( )-linalool (LIN) is a monoterpene alcohol majority in the essential oils of several aromatic species commonly found in northeastern Brazil, such as Ocimum basilicum and Cinnamomum verum. Recent studies have shown that LIN has important therapeutic properties, including its antinociceptive, anti-inflammatory and anticonvulsant. Despite the benefits described above, the LIN, such as monoterpenes generally exhibits low solubility in water, high volatility and short half-life, thus incorporation into cyclodextrins (CDs), especially β-CD, has emerged as a tool important to improve the physicochemical properties and biological isoprênico this derivative. Thus, we investigated the possible antinociceptive effects of LIN isolated and included in β-CD in chronic non-inflammatory muscle pain model, besides to the effects on the motor activity and the central action. Male Swiss mice (25-35 g) were divided into groups and subjected to induction protocols non-inflammatory nociception by injecting acid saline (pH 4; 20 μL) in the belly left of gastrocnemius muscle. Subsequently, they were treated orally (p.o.) with LIN (25 mg/kg) LIN/β-CD (25, 50 and 100 mg/kg) tramadol (TRM 4 mg/kg, i.p.) or vehicle (solution 0.9% saline), and evaluated for behavioral parameters over a period of 27 days. The animals were subjected to tests of mechanical hyperalgesia (von Frey digital), motor coordination (Rota Rod) and muscle strength (Grip Strength Metter), and to determine the possible involvement of areas of the central nervous system (CNS), the animals were treated and ninety minutes, were anesthetized, perfused, the brains extracted and cut in a cryostat. The brain sections were subjected to immunofluorescence protocol for Fos protein. The results were expressed as mean ± standard error of the mean. Differences between groups were analyzed by ANOVA test, a path followed by Bonferroni test. P values <0.05 were considered statistically significant. Treatment with LIN and LIN/β-CD showed antinociceptive activity in animal model of chronic non-inflammatory muscle hyperalgesia in mice at all doses tested, with the LIN/β-CD prolonged activity against isolated substance, confirming the possibility of significant improvement in the therapeutic action; treatment with LIN and LIN/β-CD did not induce motor incoordination or change in muscle strength. By immunofluorescence, we observed that treatment of LIN and LIN/β-CD presented action in the CNS, acting in the brain, whereas the retrosplenial cortex and periaqueductal gray central structures are strictly related to the mechanisms of pain modulatory. / O ( )-linalol (LIN) é um monoterpeno alcoólico majoritário nos óleos essenciais de várias espécies aromáticas comumente encontradas no nordeste brasileiro, tais como Ocimum basilicum e Cinnamomum verum. Estudos recentes tem demonstrado que o LIN produz importantes propriedades terapêuticas, incluindo seus efeitos antinociceptivo, anti-inflamatório e anticonvulsivante. Apesar dos benefícios supracitados, o LIN, como os monoterpenos em geral, exibe baixa solubilidade em água, alta volatilidade e curta meia vida plasmática; desta forma a incorporação em ciclodextrinas (CDs), especialmente a β-CD, vem se destacando como uma ferramenta importante para melhorar as propriedades físico-químicas e biológicas deste derivado isoprênico. Assim, foram investigados os possíveis efeitos antinociceptivos do LIN, isolado e incluso em β-CD no modelo animal de hiperalgesia muscular não inflamatória, além dos efeitos do tratamento na atividade motora e a ação central. Camundongos Swiss machos (25-35 g), foram divididos em grupos e submetidos ao protocolo de indução da nocicepção não inflamatória através da injeção de solução salina ácida (pH 4; 20 μL) no ventre do músculo gastrocnêmio esquerdo. Posteriormente, foram tratados, por via oral (v.o.) com LIN (25 mg/kg), LIN/β-CD (25, 50 e 100 mg/kg), tramadol (TRM 4 mg/kg, i.p.) ou veículo (solução salina 0,9%), e avaliados quanto à parâmetros comportamentais durante um período de 27 dias. Os animais foram submetidos aos testes da hiperalgesia mecânica (von Frey digital), da coordenação motora (Rota Rod) e da força muscular (Grip Strength Metter), e para determinar o possível envolvimento de áreas do Sistema Nervoso Central (SNC), os animais foram tratados e, noventa minutos após, foram anestesiados, perfundidos, os cérebros extraídos e cortados em criostato. As secções cerebrais foram submetidas ao protocolo de imunofluorescência para proteína Fos. Os resultados foram expressos como média ± erro padrão da média. As diferenças entre os grupos foram analisadas por meio do teste de variância ANOVA, uma via, seguido pelo teste de Bonferroni. Valores de p < 0,05 foram considerados estatisticamente significantes. O tratamento com LIN e LIN/β-CD apresentou atividade antinociceptiva no modelo animal de hiperalgesia muscular crônica não inflamatória em camundongos, em todas as doses testadas, tendo o LIN/β-CD atividade prolongada em relação a substância isolada, confirmando a possibilidade de melhora significativa na ação terapêutica; o tratamento com LIN e LIN/β-CD não induziu incoordenação motora nem alteração da força muscular. Através da imunofluorescência, observou-se que o tratamento de LIN e LIN/β-CD apresentou ação no SNC, atuando no encéfalo, visto que o córtex restrosplenial e a substância cinzenta periaquedutal são estruturas centrais intimamente relacionadas com os mecanismos modulatórios da dor.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Acute myocardial infarction (AMI), a serious public health problem, is characterized by tissue death after prolonged ischemia. Ischemia, the repercussion, triggers cellular processes, such as intracellular calcium overload and oxidative stress. There are few pharmacological intervention strategies for prevention and limitation of these lesions, and it is worth noting that the available forms of intervention are not fully effective. Recent research indicates that, as a therapeutic alternative, the use of drugs derived from natural products with antioxidant action has been shown to be an important preventive and treatment tool for the lesions resulting from the infarction. p-Cymene, a monoterpene present in essential oils of various plant species, is a substance that has a range of proven pharmacological activities, among them antinoceptive, anti-inflammatory and antioxidant activity. The aim of this study was to evaluate the effect of pre-treatment with p-cymene on ischemia and reperfusion injury (I / R) in hearts isolated from rats. For this purpose, the Langendorff-type aortic perfusion model was used to submit isolated hearts from rats to global ischemia. Wistar rats pretreated with vehicle or p-cymene at the dose of 150 mg / kg / day orally (gavage) were used for 7 days, and thereafter their isolated hearts were subjected to global ischemia and reperfusion in the Langendorff system . As a control of the adopted experimental model, hearts that were not submitted to I / R were used. Measurements of lactate dehydrogenase (LDH), indicative of tissue damage, were also performed on all groups of animals, and the contractile parameters in the isolated heart (left ventricular pressure (LVDP), time derivative of ventricular pressure (dP / Dt) and arrhythmia severity index (ASI)], as well as oxidative parameters in cardiac tissue [malonaldehyde formation (MDA), total sulfhydryl; The activity of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and glutathione peroxidase (GPx)]. We observed that the pre-treatment with p-cymene preserved the reduction of PDVE, dP / dt and caused lower ASI, improving the contractile function. It reduced the formation of MDA and LDH (62%). In addition, this monoterpene preserves the sulfhydryl groups (38%) and the activity of the antioxidant enzymes CAT (36%), SOD (62%), GPx and GR when compared to the vehicle. It can be concluded that pre-treatment with p-cymene prevents the reduction of cardiac contractile force caused by ischemia and significantly reduces the oxidative damage caused by infarction in the experimental model. / O infarto agudo do miocárdio (IAM), grave problema de saúde pública, é caracterizado pela morte tecidual após isquemia prolongada. A isquemia, seguida da reperfusão, desencadeia processos celulares, tais como sobrecarga intracelular de cálcio e estresse oxidativo, que são os eventos principais que levam a morte celular. Existem poucas estratégias de intervenção farmacológica para prevenção e limitação destas lesões, e ainda, vale destacar que as formas de intervenção disponíveis não são totalmente eficazes. Recentes pesquisas apontam que, como alternativa terapêutica, o uso de drogas derivadas de produtos naturais com ação antioxidante tem se mostrado importante ferramenta preventiva e de tratamento para as lesões decorrentes do infarto. O p-cimeno, um monoterpeno presente em óleos essenciais de várias espécies vegetais, é uma substância que apresenta uma gama de atividades farmacológicas comprovadas, dentre elas, atividade antinoceptiva, anti-inflamatória e antioxidante. O objetivo desse estudo foi avaliar o efeito do pré-tratamento com p-cimeno em lesões decorrentes da isquemia e reperfusão (I/R) em corações isolados de ratos. Para tanto, foi adotado o modelo de perfusão aórtico do tipo Langendorff para submeter corações isolados de ratos à isquemia global. Foram utilizados ratos Wistar pré-tratados com veículo ou p-cimeno na dose 150 mg/kg/dia, via oral (gavagem), durante 7 dias, e, posteriormente, seus corações isolados foram submetidos à isquemia global e reperfusão no sistema de Langendorff. Foram utilizados, como controle do modelo experimental adotado, corações que não foram submetidos à I/R. De todos os grupos de animais foram realizadas medidas de lactato desidrogenase (LDH), indicador de lesão tecidual e ainda, foram avaliados tanto os parâmetros contráteis no coração isolado [pressão desenvolvida pelo ventrículo esquerdo (PDVE), derivada temporal de pressão ventricular (dP/dt) e o índice de severidade de arritmias (ASI)], quanto os parâmetros oxidativos no tecido cardíaco [formação malonaldeído (MDA), sulfidrilas totais; a atividade das enzimas antioxidantes catalase (CAT), superóxido dismutase (SOD), glutationa redutase (GR) e glutationa peroxidase (GPx)]. Observamos que o pré-tratamento com p-cimeno preservou a redução de PDVE, dP/dt e causou menor ASI, melhorando a função contrátil. Reduziu a formação de MDA e LDH (62%). Além disso, este monoterpeno preserva os grupos sulfidrila (38%) e a atividade das enzimas antioxidantes CAT (36%), SOD (62%), GPx e GR quando comparado ao veículo. Pode-se concluir que o pré-tratamento com p-cimeno previne a redução da força contrátil cardíaca provocada pela isquemia e reduz significativamente os danos oxidativos causados pelo infarto no modelo experimental. / Aracaju

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Medicinal plants and their constituents have been shown to be a major source of new biomolecules. The (-)-borneol is an alcohol monoterpene which shows a series of biological activities with potential to accelerate the wound healing process. The present study was to conduct a systematic review that provides an overview of the characteristics of monoterpenes and derivatives iridoids with wound healing effect and its mechanisms of action. Besides, evaluate the wound healing activity of (-)-borneol incorporated to chitosan film in excisional wound model in rats. Systematic Review: articles published in LILACS, EMBASE and PubMed were examined. Seven articles were found on the wound healing effect of 3 monoterpenes and 2 derivatives iridoids. Excisional wound model: A total of 90 Wistar male rats, weighing between 250-300 g. The animals underwent fullthickness wound were treated with placebo (inert chitosan film - QUIN), chitosan films containing (-)-borneol 0.5% (QUIBO05) or chitosan films containing (-)-borneol 1% (QUIBO1) by 3, 7, 14, and 21 days. The residual wounds were photographed and evaluated by wound area, presence of edema, crust, secretion and necrosis. The tissue was removed and properly treated with histochemistry. Additional groups were submitted to excision of the skin, treated for 3 days, and the wounds were taken for performing the measurement of myeloperoxidase. Mieloperoxidase (MPO) assays: Granulation tissue samples of the wounds were submitted to MPO activity. Both QUIBO concentrations decrease the myeloperoxidase activity at 3rd day after injury. The histological sections were analyzed by the presence of polymorphonuclear and mononuclear cells, fibroblast proliferation, collagen deposition and epithelialization rates. The QUIBO05 was able to significantly reduces the wound area, accelerates the inflammatory response and it was able to increase significantly the epithelization rates at 7th day. Finally, QUIBO provided collagen proliferation when compared to QUIN group. The results suggest that QUIBO showed wound healing effect. / Plantas medicinais e seus constituintes tem se mostrado como uma grande fonte de novas biomoléculas. O (-)-borneol é um monoterpeno alcoólico que tem apresentado diversas atividades biológicas com potencial para acelerar o processo de cicatrização. O presente estudo buscou realizar uma revisão sistemática de monoterpenos e derivados iridóides com efeito cicatrizante. Além de, avaliar a atividade cicatrizante do (-)-borneol incorporado ao filme bioativo de quitosana em modelo de ferida cutânea aberta em ratos. Revisão Sistemática: foram pesquisados artigos publicados na LILACS, PubMed e EMBASE. Sete artigos foram encontrados sobre o efeito cicatrizante de 3 monoterpenos e 2 derivados iridóides. Ferida cutânea aberta: Foram utilizados 90 ratos, Wistar, machos, pesando entre 250-300 g. Os animais foram submetidos a uma excisão cutânea e tratados com filme de quitosana inerte (QUIN), filmes de quitosana contendo (-)-borneol a 0,5% (QUIBO05) ou filmes de quitosana contendo (-)-borneol a 1% (QUIBO1), por 3, 7, 14 e 21 dias. As feridas residuais foram fotografadas e avaliadas quanto à área, presença de edema, crosta, secreção, necrose e cicatriz patológica. O tecido foi retirado e devidamente tratado com técnica histoquímica. Grupos adicionais foram submetidos à excisão cutânea, tratados por 3 dias e tiveram as feridas retiradas para a realização da medida da mieloperoxidase. Medida da mieloperoxidase (MPO): amostras do tecido de granulação das feridas foram submetidas à análise da atividade da MPO. Todas as concentrações do QUIBO reduziram a atividade da enzima de MPO ao 3º dia pós-lesão. As secções foram analisadas quanto à presença de células mononucleares e polimorfonucleares, proliferação de fibroblastos, deposição de colágeno e re-epitelização. O QUIBO foi capaz de reduzir significativamente a área da ferida, acelerar a resposta inflamatória e aumentar significativamente o índice de epitelização quando comparado ao QUIN ao 7º dia pós-lesão. Além disso, todas as concentrações do QUIBO estimularam a deposição de fibras colágenas durante todos os estágios do reparo quando comparado ao QUIN. Os resultados sugerem que o QUIBO possui efeito cicatrizante.

Fundação de Apoio a Pesquisa e à Inovação Tecnológica do Estado de Sergipe - FAPITEC/SE / Cardiovascular diseases are the leading cause of death worldwide, and myocardial infarction (MI) is one of the most devastating. Cardiac ischemia-reperfusion injury represents a major threat to human health, contributing to adverse cardiovascular effects.However, although reperfusion of the ischemic heart is essential to reduce myocardial damage, restoration of blood flow may, paradoxically, amplify cellular damage.The recognition that pathological events occurring both in ischemia and reperfusion contribute to tissue injury leads to accelerated efforts to identify mechanisms of IR injury in the hope of identifying new treatments that may limit injury induced by blood reduction flow and / or damage produced by reperfusion.Myrtenol is a monoterpene with multiple pharmacological activities. However, although monoterpenes have been proposed to play beneficial roles in a variety of cardiac disorders, pharmacological actions of myrtenol in the heart are not yet reported. Hence, the aim of this study was to evaluate whether myrtenol promotes cardioprotection against myocardial ischemia-reperfusion (IR) injury, and the mechanisms involved in these effects. Male Wistar rats were orally treated for seven consecutive days with solution (NaCl 0.9% 0.20 ml + DMSO 0.05 ml), myrtenol (50 mg/kg) or N-acetyl cysteine (1.200 mg/kg, NAC).Subsequently, the hearts were submitted to cardiac IR injury, through the aortic perfusion flow system of the langedorff type, contractile parameters such as left ventricular pressure (PVE), maximum rise/down velocity of left intraventricular pressure (dP/dt), heart rate (HR), electrocardiographic parameters, HR, PR interval and duration of the QRS complex, measured the activity of Lactate Dehydrogenase (LDH), determined the severity of cardiac arrhythmias (ASI). The oxidative stress was evaluated on the basis of the determination of total hydroperoxides, total sulfhydryl (SH) groups, measurements of the reactive oxygen species (ROS) generation, the carbonylated proteins, the activity of the endogenous antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT) , glutathione peroxidase (GPx) and glutathione reductase (GR), and the infarct area. The apoptosis pathway was investigated by the expression of Bax and Bcl-2 proteins and in situ apoptosis was studied from the TUNEL assay. Here, we show that the severe impairment of contractile performance induced by IR was significantly prevented by myrtenol or NAC. Moreover, Myrtenol abolished aberrant electrocardiographic waveform (ST-segment elevation), as well as reduced life-threatening arrhythmias and infarct size induced by IR injury. Importantly, myrtenol fully prevented the massive increase of cardiac reactive oxygen species generation and oxidative stress damage. Accordingly, myrtenol restored the impairment of endogenous antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase and reductase) activities and balance of pro- and anti-apoptotic pathways (Bax and Blc-2), associated with decreased TUNEL-positive apoptotic cells. Taken together, our data show that myrtenol promotes cardioprotection against IR injury through attenuation of oxidative stress and inhibition of pro-apoptotic pathway. / A lesão de isquemia-reperfusão cardíaca representa um grande dano à saúde humana contribuindo para efeitos cardiovasculares adversos. Embora a reperfusão do coração isquêmico seja essencial para reduzir o dano miocárdico em processo de isquemia, a restauração do fluxo sanguíneo pode, paradoxalmente, amplificar o dano celular. Não existem metodologias de intervenção clínica que tratem a origem dos eventos celulares que levam ao dano celular observado. Neste contexto, o mirtenol é um monoterpeno com múltiplas atividades farmacológicas e estudos sugerem que esta molécula possa atuar em eventos que culminam na prevenção da lesão tissular. Assim, o objetivo deste estudo foi avaliar se o mirtenol promove a cardioproteção contra a lesão de isquemia-reperfusão cardíaca (IR) e os mecanismos envolvidos nesses efeitos. Para este estudo, ratos Wistar machos foram pré-tratados por via oral durante sete dias consecutivos com solução salina (NaCl 0,9% 0,20 ml + DMSO 0,05 mL), mirtenol (50 mg / kg) ou N-acetil cisteína (NAC) (1.200 mg / kg). Posteriormente, os corações foram submetidos à lesão de IR cardíaca, por meio do sistema de perfusão aórtica de fluxo constate do tipo Langedorff. Foram obtidos os parâmetros contráteis tais como pressão ventricular esquerda (PVE), derivadas máxima (+dP/dt) e mínima (-dP/dt) da PVE, frequência cardíaca (FC), parâmetros eletrocardiográficos, como FC, intervalo PR e a duração do complexo QRS. Além disso, foi mensurado a atividade da lactato desidrogenase (LDH) determinado o índice de severidade das arritmias cardíacas (ISA) e a área de infarto. O estresse oxidativo foi avaliado com base na determinação de hidroperóxidos totais, grupamento sulfidrilas totais, ainda foram mensuradas as espécies reativas de oxigênio (EROs), as proteínas carboniladas e determinada a atividade das enzimas antioxidantes endógenas: superóxido dismutase (SOD), catalase (CAT), glutationa peroxidase (GPx) e glutationa redutase (GR), e a área de infarto. A apoptose celular foi investigada mediante a expressão das proteínas Bax e Bcl-2 e a apoptose in situ foi avaliada pelo método TUNEL. Mostramos que o comprometimento severo do desempenho contrátil induzido pela IR foi prevenido de forma significativa pelo mirtenol e NAC. Além disso, que o mirtenol preveniu a forma de onda eletrocardiográfica discrepante (elevação do segmento ST), bem como, reduziu as arritmias fatais e o tamanho da área do infarto induzido pela lesão de IR. Relevantemente, o mirtenol impediu o aumento de superóxido e danos cardíacos que poderiam ser causados pelo estresse oxidativo. Consequentemente, o mirtenol restaurou a atividade das enzimas antioxidantes endógenas e o equilíbrio das vias pró e antiapoptóticas (Bax e Blc-2), associado à diminuição das células apoptóticas TUNEL-positivas. Em conjunto, nossos dados mostram que o mirtenol promove a cardioproteção contra lesão de IR através da atenuação do estresse oxidativo e inibição da via pró-apoptótica. / Aracaju

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-04-06T20:09:47Z No. of bitstreams: 1 tatianedeoliveirasouzasenra.pdf: 539017 bytes, checksum: 647e15774caf85c47318364c3b4c57e3 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-04-24T04:11:36Z (GMT) No. of bitstreams: 1 tatianedeoliveirasouzasenra.pdf: 539017 bytes, checksum: 647e15774caf85c47318364c3b4c57e3 (MD5) / Made available in DSpace on 2016-04-24T04:11:36Z (GMT). No. of bitstreams: 1 tatianedeoliveirasouzasenra.pdf: 539017 bytes, checksum: 647e15774caf85c47318364c3b4c57e3 (MD5) Previous issue date: 2013-02-07 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O presente estudo teve como objetivo avaliar a atividade carrapaticida de formulações hidroetanólicas do carvacrol, (E)-cinamaldeído, trans-anetol e linalol sobre larvas de Rhipicephalus microplus e Dermacentor nitens. Também foram realizados testes avaliando a atividade acaricida do carvacrol, timol, eugenol e (E)-cinamaldeído em diferentes concentrações sobre larvas e ninfas de Rhipicephalus sanguineus e Amblyomma cajennense. Todas as substâncias foram testadas nas concentrações de 2,5; 5,0; 10,0; 15,0 e 20,0 μl/ml, enquanto o timol foi testado nas mesmas concentrações em mg/ml; foram feitas 10 repetições para cada tratamento. Também foram feitos grupos controle com etanol na concentração necessária para solubilizar cada substância. Os grupos experimentais foram mantidos em câmara climatizada (27±1°C e UR>80±10%) por 24 horas e após este período, a mortalidade foi avaliada. Nos testes envolvendo larvas de R. microplus e D. nitens, o carvacrol e (E)-cinamaldeído causaram mortalidade superior a 98% em todas as concentrações testadas, o trans-anetol a partir da concentração de 5,0 μl/ml também causou a morte de 100% das larvas; entretanto, o linalol mesmo na maior concentração não foi eficiente, matando apenas 14,5 e 8,4% de R. microplus e D. nitens, respectivamente. Nos testes com larvas de A. cajennense a concentração de 5,0 μl/ml do carvacrol, timol e (E)-cinamaldeído causou a morte de 100% dos ixodídeos; para o eugenol este percentual só foi observado a partir da concentração de 15,0 μl/ml. Nos testes realizados com larvas de R. sanguineus, os grupos tratados com a menor concentração (2,5 μl/ml) de carvacrol e (E)-cinamaldeído apresentaram mortalidade de 100%; entretanto, para o eugenol, esse percentual só foi observado a partir da concentração de 10,0 μl/ml. Para ninfas desse mesmo ixodídeo, os monoterpenos carvacrol e timol a partir da menor concentração (2,5 μl/ml ou mg/ml) causaram letalidade de 100% em todas as concentrações, diferindo dos resultados obtidos para o eugenol e (E)-cinamaldeído, uma vez que mortalidade de 100% com a utilização dessas substâncias só foram observadas a partir da concentração de 10,0 μl/ml. Nos testes com A. cajennense a mortalidade das larvas tratadas com carvacrol, timol, eugenol e (E)-cinamaldeído na concentração de 2,5 μl/ml foi de 45, 62,7, 10,2 e 81,6%, chegando a 100% na concentração de 5,0 μl/ml para o carvacrol, timol e (E)-cinamaldeído. Para o eugenol esta taxa de mortalidade foi observada a partir da concentração de 15,0 μl/ml. Para ninfas os testes com carvacrol e timol causaram a morte de 100% dos indivíduos a partir das concentrações de 5,0 μl/ml e 10,0 mg/ml, respectivamente; o eugenol só atingiu mortalidade de 100% na concentração de 20,0 μl/ml, enquanto a xii mortalidade máxima no (E)-cinamaldeído não ultrapassou 64%. O conjunto de resultados obtidos neste estudo indica que os monoterpenos carvacrol e timol e os fenilpropanóides eugenol e (E)-cinamaldeído possuem atividade deletéria sobre estes ectoparasitos. Assim, existe a necessidade de novas pesquisas envolvendo outros estágios de desenvolvimento desses carrapatos, a investigação do sinergismo entre as substâncias, além de testes em condições naturais. Quanto a observação preliminar sobre a presença de ectoparasitos em aves silvestres, foram examinadas 72 aves de 17 espécies com prevalência de 29,2% de ectoparasitos. / The present study aimed to evaluate the efficiency of hydroethanolic formulations of carvacrol, (E)-cinnamaldehyde, trans-anetol and linalool against larval stages of Rhipicephalus microplus and Dermacentor nitens. It was also realized assays to evaluate the acaricide effect of carvacrol, thymol, eugenol and (E)-cinnamaldehyde in different concentrations in larvae and nymph of Rhipicephalus sanguineus and Amblyomma cajennense. All substances were tested in concentrations of 2.5, 5.0, 10.0, 15.0 and 20.0 μl/ml, thymol, tested in mg/ml; 10 repetitions were made for each treatment. It was also made with ethanol in a concentration necessary to solubilize each substance. The experimental groups were maintained in BOD incubator (27±1°C e UR>80±10%) during 24 hours and after this period the mortality was available. In the assays with R. microplus and D. nitens larvae, carvacrol and (E)-cinnamaldehyde caused mortality higher than 98% in all the tested concentrations, the trans-anetol caused a mortality of 100% in larvae from a concentration of 5.0 μl/ml; however, linalool was not efficien even at the higher concentration causing a mortality of only 14.5 and 8.4% in R. microplus and D. nitens, respectively. In assays realized with A. cajennense larvae, the concentration of 5.0 μl/ml of carvacrol, thymol and (E)-cinnamaldehyde cased a mortality of 100%; for eugenol this percentage was obtained from the concentration of 15.0 μl/ml. The test realized with R. sanguineus larvae, groups treated with the lower concentration (2.5 μl/ml) of carvacrol and (E)-cinnamaldehyde presented a mortality of 100%; nevertheless, for eugenol this percentage was observed only from the concentration of 10.0 μl/ml. For nymphs of the same species, the monoterpenes carvacrol and thymol from the lower concentration (2.5 μl/ml or mg/ml) caused a lethality of 100% in all concentrations, different from results obtained for eugenol and (E)-cinnamaldehyde, once mortality of 100% with these substances were observed only from the concentration of 10.0 μl/ml. In assays with A. cajennense, the mortality of larvae treated with carvacrol, thymol, eugenol and (E)-cinnamaldehyde in concentrations of 2.5 μl/ml of 45, 62.7, 10.2 and 81.6%, reaching 100% in the concentration of 5.0 μl/ml for carvacrol, thymol and (E)-cinnamaldehyde. For eugenol mortality rate was observed from the concentration of 15.0 μl/ml. For nymphs, tests with carvacrol and thymol caused a mortality of 100% of individuals from concentrations of 5.0 μl/ml and 10.0 mg/ml, respectively; the eugenol only reached this mortality rate at 20.0 μl/ml concentration, while the maximum mortality with (E)-cinnamaldehyde did not exceed 64%. The results indicate that monoterpenes and xiv phenylpropanoids, eugenol and (E)-cinnamaldehyde have deleterious activity in this ectoparasites. Thus, new researches are needed with other developmental stages of these tick species, the investigation of synergism between the substances, aside from tests in natural conditions. As regarding the preliminary observations of the presence of ectoparasites in wild birds, were examined 72 birds from 17 species different with prevalence of 29.2% ectoparasites.

Peu d’enzymes de synthèse de composés odorants sont connues chez le genre Rosa. Ce travail de thèse a permis l’identification de quelques-unes de ces protéines grâce à la technologie des puces à ADN, à l’analyse de l’expression des gènes par RT-PCR quantitative en temps réel (qPCR) et à l’analyse des parfums par chromatographie en phase gazeuse (CPG). Une puce confrontant les ADNc d’une rose parfumée à ceux d’une rose non parfumée a permis de corréler l’expression d’un gène, codant pour une Nudix hydrolase, très fortement exprimé dans la rose parfumée, avec la présence des monoterpènes dans le parfum de nombreux cultivars de rosiers. La caractérisation d’un rosier dont l’expression de ce gène est fortement réduite par ARN interférants, a permis de confirmer le rôle de celui-ci dans la synthèse des monoterpènes. La phénylacétaldéhyde synthase (PAAS) est une autre enzyme participant à la synthèse du parfum. Trois allèles de cette protéine ont précédemment été mis en évidence. Les résultats de qPCR et de CPG dans une population hybride ont permis de montrer que l’allèle a1 est le seul à pouvoir induire la synthèse et l’émission de 2-phényléthanol. Les activités respectives des différentes isoformes ont été testées in vitro chez la levure et in planta dans des feuilles de tabac et des cals de rosier : ces expériences montrent que les trois isoformes ont des activités comparables. L’absence de synthèse de 2-phényméthanol chez les plantes présentant les isoformes a2 et a3 réside donc dans la très faible expression de leurs allèles, induisant probablement une faible concentration de l’isoforme dans les cellules / Very few enzymes responsible for the biosynthesis of scent compounds in the genus Rosa are known so far. This PhD thesis aims to identify some of these proteins with DNA microarray technology, gene expression analysis by real-time quantitative PCR (qPCR) and scent analysis by gas chromatography (GC). An array comparing cDNA from a scented rose to those of a non-scented one, showed a correlation between expression of a yet-unknown gene, encoding a Nudix hydrolase, highly expressed in the scented rose, and the presence of monoterpenes in the scent of many rose cultivars. Characterization of a rose cultivar, in which expression of this gene has been decreased by RNA interference, confirmed its role in monoterpene synthesis. The phenylacetaldehyde synthase (PAAS) is another enzyme implicated in scent biosynthesis. Three alleles of this protein had been previously described. qPCR and GC experiments in a hybrid population showed that the a1 allele is the only one able to induce 2-phenylethanol biosynthesis. The respective activities of the different isoforms were tested in vitro in yeast, and in planta in tobacco leaves and rose calli: these experiments showed that the three isoforms have comparable activities. The lack of 2-phenylethanol production in plants having a2 and a3 isoforms is thus due to the very low expression of their respective alleles, probably inducing very low isoform concentration in cells

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The Citronellol (CT) is an alcoholic monoterpene present in the essential oil of some medicinal plants such as Cymbopogon citratus. Some pharmacological effects such as the antispasmodic and anticonvulsant activities have been described, however it s possible antinociceptive and anti-inflammatory effect is unknown. Thus, the objective of this study was to evaluate the possible antinociceptive and anti-inflammatory action of CT in rodents. Therefore, 362 male Swiss mice (25-35 g) with 2 to 3 months were used. The animals were divided into groups and were treated with CT (25, 50 and 100 mg/kg, i.p.), vehicle (saline solution 0.9% + Tween 80 0.2%, i.p.) or standard drug (i.p.). To evaluate the antinociceptive activity, the animals were submitted to the test of abdomnal constrictions induced by acetic acid (0.85%), the formalin test (1%) and hot plate test. In order to evaluate the effect of CT on orofacial nociception, it was conducted the orofacial test induced by formalin (2%), capsaicin and glutamate. Motor coordination was assessed using the motor coordination test (rota rod) and the spontaneous movement test. The anti-inflammatory activity was evaluated based on the model of carrageenan-induced pleurisy, making up the total leucocyte count. It was also quantified, by using ELISA, TNF-α and nitric oxide generation by macrophages. To determine the central action, the animals were treated with CT, in three doses, or vehicle and, after ninety minutes, were anesthetized, perfused, the brains removed and cut in a cryostat. The brain sections were subjected to immunofluorescence protocol for Fos protein. Results are expressed as mean ± SEM Differences between groups were analyzed by using one way ANOVA test, and followed by Tukey test. Values of p < 0.05 were considered statistically significant. Intraperitoneal administration of CT produced a significant decrease (p < 0.001) of the abdominal constrictions induced by acetic acid. In nociception induced by formalin, the pretreatment with CT caused a significant antinociceptive effect (p <0.01) in both phases of the test. In the hot plate test, the reaction time increased significantly at all doses of CT (p < 0.05 or p < 0.001), while its effect was antagonized by naloxone. In the three orofacial nociception tests, the CT produced a significant decrease (p <0.001) in the face-rubbing time of the orofacial region. No changes were observed in the motor coordination and in the spontaneous movement test. In the evaluation of anti-inflammatory activity, treatment with CT gave rise to a significant decrease (p < 0.01) in total number of leukocytes, decreasing (p < 0.05) the levels of TNF-α and nitric oxide in macrophages (p < 0.05). By immunofluorescence, it was found that CT is able to activate signicantly (p < 0.05) neurons of the olfactory bulb, the piriform cortex, the restrosplenial cortex and the periaqueductal gray. So, it can be concluded that CT has antinociceptive and anti-inflammatory activity and its action is mediated by central and peripheral mechanisms. / O Citronelol (CT) é um monoterpeno alcóolico presente no óleo essencial de algumas plantas medicinais, como o Cymbopogon citratus. Alguns efeitos farmacológicos tais como anti-espasmódico e atividade anticonvulsivante já foram descritos, sendo desconhecido seu possível efeito antinociceptivo e anti-inflamatório. Desta forma, o objetivo do presente estudo foi avaliar a possível ação antinociceptiva e anti-inflamatória do CT em roedores. Para tanto, foram utilizados 362 camundongos Swiss machos (25 a 35 g) com 2 a 3 meses. Os animais foram divididos em grupos e foram tratados com CT (25, 50 e 100 mg/kg; i.p.), veículo (solução salina 0,9% + tween 80 0,2%; i.p.) ou droga padrão (i.p.). Para avaliação da atividade antinociceptiva, os animais foram submetidos ao teste de contorções abdominais induzidas por ácido acético (0,85%), ao teste da formalina (1%) e ao teste da placa quente. Com o intuito de avaliar a ação do CT na nocicepção orofacial, foram realizados os testes de dor orofacial induzida por formalina (2%), capsaicina e glutamato. A coordenação motora dos animais foi avaliada através do teste da coordenação motora (rota rod) e do teste da movimentação espontânea. A atividade anti-inflamatória foi avaliada a partir do modelo de pleurisia induzido por carragenina, realizando-se a contagem de leucócitos totais. Foi quantificado, ainda, através do ensaio imunoenzimático ELISA, o TNF-α e a geração de óxido nítrico por macrófagos. Para determinar a ação central, os animais foram tratados com CT, nas três doses, ou veículo e, noventa minutos após, foram anestesiados, perfundidos, os cérebros extraídos e cortados em criostato. As secções cerebrais foram submetidas ao protocolo de imunofluorescência para proteína Fos. Os resultados foram expressos como média ± erro padrão da média. As diferenças entre os grupos foram analisadas por meio do teste de variância ANOVA, uma via, seguido pelo teste de Tukey. Valores de p < 0,05 foram considerados estatisticamente significantes. A administração intraperitoneal de CT produziu uma redução significativa (p < 0,001) das contorções abdominais. Na nocicepção induzida por formalina, o pré-tratamento com CT causou um efeito antinociceptivo significativo (p <0,01) em ambas as fases do teste. No teste da placa quente, o tempo de reação aumentou significativamente em todas as doses de CT (p < 0,05 ou p < 0,001), tendo seu efeito antagonizado pela naloxona. Nos três testes de nocicepção orofacial, o CT produziu uma redução significativa (p < 0,001) no tempo de fricção da região orofacial. Não foram observadas alterações no teste da coordenação motora e no teste da movimentação espontânea. Na avaliação da atividade anti-inflamatória, o tratamento com CT causou uma diminuição significativa (p < 0,01) no número total de leucócitos, diminuindo os níveis de TNF-α (p < 0,05) e de óxido nítrico em macrófagos (p < 0,05). Através da imunofluorescência, observou-se que o CT é capaz de ativar signicativamente (p < 0,05) neurônios do bulbo olfatório, do córtex piriforme, do córtex retroesplenial e da substância cinzenta periaquedutal. Conclui-se, assim, que o CT apresenta ação antinociceptiva e anti-inflamatória, tendo sua ação mediada por mecanismos centrais e periféricos.

A alelopatia pode desempenhar um papel relevante na dinâmica de campos e também no manejo de plantas daninhas em pastagens. Na região dos campos do sul do Brasil, monoculturas de Eucalyptus têm sido plantadas. A vegetação é escassa sob os plantios, o que pode estar associado à alelopatia. Em uma perspectiva aplicada, aleloquímicos de Eucalyptus poderiam ser potencialmente empregados como herbicidas naturais. Esta tese tem como objetivo avaliar se a alelopatia pode ser um fator determinante na estruturação da vegetação campestre, e se pode consistir em uma potencial ferramenta no controle de plantas daninhas. No capítulo I, uma revisão sistemática foi feita, visando evidenciar tendências gerais, antigas e atuais na pesquisa sobre alelopatia em ecossistemas campestres, com foco nos métodos utilizados. O capítulo II objetivou avaliar a fitotoxidez do extrato aquoso e do óleo essencial das folhas da serapilheira de Eucalyptus saligna Sm. sobre espécies campestres em laboratório. No capítulo III, os efeitos das folhas da serapilheira de E. saligna sobre espécies campestres foram investigados em plantios, bem como se esses efeitos estavam relacionados à alelopatia. O capítulo IV visou avaliar o potencial bioherbicida do óleo essencial das folhas da serapilheira de E. saligna e determinar quais componentes estavam associados a sua fitotoxidez. No capítulo I, evidenciou-se a potencial relevância da alelopatia na dinâmica de campos e no manejo de plantas daninhas em sistemas cultivados. Além disso, observou-se que os trabalhos recentes sobre alelopatia melhoraram em alguns aspectos de desenho experimental, mas não em outros, e que há inconsistência na terminologia utilizada. No capítulo II, observou-se que as folhas da serapilheira de E. saligna apresentaram substâncias fitotóxicas que geraram estresse oxidativo e levaram a danos nas membranas, afetando a germinação e o crescimento de plântulas. No capítulo III, evidenciou-se que as folhas da serapilheira inibiram a vegetação campestre em plantios de E. saligna, mas os efeitos foram principalmente físicos, e efeitos alelopáticos não foram detectados. No capítulo IV, foi demonstrado o potencial do óleo essencial de E. saligna como herbicida natural. O óleo foi mais fitotóxico que seus componentes majoritários, mas isso variou com o método e com as espécies receptoras. Esta tese evidenciou que generalizações sobre fitotoxidez e alelopatia devem ser evitadas. Conclui-se que a alelopatia possui um potencial maior como alternativa no manejo de plantas daninhas do que como um fator atuante sobre o estabelecimento e desenvolvimento de espécies vegetais campestres. Um maior conhecimento sobre alelopatia e seus mecanismos pode levar a avanços na ciência e em áreas aplicadas. / Allelopathy may play a relevant role in dynamics of grasslands, and also in weed management in pastures. In Southern Brazilian grasslands region, Eucalyptus monocultures have been planted. Vegetation is scarce under plantations, which may be associated with allelopathy. In an applied perspective, Eucalyptus allelochemicals may be potentially employed as natural herbicides. This thesis aimed to evaluate if allelopathy may be a key factor shaping grassland vegetation, and if it consists in a potential tool for weed control. In chapter I, a systematic review was conducted, in order to evidence general, old and current trends in allelopathy research in grassland ecosystems, focusing in used methods. Chapter II aimed to assess the phytotoxicity of Eucalyptus saligna Sm. leaf litter aqueous extract and essential oil on grassland species, in laboratory conditions. In chapter III, effects of E. saligna leaf litter on grassland species were investigated in plantations, as well as if these effects were related to allelopathy. Chapter IV aimed to evaluate the bioherbicide potential of E. saligna leaf litter essential oil, and to determine which compounds were related to the oil phytotoxicity. In chapter I, the potential relevance of allelopathy was evidenced in dynamics of grasslands and in weed management in cultivated systems. Moreover, the review demonstrated that allelopathy research has improved in some experimental design issues, but not in others, and inconsistency in terminology was observed. In chapter II, Eucalyptus saligna leaf litter showed phytotoxic compounds that generated oxidative stress and led to membrane damage, affecting seeds and seedling growth. In chapter III, leaf litter was evidenced to inhibit grassland species in E. saligna plantations, but effects were mainly physical, and allelopathic effects were not detected. In chapter IV, the essential oil showed potential as a natural herbicide. In general, E. saligna essential oil was more phytotoxic than its major compounds, but this varied according to the method and the recipient species. This thesis evidenced that generalizations should not be made, neither about phytotoxicity, nor about allelopathy. In conclusion, allelopathy showed greater potential as a tool for weed management than as a relevant factor influencing establishment and development of grassland vegetation. A better knowledge about allelopathy and its mechanisms may lead to advances in science and in applied fields.

This PhD thesis aims to improve the knowledge on the processes and chemical species in the gas- and particle-phases that are involved in the production of secondary organic aerosol (SOA) from monoterpene oxidation in the atmosphere. A denuder/filter technique that enabled the simultaneous sampling of gaseous and particulate compounds was applied in the present study. The sampling technique was comprehensively characterised and optimised using twelve atmospherically relevant carbonyl compounds. The present study improved the denuder coating procedure and the sampling performance. An additional coating with the derivatisation reagent, 2,4‑dinitrophenylhydrazine (DNPH), reduced the break-through potential (e.g., from 98% to 0.9% for methyl vinyl ketone) and the fraction of carbonyl compounds on the filter material (e.g., from 8.7% to 0% for acetone). Calibration experiments against an aerosol chamber were performed to reduce the relative standard deviation (RSD) of the calibration points in the denuder measurements. The RSDs were reduced by half for acetone, acetaldehyde, methyl vinyl ketone, glyoxal, benzaldehyde and campholenic aldehyde using a XAD‑4/DNPH denuder, and the quantification error was also reduced. This sampling technique was then applied to a series of α- and β-pinene ozonolysis experiments. The present study examined the influence of an OH radical scavenger (CO), and hence the HO2/RO2 ratio, on the SOA formation, product distribution and partitioning behaviour of selected oxidation products in conjunction with different seed particle acidities. It was shown that SOA yields increased by about 8% in α-pinene ozonolysis when CO and acidic seed particles co-existed, whereas only a marginal difference was observed (increase of 2%) for β-pinene compared to neutral seed particles. From the denuder/filter sample analysis, it was possible to tentatively identify a new compound from the α-pinene ozonolysis, i.e., terpenylic aldehyde. Gas- and particle-phase yields were estimated for the first time for this compound (i.e., 1% and 0.4%, respectively). The atmospheric relevance of terpenylic aldehyde was demonstrated based on ambient filter measurements and a possible formation pathway was suggested. Furthermore, the present study provided an additional explanation for enhanced SOA formation when acidic seed particles are used in monoterpene ozonolysis. It was demonstrated that the isomerisation of monoterpene oxides on acidic seed particles leads to the formation of highly reactive SOA precursors, whose subsequent reaction with ozone contributes significantly to SOA formation.

Les composés organiques volatils (COV) sont des composants clé en chimie atmosphérique. Ils participent à des réactions photochimiques dans la basse atmosphère et jouent ainsi un rôle majeur dans le cycle de l'ozone troposphérique et la formation d‘aérosols organiques secondaires. A l'échelle globale, ces COV sont à 10% d'origine anthropique et à 90% d'origine biotique. Les enjeux liés aux émissions de COVB sont d‘autant plus importants en région méditerranéenne en raison du potentiel d‘émission élevé ainsi que des évènements réguliers de pollution photochimique. Cette thèse propose, à travers deux campagnes de mesures intensives, la caractérisation expérimentale des COVB et de leurs produits d‘oxydation dans deux sites ruraux représentatifs de la végétation méditerranéenne.Dans le cadre du projet CANOPEE (ANR-JCJC 2011-2014) une campagne de mesure a eu lieu à l‘Observatoire du chêne pubescent de Haute Provence (O3HP). Des mesures de concentrations et de flux d‘émission ont été réalisées à l‘échelle de la canopée dans le but de quantifier l‘export des COVB hors canopée et d‘étudier la chimie intra-Canopée. Dans la forêt de l‘O3HP, des concentrations en isoprène très élevées ont été mesurées, en particulier au sein de la canopée où elles ont atteint 16 ppbv les journées les plus chaudes. Selon le facteur d‘émission de 7.2 mg m-2 h-1 déterminé, la forêt de chênes pubescents de l‘O3HP fait partie des écosystèmes les plus fortement émetteurs d‘isoprène à travers le monde. Parmi les COV oxygénés, seul le méthanol a des flux d‘émission significatifs, avec un taux d‘émission moyen égal à 0.3 mg m-2 h-1. Un des points forts de ce travail a consisté en l‘étude du transport et de la chimie intra-Canopée. D‘après nos estimations, les processus intra-Canopée sur le site de l‘OHP jouent un rôle mineur sur les flux nets d‘isoprène sortant de la canopée.La deuxième campagne de mesure s‘inscrit dans le cadre du programme ChArMEx (Chemistry-Aerosol Mediterranean Experiment). Elle a pris place sur le site du Cap Corse, situé à distance des sources anthropiques et qui abrite une diversité d'espèces végétales. Les résultats obtenus ont permis la quantification et spéciation de nombreux COVB. Un grand contraste dans la signature des émissions de COVB a été observé entre les deux sites d‘étude. Ainsi, tandis que l‘isoprène est responsable de plus de 90% des COVB en termes de concentrations à l‘OHP, il ne représente que 35% de leurs concentrations totales au Cap Corse. Les 65% restants sont composés de nombreux monoterpènes. Ces résultats ont également mis en évidence la chimie rapide des COVB et l‘importance de leur contribution à la formation de composés secondaires oxygénés dans les conditions de photochimie intense qui caractérisent le bassin méditerranéen en période estivale.Au Cap Corse, comme à l‘O3HP, les COVB représentent le puit dominant des radicaux hydroxyles. En présence de NOx et pour des concentrations équivalentes à celles de l‘OHP ou du Cap Corse, le potentiel des COVB à former de l‘ozone a été évalué à 10 ppbv en moyenne. / Volatile organic compounds (VOCs) are key components in atmospheric chemistry. They participate in photochemical reactions in the lower atmosphere and thus play a major role in the formation of tropospheric ozone and secondary organic aerosols. Biogenic VOCs (BVOCs) constitute approximately 90% of global VOC emissions. In the Mediterranean region, the emissions and reactivity of BVOCs are enhanced due to high temperatures and sunny conditions. In this context, this work proposes, through intensive field campaigns, the experimental characterization of BVOCs and their oxidation products in two rural sites where the vegetation is representative of the Mediterranean region. Within the framework of the CANOPEE program (ANR-JCJC 2011-2014) an intensive field campaign took place at the Oak Observatory of the Observatoire de Haute Provence (O3HP). Measurements of concentrations and emission fluxes were carried out in order to quantify the export of BVOC off the canopy and study in-Canopy processes. High concentrations of isoprene were measured with daily maximum ambient concentrations ranging between 2-16 ppbv inside the forest. According to the emission factor of 7.2 mg m-2 h-1 determined, downy oak forest of the O3HP is one of the strongest emitters of isoprene worldwide. Evidence of direct emission of methanol was also found exhibiting an average emission rate equal to 0.3 2 mg m-2 h-1. One of the strengths of this work consisted in the study of transport and intra-Canopy chemistry. In-Canopy chemical oxidation of isoprene was found to be weak and did not seem to have a significant impact on isoprene concentrations and fluxes above the canopy. The second field campaign is part of the ChArMEx (Chemistry-Aerosol Mediterranean Experiment) program. It took place on the remote site of Cap Corse, characterized by a strong diversity of plant species. The rich dataset obtained allowed the quantification and speciation of many BVOCs. A great contrast in the signature emissions was observed between the two study sites. Thus, while isoprene is responsible for over 90% of BVOCs in terms of concentrations at the O3HP, it represents only 35% of their total concentrations in Cap Corse. The remaining 65% are composed of many monoterpenes. These results have also highlighted the rapid chemistry of BVOCs and the importance of their contribution to the formation of secondary oxygenated compounds under intense photochemistry conditions, typical of the summertime in the Mediterranean basin. Biogenic BVOCs represented the dominant hydroxyl radicals sink at the O3HP as well as in Cap Corse. In the presence of NOx, the potential of the measured BVOCs to produce ozone has been estimated at about 10 ppbv on average.