Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Organo-synthetic insecticides are widely used to control pests in crops and stored products. The indiscriminate use of these products are responsible for many economic and environmental problems, which has been generating a growing demand for alternative control methods. The essential oils of plants have been studied as a viable alternative to pest management. Recent jobs indicate that the essential oil of alecrim-de-tabuleiro Lippia gracilis presents great potential for biopospection insecticide compounds. Thus, the aim of the present work was study the essential oils of two chemotypes of L. gracilis and their major compounds thymol and carvacrol, evaluating: i) the toxicity of these compounds against Diaphania hyalinata and selectivity over non-target organisms Apis mellifera and Polybia micans ii) the toxicity and behavioral effects of these compounds over Cryptolestes ferrugineus. The essential oils of both chemotypes of L. gracilis were toxic to D. hyalinata. The carvacrol chemotype was 29% more toxic than thymol chemotype. The monoterpenes thymol and carvacrol when applied alone, were about two and five times more toxic than their respective chemotypes. Essential oils acted very quickly causing mortality in half the population of D. hyalinata in less than three hours. All treatments were not selective in favor of A. mellifera and P. micans. The essential oils and the major compounds were highly toxic by fumigation to adults of C. ferrugineus. The concentrations of the thymol and carvacrol chemotypes required to cause 50% mortality of C. ferrugineus were 20.7 and 25.1 μL L-1, respectively. The isolated thymol and carvacrol were 1.7 and 1.9 times more toxic to C. ferrugineus. Essential oils thymol and carvacrol chemotypes caused a mortality half the population of C. ferrugineus in 63.3 and 83.1 hours. All treatments caused repellency and irritability in adult C. ferrugineus. Thus, our results show the great potential of essential oils of L. gracilis and their major compounds for the development of new products to be used in the pest control field and in stored products. / Inseticidas organossintéticos são amplamente utilizados no controle de pragas em lavouras e em produtos armazenados. O uso abusivo destes produtos é responsável por diversos problemas econômicos e ambientais, o que tem gerado uma demanda crescente por métodos alternativos de controle. Os óleos essenciais de plantas têm sido estudados como uma alternativa viável ao manejo de pragas. Trabalhos recentes apontam que o óleo essencial do alecrim-de-tabuleiro Lippia gracilis apresenta grande potencial para bioprospecção de compostos inseticidas. Desta forma, objetivou-se neste trabalho estudar os óleos essenciais de dois quimiotipos de L. gracilis e seus compostos majoritários timol e carvacrol, avaliando: i) a toxicidade destes compostos sobre Diaphania hyalinata e a seletividade sobre os organismos não-alvos Apis mellifera e Polybia micans; e ii) a toxicidade e os efeitos comportamentais destes compostos sobre Cryptolestes ferrugineus. Os óleos essenciais de ambos quimiotipos de L. gracilis se mostraram tóxicos a D. hyalinata, sendo o quimiotipo carvacrol 29% mais tóxico do que o quimiotipo timol. Os monoterpenos timol e carvacrol quando aplicados isoladamente foram cerca de duas e cinco vezes mais tóxicos que seus quimiotipos, respectivamente. Os óleos essenciais atuaram de forma rápida causando mortalidade na metade da população de D. hyalinata em menos de três horas. Todos os tratamentos não foram seletivos à A. mellífera e P. micans. Os óleos essenciais e os compostos majoritários foram altamente tóxicos por fumigação aos adultos de C. ferrugineus. As concentrações dos óleos essenciais dos quimiotipos timol e carvacrol necessárias para causar 50% de mortalidade a C. ferrugineus foram 20,7 e 25,1 μL L-1, respectivamante. O timol e o carvacrol isolados foram 1,7 e 1,9 vezes mais tóxicos a C. ferrugineus. Os óleos essenciais dos quimiotipos timol e carvacrol causaram mortalidade em metade da população de C. ferrugineus em 63,3 e 83,1 horas. Todos os tratamentos causaram repelência e irritabilidade em adultos de C. ferrugineus. Assim, nossos resultados mostram o grande potencial dos óleos essenciais de L. gracilis e seus compostos majoritários para o desenvolvimento de novos produtos a serem utilizados no controle de pragas em campo e nos produtos armazenados.

Amebic keratitis and granulomatous amebic encephalitis are diseases caused by free-living amoebas of the Acanthamoeba type. In the case of keratitis, the great problem is the relapse of this disease because of its resistance to medicines used for it, especially the cystic form of the organism. Plant essential oils have been used as potentially active agents against this protist. Therefore, the objective of this study was to determine the amebacide activity of plant essential oils of Lippia type against Acanthamoeba polyphaga trophozoites. To do this, 8 x 104 trophozoites were exposed to increased concentrations of essential oils of Lippia sidoides, Lippia gracilis, Lippia alba and Lippia pedunculosa during 24 hours, as well as of their majority compounds rotundifolona, carvone and carvacrol. Practically all the oil and compound concentrations presented amebacide activity. Based on these results, the researched oils in this study are potential candidates for a complementary healing for amebic keratitis and granulomatous amebic encephalitis. / A ceratite amebiana e a encefalite amebiana granulomatosa são doenças causadas por amebas de vida livre do gênero Acanthamoeba. No caso da ceratite, o grande problema é a recidiva da doença devido à resistência, especialmente da forma cística do organismo, aos medicamentos utilizados. Óleos essenciais de plantas têm sido utilizados como agentes potencialmente ativos contra este protista. Portanto, o objetivo deste estudo foi determinar a atividade amebicida de óleos essenciais de plantas do gênero Lippia frente à trofozoítos de Acanthamoeba polyphaga. Para isto 8 x 104 trofozoítos foram expostos durante 24 horas a concentrações crescentes dos óleos essenciais de Lippia sidoides, Lippia gracilis, Lippia alba e Lippia pedunculosa, bem como de seus compostos majoritários rotundifolona, carvona e carvacrol. Praticamente todas as concentrações dos óleos e compostos apresentaram atividade amebicida. Diante destes resultados, os óleos pesquisados neste estudo são candidatos em potencial para a terapêutica complementar na ceratite amebiana e encefalite amebiana granulomatosa.

<p>According to the Global Cancer Incidence, Mortality, and Prevention (GLOBOCAN) study for 2018, 2,089,000 women will have been diagnosed with breast cancer worldwide, with 627,000 breast cancer-related mortalities. It is estimated that between 15 – 20 % of breast cancer diagnoses are of the triple-negative subtype. Triple-negative breast cancers (TNBCs) do not express the receptors for estrogen, progesterone, and human epidermal growth factor 2, and hence cannot be treated using hormone receptor-targeted therapy. </p> <p>TNBCs are commonly of the basal-like phenotype, with high expression levels of proteins involved in epithelial-mesenchymal transition, extracellular-matrix (ECM) remodeling, cell cycle progression, survival and drug resistance, invasion, and metastasis. 5-year survival rates are significantly lower for TNBC patients, and the disease is characterized by poorer grade at the time of diagnosis as well as higher 5-year distant relapse rates, with a greater chance of lung and CNS metastases. Current treatments for TNBC take the form of aggressive cytotoxic chemotherapy regimens with multiple adverse side-effects. An important goal of on-going studies is to identify new compounds with significant TNBC-specificity, in order to improve patient survival outcomes while preserving a high quality of life during treatment.</p> <p> For several decades, compounds originally isolated from bioactive natural extracts, such as the taxanes and vinca<i> </i>alkaloids, have been at the forefront of chemotherapy. However, due to their non -specific mechanisms of action, treatment with these compounds eventually leads to significant toxicity to normal cells and tissues. Modern transcriptomics, metabolomics, and proteomics tools have greatly improved our understanding of the mechanisms governing cancer initiation and progression, and revealed the considerable heterogeneity of tumor cells. This has allowed for the identification of potential vulnerabilities in multiple cancers, including TNBCs. By leveraging these new technologies and insights with the tremendous diversity of bioactive compounds from organisms that remain unstudied, new classes of onco-drugs targeting pathways specific to TNBC cells could be identified in the near future.</p> <p>Here, we describe the cytotoxic effects of extracts from <i>Lippia origanoides </i>- a species of medicinal shrub native to Central and South America - on TNBC cells. We report that these extracts induce rapid, sustained, and irreversible apoptosis in TNBC cells <i>in vitro</i>, with significantly reduced cytotoxicity against normal mammary epithelial cells. The <i>L. origanoides </i>extracts LOE and L42 exploited two TNBC-specific characteristics to induce apoptosis in these cells: i) inhibiting the constitutively active survival and inflammatory NF-kB signaling pathway, and ii) significantly dysregulating the expression levels of mitochondrial enzymes required to maintain the TCA cycle and oxidative phosphorylation; metabolic pathways that are required for the maintenance of TNBC cell growth and proliferation.</p> <p>Finally, to lay the foundations for future studies on the abilities of these extracts to prevent tumor initiation and inhibit tumor growth <i>in vivo</i>, we also show that the <i>L. origanoides </i>extract, L42, is non-toxic<i> </i>to immunocompetent C57BL/6 mice, and have developed an <i>in vivo </i>model of human TNBC in athymic <i>nu/nu</i> mice. </p> <p>Collectively, our studies are the first to identify the anti-TNBC-specific properties of bioactive extracts from the <i>Lippia </i>species, and reveal that targeting NF-kB signaling and mitochondrial metabolism are potential avenues to new therapeutics against this subtype of breast cancer. Future work in our lab will focus on identifying the bioactive components (BACs) of the extract mediating its apoptotic effects, and shedding light on their protein binding partners within the cell.</p>

The aim of this study was to determine the influence of harvesting time and water stress on the chemical composition of the essential oil and to test the activities against ticks and fungus of the essential oil of L. gracilis. For the analysis of harvest season the plant material was collected from seven genotypes of L. gracilis at the Research Farm "Campus Rural da UFS", in the rainy and dry seasons. The experiment testing water stress was conducted in the dry season. The extraction of essential oils was performed in the Laboratory of Phytotechnology of the UFS through hydrodistillation. Chemical analysis of the essential oil performed using GC-MS in the Laboratory of Chromatography of the UFS. For the activity tests against ticks package larvae and immersion of engorged tick Rhipicephalus microplus in different concentrations of essential oil, thymol or carvacrol. To test of fungicidal activity, the essential oil at different concentrations was added to PDA medium. Each plate was inoculated with mycelia culture of Thielaviopis paradoxa. The essential oil of L. gracilis presented two distinct chemotypes, one genotype LGRA-106 presenting as major compound thymol and the other genotypes presenting carvacrol as major compound. The leaves provided essential oil with an average grade of 1.55% in the rainy season and 2.09% in the dry season. In the rainy season there was no significant difference in both the yield and the content. The chemical composition of essential oils L. gracilis showed high levels of terpenes, 92% in the rainy season and 96% in the dry season. In the experiment with irrigation the values of content and yield of all genotypes were smaller when compared without irrigation. In general, the species L. gracilis, for the presence of water in the soil, provides stability in the chemical composition of the essential oil regardless of season, since plants subjected to irrigation, even in the dry season, the essential oil produced in quantity and quality similar to the rainy season. The essential oil of L. gracilis exhibits high activity against ticks, proven by lethal concentrations of genotypes LGRA-201 (1.31 mg.mL-1) and LGRA-106 (4.66 mg.mL-1), demonstrating efficiency in the control of this parasite. The tests showed that the concentrations 0.45; 0.91 and 2.75 mg.mL-1 of all genotypes of L. gracilis completely inhibited the development of the pathogen T. paradoxa, corresponding to a percentage of mycelium growth inhibition of 100%. The concentration of 0.18 mg.mL-1 of essential oil was sufficient to significantly reduce the number of spores of T. paradoxa. The minimal fungicidal concentration T. paradoxa was found between concentrations from 0.80 to 0.98 mg.mL-1 for the essential oils and 0.26 mg.mL-1 for carvacrol and 0.35 mg.mL-1 to thymol. / O presente trabalho teve como objetivo determinar a influencia da época de colheita e do estresse hídrico na composição química do óleo essencial e testar a atividade carrapaticida e fungicida do óleo essencial de L. gracilis. Para as análises de época de colheita o material vegetal foi colhido de sete genótipos de L. gracilis no Campus Rural da UFS , nas épocas chuvosa e seca. Já para o ensaio de estresse hídrico o experimento foi conduzido na época seca. A extração de óleos essenciais foi realizada no Laboratório de Fitotecnia da UFS, por meio de hidrodestilação. A análise química do óleo essencial foi realizada utilizando CG-EM, no Laboratório de Cromatografia da UFS. Para a atividade carrapaticida foram realizadas os testes de pacote de larvas e de imersão de fêmeas ingurgitadas do carrapato Rhipicephalus microplus em diversas concentrações de óleo essencial, timol ou carvacrol. Para o teste de atividade fungicida, o óleo essencial, em diferentes concentrações foi adicionado ao meio BDA. Cada placa foi inoculada com micélios da cultura monospórica de Thielaviopis paradoxa. O óleo essencial de L. gracilis apresentou dois quimiotipos distintos um com o genótipo LGRA-106 apresentando o composto timol como majoritário e os demais o carvacrol. As folhas forneceram óleos essenciais amarelados com teor médio de 1,55% na época chuvosa e 2,09% na seca. Na época chuvosa não houve diferença significativa tanto no rendimento como no teor. A composição química dos óleos essenciais de L. gracilis apresentou altos níveis de terpenos, 92% na época chuvosa e 96% na época seca. No ensaio com irrigação os valores de teor e rendimento de todos os genótipos são menores quando comparado com o ensaio sem irrigação. De maneira geral a espécie L. gracilis, quanto à presença de água no solo, apresenta estabilidade na composição química do óleo essencial independente da época do ano, uma vez que as plantas submetidas à irrigação, mesmo no verão, produziram óleo essencial em quantidade e qualidade semelhantes à época do inverno. O óleo essencial de L. gracilis apresenta alta atividade carrapaticida, comprovados pelas concentrações letais dos genótipos LGRA-201 (1,31 mg.mL-1) e LGRA-106 (4,66 mg.mL-1), demonstrando eficiência no controle desse parasita. Os ensaios demonstraram que as concentrações 0,45; 0,91 e 2,75mg.mL-1 de todos os genótipos de L. gracilis inibiram completamente o desenvolvimento do patógeno T. paradoxa, correspondendo a uma porcentagem de inibição do crescimento micelial de 100%. A concentração de 0,18mg.mL-1 de óleo essencial foi suficiente para reduzir significativamente o número de esporos de T. paradoxa. A concentração fungicida mínima de T. paradoxa foi encontrada entre as concentrações de 0,80 a 0,98mg.mL-1 para os óleos essenciais e 0,26mg.mL-1 para o carvacrol, e 0,35mg.mL-1 para o timol.

The aim of this study was to determine the influence of harvesting time and water stress on the chemical composition of the essential oil and to test the activities against ticks and fungus of the essential oil of L. gracilis. For the analysis of harvest season the plant material was collected from seven genotypes of L. gracilis at the Research Farm "Campus Rural da UFS", in the rainy and dry seasons. The experiment testing water stress was conducted in the dry season. The extraction of essential oils was performed in the Laboratory of Phytotechnology of the UFS through hydrodistillation. Chemical analysis of the essential oil performed using GC-MS in the Laboratory of Chromatography of the UFS. For the activity tests against ticks package larvae and immersion of engorged tick Rhipicephalus microplus in different concentrations of essential oil, thymol or carvacrol. To test of fungicidal activity, the essential oil at different concentrations was added to PDA medium. Each plate was inoculated with mycelia culture of Thielaviopis paradoxa. The essential oil of L. gracilis presented two distinct chemotypes, one genotype LGRA-106 presenting as major compound thymol and the other genotypes presenting carvacrol as major compound. The leaves provided essential oil with an average grade of 1.55% in the rainy season and 2.09% in the dry season. In the rainy season there was no significant difference in both the yield and the content. The chemical composition of essential oils L. gracilis showed high levels of terpenes, 92% in the rainy season and 96% in the dry season. In the experiment with irrigation the values of content and yield of all genotypes were smaller when compared without irrigation. In general, the species L. gracilis, for the presence of water in the soil, provides stability in the chemical composition of the essential oil regardless of season, since plants subjected to irrigation, even in the dry season, the essential oil produced in quantity and quality similar to the rainy season. The essential oil of L. gracilis exhibits high activity against ticks, proven by lethal concentrations of genotypes LGRA-201 (1.31 mg.mL-1) and LGRA-106 (4.66 mg.mL-1), demonstrating efficiency in the control of this parasite. The tests showed that the concentrations 0.45; 0.91 and 2.75 mg.mL-1 of all genotypes of L. gracilis completely inhibited the development of the pathogen T. paradoxa, corresponding to a percentage of mycelium growth inhibition of 100%. The concentration of 0.18 mg.mL-1 of essential oil was sufficient to significantly reduce the number of spores of T. paradoxa. The minimal fungicidal concentration T. paradoxa was found between concentrations from 0.80 to 0.98 mg.mL-1 for the essential oils and 0.26 mg.mL-1 for carvacrol and 0.35 mg.mL-1 to thymol. / O presente trabalho teve como objetivo determinar a influencia da época de colheita e do estresse hídrico na composição química do óleo essencial e testar a atividade carrapaticida e fungicida do óleo essencial de L. gracilis. Para as análises de época de colheita o material vegetal foi colhido de sete genótipos de L. gracilis no Campus Rural da UFS , nas épocas chuvosa e seca. Já para o ensaio de estresse hídrico o experimento foi conduzido na época seca. A extração de óleos essenciais foi realizada no Laboratório de Fitotecnia da UFS, por meio de hidrodestilação. A análise química do óleo essencial foi realizada utilizando CG-EM, no Laboratório de Cromatografia da UFS. Para a atividade carrapaticida foram realizadas os testes de pacote de larvas e de imersão de fêmeas ingurgitadas do carrapato Rhipicephalus microplus em diversas concentrações de óleo essencial, timol ou carvacrol. Para o teste de atividade fungicida, o óleo essencial, em diferentes concentrações foi adicionado ao meio BDA. Cada placa foi inoculada com micélios da cultura monospórica de Thielaviopis paradoxa. O óleo essencial de L. gracilis apresentou dois quimiotipos distintos um com o genótipo LGRA-106 apresentando o composto timol como majoritário e os demais o carvacrol. As folhas forneceram óleos essenciais amarelados com teor médio de 1,55% na época chuvosa e 2,09% na seca. Na época chuvosa não houve diferença significativa tanto no rendimento como no teor. A composição química dos óleos essenciais de L. gracilis apresentou altos níveis de terpenos, 92% na época chuvosa e 96% na época seca. No ensaio com irrigação os valores de teor e rendimento de todos os genótipos são menores quando comparado com o ensaio sem irrigação. De maneira geral a espécie L. gracilis, quanto à presença de água no solo, apresenta estabilidade na composição química do óleo essencial independente da época do ano, uma vez que as plantas submetidas à irrigação, mesmo no verão, produziram óleo essencial em quantidade e qualidade semelhantes à época do inverno. O óleo essencial de L. gracilis apresenta alta atividade carrapaticida, comprovados pelas concentrações letais dos genótipos LGRA-201 (1,31 mg.mL-1) e LGRA-106 (4,66 mg.mL-1), demonstrando eficiência no controle desse parasita. Os ensaios demonstraram que as concentrações 0,45; 0,91 e 2,75mg.mL-1 de todos os genótipos de L. gracilis inibiram completamente o desenvolvimento do patógeno T. paradoxa, correspondendo a uma porcentagem de inibição do crescimento micelial de 100%. A concentração de 0,18mg.mL-1 de óleo essencial foi suficiente para reduzir significativamente o número de esporos de T. paradoxa. A concentração fungicida mínima de T. paradoxa foi encontrada entre as concentrações de 0,80 a 0,98mg.mL-1 para os óleos essenciais e 0,26mg.mL-1 para o carvacrol, e 0,35mg.mL-1 para o timol.

Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The methodology developed in the present work, through chromatographic fingerprint by LC-DAD and Principal Component Analysis (PCA), was adequate for the chemical differentiation of Lippia alba (Mill) NE Brown accessions, considering the extraction method (infusion and maceration hydroalcoholic) and the collection season (summer and winter), allowing to evaluate the influence of these variables on the chemical composition of extracts obtained. In addition, all extracts were submitted to cytotoxic tests against two tumor cell lines, HL-60 (human leukemia) and HepG2 (human hepatocellular carcinoma), and to inhibition tests against the enzyme acetylcholinesterase (AChE) in order to evaluate the form of extraction and which access is the most promising from a pharmacological point of view. Among the extracts analyzed against tumor cells, eight of them, two from the infusion: 01IW and 02IW and six from the hydroalcoholic extraction: 02HW, 24HW, 39HW, 54HW, 01HS and 24HS, presented better cell inhibition performances against HL-60, with percentage of inhibition between 40.0 and 52.0%, suggesting that these extracts are potentially promising against this lineage. With respect to the anticholinesterase tests, all extracts studied had low AChE inhibitory effect when compared to galantamine (87.4%), highlighting only the samples 01HW (28.4%), 02HW (27.2%), 54HW (29.3%), 01HS (32.7%), 02HS (27.4%) and 24HS (30.3%). Through the analyzes performed by Nuclear Magnetic Resonance (NMR) it was possible to identify the structures of some compounds isolated from this species: flavone tricin and phenylpropanoid verbascoside/isoverbascoside. In addition, the classes of three other metabolites were proposed: two phenylpropanoid derivatives and one flavonoid diglucoside. / A metodologia desenvolvida no presente trabalho, utilizando fingerprint cromatográfico por CL-DAD e Análise de Componentes Principais (PCA), se mostrou adequado para a diferenciação química de acessos de Lippia alba (Mill) N. E. Brown, considerando o método de extração (infusão e maceração hidroalcoólica) e a época de coleta (verão e inverno), permitindo, ainda, avaliar a influência destas variáveis na composição química dos extratos obtidos. Adicionalmente, todos os extratos foram submetidos a testes citotóxicos frente a duas linhagens de células tumorais, HL-60 (leucemia humana) e HepG2 (carcinoma hepatocelular humano), e de inibição enzimática frente a acetilcolinesterase (AChE) com o intuito de avaliar qual a forma de extração e qual acesso é o mais promissor do ponto de vista farmacológico. Entre todos os extratos analisados frente as células tumorais, dois provenientes da infusão: 01II e 02II e seis da extração hidroalcoólica: 02HI, 24HI, 39HI, 54HI, 01HV e 24HV, apresentaram melhores desempenhos de inibição frente a HL-60, com percentual de inibição entre 40,0 e 52,0%, sugerindo que estes extratos são potencialmente promissores contra esta linhagem. Com relação aos testes anticolinesterásicos, todos os extratos estudados apresentaram baixo efeito inibidor da AChE quando comparados a galantamina (87,4%), com destaque para os extratos 01HI (28,4%), 02HI (27,2%), 54HI (29,3%), 01HV (32,7%), 02HV (27,4%) e 24HV (30,3%). Através das análises realizadas por Ressonância Magnética Nuclear (RMN) foi possível identificar as estruturas de alguns dos compostos isolados a partir desta espécie: a flavona tricina e o fenilpropanóide verbascosídeo/isoverbascosídeo. Além disso, foram propostas as classes de outros três metabólitos: dois derivados de fenilpropanóides e um flavonóide diglicosilado. / São Cristóvão, SE

Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Sitophilus zeamais is a global important pest-insect, known for the losses that it causes during production and grain storage. One of the main problems caused by synthetic pesticides use is the development of resistant insect populations. Essential oils of plants have been considered a promising alternative to the conventional management of pests insects, however, its direct application has limitations due to the instability of its composition and high volatility. According to this, the technology of nanoformulations production can increase the potential of essential oil use, and its compounds, by increasing the efficiency, and stability of these compounds. The aim of this study was evaluate the lethal and sub lethal effects of Lippia sidoides essential oil, its majoritary compound (thymol), and nanoformulations of essential oil based prototypes, under S. zeamais populations (N = 5) from different regions of Brazil. The chemical composition of the essential oil, and nanoformulation was analyzed by GC/MS/DIC. The completely random design consisted in four replications for each combination of treatment x populations of S. zeamais. Bioassays of acute toxicity were conducted in Petri dishes (6 x 1,5 cm) with 10 unsexed adults. The lethal doses (LD50 and LD90), and the lethal time (LT50) were obtained through topic application bioassays. To the nanoformulation storage efficiency was used the LD95. On the population growth, and grain consume bioassays were used different concentrations of nanoformulations to mass grain treatment. The major compound present in the L. sidoides essential oil was thymol (68.5%). During nanoformulation storage, the concentration of these compound increased, getting to 82% of the essential oil composition. L. sidoides essential oil lethal doses, needed to kill 50% of S. zeamais populations, vary between 7.1 to 19.9 μg mg-1. Jacarezinho, and Maracaju populations were, respectively, the most tolerant and susceptible populations to L. sidoides essential oil. Intermediates susceptibilities were observed to Aracaju, Rio Branco, and Sete Lagoas populations. To thymol LD50 were observed variations between 17.1 to 25.7 μg mg-1. To nanoformulations prototypes (18%) containing L. sidoides essential oil, and thymol were observed LD50 variations between 26.4 to 36 e 20.8 a 27.7 μg mg-1, respectively. L. sidoides essential oil, thymol, and their nanoformulations had a fast action under S. zeamais populations. Lethal times needed to kill 50% of the population vary between 5.8 to 62 hours. In all of those cases, the nanoformulations took longer to cause the same mortality. The nanoformulations caused mortalities higher than 80% of S. zeamais populations of over 210 storage days. Nanoformulations increasing concentrations resulted on intrinsic tax (ri), and grain consume reduction of S. zeamais populations. This study prove that nanoformulations of L. sidoides oil essential based prototypes and its majoritary compound are promising alternatives to the management of S. zeamais once these demonstrated high efficacy, low liberation of the substances, and good storage stability. / Sitophilus zeamais é uma praga de importância mundial que ocasiona danos em grãos durante sua produção e estocagem. Um dos principais problemas da utilização de inseticidas organosintéticos, usados para o controle desta praga, consiste no desenvolvimento de populações de insetos resistentes. Óleos essenciais de plantas tem sido considerados uma boa alternativa ao manejo convencional, porém sua utilização direta apresenta limitações devido à instabilidade de sua composição e a alta volatilidade. Neste sentido, a tecnologia de produção de nanoformulações pode aumentar a potencialidade do uso dos óleos essenciais e seus compostos por aumentar a eficiência e a estabilidade destes compostos. No presente estudo analisamos os efeitos letais e subletais do óleo essencial de Lippia sidoides, de seu composto majoritário (timol) e de protótipos de nanoformulações a base destes compostos sobre populações de S. zeamais (N = 5) provenientes de diferentes regiões do Brasil. A composição química do óleo essencial e da nanoformulação foram analisadas por CG/EM/DIC. O delineamento experimental inteiramente casualizado, consistiu em quatro repetições para cada combinação de tratamento x população de S. zeamais. Bioensaios de toxicidade aguda foram conduzidos em placas de Petri (6 x 1,5 cm) contendo 10 adultos não sexados. As doses letais (DL50 e DL90) e o tempo letal (TL50) foram determinados em bioensaios de aplicação tópica. Para a eficiência do armazenamento das nanoformulações utilizou-se a DL95. Nos bioensaios de crescimento populacional e consumo de grãos foram utilizadas diferentes concentrações das nanoformulações para o tratamento da massa de grãos. O composto majoritário presente no óleo essencial de L. sidoides foi o timol (68,5%). Durante o armazenamento da nanoformulação esse composto aumentou, chegando a até 82% da composição do óleo. As doses letais do óleo essencial de L. sidoides necessárias para matar 50% das populações de S. zeamais variaram de 7,1 a 19,9 μg mg-1. As populações de Jacarezinho e Maracaju foram, respectivamente, a mais tolerante e susceptível ao óleo essencial de L. sidoides. Susceptibidades intermediárias foram observadas para as populações de Aracaju, Rio Branco e Sete Lagoas. O timol apresentou DL50 que variou de 17,1 a 25,7 μg mg-1. Os protótipos das nanoformulações (18%) contendo óleo essencial de L. sidoides e timol apresentaram DL50 que variaram de 26,4 a 36 e 20,8 a 27,7 μg mg-1, respectivamente. O óleo essencial de L. sidoides, o timol e suas nanoformulações atuaram rapidamente sobre as populações de S. zeamais. Os tempos letais necessários para matar 50% das populações variaram de 5,8 a 62 horas. Em todos os casos as nanoformulações demoram mais tempo para causar a mesma mortalidade. As nanoformulações causaram mortalidades superiores a 80% nas populações de S. zeamais até 210 dias de armazenamento. Concentrações crescentes das nanoformulações ocasiononaram redução da taxa intrínseca de crescimento (ri) e do consumo de grãos das populações de S. zeamais. Este trabalho comprova que os protótipos de nanoformulação a base do óleo essencial de L. sidoides e seu composto majoritário são promissores para o manejo de populações de S. zeamais uma vez que demonstraram alta eficácia, liberação lenta das substâncias e boa estabilidade no armazenamento.

The evolution of aerobic metabolic processes unavoidably led to the production of reactive oxygen species (ROS). ROS have the ability to cause harmful oxidative damage to biomolecules. Increased ROS generation and subsequent oxidative stress have been associated with aging and neurodegenerative disorders such as Parkinson’s and Alzheimer’s diseases as a result of the extreme sensitivity of the central nervous system to damage from ROS. Antioxidant defence systems have co–evolved with aerobic metabolic processes to counteract oxidative damage inflicted by ROS. The impact of neurodegenerative disorders on society is increasing rapidly as the life expectancy of the global population increases. In this day and age, a much younger group of the population is also experiencing neurodegenerative symptoms as a result of the harmful effect of the human immunodeficiency virus (HIV) on the central nervous system. Plants are an invaluable source of medicinal compounds. The use of plants for their healing properties is rooted in ancient times. The aim of this study was to select from twenty one plants, the plant with the most promising antioxidant activity and to determine whether extracts of this plant could act as free radical scavengers, comparing the results to Trolox, a known free radical scavenger. The next step was to isolate and characterize a compound from an extract exhibiting promising antioxidant activity. Bioassay–guided fractionation was followed to achieve this. During screening trials, twenty one plants, namely Berula erecta, Heteromorpha arborescens, Tarchonanthus camphoratus, Vernonia oligocephala, Gymnosporia buxifolia, Acacia karroo, Elephantorrhiza elephantina, Erythrina zeyheri, Leonotis leonurus, Plectranthus ecklonii, P. rehmanii, P. venteri, Salvia auretia, S. runcinata, Solenostemon latifolius, S. rotundifolius, Plumbago auriculata, Clematis brachiata, Vangueria infausta, Physalis peruviana and Lippia javanica were selected from literature, based on reported antioxidant activity within the plant families, for screening of their antioxidant activity. One hundred and ten extracts were prepared from the leaves, using Soxhlet extraction and the solvents petroleum ether (PE), dichloromethane (DCM), ethyl acetate (EtOAc) and ethanol (EtOH), consecutively. The focus during initial screening trials was on chemistry–based assays. The oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays were employed for the primary screening of the one hundred and ten leaf extracts. The ORAC assay was used to determine whether the plant extracts were able to scavenge peroxyl radicals and the FRAP assay was used to determine the reducing abilities of the extracts. Quantification of the peroxyl radical scavenging activity by the ORAC assay revealed that activity was observed for most of the extracts, with the ethyl acetate and ethanol extracts of L. javanica exhibiting the most promising activity. This pattern of activity was also found with the reducing capacity evaluated by the FRAP assay in which the EtOAc and EtOH extracts of L. javanica also exhibited the most promising activity. L. javanica was selected for further study by screening for biological activity, employing the nitro–blue tetrazolium (NBT) assay and thiobarbituric acid reactive substances (TBARS) assay. Using a cyanide model to induce neurotoxic effects in rat brain homogenate, the neuroprotective properties of the extracts of L. javanica leaves were examined using the NBT assay and compared to that of Trolox. The NBT assay determines the level of superoxide anions. All the extracts of L. javanica significantly reduced superoxide anion generation at all concentrations used. The petroleum ether and ethyl acetate extracts, at all concentrations, reduced superoxide anion generation to values lower than that of the control, suggesting that these extracts may be able to attenuate normal free radical processes in the brain. The petroleum ether extract exhibited the most promising activity at a concentration of 1.25 and 2.5 mg/ml and also exhibited similar results as the ethyl acetate extract at a lower concentration than the ethyl acetate extract (2.5 mg/ml compared to 5 mg/ml). A toxin–solution consisting of hydrogen peroxide (H2O2), iron(III)chloride (FeCl3) and ascorbic acid was used to induce lipid peroxidation and the ability of the extracts of the leaves of L. javanica to attenuate lipid peroxidation was investigated in rat brain homogenate and compared to that of Trolox. All of the extracts of L. javanica significantly attenuated toxininduced lipid peroxidation at all concentrations used. All of the extracts were also able to significantly attenuate toxin–induced lipid peroxidation to values lower than that of the control. These results suggest that all of the extracts of L. javanica possess the ability to attenuate not only toxin–induced lipid peroxidation, but also lipid peroxidation that occurs during normal processes in the brain. The petroleum ether extract was subjected to bioassay–guided fractionation using column and thin–layer chromatography and the NBT and TBARS assays. Fraction DD1 was investigated by means of nuclear magnetic resonance, infrared and mass spectrometry. The exact structure of fraction DD1 was not elucidated. Considering all the results, it is clear that L. javanica shows great potential as a medicinal plant with antioxidant activity and may therefore be beneficial in diminishing the destructive oxidative effects inflicted by free radicals. There are however still many compounds to be isolated from L. javanica. Key words: Verbenaceae, Lippia javanica, antioxidant, neurodegeneration, oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), nitro–blue tetrazolium assay (NBT), thiobarbituric acid reactive substances assay (TBARS). / Thesis (M.Sc. (Pharmaceutical Chemistry))--North-West University, Potchefstroom Campus, 2011.

The evolution of aerobic metabolic processes unavoidably led to the production of reactive oxygen species (ROS). ROS have the ability to cause harmful oxidative damage to biomolecules. Increased ROS generation and subsequent oxidative stress have been associated with aging and neurodegenerative disorders such as Parkinson’s and Alzheimer’s diseases as a result of the extreme sensitivity of the central nervous system to damage from ROS. Antioxidant defence systems have co–evolved with aerobic metabolic processes to counteract oxidative damage inflicted by ROS. The impact of neurodegenerative disorders on society is increasing rapidly as the life expectancy of the global population increases. In this day and age, a much younger group of the population is also experiencing neurodegenerative symptoms as a result of the harmful effect of the human immunodeficiency virus (HIV) on the central nervous system. Plants are an invaluable source of medicinal compounds. The use of plants for their healing properties is rooted in ancient times. The aim of this study was to select from twenty one plants, the plant with the most promising antioxidant activity and to determine whether extracts of this plant could act as free radical scavengers, comparing the results to Trolox, a known free radical scavenger. The next step was to isolate and characterize a compound from an extract exhibiting promising antioxidant activity. Bioassay–guided fractionation was followed to achieve this. During screening trials, twenty one plants, namely Berula erecta, Heteromorpha arborescens, Tarchonanthus camphoratus, Vernonia oligocephala, Gymnosporia buxifolia, Acacia karroo, Elephantorrhiza elephantina, Erythrina zeyheri, Leonotis leonurus, Plectranthus ecklonii, P. rehmanii, P. venteri, Salvia auretia, S. runcinata, Solenostemon latifolius, S. rotundifolius, Plumbago auriculata, Clematis brachiata, Vangueria infausta, Physalis peruviana and Lippia javanica were selected from literature, based on reported antioxidant activity within the plant families, for screening of their antioxidant activity. One hundred and ten extracts were prepared from the leaves, using Soxhlet extraction and the solvents petroleum ether (PE), dichloromethane (DCM), ethyl acetate (EtOAc) and ethanol (EtOH), consecutively. The focus during initial screening trials was on chemistry–based assays. The oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays were employed for the primary screening of the one hundred and ten leaf extracts. The ORAC assay was used to determine whether the plant extracts were able to scavenge peroxyl radicals and the FRAP assay was used to determine the reducing abilities of the extracts. Quantification of the peroxyl radical scavenging activity by the ORAC assay revealed that activity was observed for most of the extracts, with the ethyl acetate and ethanol extracts of L. javanica exhibiting the most promising activity. This pattern of activity was also found with the reducing capacity evaluated by the FRAP assay in which the EtOAc and EtOH extracts of L. javanica also exhibited the most promising activity. L. javanica was selected for further study by screening for biological activity, employing the nitro–blue tetrazolium (NBT) assay and thiobarbituric acid reactive substances (TBARS) assay. Using a cyanide model to induce neurotoxic effects in rat brain homogenate, the neuroprotective properties of the extracts of L. javanica leaves were examined using the NBT assay and compared to that of Trolox. The NBT assay determines the level of superoxide anions. All the extracts of L. javanica significantly reduced superoxide anion generation at all concentrations used. The petroleum ether and ethyl acetate extracts, at all concentrations, reduced superoxide anion generation to values lower than that of the control, suggesting that these extracts may be able to attenuate normal free radical processes in the brain. The petroleum ether extract exhibited the most promising activity at a concentration of 1.25 and 2.5 mg/ml and also exhibited similar results as the ethyl acetate extract at a lower concentration than the ethyl acetate extract (2.5 mg/ml compared to 5 mg/ml). A toxin–solution consisting of hydrogen peroxide (H2O2), iron(III)chloride (FeCl3) and ascorbic acid was used to induce lipid peroxidation and the ability of the extracts of the leaves of L. javanica to attenuate lipid peroxidation was investigated in rat brain homogenate and compared to that of Trolox. All of the extracts of L. javanica significantly attenuated toxininduced lipid peroxidation at all concentrations used. All of the extracts were also able to significantly attenuate toxin–induced lipid peroxidation to values lower than that of the control. These results suggest that all of the extracts of L. javanica possess the ability to attenuate not only toxin–induced lipid peroxidation, but also lipid peroxidation that occurs during normal processes in the brain. The petroleum ether extract was subjected to bioassay–guided fractionation using column and thin–layer chromatography and the NBT and TBARS assays. Fraction DD1 was investigated by means of nuclear magnetic resonance, infrared and mass spectrometry. The exact structure of fraction DD1 was not elucidated. Considering all the results, it is clear that L. javanica shows great potential as a medicinal plant with antioxidant activity and may therefore be beneficial in diminishing the destructive oxidative effects inflicted by free radicals. There are however still many compounds to be isolated from L. javanica. Key words: Verbenaceae, Lippia javanica, antioxidant, neurodegeneration, oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), nitro–blue tetrazolium assay (NBT), thiobarbituric acid reactive substances assay (TBARS). / Thesis (M.Sc. (Pharmaceutical Chemistry))--North-West University, Potchefstroom Campus, 2011.

Submitted by Johnny Rodrigues (johnnyrodrigues@ufcg.edu.br) on 2018-05-03T18:31:57Z No. of bitstreams: 1 ANDRESSA LACERDA NÓBREGA UGULINO - DISSERTAÇÃO PPGSA ACADÊMICO 2017..pdf: 676544 bytes, checksum: 0dd2e41c1011a60929ca0702449f6287 (MD5) / Made available in DSpace on 2018-05-03T18:31:57Z (GMT). No. of bitstreams: 1 ANDRESSA LACERDA NÓBREGA UGULINO - DISSERTAÇÃO PPGSA ACADÊMICO 2017..pdf: 676544 bytes, checksum: 0dd2e41c1011a60929ca0702449f6287 (MD5) Previous issue date: 2017 / O fungo Macrophomina phaseolina (Tassi.). Goid é o agente etiológico de uma das principais doenças na cultura do feijão-caupi, vulgarmente conhecida como podridão cinzenta do caule. Em decorrência da falta de produtos fitossanitários registrados para o controle desta doença um aumento na busca por métodos de controle alternativo é cada vez mais comum. O objetivo do presente trabalho foi avaliar o efeito de óleos essenciais sobre o crescimento micelial do fungo M. phaseolina. Os óleos testados foram de hortelã (Mentha sp.), eucalipto (Eucalyptus spp.), copaíba ( Copaifera sp.) e alecrim-da-chapada (Lippia gracilis), nas concentrações 0,4; 0,6; 0,8 e 1,0%. Os óleos foram incorporados ao meio de cultura do tipo BDA e posteriormente vertidos em placas de Petri de 90x15mm, após a solidificação do meio, discos de 8mm contendo o micélio do patógeno foram repicados para as placas com os respectivos tratamentos. O experimento foi conduzido no delineamento experimental inteiramente casualizado (DIC) em arranjo fatorial 4 x 4 + 1 +1, sendo dezesseis tratamentos mais uma testemunha negativa e uma testemunha positiva que constou da aplicação suplementar no meio (BDA) do fungicida Sportak 450 EC (Procloraz), e cinco repetições, totalizando 90 parcelas experimentais. Posteriormente todas as placas foram transferidas para incubadora do tipo B.O.D. (biochemical oxygen demand) a 27 ± 2 ºC. Foram realizadas medições diárias do diâmetro da colônia em dois sentidos perpendiculares até que tomasse toda a superfície do meio de cultura de uma das placas. Os dados obtidos foram interpretados por meio de análise de variância não paramétrica e as médias foram comparadas pelo teste de Kruskal-wallis, a 5% de probabilidade de erro e Scott Knott a 5% de probabilidade nos casos em que existiram diferenças significativas e normalidade dos dados. Os óleos essenciais de hortelã e alecrimda-chapada apresentaram melhores resultados quanto a inibição do crescimento micelial, enquanto que os óleos de eucalipto e copaíba apesar de potencialmente promissores, apresentaram inibição intermediária do crescimento micelial do fungo. / Macrophomina phaseolina (Tassi.). Goid is the etiological agent of one of the major diseases in cowpea bean crop, commonly known as gray rot stem. Due to the lack of registered phytosanitary products to control this disease, the search for alternative control methods is increasingly common. The objective of this study was to evaluate the effect of essential oils on the mycelial growth of M. phaseolina. The oils tested were Mentha sp.,Eucalyptus spp., Copaifera sp., and Lippia gracilis at concentrations of 0.4; 0.6; 0.8 and 1.0%. The work was conducted in Phytopathology Laboratory of the Federal University of Campina Grande. Daily measurements of the colony diameter were performed in two perpendicular directions until it filled the entire surface of the culture medium of one of the plates. The experiment was conducted in a completely randomized experimental design in a factorial arrangement 4 x 4 + 1+1, with sixteen treatments plus one negative control and one positive control, which consisted of the supplemental application in the medium (BDA) of the fungicide Sportak 450 EC (Procloraz), and five replications, totaling 90 experimental plots. The data were interpreted through non-parametric analysis of variance and the means were compared by the Kruskal-wallis test, with a 5% probability of error and Scott Knott at 5% probability in cases where there were significant differences and data normality. The Mentha sp. essential oil and Lippia gracilis showed better results in inhibiting mycelial growth, while the Eucalyptus essential oil and Copaifera, although potentially promising, showed intermediate inhibition of fungal mycelial growth.