Breast cancer is a growing international health epidemic, and patients in low and middle income countries (LMCs) have worse outcomes than those in high income countries. High quality, well-implemented guidelines help improve patient outcomes, but are often not resource-sensitive, and support therapies that may not be feasible in LMCs. A systematic review to address the content, quality, and resource-sensitivity of international breast cancer guidelines was completed. Also, a survey of global physicians evaluated the impact of resource setting on practice patterns and guideline use. Guideline use did not appear to be directed by quality (which was variable across guidelines) or resource-sensitivity (found in few guidelines). However, practice patterns were found to vary by resource setting and by continent, often due to the cost of certain therapies. In order for guidelines to better impact global breast cancer outcomes, they need to be of higher quality, more resource-sensitive, and better implemented.

Breast cancer is a growing international health epidemic, and patients in low and middle income countries (LMCs) have worse outcomes than those in high income countries. High quality, well-implemented guidelines help improve patient outcomes, but are often not resource-sensitive, and support therapies that may not be feasible in LMCs. A systematic review to address the content, quality, and resource-sensitivity of international breast cancer guidelines was completed. Also, a survey of global physicians evaluated the impact of resource setting on practice patterns and guideline use. Guideline use did not appear to be directed by quality (which was variable across guidelines) or resource-sensitivity (found in few guidelines). However, practice patterns were found to vary by resource setting and by continent, often due to the cost of certain therapies. In order for guidelines to better impact global breast cancer outcomes, they need to be of higher quality, more resource-sensitive, and better implemented.

Subtilase cytotoxin (SubAB) is the prototype of a novel class of AB₅ cytotoxins produced by Shiga-toxigenic Escherichia coli (STEC). The A subunit (SubA) is a serine protease that cleaves the ER chaperone BiP causing cell death by a previouslyundetermined mechanism. The B subunits of AB₅toxins typically recognise host cell glycan receptors and direct the subcellular transport of the A subunit. Although the function of SubA and its intracellular substrate have been elucidated, the B subunit (SubB) is relatively uncharacterised. The subcellular trafficking pathway of SubAB was initially examined. SubAB conjugated to Oregon Green 488 (SubAB-OG) was internalised by Vero cells by 5 min, and co-localised with its ER target BiP within 30 min. When Vero cells were incubated with SubAB-OG and either Alexa Fluor 594-conjugated Cholera toxin B subunit (CtxBAF594) or Texas Red-conjugated Shiga toxin B subunit (StxB-TR), individual cells exhibited differential toxin uptake. This was shown to be cell cycle-dependent, in which, SubAB-OG was preferentially internalised by cells migrating through G1 and early S phases. In contrast, CtxB-AF594 was taken up by cells in S through M phases and by a majority of cells in G1, while StxB-TR endocytosis occurred in cells traversing G1. Fluorescent SubAB co-localised with the clathrin marker transferrin, but not with Caveolin-1 (a marker for cholesterol-associated caveolae) and was subsequently trafficked via a retrograde pathway to the TGN, Golgi and ER. The clathrin inhibitor phenylarsine oxide prevented SubAB entry and BiP cleavage in SubAB-treated Vero, HeLa and N2A cells, while cholesterol depletion did not, demonstrating that, unlike either Stx or Ctx, SubAB internalisation is exclusively clathrin-dependent. Identification of the SubB receptor was initially approached using toxin overlay assays in which Vero cell glycolipid extracts were separated by thin-layer chromatography and overlaid with SubAB. SubAB exhibited a high affinity for particular acidic species in the ganglioside fraction. However, none co-migrated with commercial glycolipid standards. SubAB-OG also exhibited an affinity for the oligosaccharide structures of chimeric LPS from GM₂ and GM₃ bacterial receptor mimic constructs in an LPS toxin overlay assay. Glycan array analysis revealed that SubB possessed a unique affinity for carbohydrate receptors with a terminal Neu5Gcα(2→3)Galβ disaccharide. Monovalent receptor analogues with distal Neu5Gc or Neu5Gcα(2→3)Galβ and highly-sialylated α₁-AGP did not prevent endocytosis of SubAB-OG, BiP cleavage or cytotoxicity in Vero cells. This indicated that SubAB has a greater affinity for the host cell receptors than the receptor analogues and may engage multiple receptors displayed on a lipid bilayer. In addition to mediating toxin binding and subcellular trafficking, CtxB and StxB can also potentiate the immune response to co-administered antigen. Accordingly, the systemic immunomodulatory properties of SubB administered by the i.p. route were assessed in mice. Using SubAA₂₇₂ as a bystander antigen, SubB significantly increased mouse anti-SubAA₂₇₂ titres to levels that were comparable to those obtained using Alum adjuvant. However, when admixed with structurally-unrelated OVA, SubB did not significantly affect anti-OVA titres whereas Alum and CtxB did. This indicated that SubB may function as a systemic carrier protein (rather than an adjuvant) for particular antigens. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1363363 / Thesis (Ph.D.) - University of Adelaide, School of Molecular and Biomedical Science, 2009

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 6 uppsatser.

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 112-123).

Made available in DSpace on 2014-06-11T19:28:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-18Bitstream added on 2014-06-13T19:58:10Z : No. of bitstreams: 1 aguiarjunior_ho_me_botfca.pdf: 487083 bytes, checksum: cf6b43776665fed2b2912c525547c0ba (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Com o intuito de melhorar a eficiência da cobertura foliar e diminuir as perdas na produtividade da cultura da soja, o presente trabalho teve como objetivos: avaliar a deposição da pulverização, perdas por deriva, produção (peso de 1000 sementes), produtividade da cultura (kg/ha), tensão superficial estática e retenção foliar da calda de pulverização. Três experimentos, dois a campo e um em laboratório foram conduzidos nesse estudo; os experimentos 1 e 2 foram conduzidos na safra 2008/09, em área experimental da fazenda de Ensino, Pesquisa e Produção (FEPP) da FCA/UNESP – Campus de Botucatu, Gleba Cascalheira na cultura da soja, cultivar Conquista e o experimento 3 foi realizado no Laboratório de Tecnologia de Aplicação de Defensivos do Departamento de Produção Vegetal – Defesa Fitossanitária da Faculdade de Ciências Agronômicas FCA/UNESP, campus de Botucatu. O experimento 1 foi conduzido no delineamento experimental em blocos ao acaso com três dosagens de surfatantes associados ao marcador oxicloreto de cobre combinadas a dois níveis de assistência de ar na barra de pulverização (0 e 29 km h-1), totalizando 6 tratamentos e 4 repetições. O pulverizador foi equipado com barra de pulverização de 18,5 m de comprimento e 37 pontas de pulverização de jato plano XR 8002, à pressão de 400 kPa e volume de 150 L ha-1. Para a quantificação dos depósitos da pulverização foram colocados em cada parcela alvos artificiais (papel filtro com dimensões de 3 x 3 cm) distribuídos em 10 plantas tomadas ao acaso. Em cada uma das 10 plantas amostradas foram fixados quatro coletores (papel filtro), um na superfície adaxial e outro na superfície abaxial do mesmo folíolo nas partes superior e inferior da mesma planta. Para avaliação do potencial de deriva, nas diferentes tecnologias de aplicação, foram colocados externamente às parcelas duas hastes de madeira... / With the intention to improve the leaf coverage efficiency and reduce in losses productivity of soybean crop, the present study aimed to evaluate the spray deposition and drift losses, production (weight of 1000 seeds), soybean crop productivity (kg / ha), static surface tension and spray leaf retention. Three experiments, two in the field and another one the laboratory, were conducted in this study. The experiments 1 and 2 were carried out during 2008/2009 crop season in experimental area of Experimental Farm, FCA / UNESP – campus of Botucatu, soybean crop, Conquista variety, and the third experiment was set at the of Laboratory Technology Application of Pesticides, in Plant Production Department, College of Agricultural Sciences of the São Paulo State University FCA/UNESP, campus of Botucatu. The Experiment 1 was conducted in randomized block design with three doses of surfactants associated with one oxychloride cooper tracer, combined at two levels of air assistance on spray boom (0 and 29 km h-1), 6 treatments in 4 repetitions. The sprayer was equipped with spray boom 18.5 m of length and 37 flat fan spray nozzle XR 8002 type at working, pressure of 400 kPa, resulting in the volume of 150 L ha-1. For the quantification of spray deposits were placed in each plot artificial targets (filter paper with dimensions of 3 x 3 cm) distributed in 10 plants randomly selected. In each one of the 10 plants were fixed four collectors (filter paper), one on adaxial surface and other on abaxial surface in the same leaflet at the top and bottom of the same plant. To evaluate the spray drift potential in different application technologies, were placed outside the plots two stems of wood at 1.0 m and 3.0 m away at the end of crop. In the stems positioned at 1.0 m of the crop were fixed artificial targets, pipe cleaners, positioned at 0.5 and 1.0 m above ground and stems distanciated at 3 m of the crop... (Complete abstract click electronic access below)

O câncer de pulmão permanece a neoplasia mais letal com cerca de 1,59 milhões de mortes por ano em todo o mundo, com eficácia terapêutica limitada e mau prognóstico. Aproximadamente 80 % dos casos são de câncer de pulmão de não pequenas células (CPNPC), desses, cerca de 50 % são os adenocarcinomas (AdC). Atualmente o tratamento padrão-ouro no tratamento de AdC pulmonar é baseado em agentes de platina (cisplatina, carboplatina) normalmente administrados em combinação com outros agentes, entretanto a doença é raramente curável. Vários fatores contribuem para a alta taxa de mortalidade e um dos mais comuns é a quimioresistência. Portanto, existe uma necessidade urgente de terapias mais eficazes em aumentar a sobrevida global dos pacientes com essa patologia. Tendo em vista que o desenvolvimento de novos medicamentos requer muito tempo e investimento financeiro, o uso de drogas existentes como terapia combinada torna-se uma boa abordagem. Estudos anteriores de nosso grupo demonstraram que a agressividade do AdC de pulmão está associada com elevado estresse oxidativo onde o peróxido de hidrogênio (H2O2) tem um papel crucial, uma vez que o tratamento com a enzima antioxidante catalase (CAT) atenuou a agressividade do tumor. Assim, este estudo avaliou a eficácia da terapia combinada com CAT na linhagem celular A549 de AdC humano. Primeiramente, a adição de CAT causou uma inibição dose-dependente da proliferação celular com um efeito máximo na dose de 1000 U/mL. Essa inibição foi relacionada a um efeito citostático, não citotóxico, do consumo de H2O2 intracelular, já que a retirada da CAT restaurou a taxa proliferativa celular a níveis de controle. Após, a avaliação do “status” de ativação do NFB mostrou um aumento de 2,9 vezes no imunoconteúdo citosólico da subunidade p65, sugerindo uma diminuição da ativação do NFB em células tratadas com CAT. A análise do efeito do tratamento de CAT em parâmetros redox mostrou uma redução nos níveis intracelulares de tióis (-SH) e no potencial antioxidante total (TRAP) e um aumento na produção de H2O2 e nos níveis de glutationa (GSH). Entre as drogas testadas (Cisplatina, 5-Fluorouracil, Paclitaxel, Hidroxiuréia e Daunorrubicina), apenas o tratamento com paclitaxel e hidroxiureia mostraram aumento da produção de H2O2 quando comparados com o veículo. No entanto, o co-tratamento com CAT e paclitaxel não alterou a produção de H2O2. Hidroxiuréia e CAT promoveu uma redução na produção de H2O2, quando comparado com a droga sozinha. A cisplatina, por si só não teve efeito na produção de H2O2, mas cisplatina e CAT promoveram um aumento na produção de H2O2. A eficácia da terapia combinada com CAT em potencializar a citotoxicidade dos quimioterápicos, foi analisada pelo ensaio SRB e pelo software CalcuSyn®. Pela análise dos valores de “Combination Index” (CI) observamos que, com a exceção da combinação de CAT e paclitaxel, que gerou antagonismo, as combinações de CAT com cisplatina, 5-fluorouracil e hidroxiuréia exibiram um efeito sinérgico na eliminação de células da linhagem de AdC humano A549. Dessa forma, os dados aqui apresentados sugerem que a terapia combinada de CAT com cisplatina, 5-fluorouracil e hidroxiuréia pode surgir como uma nova estratégia terapêutica para o AdC. / Lung cancer remains the most lethal malignant disease with nearly 1.59 million deaths annually worldwide, limited efficacy of current therapeutics and dismal prognostic. Approximately 80% of the cases are non-small cell lung cancer (NSCLC), of these, roughly 50% are adenocarcinomas (AdC). Currently, the gold standard treatment for AdC is based on platinum agents, usually given in combination with other agents. Despite these therapies, the disease is rarely curable. Several factors contribute to the high mortality rate and one of the most common includes tumor cell chemoresistance to cytotoxic drugs. Therefore, there is an urgent need for more effective therapies that could increase the overall survival of lung AdC patients. With the notion that the development of novel drugs require much time and financial investment, the use of existing drugs as adjuvant treatment becomes a good approach. Previous studies of our group demonstrated that lung AdC aggressiveness is associated with elevated intracellular oxidative stress, in which hydrogen peroxide (H2O2) plays a crucial role, since the exogenous treatment with the antioxidant enzyme catalase (CAT) attenuated tumor aggressiveness. Then, this study evaluated the efficacy of CAT adjuvant treatment in the human AdC cell line A549. Firstly, exogenous addition of CAT caused a dose dependent inhibition on cellular proliferation with a maximal dose effect of 1000 U/mL. Growth inhibition was related to a cytostatic, not cytotoxic, effect of intracellular H2O2 consumption, since CAT washout readily restored cellular proliferative rate similar to control. After that, evaluation of NFB activation status showed a 2.9-fold increase in the cytosolic immunocontent of the NFB subunit p65, suggesting a decreased NFB activation in CAT-treated cells. Analysis of CAT treatment effect in redox parameters showed decreased intracellular thiol levels (-SH) and non-enzymatic antioxidant potential (TRAP) and increases the H2O2 production and glutathione levels (GSH). Among drugs tested (cisplatin, paclitaxel, 5-fluorouracil, daunorubicin, hydroxurea) (GI50), only paclitaxel and hydroxyurea showed increased production of H2O2 when compared with vehicle. However, co-treatement with CAT and paclitaxel had no alteration in the H2O2 production. Hydroxyurea plus CAT had a decreased in H2O2 production when compared with the drug alone. Cisplatin, alone had no effect in H2O2 production, but cisplatin plus CAT, had an increased in H2O2 production. Regarding the effectiveness of adjuvant CAT treatment in potentiate chemotherapeutic drugs cytotoxicities, we used de SRB assay and Calcusyn Software to access this interaction. From analyzes of combination index (CI) values, generated by CalcuSyn, we observed that with the exception of CAT plus paclitaxel, all combinations exhibited a synergistic effect. Taken together, data presented here suggest that adjuvant CAT treatment can act synergistically with chemotherapeutics and modulate tumorassociated signaling pathways providing a new therapeutic strategy for AdC therapy.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / In cats, cutaneous healing presents qualitative and quantitative differences in comparison with another species, that seems predispose them to healing problems. Wounds that heal by first intention present less resistance, rate of contraction and epithelization. In the second intention healing, the inflammatory phase is bland and the proliferative phase has slow appearing and progression. Considering these aspects, it becomes important to search ways to promote and conduce cutaneous wound healing in cats. The aims of this study were standardize a technique to obtain autologous platelet-rich plasma (PRP) in cats and evaluate your effect on cutaneous healing of experimentally produced wounds in this species. The experiment was developed in two stages: three manual protocols to produce autologous platelet-rich plasma in cats were compared in the first stage. Fifteen adult healthy indoor cats, randomly divided into three groups, were utilized. Five samples of 4.5 ml of jugular blood were collect, with a vacuum system, sequentially in citrated tubes from each cat. The samples were processed using double centrifugation protocols (P): P1 (113g 10 minutes; 652g 5 minutes); P2 (400g 10 minutes; 800g 10 minutes); P3 (400g 10 minutes; 600g 10 minutes). Platelet concentration obtained in PRP obtained in different protocols and the ratio of PRP and platelet counts were compared by t-test. The feasibility of the collect method to the species and ease of execution of techniques was evaluated too. In the second stage, ten healthy adult indoor cats were used. In each animal were surgically produced four circular skin defects, with a diameter of 1 cm, on left and right sides of trunk dorsal midline. The lesions on one side were treated with fluid PRP, obtained of animal itself, according to the best protocol defined in the first stage of the experiment. The control defects in the opposite side were filled with NaCl 0,9% sterile solution. Three observers evaluated the wounds, every three days, for 28 days. The presence of crusts, pain, swelling, exudate, granulation tissue and the color were considered. The results were statistically evaluated by Mann-Whitney rank test. The area of each lesion was analyzed by morphometry every seven days and the data compared by t-test. Skin fragments with normal and scar tissue, from two more cranial lesions (treated and control), were obtained weekly. The fragments were stained with hematoxylin-eosin (HE) and Masson's trichrome, and qualitatively evaluated for the presence of crust, ulcer, acanthosis, fibroplasia, collagen deposition and neovascularization. These data were analyzed using the Mann-Whitney rank test. The collect method was satisfactory and simplified the procedure. Two techniques (P2 and P3) were effective to produce PRP, yielding concentrations higher than 1x106 platelets.μL-1. The P3 protocol was statistically higher in capacity increase, and was selected for use in experimental wounds. In the second stage, the techniques for blood collection, and the selected protocol, were satisfactory. The use of PRP in liquid form, deposited directly on the lesion, made it easy to use. Clinically, there was a statistically significant difference (p = 0.025) and higher exudation in wounds treated to three days. In morphometric analysis we noticed a greater contraction in wounds treated with PRP, especially of the first seven days (p = <0.001). Histopathological evaluation was not significant in any compared parameters. Therefore, it is concluded that the sampling method is efficient and P2 and P3 protocols can be considered feasible to obtain PRP from cats. The protocol P3 was superior in the platelet concentration ability, producing a better quality PRP. Activation of PRP liquid by tissue injury is possible. The increased exudation suggests enhancement of the inflammatory process in treated wounds. Therapeutic use of liquid PRP improves contraction capacity on experimental wounds in cats, despite the absence of histological evidence by way the evaluation adopted. / A cicatrização cutânea em gatos possui diferenças qualitativas e quantitativas quando comparada à de outras espécies, fato que parece predispor a problemas. Feridas que cicatrizam por primeira intenção apresentam menor resistência, taxa de contração e epitelização. Já na cicatrização por segunda intenção, a fase inflamatória é branda e a fase proliferativa apresenta surgimento e progressão lenta. Tendo em vista esses aspectos, torna-se importante a busca de meios que ajudem a favorecer e conduzir o reparo de feridas cutâneas em gatos. Este estudo objetivou a padronização de uma técnica para obtenção de plasma rico em plaquetas (PRP) autólogo em gatos e a avaliação de sua ação sobre a cicatrização de feridas cutâneas produzidas experimentalmente nessa espécie. Ele desenvolveu-se em duas etapas distintas: na primeira foram comparados três protocolos manuais para produção de PRP em gatos. Utilizaram-se 15 animais adultos hígidos e domiciliados, divididos aleatoriamente em três grupos. Cinco amostras de 4,5ml de sangue foram colhidas da jugular de cada animal, por sistema a vácuo, sequencialmente, em tubos com citrato de sódio. As amostras de cada grupo foram processadas por meio de protocolos de dupla centrifugação: P1 (113g 10 minutos; 652g 5 minutos); P2 (400g 10 minutos; 800g 10 minutos); P3 (400g 10 minutos; 600g 10 minutos). As concentrações plaquetárias obtidas no PRP dos diferentes protocolos e a relação entre o PRP e contagem de plaquetas foram comparadas por teste-t. A viabilidade do modo de coleta a vácuo para a espécie e a facilidade de execução das técnicas também foram avaliadas. Na segunda etapa, foram utilizados dez gatos adultos hígidos, domiciliados. Em cada animal foram produzidos, cirurgicamente, quatro defeitos circulares, pareados, com diâmetro de 1 cm, na pele, nas laterais esquerda e direita da linha média dorsal do tronco. As lesões de um lado foram tratadas com PRP líquido, obtido do próprio animal, de acordo com o melhor protocolo definido na primeira etapa do experimento. Os defeitos do lado oposto, considerados controle, foram preenchidos com solução de NaCl 0,9% estéril. As feridas foram avaliadas por três observadores, a cada três dias, por 28 dias. Considerou-se a presença de crostas, dor, edema, exsudato, tecido de granulação e a coloração. Os resultados foram avaliados estatisticamente pelo teste de Mann-Whitney em ranks. A área de cada lesão foi analisada por morfometria a cada sete dias e os dados comparados pelo teste-t. Também, semanalmente, foram obtidos fragmentos de pele contendo tecido normal e cicatricial a partir das duas lesões (tratada e controle) mais craniais. Os fragmentos foram corados por hematoxilina-eosina (HE) e tricrômico de Masson, e avaliados qualitativamente quanto a presença de crosta, úlcera, acantose, fibroplasia, colagenização e neovascularização. Esses dados foram analisados pelo teste de Mann-Whitney em ranks. O sistema de coleta de sangue a vácuo mostrou-se satisfatório e simplificou o procedimento. Duas das técnicas (P2 e P3) mostraram-se factíveis para a produção do PRP, proporcionando concentrações superiores a 1x106 plaquetas.μl-1. O protocolo P3 mostrou-se estatisticamente superior na capacidade de incremento, e foi selecionado para uso nas feridas experimentais. Na segunda etapa, a técnica para coleta de sangue e o protocolo selecionado, mostraram-se satisfatórios. A utilização do PRP na forma líquida, depositado diretamente sob a lesão, tornou fácil sua utilização. Clinicamente, houve diferença estatisticamente significativa (p=0,025) e maior exsudação nas feridas tratadas no dia três. Na análise morfométrica, notou-se maior contração nas feridas tratadas com PRP, principalmente nos primeiros sete dias (p=<0,001). Na avaliação histopatológica semi-quantitativa não houve significância em nenhum dos parâmetros comparados. Logo, conclui-se que o método de coleta é eficiente, e podem-se considerar exequíveis as técnicas para obtenção do PRP em gatos, a partir dos protocolos P2 e P3. O protocolo P3 demonstra superior eficiência na capacidade de concentrar plaquetas, gerando um PRP de melhor qualidade. É possível a ativação do PRP líquido por meio da lesão tecidual. O aumento da exsudação sugere intensificação do processo inflamatório nas feridas tratadas. O uso terapêutico do PRP na forma líquida melhora a capacidade de contração em feridas produzidas experimentalmente em gatos, apesar da ausência de evidências histopatológicas pela avaliação adotada.

Made available in DSpace on 2014-06-11T19:31:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-12-11Bitstream added on 2014-06-13T18:42:24Z : No. of bitstreams: 1 oliveira_map_dr_botfca.pdf: 804551 bytes, checksum: 700238b1422e359e397bc2e36724e08e (MD5) / Outros / O presente trabalho teve por objetivos verificar o efeito da remoção pela chuva de diferentes formulações de flutriafol, em condições de laboratório, aplicadas com e sem a adição de óleo na calda de pulverização, assim como verificar a influencia do óleo no espectro de gotas. O experimento considerou dez caldas resultantes da interação de cinco tratamentos contendo o flutriafol (quatro formulações de flutriafol isolado e uma mistura de flutriafol com tebuconazole), todas aplicadas com e sem óleo mineral. As dez caldas foram aplicadas sobre plantas de soja que receberam posteriormente lavagem por chuva simulada em laboratório com quatro intervalos de tempo entre a aplicação e a chuva. As chuvas foram simuladas em duas etapas, sendo uma lâmina inicial de 5 mm seguida de uma lâmina complementar de 15 mm. As coletas foram realizadas com quatro repetições, sendo cada uma delas constituída de um vaso com quatro plantas. A avaliação da concentração de flutriafol nas soluções resultantes da lavagem das plantas foi realizada através da quantificação dos resíduos do fungicida na água pelo método de cromatografia gasosa e espectrometria de massa (GCMS). Após a aplicação das caldas, as plantas foram retiradas do laboratório e deixadas à temperatura ambiente, a espera do tempo decorrido para a simulação da chuva, de acordo com cada tratamento. Após a simulação das lâminas de chuva as plantas foram cortadas dos vasos e lavadas em saco plástico contendo 200 mL de água destilada. Todas as lavagens foram realizadas em 4 intervalos: 0 h (no momento da aplicação), 1 h, 2 h e 48 h após a aplicação. Para o cálculo do percentual extraído, as quantidades de flutriafol nas soluções de lavagem foram comparadas à quantidade total depositada nas plantas, a qual foi representada... / The present study had for objectives to verify the effect of rainfastness of different flutriafol formulations, in laboratory conditions, applied with and without the oil adjuvant in the spray solution, as well as to verify the influence of the oil on the droplet spectrum. The experiment considered ten spray solutions related to five treatments containing flutriafol (four formulations of flutriafol and a flutriafol mixture with tebuconazole), all applied with and without mineral oil. The ten solutions were applied on soybean plants that were washed by simulated rainfall in laboratory with four time intervals between the application and the rainfall. Rainfall was simulated by one initial precipitation of 5 mm followed by a complementary 15 mm rainfall. Each vase with four plants was considered one replication. The washing solutions from the vases were collected and the flutriafol concentration on those solutions was evaluated through the quantification of the residues of the fungicide in the water using the Gas Chromatography-Mass Spectrometry (GC-MS). After the spray application the plants were removed of the laboratory and left to room temperature, the wait of the time elapsed for the simulation of the rain, in agreement with each treatment.After the simulated rain the plants were cut out of the vases and washed by immersion in plastic sack containing 200 mL of distilled water. All of the washing solutions (from the rain and the immersion process) were accomplished in 4 intervals: 0 h (in the moment of the spray application), 1 h, 2 h and 48 h after the application. For the calculation of the flutriafol extracted, the amounts of fungicide in the wash solutions were compared to the total amount deposited in the plants, which was represented by the sum of the amounts of the assets recovered in the washing... (Complete abstract click electronic access below)

Made available in DSpace on 2016-12-08T16:24:21Z (GMT). No. of bitstreams: 1 PGCA15MA170.pdf: 1407687 bytes, checksum: 6d77197fef831a6cbdc6fcf9e39c8629 (MD5) Previous issue date: 2015-07-23 / For a long time, Pasterella multocida has been considered only as a secondary agent in pneumonias in swine. However, studies developed in Brazil identified different isolates of P. multocida able to operate as primary disease agent in swine. The objective of his project was to study protection aspects of a vaccine with pathogenic samples of P. multocida in experimental model with high sanity status pigs. In addition, it aims to describe the expression of immune response in swine induced by P. multocida infection, and determinate the prevalence of P. multocida in tonsils of vaccinated and not vaccinated pigs through immunohistochemistry. The study was divided into three experiments: Immunization of mice with vaccines prepared with different adjuvants , Test of two adjuvants used in vaccine with a P. multocida homologous strain in swine and Experimental challenge in swine with a P. multocida heterologous strain to the vaccine . Four adjuvants were tested in mice: Al(OH)3, ISA 206, Montanide GEL 01 and ISA 760. Two of them were selected for testing in swine: Al(OH)3 which showed better results in mice-, and ISA 760 for being an oily adjuvant. At the second experiment, although two vaccines have been effective in the disease protection, the one with the adjuvant Al(OH)3 was selected, because showed lower vaccine reaction. Eighteen pigs were distributed into three groups for the specific test of the vaccine for pathogenic samples of P. multocida. Group 1 (G1) vaccine with Al(OH)3; group 2 (G2) infection control; and group 3 (G3) negative control. After the experiment, the animals were euthanized, and samples were collected in ice - for microbiological cultivation and relative quantification of cytokines gene expression (IL1 &#946;, IL2, TNF&#945;) and TLR4 , and in formaldehyde, for routine histopathology and immunohistochemistry. Results showed that the vaccine prototype with adjuvant Al(OH)3 was efficient in controlling pulmonary pasteurellosis. The immune response was mediated by TLR4, with increased expression of TNF&#945;. IL-1&#946; and IL2 were also expressed, however, there was no significant difference between the groups. The immunostaining of P. multocida in tonsil was positive in all pigs challenged with the bacteria / Por muito tempo, Pasteurella multocida foi considerada apenas como agente secundário nas pneumonias em suínos, todavia, estudos realizados no Brasil identificaram diferentes isolados de P. multocida capazes de atuar como agente primário de doença em suínos. Esse projeto teve como objetivo estudar aspectos de proteção de uma vacina com amostras patogênicas da P. multocida em modelo experimental com suínos de alto status sanitário . E de forma paralela descrever expressão da resposta imune dos suínos induzida pela infecção da P. multocida e determinar a prevalência dessa bactéria em tonsilas de suínos vacinados e não vacinados através de imunohistoquímica. O trabalho foi dividido em três experimentos: Imunização de camundongos com vacinas preparadas com diferentes adjuvantes ; Teste de dois adjuvantes utilizados na vacinação de suínos frente ao desafio com uma cepa homóloga de P. multocida ; e Desafio experimental em suínos com uma cepa de P. multocida heteróloga à da vacina . Em camundongo foram testados quatro adjuvantes: Al(OH)3, ISA 206, Montanide GEL 01 e ISA 760, dos quais, foram selecionados dois para teste em suínos: Al(OH)3, que apresentou melhor resultado nos camundongos e ISA 760, por ser um adjuvante oleoso. Embora as duas vacinas tenham sido efetivas na proteção da doença, no segundo experimento, selecionou-se àquela com adjuvante Al(OH)3, que apresentou menor reação vacinal. No terceiro experimento realizado em suínos, foram utilizados 18 suínos em três grupos: grupo 1 (G1) - vacina com Al(OH)3, e grupo 2 (G2) controle de infeção e grupo 3 (G3) controle negativo. Após o experimento os animais foram eutanasiados para avaliação patológica, coleta de amostras em gelo para cultivo microbiológico e quantificação relativa da expressão gênica das citocinas (IL1 &#946;, IL2, TNF&#945;) e TLR4, para histopatologia de rotina e imunohistoquimica. Os resultados demostraram que o protótipo de vacina com adjuvante Al(OH)3 foi eficiente no controle de pasteurelose pulmonar. O perfil da expressão da resposta imune foi mediado por TLR4, com aumento da expressão de TNF&#945;. Também foram expressos IL-1&#946; e IL2, porém sem diferença significativa entre os grupos. A imunomarcação de P. multocida na tonsila foi positiva em todos os suínos desafiados com a bactéria, independente de serem ou não vacinados