Objectives The objective of the present study was to investigate the usefulness of time-frequency analysis (TFA) of olfactory-induced EEG change with a low-cost, portable olfactometer in the clinical investigation of smell function. Materials & methods A total of 78 volunteers participated. The study was composed of three parts where olfactory stimuli were presented using a custom-built olfactometer. Part I was designed to optimize the stimulus as well as the recording conditions. In part II EEG-power changes after olfactory/trigeminal stimulation were compared between healthy participants and patients with olfactory impairment. In Part III the test-retest reliability of the method was evaluated in healthy subjects. Results Part I indicated that the most effective paradigm for stimulus presentation was cued stimulus, with an interstimulus interval of 18-20s at a stimulus duration of 1000ms with each stimulus quality presented 60 times in blocks of 20 stimuli each. In Part II we found that central processing of olfactory stimuli analyzed by TFA differed significantly between healthy controls and patients even when controlling for age. It was possible to reliably distinguish patients with olfactory impairment from healthy individuals at a high degree of accuracy (healthy controls vs anosmic patients: sensitivity 75%; specificity 89%). In addition we could show a good test-retest reliability of TFA of chemosensory induced EEG-power changes in Part III. Conclusions Central processing of olfactory stimuli analyzed by TFA reliably distinguishes patients with olfactory impairment from healthy individuals at a high degree of accuracy. Importantly this can be achieved with a simple olfactometer.

Reconhecida pela sua megadiversidade, mas também pelo alto nível de desmatamento e perda de espécies que vem ocorrendo nas últimas décadas, a Mata Atlântica é um dos biomas mais ameaçados do planeta, recebendo a denominação de hotspot mundial. As abelhas exercem papel fundamental para a manutenção da vegetação por meio da polinização e o conhecimento sobre a composição do táxon contribui para ações de conservação. Abelhas da tribo Euglossini podem atuar como bioindicadoras da qualidade ambiental e exercem uma estreita relação com determinadas plantas, principalmente da família Orchidaceae, sendo fundamentais para a perpetuação de muitas espécies desta família. A tribo Euglossini apresenta riqueza de espécies nas florestas tropicais decaindo em direção às latitudes altas. Este estudo propôs investigar e reconhecer a fauna de Euglossini presente na região norte do estado de Santa Catarina, no sul do Brasil, a partir de coletas mensais em seis localidades, distribuídas ao longo de um gradiente altitudinal entre o nível do mar e 800 metros, nos municípios de Joinville e São Francisco do Sul. Duas localidades são constituídas por morros remanescentes de Mata Atlântica, inseridos na malha urbana de Joinville, com extensões de aproximadamente 390 e 525 hectares. As demais localidades compõem áreas contínuas de floresta na costa litorânea e Serra do Mar. Em cada local houve um dia de coleta por mês nos períodos de setembro/2013 até abril/2014 e de setembro/2014 até abril/2015. As iscas odoríferas disponibilizadas foram: benzoato de benzila, cineol, eugenol e vanilina. Nos últimos seis meses do estudo foi testada uma nova fragrância à base de menta: o mentol. O estudo registrou 794 machos de Euglossini dos gêneros Eufriesea, Euglossa e Eulaema, distribuídos em dez espécies válidas e uma potencialmente nova. As localidades com maiores riquezas foram os remanescentes no Morro do Finder e Morro do Boa Vista, e as maiores diversidades ocorreram na Vila da Glória e Mutucas. As espécies Eufriesea violacea Blanchard 1840, Euglossa annectans Dressler 1982 e Eulaema nigrita Lepeletier, 1841 foram registradas nas seis localidades estudadas. Ef. violacea, Eg. annectans Eg. stellfeldi representaram 91,56% da comunidade amostrada, sendo 66,87% somente da primeira espécie. O cineol atraiu 299 abelhas, o eugenol 190 e vanilina 165, mas em relação à riqueza, os mais atrativos foram o benzoato de benzila, cineol e vanilina, com seis espécies cada. O mentol mostrou eficácia, atraindo 75 abelhas de quatro espécies. A distribuição das abelhas em relação a variação altitudinal demonstrou que as maiores riqueza e abundância ocorreram entre o nível do mar e 200 metros de altitude com diminuição do número de indivíduos e espécies acima desta cota. Interações entre as abelhas e oito espécies de orquídeas foram registradas a partir de polinários aderidos às abelhas. Apesar da ocupação humana no entorno dos fragmentos da área urbana, os Morros do Finder e do Boa Vista apresentaram bom estado de conservação para a comunidade de Euglossini. Novos esforços e utilização de outras metodologias podem enriquecer os dados obtidos contribuindo para uma melhor caracterização ambiental da região. A partir dos dados existentes na literatura sobre a tribo Euglossini em Santa Catarina, nota-se uma riqueza significativa nos municípios de Joinville e São Francisco do Sul. / Recognized for its megadiversity, but also the high level of deforestation and species loss that has occurred in recent decades, the Atlantic Forest is one of the most threatened biomes on the planet, getting the global name hotspot. Bees play a crucial role in the maintenance of vegetation through pollination and knowledge about the taxon composition contributes to conservation actions. The tribe Euglossini can act as bioindicators of environmental quality and exert a close relationship with certain plants, particularly the orchid family, being fundamental to the perpetuation of many species of this family. The tribe Euglossini has a species richness in tropical forests which decreases in the higher latitudes. This study aimed to investigate and identify the Euglossini fauna present in the northern state of Santa Catarina, in southern Brazil, from monthly collections in six locations distributed along an altitudinal gradient between sea level and 800 meters, in the cities of Joinville and São Francisco do Sul. Two locations are composed of remnants of Atlantic Forest hills, situated in the urban area of Joinville, with extensions of approximately 390 and 525 hectares. Other locations comprise continuous areas of forest on the coastline and Serra do Mar. At each site there was one collection day per month in the period from September / 2013 to April / 2014 and September / 2014 to April / 2015. The available scent baits were: benzyl benzoate, cineol, eugenol and vanillin. In the last six months of the study a new mint-based fragrance was tested : menthol. The study recorded 794 Euglossini males of the genera Eufriesea, Euglossa and Eulaema, distributed in ten valid species and one potentially new species. The localities with the greatest richness were in Morro do Finder and Morro do Boa Vista, and the largest diversity occurred in the Vila da Glória and Mutucas. The species Eufriesea violacea Blanchard 1840, Euglossa annectans Dressler 1982 e Eulaema nigrita Lepeletier, 1841 were recorded in the six studied locations. Ef. violacea, Eg. annectans and Eg. stellfeldi represented 91.56% of the sampled community, with the first species making up 66.87% of that figure. The cineole attracted 299 bees, eugenol 190 and vanillin 165, but in relation to richness, the most attractive were the benzyl benzoate, cineole and vanillin, with six species each. The menthol was effective, attracting 75 bees from four species. The distribution of bees in relation to altitudinal variation showed that the greatest richness and abundance occured between sea level and an altitude of 200 metres, with a decrease in the number of individuals and species above this altimetry quota. Interactions between bees and eight species of orchids were recorded from pollinaria adhered to the bees. Despite the surrounding human occupation of urban fragments, Morros Finder and Boa Vista showed good conditions for the community of Euglossini. New efforts and using other methodologies can enrich the data, contributing to a better environmental characterization of the area. From the literature data on the tribe Euglossini in Santa Catarina, there is a significant richness in the cities of Joinville and São Francisco do Sul.

Reconhecida pela sua megadiversidade, mas também pelo alto nível de desmatamento e perda de espécies que vem ocorrendo nas últimas décadas, a Mata Atlântica é um dos biomas mais ameaçados do planeta, recebendo a denominação de hotspot mundial. As abelhas exercem papel fundamental para a manutenção da vegetação por meio da polinização e o conhecimento sobre a composição do táxon contribui para ações de conservação. Abelhas da tribo Euglossini podem atuar como bioindicadoras da qualidade ambiental e exercem uma estreita relação com determinadas plantas, principalmente da família Orchidaceae, sendo fundamentais para a perpetuação de muitas espécies desta família. A tribo Euglossini apresenta riqueza de espécies nas florestas tropicais decaindo em direção às latitudes altas. Este estudo propôs investigar e reconhecer a fauna de Euglossini presente na região norte do estado de Santa Catarina, no sul do Brasil, a partir de coletas mensais em seis localidades, distribuídas ao longo de um gradiente altitudinal entre o nível do mar e 800 metros, nos municípios de Joinville e São Francisco do Sul. Duas localidades são constituídas por morros remanescentes de Mata Atlântica, inseridos na malha urbana de Joinville, com extensões de aproximadamente 390 e 525 hectares. As demais localidades compõem áreas contínuas de floresta na costa litorânea e Serra do Mar. Em cada local houve um dia de coleta por mês nos períodos de setembro/2013 até abril/2014 e de setembro/2014 até abril/2015. As iscas odoríferas disponibilizadas foram: benzoato de benzila, cineol, eugenol e vanilina. Nos últimos seis meses do estudo foi testada uma nova fragrância à base de menta: o mentol. O estudo registrou 794 machos de Euglossini dos gêneros Eufriesea, Euglossa e Eulaema, distribuídos em dez espécies válidas e uma potencialmente nova. As localidades com maiores riquezas foram os remanescentes no Morro do Finder e Morro do Boa Vista, e as maiores diversidades ocorreram na Vila da Glória e Mutucas. As espécies Eufriesea violacea Blanchard 1840, Euglossa annectans Dressler 1982 e Eulaema nigrita Lepeletier, 1841 foram registradas nas seis localidades estudadas. Ef. violacea, Eg. annectans Eg. stellfeldi representaram 91,56% da comunidade amostrada, sendo 66,87% somente da primeira espécie. O cineol atraiu 299 abelhas, o eugenol 190 e vanilina 165, mas em relação à riqueza, os mais atrativos foram o benzoato de benzila, cineol e vanilina, com seis espécies cada. O mentol mostrou eficácia, atraindo 75 abelhas de quatro espécies. A distribuição das abelhas em relação a variação altitudinal demonstrou que as maiores riqueza e abundância ocorreram entre o nível do mar e 200 metros de altitude com diminuição do número de indivíduos e espécies acima desta cota. Interações entre as abelhas e oito espécies de orquídeas foram registradas a partir de polinários aderidos às abelhas. Apesar da ocupação humana no entorno dos fragmentos da área urbana, os Morros do Finder e do Boa Vista apresentaram bom estado de conservação para a comunidade de Euglossini. Novos esforços e utilização de outras metodologias podem enriquecer os dados obtidos contribuindo para uma melhor caracterização ambiental da região. A partir dos dados existentes na literatura sobre a tribo Euglossini em Santa Catarina, nota-se uma riqueza significativa nos municípios de Joinville e São Francisco do Sul. / Recognized for its megadiversity, but also the high level of deforestation and species loss that has occurred in recent decades, the Atlantic Forest is one of the most threatened biomes on the planet, getting the global name hotspot. Bees play a crucial role in the maintenance of vegetation through pollination and knowledge about the taxon composition contributes to conservation actions. The tribe Euglossini can act as bioindicators of environmental quality and exert a close relationship with certain plants, particularly the orchid family, being fundamental to the perpetuation of many species of this family. The tribe Euglossini has a species richness in tropical forests which decreases in the higher latitudes. This study aimed to investigate and identify the Euglossini fauna present in the northern state of Santa Catarina, in southern Brazil, from monthly collections in six locations distributed along an altitudinal gradient between sea level and 800 meters, in the cities of Joinville and São Francisco do Sul. Two locations are composed of remnants of Atlantic Forest hills, situated in the urban area of Joinville, with extensions of approximately 390 and 525 hectares. Other locations comprise continuous areas of forest on the coastline and Serra do Mar. At each site there was one collection day per month in the period from September / 2013 to April / 2014 and September / 2014 to April / 2015. The available scent baits were: benzyl benzoate, cineol, eugenol and vanillin. In the last six months of the study a new mint-based fragrance was tested : menthol. The study recorded 794 Euglossini males of the genera Eufriesea, Euglossa and Eulaema, distributed in ten valid species and one potentially new species. The localities with the greatest richness were in Morro do Finder and Morro do Boa Vista, and the largest diversity occurred in the Vila da Glória and Mutucas. The species Eufriesea violacea Blanchard 1840, Euglossa annectans Dressler 1982 e Eulaema nigrita Lepeletier, 1841 were recorded in the six studied locations. Ef. violacea, Eg. annectans and Eg. stellfeldi represented 91.56% of the sampled community, with the first species making up 66.87% of that figure. The cineole attracted 299 bees, eugenol 190 and vanillin 165, but in relation to richness, the most attractive were the benzyl benzoate, cineole and vanillin, with six species each. The menthol was effective, attracting 75 bees from four species. The distribution of bees in relation to altitudinal variation showed that the greatest richness and abundance occured between sea level and an altitude of 200 metres, with a decrease in the number of individuals and species above this altimetry quota. Interactions between bees and eight species of orchids were recorded from pollinaria adhered to the bees. Despite the surrounding human occupation of urban fragments, Morros Finder and Boa Vista showed good conditions for the community of Euglossini. New efforts and using other methodologies can enrich the data, contributing to a better environmental characterization of the area. From the literature data on the tribe Euglossini in Santa Catarina, there is a significant richness in the cities of Joinville and São Francisco do Sul.

Os mecanismos fisiológicos de detecção e controle da sede no paciente cirúrgico idoso ocorrem de forma diferenciada do que em adultos e crianças o que justifica o investimento na condução de estudos sobre estratégias que podem proporcionar alívio deste sintoma com o uso de temperatura fria e volume pequeno. O objetivo do estudo foi comparar a intensidade e o desconforto da sede de pacientes idosos que degustaram o picolé mentolado com a sede de pacientes idosos que receberam o cuidado usual, no pós-operatório imediato. Trata-se de ensaio clínico randomizado em paralelo com dois grupos, realizado em sala de recuperação pós-anestésica de hospital público de ensino no sul do Brasil. A amostra foi composta de 50 pacientes idosos em pós-operatório imediato, os quais foram alocados em dois grupos, sendo 25 no grupo controle e 25 no grupo experimental, sem perda de seguimento. Os critérios de inclusão foram: pacientes com idade >= 60 anos; estar em jejum; verbalizar sede e ter sido aprovado na avaliação do Protocolo de Segurança no Manejo da Sede. Os pacientes com restrições à ingesta ou deglutição, bem como os que autorreferiram alergia à menta, foram excluídos. Os pacientes alocados no grupo controle receberam o cuidado usual de rotina da instituição, onde o estudo foi conduzido, que consiste na manutenção do jejum. Já os pacientes alocados no grupo experimental receberam um picolé mentolado de 20 ml. Os desfechos primários de interesse foram a intensidade e o desconforto da sede, sendo que ambos foram avaliados inicialmente (T0) e novamente (T1) após 20 minutos da intervenção ou cuidado usual. A intensidade da sede foi mensurada por meio da Escala Numérica (zero significa nenhuma sede e 10 a maior sede já vivenciada pelo paciente), e o desconforto da sede pela Escala de Desconforto da Sede Perioperatória (pontuação de zero a 14 pontos, sendo zero a ausência de desconforto e 14 corresponde ao desconforto mais intenso relacionado à sede). A mediana da intensidade e do desconforto da sede inicial (T0) foi de 6,0 para os participantes alocados no grupo experimental, e 5,0 para a intensidade e 6,0 para o desconforto da sede para os participantes alocados no grupo controle. Para a comparação das mudanças nos escores da Escala Numérica e da Escala de Desconforto da Sede Perioperatória entre o grupo experimental e o grupo controle (valor final menos valor inicial), o teste de Mann-Whitney foi empregado para diferenças de medianas. Os resultados do ensaio clínico randomizado demonstraram que houve diminuição estatisticamente significante na intensidade (p<0,001) e no desconforto da sede (p<0,001) dos pacientes cirúrgicos idosos, após vinte minutos da degustação do picolé mentolado quando comparados à intensidade e desconforto da sede dos pacientes que receberam o cuidado usual, no pósoperatório imediato. Considerando a escassez de evidências sobre estratégias para manejo da sede do paciente cirúrgico idoso, o picolé mentolado é uma estratégia inovadora e segura, podendo se tornar método de escolha para uso na prática clínica / Physiological mechanisms of detection and control of thirst in the elderly surgical patient occur differently than in adults and children, which justifies the investment in conducting studies on strategies that can provide relief of this symptom with use of cold temperature and small volume. The aim of the study was to compare thirst intensity and discomfort of elderly patients who savoured mentholated popsicle with thirst of elderly patients who received the usual care in the immediate postoperative period. This is a randomized controlled clinical trial in parallel with two groups, conducted in a post anesthesia care unit of a public teaching hospital in southern Brazil. The sample consisted of 50 elderly patients in the immediate postoperative period, who were allocated in two groups, 25 in the control and 25 in the experimental group, without follow-up losses. Inclusion criteria were: patients aged >= 60 years; being fasting; verbalize thirst and having been approved in the assessment of Safety Protocol of Thirst Management. Patients with restrictions to ingestion or swallowing, as well as those who self-referred peppermint allergy, were excluded. Patients assigned to the control group received usual routine care of the institution where the study was conducted, which consists in maintenance of fasting. Patients assigned to the experimental group received a 20 ml mentholated popsicle. Primary outcomes of interest were thirst intensity and discomfort, both of which were assessed initially (T0) and again (T1) after 20 minutes of the intervention or usual care. Thirst intensity was measured using a Numerical Scale (zero means no thirst and 10 the greatest thirst ever experienced by patient), and thirst discomfort by Perioperative Thirst Discomfort Scale (score of zero to 14 points, being zero the absence of discomfort and 14 corresponds to the most intense discomfort related to thirst). Median of initial thirst intensity and discomfort (T0) was 6.0 for the participants assigned to the experimental group, 5.0 for thirst intensity and 6.0 for thirst discomfort for the participants assigned to the control group. Mann-Whitney test was used evaluating differences in medians, comparing changes in Numerical Scale and in Perioperative Thirst Discomfort Scale between the experimental group and the control group (final value minus initial value). Results of the randomized clinical trial demonstrated that there was a statistically significant decrease in thirst intensity (p <0.001) and discomfort (p <0.001) of elderly surgical patients, after twenty minutes of tasting the mentholated popsicle when compared to thirst intensity and discomfort of the patients who received the usual care, in the immediate postoperative period. Considering the shortage of evidence on strategies for thirst management of elderly surgical patient, mentholated popsicle is an innovative and safe strategy, and may become a method of choice for use in clinical practice

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-09-12T11:16:41Z No. of bitstreams: 1 arquivototal.pdf: 71034772 bytes, checksum: 9c21531ff7d7e2de79d846d056ac6485 (MD5) / Made available in DSpace on 2017-09-12T11:16:41Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 71034772 bytes, checksum: 9c21531ff7d7e2de79d846d056ac6485 (MD5) Previous issue date: 2015-02-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The monoterpenes found in essential oils from plants act on transient receptor potential channels (TRP). Some TRP channels with altered expression in hypertensive rats may be new therapeutic targets for the control of hypertension. Aim: Compare the responses induced by Essential Oil of Mentha x villosa Hudson (OEMV), rotundifolone and menthol in Spontaneously Hypertensive rats (SHR) and normotensive Wistar Kyoto (WKY), evaluating the role of TRP channels. Methods and Results: In vivo (blood pressure measurement and heart rate), in vitro (measure of the frequency and force of contraction in the atria and the isometric tension in superior mesenteric arteries) and biochemical (PCR and Western blot) studies were used. The OEMV (3, 5, 10 and 20 mg/kg), the rotundifolone (10, 20 and 30 mg/kg) and the menthol (3, 5, 10 and 20 mg/kg) induced significant hypotensive and bradycardic response in non-anesthetized SHR and WKY rats. The reduction in the diastolic blood pressure was significantly greater than the decrease in the systolic blood pressure, suggesting a greater action on the vascular component of blood pressure. However, the significant bradycardic effect and reduction in the systolic blood pressure also suggest an action on the cardiac component. Furthermore, the decrease in the blood pressure and heart rate induced by rotundifolone and by menthol were significantly more potent in SHR. The action of OEMV, the rotundifolone and menthol in the right atrium (with spontaneous activity) and left (electrically stimulated) showed negative inotropic and chronotropic effects and culminating in complete inhibition of cardiac activity. Moreover, the negative inotropic effect was more potent in SHR and protein TRPM8 channel showed increased expression in the ventricles (left > right) and atria (left > right) of SHR rats. Also, OEMV, rotundifolone and menthol induced vasorelaxant response in superior mesenteric arteries of SHR and WKY rats, precontracted with PHE. The major mechanism involves the endothelium-independent route, which was more potent in SHR. The mechanism of the endothelium-independent vasorelaxant response induced by rotundifolone and menthol probably involves TRPM8 channels, which showed increased expression in SHR, and TRPC1, TRPC3 and TRPC6 channels. However, the response induced by menthol in WKY rats involves other TRP channels (probably TRPM6 and TRPM7). In addition, the flow cytometry showed an increase in [Ca2+]i induced by rotundifolone in SHR vascular myocytes, probably by activating of the TRPM8 channel. Conclusions: The hypotensive, bradycardia, negative inotropic and vasorelaxant responses induced by OEMV, rotundifolone and menthol were significantly more potent in SHR than in WKY rats. The mechanism of the endothelium-independent vasorelaxant response induced by rotundifolone and menthol involves TRPM8, TRPC (probably TRC1, TRPC3 and TRPC6), BKCa and CaV channels, but menthol may be acting in other TRP channels (probably TRPM6 and TRPM7) in WKY rats. The TRPM8 channel showed increased expression in SHR rats. Thus, the action of OEMV, rotundifolone and menthol on these channels can be related with the higher potency observed in SHR rats. / Os monoterpenos presentes em óleos essenciais de plantas atuam sobre canais Potencial Receptor Transiente (TRP). Alguns canais TRP com expressão alterada em ratos hipertensos podem ser novos alvos terapêuticos para o controle da hipertensão arterial. Objetivo: Comparar as respostas induzidas pelo Óleo Essencial de Mentha x-villosa Hudson (OEMV), pela rotundifolona e pelo mentol em Ratos Espontaneamente Hipertensos (SHR) e normotensos Wistar Kyoto (WKY), avaliando o papel de canais TRP. Métodos e Resultados: Estudos in vivo (medida de pressão arterial e freqüência cardíaca), in vitro (medida da freqüência e força de contração em átrios e da tensão isométrica em artérias mesentéricas superiores) e bioquímicos (PCR e Western blot) foram usados. O OEMV (3, 5, 10, 20 mg/kg), a rotundifolona (10, 20 e 30 mg/kg), e o mentol (3, 5, 10 e 20 mg/kg) induziram significativa resposta hipotensora e bradicárdica em ratos SHR e WKY não-anestesiados. A redução na pressão arterial diastólica foi significativamente maior do que a redução na pressão arterial sistólica, sugerindo uma maior ação sobre o componente vascular da pressão arterial. Entretanto, o significativo efeito bradicárdico e a redução na pressão arterial sistólica sugerem também uma ação sobre o componente cardíaco. Além disso, a diminuição na pressão arterial e freqüência cardíaca induzida por rotundifolona e por mentol foram significativamente mais potentes em ratos SHR. A ação do OEMV, da rotundifolona e do mentol em átrios direito (com atividade espontânea) e esquerdo (estimulado eletricamente) mostrou efeitos cronotrópico e inotrópico negativos e culminando na completa inibição da atividade cardíaca. Além disso, o efeito inotrópico negativo foi mais potente em ratos SHR e a proteína do canal TRPM8 mostrou expressão aumentada nos ventrículos (esquerdo > direito) e nos átrios (esquerdo > direito) de ratos SHR. O OEMV, a rotundifolona e o mentol também induziram resposta vasorrelaxante em artérias mesentéricas superiores de ratos SHR e WKY, pré-contraídos com FEN. O mecanismo majoritário envolve a via independente do endotélio, que foi mais potente em ratos SHR. O mecanismo da resposta vasorrelaxante independente do endotélio induzida por rotundifolona e mentol envolve provavelmente canais TRPM8, que apresentaram expressão aumentada em ratos SHR, e canais TRPC1, TRPC3 e TRPC6. Entretanto, a resposta induzida por mentol em ratos WKY envolve outros canais TRP (provavelmente TRPM6 e TRPM7). Além disso, a citometria de fluxo mostrou um aumento na [Ca2+]i induzido por rotundifolona em miócitos vasculares de ratos SHR, provavelmente por ativação de canais TRPM8. Conclusões: As respostas hipotensora, bradicárdica, inotrópica negativa e vasorrelaxante induzidas por OEMV, rotundifolona e mentol foram significativamente mais potentes em ratos SHR do que em ratos WKY. O mecanismo da resposta vasorrelaxante independente de endotélio induzida por rotundifolona e mentol envolve canais TRPM8, TRPC (provavelmente TRPC1, TRPC3 e TRPC6), BKCa e CaV, porém o mentol pode estar atuando em outros canais TRP (provavelmente TRPM6 e TRPM7) em ratos WKY. Os canais TRPM8 mostraram expressão aumentada em ratos SHR. Dessa forma, a ação do OEMV, da rotundifolona e do mentol sobre esses canais pode estar relacionada com a maior potência observada em ratos SHR.

Submitted by (edna.saturno@ufrpe.br) on 2016-12-01T14:41:32Z No. of bitstreams: 1 Flavia de Souza Born.pdf: 858418 bytes, checksum: 5785147d20d21fb9c69b74322ff6448b (MD5) / Made available in DSpace on 2016-12-01T14:41:32Z (GMT). No. of bitstreams: 1 Flavia de Souza Born.pdf: 858418 bytes, checksum: 5785147d20d21fb9c69b74322ff6448b (MD5) Previous issue date: 2012-02-01 / Two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), has been controlled mainly chemical synthetic acaricides. The use of essential oils can be an alternative to this method of control. The aim of this study was to determine the effect of essential oils from plants of Families Burseraceae, Lamiaceae, Rutaceae and Verbenaceae against T. urticae and the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae). Repellent and toxic activities of the oils were determined and compared to eugenol. Toxicity was evaluated by fumigation and residual effect in the laboratory and by direct contact in a greenhouse. The results showed that the oils of Ocimum basilicum L. and Thymus vulgaris L. were the most repellent, with RC50 = 3.43 x 10-7 and 2 x 10-4 μL/mL, respectively. The toxicity against eggs of T. urticae revealed T. vulgaris, Rosmarinus officinalis L., Citrus reticulata Blanco and O. basilicum oils as more active. Estimates of mortality concentration curves for fumigation of adult females of T. urticae revealed that was more toxic eugenol, with LC50 = 4 x 10-3 μL/L air. The test results showed that residual C. reticulata, T. vulgaris and O. basilicum oils present the same toxicity observed for eugenol. The toxicity tests to N. californicus indicated that all the oils are selective fumigation and that the O. basilicum, R. officinalis and C. reticulata oils were more selective by residual effect. The relative toxicity of the individual constituents of the essential oils and their contribuition to the acaricidal activity on T. urticae are also discussed. The highest average mortality on T. urticae in the greenhouse after 72 h of exposure were 100%, 98.85% and 96.59%, promoted by Citrus sinensis Osbek x Citrus reticulata Blanco, R. officinalis and Lippia gracilis Schauer oils, respectively. / O ácaro rajado, Tetranychus urticae Koch (Acari: Tetranychidae), tem sido controlado principalmente com acaricidas químicos sintéticos. O uso de óleos essenciais pode ser uma alternativa a esse método de controle. O objetivo deste trabalho foi determinar o efeito de óleos essenciais de plantas das Famílias Burseraceae, Lamiaceae, Rutaceae e Verbenaceae sobre T. urticae e o ácaro predador Neoseiulus californicus (McGregor) (Acari: Phytoseiidae). As atividades repelente e tóxica dos óleos foram determinadas e comparadas ao eugenol. A toxicidade foi avaliada por fumigação e efeito residual, em laboratório e por contato direto em casa de vegetação. Os resultados demonstraram que os óleos de Ocimum basilicum L. e Thymus vulgaris L. foram mais repelentes, com CR50 de 3,43 x 10-7 e 2 x 10-4 μL/mL, respectivamente. A toxicidade sobre ovos de T. urticae revelou os óleos de T. vulgaris, Rosmarinus officinalis L., Citrus reticulata Blanco e O. basilicum como mais ativos. As estimativas das curvas de concentração mortalidade para fumigação sobre fêmeas adultas de T. urticae revelaram que eugenol foi mais tóxico, com CL50 de 4 x 10-3 μL/L de ar. Os resultados dos testes de efeito residual demonstraram que os óleos de C. reticulata, T. vulgaris e O. basilicum apresentaram a mesma toxicidade observada para o eugenol. Os testes de toxicidade a N. californicus indicaram que todos os óleos foram seletivos por fumigação e os óleos de O. basilicum, R. officinalis e C. reticulata foram os mais seletivos por efeito residual. A toxicidade relativa de constituintes individuais dos óleos e sua contribuição para a atividade sobre T. urticae também são discutidos. As maiores médias de mortalidade sobre T. urticae em casa de vegetação após 72 h de exposição foram de 100%, 98,85% e 96,59%, promovidas pelos óleos de Citrus sinensis Osbek x Citrus reticulata Blanco, R. officinalis e Lippia gracilis Schauer, respectivamente.

Abstract Peppermint (Mentha x piperita L.) oil is one of the most popular and widely used essential oils, mostly because of its main components menthol and menthone. Peppermint oil is used for flavouring pharmaceuticals and oral preparations. Corn mint is the richest source of natural menthol. Carvone-scented mint plants, such as spearmint (M. spicata), are rich in carvone and are widely used as spices, and they are cultivated in several countries. Studies were made into the yield and essential oil content of several mint species and the original. The general aim of the work was to examine the optimal conditions for cultivating mint in Northern Finland. The specific aims of the study were (first) to investigate the differences in the oil content for several mint species and (secondly) to compare the effect of various factors such as plant spacing (10, 20 and 30 × 50 cm), liming (0, 4, 8, 12 and 16 tons ha-1), propagation methods (micropropagated and conventionally propagated plants) and harvest date (once at the end of August in comparison with first cut at the beginning of August and second cut in mid September) on the cultivation success, quality and quantity of the plants. The constituents of the essential oil were analysed from leaf samples using GC-MS. Among the peppermints of different origins studied, peppermint of USA and Egypt origin ('Black Mitcham') contain the highest menthol and optimum oil yield. Corn mint and Sachalin mints both had high menthol content. Due to several reasons, such as no significant differences between the different densities and oil composition, markedly higher amount of weeds at 30 × 50 cm than at 10 × 50 and 20 × 50 cm spacing and the high seedling costs and the danger of fungi and disease at a 10 × 50 cm spacing, a plant optimum of 20 × 50 cm spacing is recommended for Northern Ostrobothnia. If the pH value is lower than 6, or levels of Mg and Ca are low, liming at a rate of 4–8 t ha-1 for sandy soils in Finland is recommended in order to achieve higher fresh and oil yields. In the first year, there were no differences in the dry leaf yield of micropropagated and conventionally propagated plants, but the menthol content was significantly higher in conventionally than in micropropagated plants. In the second year, only the dry leaf yield of micropropagated plants was higher than that of their conventionally propagated counterparts. Cutting peppermint only once during full bloom (the end of August) gives the maximum oil yield of good quality. In conclusion, it is possible to achieve as high as or even higher oil quality and dry yield in North Ostrobothnia than it is in central Europe or south Asia. However, this requires observing certain cultivation factors such as having the right type of mint, soil pH, planting density, harvesting time and propagation method In addition, mints must be cultivated in the same place for only two and a maximum for three years.

Der Hen´s Egg Test for Micronucleus-Induction (HET-MN) wurde bereits mehrfach als ein alternatives, versuchstierfreies Testsystem für in-vivo Mutagenitätstests am Nager unter Verwendung von angebrüteten Hühnereiern vorgestellt (Wolf and Luepke 1997; Wolf, Niehaus-Rolf et al. 2002; Wolf, Niehaus-Rolf et al. 2003; Wolf, Niehaus-Rolf et al. 2008). Während der häufig verwendete in-vivo Micronucleustest am Nager aufgrund seines komplexen in-vivo Stoffwechsel für die Bewertung von Mutagenen sehr gute Ergebnisse liefert (Wolf 2003), können Alternativen zur Testung am Tier, zum Beispiel in-vitro Micronucleustests, die Vorteile der in-vivo Testung nicht erreichen, weshalb sie häufig zu sensitiv sind. Gegenüber herkömmlichen in-vitro Tests stellt die Komplexität des angebrüteten Hühnereis, welches sowohl toxikodynamische als auch toxikokinetische Wirkungen der Prüfsubstanz aufzeigen kann, einen besonderen Vorteil dar, so dass der HET-MN unter anderem nicht nur direkte Mutagene sondern auch Promutagene detektieren kann. Mit Hilfe von zehn ausgewählten Substanzen wurde in dieser Arbeit die Prädiktivität des HET-MN weiter untersucht. Hierzu gehörten, neben den in-vitro und in-vivo negativen Stoffen Ephedrin-HCl und Ampicillin-Natrium und den in-vitro und in-vivo positiven Substanzen para-Chloranilin, Carbendazim und Vinorelbintartrat, auch die fünf so genannten irrelevant positiven Substanzen Isophoron, 2,4-Dichlorphenol, Phthalsäureanhydrid, Menthol und Malathion, für die uneinheitliche in-vivo und in-vitro Daten vorliegen, die aber aufgrund validierter Testverfahren als in-vivo negativ eingestuft werden. Die im HET-MN ermittelten Ergebnisse zeigen für alle zehn Substanzen, im Vergleich zu den gängigen Einstufungen auf Grundlage validierter Tests, eine vollständige Übereinstimmung und damit eine sehr hohe Prädiktivität des HET-MN. Die Ergebnisse stärken daher die Position des HET-MN als einen zuverlässigen in-vitro Test mit besonderen metabolischen Fähigkeiten, der darüber hinaus dazu beitragen kann, so genannte irrelevant positive Substanzen besser einzustufen.

Doctor of Philosophy(PhD) / Two recently identified transient receptor potential (TRP) channels, TRPM8 and TRPA1, have been proposed to play an important role in mammalian cool and cold peripheral sensory transduction. When expressed in cell-lines the cloned TRPM8 and TRPA1 receptors have distinct pharmacological and temperature response characteristics. Although these receptors are also transported to the central terminals of primary afferents, little is known about their centrally mediated actions. In this thesis, I use an in vitro electrophysiological approach to investigate the dorsal horn processing of cool afferent modalities and the role of TRP ion channels. The results of this thesis provide further information on thermal processing, indicate direction for further research and suggest possible therapeutic targets for the management of abnormal cold sensory processing. Initial experiments demonstrate that the cooling agents and known TRPM8 and TRPA1 agonists, menthol and icilin, inhibit primary afferent evoked excitatory postsynaptic currents (EPSCs) in rat spinal cord dorsal horn neurons. In addition, temperature reduction, menthol and icilin increase the frequency of miniature EPSCs without affecting amplitude distribution or kinetics. Little or no direct postsynaptic effect on dorsal horn neurons, GABAergic or glycinergic transmission was found. In combination, these observations demonstrate that temperature reduction, menthol and icilin act presynaptically to increase the probability of glutamate release from primary afferent fibres. Further examination of the changes in glutamatergic synaptic transmission induced by temperature reduction, menthol and icilin reveals a subset of neurons sensitive to innocuous cool (< 29 oC) and low concentrations of icilin (3-10 µM) which closely match the temperature activation and pharmacological profile of TRPM8. In addition, the majority of lamina I and II neurons displayed characteristics partly consistent with TRPA1-activation, including a concentration-dependent response to icilin and blockade by ruthenium red. The present experiments did not allow thermal characterisation of these TRPA1-like responses. Together these observations indicate that the effects of menthol and icilin on glutamatergic synaptic transmission in the superficial dorsal horn are mediated by TRPM8 and possibly by TRPA1. Examination of the anatomical location of neurons activated by temperature reduction, menthol, icilin and capsaicin allowed the central termination pattern of thermoreceptive primary afferent fibres with specific TRP-like response characteristics to be determined. TRPM8-like presynaptic activation was confined to a subpopulation of neurons located in lamina I and outer lamina II, while the majority of neurons throughout laminae I and II received inputs sensitive to menthol, high concentrations of icilin and capsaicin. These findings suggest that innocuous cool sensation projects to a specific subpopulation of superficial dorsal horn neurons unlike other modalities (mediated by TRPV1, possibly TRPA1 and other receptors), which non-selectively engage circuits within the entire superficial dorsal horn. No morphological specificity was identified for recovered neurons after electrophysiological characterisation. Finally, mu-opioids were shown to inhibit basal glutamatergic synaptic transmission as well as menthol- and icilin-induced transmission in the superficial dorsal horn. Of particular interest, delta-opioids selectively inhibited icilin-induced synaptic transmission within the same location. The selective effect of delta-opioids suggests a possible role in modulating receptors activated by icilin (TRPM8 and TRPA1). Overall, this thesis provides further evidence that TRPM8 is responsible for the transduction of innocuous cold sensation in mammals and is a potential therapeutic target in humans with cold hyperaesthesia secondary to abnormal thermal processing. The use of delta-opioid agonists warrants further investigation in cold hypersensitivity states and potentially other forms of pain.

Doctor of Philosophy(PhD) / Two recently identified transient receptor potential (TRP) channels, TRPM8 and TRPA1, have been proposed to play an important role in mammalian cool and cold peripheral sensory transduction. When expressed in cell-lines the cloned TRPM8 and TRPA1 receptors have distinct pharmacological and temperature response characteristics. Although these receptors are also transported to the central terminals of primary afferents, little is known about their centrally mediated actions. In this thesis, I use an in vitro electrophysiological approach to investigate the dorsal horn processing of cool afferent modalities and the role of TRP ion channels. The results of this thesis provide further information on thermal processing, indicate direction for further research and suggest possible therapeutic targets for the management of abnormal cold sensory processing. Initial experiments demonstrate that the cooling agents and known TRPM8 and TRPA1 agonists, menthol and icilin, inhibit primary afferent evoked excitatory postsynaptic currents (EPSCs) in rat spinal cord dorsal horn neurons. In addition, temperature reduction, menthol and icilin increase the frequency of miniature EPSCs without affecting amplitude distribution or kinetics. Little or no direct postsynaptic effect on dorsal horn neurons, GABAergic or glycinergic transmission was found. In combination, these observations demonstrate that temperature reduction, menthol and icilin act presynaptically to increase the probability of glutamate release from primary afferent fibres. Further examination of the changes in glutamatergic synaptic transmission induced by temperature reduction, menthol and icilin reveals a subset of neurons sensitive to innocuous cool (< 29 oC) and low concentrations of icilin (3-10 µM) which closely match the temperature activation and pharmacological profile of TRPM8. In addition, the majority of lamina I and II neurons displayed characteristics partly consistent with TRPA1-activation, including a concentration-dependent response to icilin and blockade by ruthenium red. The present experiments did not allow thermal characterisation of these TRPA1-like responses. Together these observations indicate that the effects of menthol and icilin on glutamatergic synaptic transmission in the superficial dorsal horn are mediated by TRPM8 and possibly by TRPA1. Examination of the anatomical location of neurons activated by temperature reduction, menthol, icilin and capsaicin allowed the central termination pattern of thermoreceptive primary afferent fibres with specific TRP-like response characteristics to be determined. TRPM8-like presynaptic activation was confined to a subpopulation of neurons located in lamina I and outer lamina II, while the majority of neurons throughout laminae I and II received inputs sensitive to menthol, high concentrations of icilin and capsaicin. These findings suggest that innocuous cool sensation projects to a specific subpopulation of superficial dorsal horn neurons unlike other modalities (mediated by TRPV1, possibly TRPA1 and other receptors), which non-selectively engage circuits within the entire superficial dorsal horn. No morphological specificity was identified for recovered neurons after electrophysiological characterisation. Finally, mu-opioids were shown to inhibit basal glutamatergic synaptic transmission as well as menthol- and icilin-induced transmission in the superficial dorsal horn. Of particular interest, delta-opioids selectively inhibited icilin-induced synaptic transmission within the same location. The selective effect of delta-opioids suggests a possible role in modulating receptors activated by icilin (TRPM8 and TRPA1). Overall, this thesis provides further evidence that TRPM8 is responsible for the transduction of innocuous cold sensation in mammals and is a potential therapeutic target in humans with cold hyperaesthesia secondary to abnormal thermal processing. The use of delta-opioid agonists warrants further investigation in cold hypersensitivity states and potentially other forms of pain.