedes aegypti baseadas em compostos de baixa toxidade ao ser humano e ao meio ambiente

Orientador : Prof. Dr. Francisco de Assis MarquesTese (doutorado) - Universidade Federal do Paraná, Setor de Ciências Exatas, Programa de Pós-Graduação em Química. Defesa: Curitiba, 26/04/2017Inclui referências : f. 77-83Resumo: A fêmea do mosquito Aedes aegypti é o principal vetor responsável pela transmissão do vírus da dengue, zika, chikungunya e febre amarela. Enquanto não existe uma vacina eficaz para controle dessas doenças, a única forma de reduzir sua incidência se restringe ao controle do A. aegypti. Desta forma se faz necessário a busca por novos métodos de controle baseados em substâncias com propriedades inseticidas que reduzam o impacto ambiental e que sejam menos tóxicas ao ser humano, bem como minimizem desenvolvimento de resistência por parte do mosquito. Este trabalho teve como objetivo avaliar a atividade larvicida de compostos sintéticos, para o controle do A. aegypti bem como buscou-se determinar a atividade adulticida e repelente de compostos de origem vegetal e ou de derivados sintéticos frente a adultos de A. aegypti. Os resultados dos estudos nos levaram a identificar um composto com forte ação larvicida, pertencente a uma classe de compostos ainda não empregada para o controle de pragas a nível mundial. Os estudos também nos possibilitaram determinar alguns compostos responsáveis por uma ação adulticida bem como foi possível determinar a relação entre os compostos, necessária para que se observe o efeito adulticida de forma mais pronunciada. No que tange ao efeito de repelência, foi sintetizada uma substância, a partir de um produto natural, que apresentou forte ação repelente. Vale a pena ressaltar que essa substância é produzida em apenas uma etapa e utilizando os conceitos da química verde, como solventes atóxicos e elevado rendimento. Esses resultados abrem perspectivas para pesquisa no sentido de desenvolver um novo larvicida para substituir o temefós, pesticida com amplo espectro de atuação, bem como para o desenvolvimento de um novo adulticida, baseado na presença de compostos naturais, de baixa toxicidade ao ser humano, que poderá ser usado para substitui os piretroides. Também se tem como resultado desse trabalho a possibilidade de se fornecer ao poder público um repelente que pode ser sintetizado em grande escala, a partir de produto natural de baixa toxidade, podendo atender às demandas nacionais sem a necessidade de se importar princípios ativos com ação repelente. Palavras-chave: Aedes aegypti. Larvicida. Adulticida. Repelente.Abstract: The female of the Aedes aegypti mosquito is the main vector responsible for the transmission of the dengue virus, zika, chikungunya and yellow fever. While there is no effective vaccine to control these diseases, the only way to reduce their incidence is to control A. aegypti. In this way it is necessary to search for new methods of control based on substances with insecticidal properties that reduce the environmental impact and that are less toxic to the human being, as well as minimize the development of resistance by the mosquito. The objective of this work was to evaluate the larvicidal activity of synthetic compounds for the control of A. aegypti as well as to determine the adulticidal and repellent activity of compounds of plant origin and of synthetic derivatives against adults of A. aegypti. The results of the studies led us to identify a compound with strong larvicidal action, belonging to a class of compounds not yet used for pest control worldwide. The studies also allowed us to determine some compounds responsible for an adulticidal action as well as it was possible to determine the relationship between the compounds, necessary to observe the adulticidal effect in a more pronounced way. As regards the effect of repellency, a substance was synthesized from a natural product, which presented a strong repellent action. It is worth mentioning that this substance is produced in only one step and using the concepts of green chemistry, such as non-toxic solvents and high yield. These results open perspectives for research to develop a new larvicide to replace temephos, a pesticide with a broad spectrum of action, as well as for the development of a new adulticide, based on the presence of natural compounds with low toxicity to humans. Can be used to replace the pyrethroids. The result of this work is also the possibility of providing the public authorities with a repellent that can be synthesized on a large scale from a natural product of low toxicity and can meet national demands without the need to import active ingredients with repellent action. Key-words: Aedes aegypti. Larvicida. Adulticide. Repellent

Determinação estrutural, síntese e aplicação de compostos de origem botânica e ou derivados sintéticos visando o controle populacional e o efeito de repelência do mosquito Aedes aegypti

Orientador : Prof. Dr. Francisco de Assis MarquesDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Exatas, Programa de Pós-Graduação em Química. Defesa: Curitiba, 31/10/2012Inclui referências : f. 80-88Resumo: A fêmea do mosquito Aedes aegypti é o principal vetor responsável pela transmissão do vírus da dengue. Enquanto não existe uma vacina eficaz para controle dessa doença, a única forma de reduzir sua incidência se restringe ao controle do A. aegypti. Tendo em vista à necessidade da busca por novos métodos de controle que reduzam o impacto ambiental e que não permitam o desenvolvimento de resistência por parte do mosquito, a utilização de extratos vegetais como alternativa para se encontrar substâncias com propriedades inseticidas para o controle de mosquitos em geral, podendo atuar como um método alternativo aos inseticidas sintéticos de amplo espectro de ação. Este trabalho teve como objetivo avaliar a atividade larvicida de compostos sintéticos, pouco tóxicos ao ser humano e ao meio ambiente, para o controle do A. aegypti bem como buscou-se determinar a atividade repelente de compostos de origem vegetal frente a adultos de A. aegypti. Os resultados do estudo nos levaram a descobrir um composto com forte ação larvicida, pertencente a uma classe de compostos ainda não empregada para o controle de pragas a nível mundial. Também foi possível determinar quais os compostos identificados do extrato de Ocimum basilicum que são os responsáveis pela ação de repelência já descrita em literatura para o óleo essencial desta planta. Uma surpreendente ação adulticida de uma mistura de compostos presentes no óleo essencial de Ocimum basilicum foi observada durante os testes de repelência. Os estudos nos possibilitaram determinar os compostos responsáveis por essa ação adulticida bem como a relação entre os compostos, necessária para que se observe o efeito adulticida. Esses resultados abrem perspectivas para que se trabalhe no sentido de desenvolver um novo larvicida para substituir o temefós, pesticida com amplo espectro de atuação e que deverá ser banido do mercado em poucos anos, bem como para o desenvolvimento de um novo adulticida, baseado na presença de compostos naturais, de baixa toxicidade ao ser humano, que poderá ser usado para substitui os piretróides que também deverão ser retirados do mercado mundial em poucos anos. Palavras-chave: Aedes aegypti. Larvicida. Repelente. Adulticida.Abstract: The female mosquito Aedes aegypti is the main vector responsible for transmission of dengue virus. While there is no effective vaccine to control the disease, the only way to reduce their incidence is restricted to the control of A. aegypti. Given the need to search for new control methods that reduce environmental impact and not allow the development of resistance in the mosquito, the use of plant extracts are an alternative to find substances with insecticidal properties to control mosquitoes in general, can act as an alternative method to chemical insecticides broad spectrum of action. This study aimed to evaluate the larvicidal activity of synthetic compounds, some toxic to humans and the environment, for the control of A. aegypti and we sought to determine the repellent activity of compounds of plant origin against adults of A. aegypti. The study results led us to discover a compound with strong larvicidal action, belongs to a class of compounds not used to control pests worldwide. It was also possible to determine which of the identified compounds Ocimum basilicum extract that are responsible for the repellent action already described in the literature for the essential oil of this plant. A surprising adulticide action of a mixture of compounds in the essential oil of Ocimum basilicum was observed during testing repellency. The studies allowed us to determine the compounds responsible for this action adulticide well as the relationship between the compounds necessary to note that the adulticide effect. These results open perspectives in order to work towards developing a new larvicide temephos to replace pesticide with broad spectrum of activity and should be banned from the market in a few years, as well as for the development of a new adulticide, based on the presence of natural compounds of low toxicity to humans, which can be used to replace pyrethroids which should also be removed from the world market in a few years. Key-words: Aedes aegypti. Larvicida. Repellent. Adulticid

Experiências dançantes: corporeidade de sujeitos nômades

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências da Educação, Programa de Pós-Graduação em Educação, Florianópolis, 2011Esta dissertação teve como objetivo analisar os significados atribuídos à experiência de viver nas ruas pelos acolhidos na Associação Vida Nueva (AVN) localizada no município da Palhoça/SC, e como estes participam na constituição da sua corporeidade. A concepção metodológica configura-se como estudo de caso de abordagem etnográfica numa relação aproximada entre pesquisadora e os atores do contexto investigado, sendo utilizado como instrumentos de coleta de dados, diário de campo, documentos disponibilizado pela instituição, observações e entrevistas. No decorrer dapesquisa procurei legitimar os fios que configuram os modos de ser e estar nas ruas, marcado pelos deslocamentos infinitos. Os desdobramentos que compõem a caminhada tecem olhar sobre breve panorama da população em situação de rua; a estética das ruas e seus adereços, assim como o movimento das piruetas no trecho pelos protagonistas da pesquisa. Para análise foi privilegiado as narrativas de sujeitos que passaram e/ou estão na AVN, em especial dois deles, Val e Jota, denominados protagonistas desta pesquisa. Atítulo de considerações, esse estudo possibilita defender as imprevisibilidades inscritas na corporeidade dos sujeitos que passaram pela experiência de viver nas ruas e apropriam-se dos deslocamentos como legítimos de suas existências. As relações estabelecidas nas e com as ruas são afetadas pelas adversidades que a compõem pelos bons e ruins momentos. E a permanência em habitar as ruas, torna-se, algumas vezes, in-evitável pelo desejo-escolha de ali estar.The purpose of this dissertation is to analyze the meanings that individuals, who have been taken in by the Associação Vida Nueva (AVN) institution located in Palhoça SC, assign to the experience of living on the streets, and how these meanings take part in the construction of their corporeality. The methodology used for the research was a case study, with an ethnographic approach, that resulted from the relationship established by the researcher, the actors and the investigated context. The tools used for data collection includes a field journal, documents made available by the institution, observations, and information brought up in conversations with participants in different situations and opportunities. During this journey, I sought to legitimize the threads that shape the ways of being on the streets, marked by unlimited movements that place the individuals on the streets and stretches. I emphasized the developments that arrange the treks of each one of the participants, with analogies taken from a brief overview on street population. I added to my arguments a look at the esthetics adorning these streets, at the accessories decorating this lifestyle, and at the juggling movements that happen on the stretches traveled by the leading characters of this research. In the analysis, I highlighted the narratives by two individuals who are and/or were staying at AVN. They were called Val and Jota. As for the considerations, this study ventures to defend the idea that the unpredictable elements inscribed in the corporeality of individuals who lived on the streets are dimensions that encourage the movements and that make the self-perceptions, as well as the perceptions of the world, authentic. The affinities established on and with the streets are affected by the adversities they experience in their everyday lives, in which they dance between good and bad moments. This dissertation understands that the stay on the streets results in a living style that becomes an in-evitable welcoming made of desires, choices, addictions, sense of being there

DNA methylation dynamics and epigenetic diversity in development

Epigenetics refers to heritable changes in phenotype without alterations to the genotype. Epigenetic changes involve two main mechanisms: DNA methylation and histone modification. Methylation of DNA at cytosine bases is the best-studied epigenetic process to date. CpG methylation states are thought to be maintained throughout cell divisions. However, loss of DNA methylation or DNA demethylation has been observed in specific stages of mammalian development. Such prominent examples of developmental DNA demethylation processes occur in developing primordial germ cells and in preimplantation embryos. However, little is known about DNA methylation changes of other tissues in mammalian development. Therefore, the first aim of this PhD study was to investigate changing nuclear distributions and levels of DNA methylation during development in order to discover dynamic variations amongst developing mouse tissues. In addition, a transgenic MBD-GFP mouse was employed to visualise DNA methylation in tissues. Several hypothetical mechanisms for the enzymatic removal of 5mC have been proposed. One of the proposed candidates is Tet-mediated successive oxidation of 5mC to generate 5hmC, 5fC and 5caC. 5hmC has therefore been considered as a transient intermediate in an active cytosine demethylation pathway. Nevertheless, some studies suggest that 5hmC may also function as an epigenetic modification in its own right. Thus, the second aim of this study was to address the research question of how and where 5hmC originates during development. In order to be able to identify tissues undergoing dynamic nuclear changes in DNA methylation and hydroxymethylation states during early mouse development, new working protocols for immunodetection of 5mC and 5hmC on tissue cryosections were required. The protocol optimisation for 5mC immunodetection is discussed in greater detail in Chapter 3. It was found that DNA methylation immunostaining of cryosections required heat-mediated DNA denaturation, which was partly compatible with protein immunostaining. Next, Chapter 4 focuses on identifying tissues undergoing dynamic changes in 5mC and 5hmC patterns during development from E9.5 to E14.5 mouse embryonic stages, using optimised immunohistochemistry protocols. These protocols revealed interesting dynamic observations of 5mC and 5hmC in the developing cerebral neocortex, surface ectoderm, liver, red blood cells, diaphragm and heart. These findings suggested that dynamic changes of 5mC and 5hmC during neocortical and compact myocardial development were in good agreement with a model where the formation of 5hmC may correlate with the loss of old 5mC, but the observations were also consistent with an involvement of de novo methylation in the generation of 5hmC. In other developing tissues, including surface ectoderm, liver, red blood cells, diaphragm and cardiac trabeculae, dynamic changes in 5mC and 5hmC levels were in line with a model where the 5hmC may act as a new epigenetic mark that functions independently. The optimised protocol also confirmed DNA demethylation of the germ cells at E12.5. The presence of three Tet family enzymes (Tet1, Tet2, Tet3) and de novo methyltransferase DNMT3A in mouse E12.5 tissues is reported in the second part of Chapter 4. It was found that Tet1, Tet2, Tet3 and Dnmt3a were present at detectable levels in neocortex, liver, diaphragm and heart. Contrastingly, no apparent signals for Tet1, Tet2, Tet3 and Dnmt3a were observed in red blood cells. This result was expected due to the very low levels of 5hmC staining in E12.5 red blood cells. The third aim of the present study was to investigate the existence of crosstalk between various epigenetic mechanisms. Thus, Chapter 5 focuses on exploring the relationship between 5mC and repressive histone marks, H3K9me3 and H3K27me3. Histone methylation dynamics at H3K9 and H3K27 were observed during mouse fetal development in neocortex and heart. The overall distribution patterns of H3K9me3 and H3K27me3 demonstrated strong association with developmental changes in 5mC, suggesting that these three repressive epigenetic marks work in concert to establish a silenced state of heterochromatin. Chapter 6, on the other hand, focuses on visualising DNA methylation in tissues using mouse transgenic tools. It was found that brain, liver, heart and neural tube expressed high levels of GFP. But no apparent developmental dynamics of GFP was observed. In conclusion, this study will contribute scientific understanding of dynamic DNA methylation and nuclear heterochromatin organisation during mammalian development, and its role in the specification and maintenance of cell lineages forming tissues and organs. This knowledge will provide insight into current barriers to cell fate reprogramming, which will be of benefit to cell regenerative biomedical technologies

Hubungan Antara Beban Kerja Fisik Dengan Kelelahan Kerja Pada Tenaga Kerja Bagian Produksi Tulangan Beton di PT Wijaya Karya Beton Tbk PPB Majalengka

The workload is one of the factors that may affect workers experiencing job burnout. Workers who have jobs that are too heavy and excessive muscle contraction will accelerate the body which will accelerate the occurrence of fatigue. Physical workload is excessive and uncontrolled can increase work exhaustion, as well as the level of fatigue on the production of concrete reinforcement that has a significant workload than any other production site. The purpose of this study was to determine the relationship of physical workload with fatigue on the part of workers in the production of concrete reinforcingbPT Wijaya Karya Beton Tbk PPB Majalengka. The research method used observational analytic research with cross sectional approach. The sampling technique using purposive sampling, the sample was 30 workers. Analysis of data used statistical test of Spearman-Rho correlation with a significant level (α = 0.05). Statistical tests using the Spearman-Rho found any significant relationship between physical workload with fatigue (p = 0.000) (r = 0.714) with the level of the relationship strong and indicate the direction of positive correlation. From the results study concluded there is the correlation between work load and fatigue

Effects of over-expressing ethylene responsive transcription factor on expression of selected fruit ripening-related genes in oil palm (Elaeis guineensis Jacq.) mesocarp

Gene expression is regulated mainly at the transcriptional level through binding of transcription factors to specific promoter regulatory elements. The expression profile of EgDREB, a transcription factor belonging to the AP2 family, was determined by reverse transcriptase (RT)-PCR and it was found that it was expressed at different stages of oil palm mesocarp development as well as in vegetative tissues (roots and leaves) but not in the mesocarp at the early ripening stage, which is 12 WAA (weeks after anthesis). Thus, the effects of over-expressing EgDREB on the transcriptional regulation of genes from five functional groups related to ripening were investigated in 12 WAA mesocarp of oil palm. Co-bombardment of 12 WAA mesocarp tissues with recombinant vector construct harbouring EgDREB and plasmid containing the GFP reporter gene was carried out. Fluorescent detection of GFP and verification via RT-PCR using GFP-specific primers enabled selection of successfully transformed tissues. Using transient expression assay, it was demonstrated that over-expression of EgDREB results in up-regulation of translationally controlled tumor protein (TCTP) and type 2 metallothionein-like genes (MET2a and MET2b). These proteins are categorised under biogenesis of the cellular component and proteins with binding functions or cofactor requirements. More specifically, the roles of metallothioneins are in homeostasis of essential metal ions and oxidative stress response. This may suggest that EgDREB is involved in regulating cellular processes related to the roles of these three proteins

A concepção das mulheres de Mirandópolis-São Paulo acerca do exame de papanicolau

http://dx.doi.org/10.5902/217976929647Aim: To verify the knowledge of women Mirandópolis- São Paulo about on the Pap smear. Method: Field research exploratory descriptive quantitative approach, performed in the city of Mirandópolis - São Paulo, with a sample of a hundred women, who answered a self-administered questionnaire. Data were analyzed with SPSS software, and arranged in percentage. Results: The data showed that 83% of women have performed the Pap smear, 42% of them perform the test once a year, with a loss of periodicity recommended by the Ministry of health. The embarrassment and lack of time presented as the main barrier for not performing the Pap smear. Conclusion: CCU screening should be focused on knowledge, and the reduction of the factors contributing to not realization of the pap smear, which comes from a relationship of trust and empathy among users and health professionals.Objetivo: Verificar el conocimiento que las mujeres presentan sobre el examen Papanicolaou. Método: investigación exploratoria de abordaje cuantitativo descriptivo, realizada en Mirandópolis – SP, con una muestra de cien mujeres, que respondieron un cuestionario auto-aplicable. Los datos fueron analizados con el software SPSS, y dispuestos en porcentaje. Resultados: Los datos mostraron que el 83% de las mujeres han realizado el examen de Papanicolaou, el 42% realizaron la prueba una vez al año, con una pérdida de la periodicidad recomendada por el Ministerio de la Salud. La vergüenza y la falta de tiempo se presentan como el principal obstáculo para no realizar el examen Papanicolaou. Conclusión: La detección del Cáncer de Cuello Uterino debe centrarse en el conocimiento, y la reducción de los factores que contribuyen para no realización del examen Papanicolaou, que viene de una relación de confianza y empatía entre las usuarias y el profesional de la salud.http://dx.doi.org/10.5902/217976929647Objetivo: verificar o conhecimento das mulheres de Mirandópolis-São Paulo apresentam sobre o exame de papanicolau. Método: pesquisa de campo exploratória de abordagem quantitativa descritiva, realizada no município de Mirandópolis – São Paulo, com amostra de cem mulheres, que responderam a um questionário autoaplicável. Os dados foram analisados com o auxílio do software SPSS, e dispostos em porcentagem. Resultados: os dados evidenciaram que, 83% das mulheres, já realizaram o exame Papanicolau, 42% delas realizam o exame uma vez ao ano, com perda da periodicidade preconizada pelo Ministério da Saúde. A vergonha e a falta de tempo apresentaram como a principal barreira para a não realização do exame de papanicolau. Conclusão: o rastreamento do Câncer do colo uterino deve estar focado no conhecimento, e na diminuição dos fatores contribuintes para a não realização do Exame de Papanicolau, que advém de um relacionamento empático e de confiança entre as usuárias e o profissional de saúde

Agrin Binds BMP2, BMP4 and TGFβ1

The C-terminal 95 kDa fragment of some isoforms of vertebrate agrins is sufficient to induce clustering of acetylcholine receptors but despite two decades of intense agrin research very little is known about the function of the other isoforms and the function of the larger, N-terminal part of agrins that is common to all isoforms. Since the N-terminal part of agrins contains several follistatin-domains, a domain type that is frequently implicated in binding TGFβs, we have explored the interaction of the N-terminal part of rat agrin (Agrin-Nterm) with members of the TGFβ family using surface plasmon resonance spectroscopy and reporter assays. Here we show that agrin binds BMP2, BMP4 and TGFβ1 with relatively high affinity, the KD values of the interactions calculated from SPR experiments fall in the 10−8 M–10−7 M range. In reporter assays Agrin-Nterm inhibited the activities of BMP2 and BMP4, half maximal inhibition being achieved at ∼5×10−7 M. Paradoxically, in the case of TGFβ1 Agrin N-term caused a slight increase in activity in reporter assays. Our finding that agrin binds members of the TGFβ family may have important implications for the role of these growth factors in the regulation of synaptogenesis as well as for the role of agrin isoforms that are unable to induce clustering of acetylcholine receptors. We suggest that binding of these TGFβ family members to agrin may have a dual function: agrin may serve as a reservoir for these growth factors and may also inhibit their growth promoting activity. Based on analysis of the evolutionary history of agrin we suggest that agrin's growth factor binding function is more ancient than its involvement in acetylcholine receptor clustering