Gastric ulcer is a disease that affects a large part of the world population, with a prevalence of 2.4% in the Western population and with annual incidence rates of 0.10% to 0.19%, with a high cure rate, but with a high recurrence rate. The essential oil (OE) of Lippia gracilis Schauer (Verbenaceae) is popularly used in the treatment of various disorders and presents antinociceptive, anti-inflammatory, antioxidant and antimicrobial activities, but its gastroprotective effect has not been studied yet. The animals were treated with OE at doses of 20, 60 and 180 mg/kg). The acute ethanol/HCl induced ulcer model was used to evaluate the gastroprotective effect of OE and to investigate some possible mechanisms involved. The animals were pretreated with yohimbine -adrenergic antagonist), glibenclamide (KATP channel blocker), L-NAME (NO synthesis inhibitor), NEM (sulfhydryl grouping blocker), indomethacin (nonsteroidal anti-inflammatory) And all were treated with OE (180 mg/kg). The pylorus ligature model was used to evaluate the effect of OE on mucus secretion and gastric acidity. And the action of OE on the activity of H +, K +-ATPase evaluated by in vitro assay. We observed that OE dose-dependently prevented lesions caused by ethanol/HCl, with decreased edema and loss of epithelial cells. The gastroprotective effect of OE was partially diminuided by pre-treatments with LNAME, glibenclamide, indomethacin and yohimbine, and total with NEM. In the pylori ligation model OE increased mucus production, it was able to increase the pH of the gastric juice, decrease the volume of secretion and [H +]. OE inhibited the activity of H +, K + -ATPase. These results confirm that Lippia gracilis OE has a gastroprotective effect, decreasing gastric secretion and stimulating protective factors, w -adrenergic receptors, prostaglandins and KATP channels. / A úlcera gástrica é uma doença que acomete grande parcela da população mundial, com prevalência de 2,4% na população ocidental e com taxas de incidência anuais de 0,10% a 0,19%, com alta taxa de cura, porém com elevado índice de recorrência. O óleo essencial (OE) da Lippia gracilis Schauer (Verbenaceae) é utilizado popularmente no tratamento de diversos distúrbios e apresenta atividades antinociceptiva, anti-inflamatória, antioxidante e antimicrobiana, mas seu efeito gastroprotetor ainda não foi estudado. Os animais foram tratados com OE nas doses de 20,60 e 180 mg/kg). Foi utilizado o modelo de úlceras agudas induzidas por etanol/HCl para avaliar o efeito gastroprotetor do OE e investigar alguns possíveis mecanismos envolvidos. Os animais foram pré- - adrenérgico), glibenclamida (bloqueador dos canais KATP), L-NAME (inibidor da síntese de NO), NEM (bloqueador de grupamento sulfidrílicos), indometacina (antiinflamatório não esteroidal) e todos foram tratados com OE (180mg/kg). O modelo de ligadura de piloro foi utilizado para avaliar efeito do OE sobre a secreção de muco e acidez gástrica. E a ação do OE sobre a atividade da H+, K+-ATPase avaliada por ensaio in vitro. Observamos que o OE preveniu de forma dose-dependente as lesões provocadas pelo etanol/HCl, com diminuição de edema e perdas de células epiteliais. O efeito gastroprotetor do OE foi diminuido de forma parcial pelos prétratamentos com L-NAME, glibenclamida, indometacina e ioimbina, e total com NEM. No modelo de ligadura de piloro o OE aumentou a produção de muco, foi capaz de aumentar o pH do suco gástrico, diminuir o volume de secreção e a concentração de H+. O OE inibiu a atividade da H+, K+-ATPase. Estes resultados comprova que o OE de Lippia gracilis possui efeito gastroprotetor, diminuindo a secreção gástrica e estimulando fatores protetores, com participação de NO, grupamentos sulfidrílicos, receptores -adrenérgicos, prostaglandinas e canais KATP.

Os ciclos de vida de Clytia linearis (Thornely, 1899) e de duas espécies apresentando caracteres considerados diagnósticos de Clytia gracilis (M. Sars, 1850) – aqui denominadas Clytia cf. gracilis sp. 1 e Clytia cf. gracilis sp. 2 – foram estudados com base em espécimes coletados no infralitoral raso da costa de São Sebastião e Ilhabela, sudeste do Brasil, entre fevereiro de 1999 e abril de 2000. Medusas foram cultivadas em laboratório, a temperatura de 22-24oC. Colônias de C. linearis são monossifônicas, simpodiais, com até 21,5mm de altura e portando até 26 hidrantes e 10 gonângios. Medusas adultas, alcançando 2,5-3,6mm de diâmetro e até 29 tentáculos e 28 estatocistos, podem ser distinguidas de outras espécies de Clytia pela presença de nematocistos microbásicos mastigóforos do tipo C. Medusas adultas de Clytia cf. gracilis spp. 1 e 2 podem ser distinguidas das demais espécies do gênero estudadas até o momento pela presença de uma fileira de nematocistos microbásicos mastigóforos do tipo A na umbrela, no nível do canal circular. Medusas adultas de C. cf. gracilis sp. 1 e C. cf. gracilis sp. 2 apresentam até 16 tentáculos e podem ser distinguidas entre si pelo diâmetro da umbrela: 6,6-10,1mm e 3,6-5,5mm, respectivamente. Quanto ao estágio de pólipo, C. cf. gracilis sp. 1 apresenta usualmente colônias dicotômicas eretas, hidrotecas alongadas, e gonotecas na hidrorriza e pedículos. Estes caracteres concordam com a descrição de C. gracilis, mas as espécies diferem entre si pela morfometria das gonotecas e dos nematocistos microbásicos mastigóforos do tipo B: aproximadamente 15mm de comprimento para C. gracilis e 9-10mm para C. cf. gracilis sp. 1. Por outro lado, nematocistos do tipo B de C. cf. gracilis sp. 2, com aproximadamente 14,5mm de comprimento, em média, são morfometricamente semelhantes aos de C. gracilis. No entanto, C. cf. gracilis sp. 2 difere de C. gracilis pela forma da hidroteca, por apresentar gonotecas apenas na hidrorriza, e pelo hábito polissifônico do colônias bem desenvolvidas. Uma terceira espécie, C. cf. gracilis sp. 3, é descrita com base em uma colônia sem gonângios. Aspectos da sistemática de Clytia são discutidos. / he life-cycles of Clytia linearis (Thornely, 1899) and two species with characters considered diagnostic of Clytia gracilis (M. Sars, 1850) – Clytia cf. gracilis sp. 1 and Clytia cf. gracilis sp. 2 – have been studied based on specimens collected in the shallow subtidal coast of São Sebastião and Ilhabela, southeast Brazil, between February 1999 and April 2000. Medusae were cultured in the laboratory (22-24oC). Colonies of C. linearis are monosiphonic, sympodial, up to 21.5mm high and bearing up to 26 hydranths and 10 gonangia. Adult medusae reached 2.5-3.6mm in diameter, and up to 29 tentacles and 28 statocysts. The presence of microbasic mastigophore type C nematocysts distinguishes adult medusae of C. linearis from other species of Clytia. A band of microbasic mastigophore type A nematocysts in the umbrella, at the level of the circular canal, distinguishes adult medusae of Clytia cf. gracilis spp. 1 and 2 from other species of the genus. Adult medusae of C. cf. gracilis sp. 1 and C. cf. gracilis sp. 2 have up to 16 tentacles, and can be distinguished by the diameter of the umbrella: 6.6-10.1mm and 3.6-5.5mm, respectively. Colonies of C. cf. gracilis sp. 1 are usually erect and dichotomous, the hydrothecae are elongated and the gonothecae present in the hydrorhiza and pedicels. These features closely match with the description of C. gracilis, but both species differ in the morphometry of the gonothecae and microbasic mastigophore type B nematocysts: about 15mm (length) for C. gracilis and 9-10mm for C. cf. gracilis sp. 1. Type B nematocysts of C. cf. gracilis sp. 2 (about 14.5mm in length) are more similar in size to those of C. gracilis. However, the former species differs from the latter in the shape of the hydrothecae, by having gonothecae only at the hydrorhiza and polysiphonic well-developed colonies. A third species, C. cf. gracilis sp. 3, is described based on an unfertile colony. Aspects of the systematics of Clytia are discussed.

Diplomová práce popisuje fyzikální principy mikroskopů TEM, SEM a STEM spolu s jejich vhodností pro pozorování vzorků citlivých na elektrony, jako jsou zalévací média či biologické vzorky. Dále je popsána příprava vzorků pro STEM (TEM) a popis interakcí, ke kterým dochází mezi primárním elektronovým svazkem a vzorkem. Součástí práce je také pojednání o zpracování mikroskopických obrazů s podkapitolou o metodách kvantitativního porovnání obrazů ze STEM. Praktická část práce je zaměřena zejména na měření úbytku hmoty zalévacích médií (Epon, Spurr, LR White) způsobený primárním svazkem elektronů v nízkonapěťovém STEM. Ultratenké řezy několika tlouštěk byly zkoumány při různých nastaveních mikroskopu (urychlovací napětí, celková dávka, proud svazku, čištění povrchu vzorku a komory mikroskopu) a zobrazovacích módech (světlé a tmavé pole). Dále jsou zkoumány také biologické vzorky Krásnoočka štíhlého (Euglena gracilis) zalitého v pryskyřicích Epon a Spurr. Nasbírané snímky, vytvořené algoritmy a získané výsledky jsou diskutovány a zhodnoceny.

Die lichtaktivierte Kinase Phototropin aus Chlamydomonas reinhardtii, die photoaktivierte Adenylatcyclase (PAC) aus Euglena gracilis und das BLUF-Protein Slr1694 aus Synechocystis sp. PCC 6803 wurden in Hinblick auf die molekularen Details der primären photochemischen Prozesse sowie der Signalweiterleitung untersucht. Phototropin wurde mit Hilfe von Arginin aus Escherichia coli in Milligramm Mengen isoliert. Ohne Arginin wurde E. coli cAMP Rezeptorprotein assoziiert aufgefunden, welches eine hohe Homologie zu einer cAMP aktivierten Kinase aus C. reinhardtii besitzt. Volllängen Phototropin bildet wie einzelne LOV-Domänenkonstrukte ohne Kinasedomäne den Flavin-Triplettzustand und das kovalente Cysteinyl-Addukt. Der Zerfall des Signalzustandes ist in Anwesenheit von ATP beschleunigt und deutet auf Photorezeptor-Kinase Interaktion hin. Strukturelle Änderungen in der Kinasedomäne wurden durch FTIR-Differenzspektroskopie gezeigt. Über ELDOR-Spektroskopie wurde der Abstand der Photorezeptordomänen auf etwa 25 Angstrom bestimmt. Mutationen in Slr1694 an S28, N31 und W91 zeigten keine konservierten Einfluss auf die Dynamik des Signalzustands. Die Entfernung der Seitenkette von S28 führte zu einer 15 nm Rotverschiebung des Absorptionsspektrums aufgrund veränderter Wasserstoffbrückenkoordination des Kofaktors. Die Einführung von positiv geladenen Seitenketten an Stelle von N31 erhöhte die Kofaktorbindung von phosphorylierten Flavinen. Künstliche Kofaktoren wie Roseoflavin konnten in Slr1694 durch Koexpression eines prokaryotischen Flavintransporters erreicht werden. Die Rolle von M152 in PAC für die Signalweiterleitung wurde anhand der lichtaktivierten cAMP Synthese-Aktivität gezeigt. Durch ultraschnelle IR-Spektroskopie wurde die Beteiligung der Seitenketten von Y8 sowie Q50 bestätigt und eine genauere Beschreibung der Wasserstoffbrücken im langlebigen Signalzustand ermöglicht. / The light activated kinase Phototropin from Chlamydomonas reinhardtii, the photoactivated adenylylcyclase (PAC) from Euglena gracilis and the BLUF protein Slr1694 from Synechocystis sp. PCC 6803 were investigated concerning the molecular details of the primary photochemistry as well as signal transduction. Phototropin was isolated from Escherichia coli in mg amounts after solubilization with arginine. Without arginine E. coli cAMP receptor protein, which shows high homology to a cAMP activated kinase from C. reinhardtii, was copurified. Full length Phototropin shows similar photochemistry to LOV-domain containing proteins without the kinase including triplet and covalent cysteinyl adduct formation. Signaling state decay is accelerated in the presence of ATP and suggests photoreceptor-kinase interaction. FTIR spectroscopy showed light induced structural changes in the kinase domain. The distance of the photoreceptor domains of 25 Angstrom was determined by ELDOR spectroscopy. Mutation of the side chains of S28, N31 and W91 in Slr1694 showed no conserved influence on the dynamic of the signaling state. Removal of the hydroxyl group of S28 lead to a 15 nm red shift of the absorption spectrum as a result of altered hydrogen bond coordination of the cofactor. Introduction of positively charged side chains at the position of N31 strengthened the binding of phosphorylated flavins. An artificial flavin like roseoflavin was introduced in Slr1694 by coexpression of a bacterial flavin transporter. The essential role of M152 in PAC for signal transduction was shown by determination of light activated cAMP synthesis activity. Ultrafast IR spectroscopy confirmed the contribution of Y8 and Q50 in the photocycle and gave a more detailed description of the hydrogen bonding situation in the signaling state.

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.

Musculoskeletal injuries are commonly associated with muscle atrophy as a function of immobilization or change of normal function. For example, injuries to the anterior cruciate ligament (ACL) which may involve ligament reconstruction, results in the "quadriceps avoidance" gait which leads to atrophy of the knee extensormuscles. In these situations it is not clear whether or not the atrophy is associated with loss of specific muscle fibre types with accompanying functional deficits. Such knowledge would be helpful in implementing exercise regimes designed to compensate for loss of particular fibre types. It is believed that isokinetic exercise performed at speeds below 180° per second strengthens type I muscle fibres, and type II fibres at fast speeds. However, there is no evidence to indicate the specific muscle fibre response to different rates of muscle contraction. Identification of muscle fibre type is most directly determined by biopsy technique but is too invasive for a routine measurement. Electromyography median frequency has been used as a non-invasive measure to infer muscle fibre composition in various studies. However, the reliability and accuracy of this technique has been questioned and improvement is necessary. This research was designed to provide a more accurate and reliable protocol for the determination of EMG median frequency which may be used, after validation against more direct biopsy techniques, as a routine method for inferring muscle fibre composition. The investigation also explored the muscular response as measured by EMG median frequency to varying speeds of muscle contraction, fatiguing exercise and atrophy following ACL reconstruction. The ultimate aim of this research was to improve the reliability of the determination of EMG median frequency to enhance its application as a predictor of muscle fibre composition. This provides information which may improve ACL rehabilitation programs designed to restore and prevent specific muscle fibre types loss that have not previously been targeted by current rehabilitation programs. This research was conducted in three studies. Study one determined the stability of the EMG median frequency bilaterally for the quadriceps and hamstrings muscles and identified the mode of contraction associated with the greatest reliability. The strength and EMG median frequency of the vastus lateralis, medial hamstrings and vastus medialis of 55 subjects was determined across 5 speeds from 0° to 240° per second using a Kin-Com isokinetic dynamometer and an EMG data acquisition system. Isometric contraction was found to have the least bilateral discrepancy (4.01% ±3.06) and between trials standard deviation (4.50) in the vastus lateralis, medial hamstrings and vastus medialis. Study two investigated the EMG median frequency changes in the vastus lateralis which occur immediately following different speeds of isokinetic exercise to the point of fatigue in normal subjects. Thirty-four subjects participated in the study, and performed a 90-second period of isokinetic exercise to activate the knee extensors at either 30° or 300° per second. EMG median frequency of the vastus lateralis was determined before, immediately after and 7 minutes after the fatiguing exercise. The percentage drop in EMG median frequency of the vastus medialis was gnificantly (p<0.05) greater after slow speed (27.9%) than fast speed (20.25%) exercise, while no significant difference was found for the percentage drop in extension torque. Full recovery was found 7 minutes after the fatiguing exercise. By reference to previous research showing a relationship between EMG median frequency and muscle fibre type, an increase in activation of type I muscle fibres with slow speed exercise and an increase in type II muscle fibres with fast speed exercise was observed. Study three identified the changes in EMG median frequency following ACL reconstruction and evaluated the bilateral differences in EMG median frequency of the knee muscles. The relationships between EMG median frequency and the measures of knee functional ability, knee muscle strength, age and time since surgery were also investigated. Twelve subjects who had undergone ACL reconstruction using a semitendinosus and gracilis graft 6 to 12 months earlier, participated in the study. EMG median frequency was determined from an 8-second isometric contraction and knee functional ability was assessed using the Cincinnati Rating Scale. Bilateral EMG median frequency shifts were inconsistent among subjects. On the basis of previous research which indicated a relationship between EMG median frequency and fibre type, no consistent pattern of muscular fibre type atrophy subsequent to ACL reconstruction occurred within 6 to 12 months (ranged from -43 to 57 Hz). Additionally, no significant correlations were found between the EMG median frequency and the knee functional score and knee extension and flexion torques, age, time since operation and the bilateral differences in EMG median frequency. The results of this investigation will serve to improve the reliability of EMG median frequency across a range of conditions in which it has been evaluated. Further research is needed to confirm the relationship between EMG median frequency and direct observations of muscle fibre composition to improve the predictive value of this measure. Following this validation it will be possible to evaluate the bilateral EMG median frequency shift to infer the type of muscle fibre atrophy, and use this measure in determining the efficacy of specific rehabilitation programs. In conclusion * An 8-second isometric contraction is recommended for determining EMG median frequency. * EMG median frequency of a muscle decreases significantly more after slow fatiguing exercise than after fast speed fatiguing exercise. * There was no generalised bilateral EMG median frequency shift found in a group of subjects 6 to 12 months following semitendinosus and gracilis graft ACL reconstruction. * The results of this study will serve to improve the reliability of procedures used to determine EMG median frequency under a range of different contractile conditions. The EMG median frequency changes in response to these conditions require further validations with muscle biopsy in future.

This dissertation analyses the effects of current fisheries practices and management regulations of the green spiny lobster (Panulirus gracilis) in the Eastern Tropical Pacific Region (ETP). P. gracilis has reached a critical state in the ETP. Country-based studies report that between 60 and 98% of lobsters caught in the wild are under the minimum landing size (MLS). This means that spiny lobsters are being extracted before reproducing and contributing to the replenishment of interconnected populations. The recovery of green spiny lobster populations in the ETP and the future maintenance of a sustainable fishery will depend on effective management decisions and on taking in account environmental factors that influence the population dynamics of the lobsters. In the first study (Chapter 2), the B52 Spiny Lobster individual based simulation model was used for conducting a population viability analysis to quantify the effect of current fishing practices and the effect of varying management regulations on minimum landing size (MLS) and fishing effort. The best suit of regulations to maintain the highest abundance, production of offspring and catch is to protect juveniles and egged females, and to establish a MLS that assures the reproduction of individuals before being extracted. This study revealed regional variations, however the patterns and the causes for variation were not yet clear. This led to the next chapters in this dissertation. In Chapter 3, I used a meta-analysis to explore regional lobster variability by comparing published studies from the ETP. The objective was to identify patterns of variation related to geographic and environmental factors of the region that can inform the establishment and evaluation of coordinated regulations. Morphological relationships showed to be more variable at northern latitudes, where the mean annual sea surface temperatures are higher than at lower temperatures at the Equator. In terms of management, MLS regulations should be adapted accounting for the effect of sea surface temperature and its variation. Additionally, it was observed that monitoring methodologies are not standardized within the region and even in some cases, neither within countries. Furthermore, in most places monitoring of the spiny lobster fishery happens sporadically, only in Galapagos takes place every year. Identifying patterns of variations can improve the accuracy of prediction models which can help to explore, design, and apply more effective management measures, as well as promote regional coordination to support the recovery and maintenance of spiny lobsters. In Chapter 4, I contrast current Ecuadorian minimum landing size (MLS) regulation to lobster empirical measurements within Ecuador in order to recognize potential pitfalls for management enforcement. I used linear regression and multiple regression models with the objective of identifying potential relative size variations of the individuals caught in the wild over time and in the different fishing areas in Ecuador, as well as to analyse the effect of locality, sex, age and mean SST on the tail length/total length ratio. Morphological relations were significantly different among sexes, in time and by all sites. Most importantly, this study shows that current minimum size regulations are not applicable to all sites. Additionally, I found that water temperature has a significant effect on morphological relationship variations. However, it was not the main site-specific variable responsible for explaining such variations. In general, this work emphasizes the need for length data collection standardization and the consideration of temporal and spatial variation implications in national and regional fishery management planning, enforcement and evaluation.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Lippia gracilis Schauer is a plant that belongs to the Verbenaceae family, is known as “alecrim-da-chapada” or “alecrim-de-tabuleiro”, and occure in the caatinga biome. Because of its bioactive potential, the essential oil of L. gracilis is been tested for control of different agriculture and livestock pests and diseases. One of these pests is the coconut mite, Aceria guerreronis (Acari: Eriophyidae), an important pest of the coconut crop in Brazil. The objective of this study was to evaluate the toxicity and repellency of the essential oil of two genotypes of L. gracilis and their major compounds thymol and carvacrol against the coconut mite (A. guerreronis). Adults of the coconut mite were subjected to increasing concentrations of the essential oils of the genotypes LGRA-106 and LGRA-109, of thymol and carvacrol, allowing the estimation of the lethal concentrations (LC). In addition to toxicity, the repellency of essential oils of L. gracilis, thymol and carvacrol was evaluated by releasing mites in the center of an arena with half area sprayed and the other half unsprayed. The analysis of the chemical composition of the essential oils of L. gracilis identified thymol (52.41%) and carvacrol (49.36%) as the major compounds for genotypes LGRA-106 and LGRA-109, respectively. The genotype LGRA-106 showed higher toxicity (LC50 = 4.73 μL/mL) to the coconut mite compared to the genotype LGRA-109 (LC50 = 30.99 μL/mL). The toxicity of LGRA-106 did not differ from its major compound thymol (LC50 = 5.34 μL/mL). The essential oil of LGRA-106 was 2.3 times more toxic than its major compound thymol at LC90. The LC50 and LC99 of genotypes LGRA-106 (4.73 and 13.18 μL/mL), LGRA-109 (30.99 and 54.33 μL/mL) and carvacrol (6.84 and 45.97 μL/mL) repelled the coconut mite after 24 h of exposure. The LC50 and LC99 of thymol (5.34 and 56.35 μL/mL) presented a neutral effect on the coconut mite after 24 h of exposure. We can conclude that the essential oil of genotype LGRA-106 has potential for development of a product to control the coconut mite. / Lippia gracilis Schauer é uma planta pertencente à família Verbenaceae popularmente conhecida como “alecrim-da-chapada” ou “alecrim-de-tabuleiro” e própria do bioma caatinga. Devido ao potencial bioativo, o óleo essencial de L. gracilis vem sendo testado para o controle de diferentes pragas e doenças na agricultura e pecuária. Uma destas pragas é o ácaro-da-necrose Aceria guerreronis (Acari: Eriophyidae), importante praga da cultura do coqueiro no Brasil. O objetivo deste trabalho foi avaliar a toxicidade e repelência do óleo essencial de dois genótipos de L. gracilis e seus compostos majoritários timol e carvacrol ao ácaro-da-necrose (A. guerreronis). Adultos do ácaro-da-necrose foram submetidos a concentrações crescentes dos óleos essenciais dos genótipos LGRA-106 e LGRA-109 de L. gracilis, de timol e carvacrol, possibilitando a estimativa das concentrações letais (CL). Adicionalmente à toxicidade, foi avaliada a repelência dos óleos essenciais de L. gracilis, timol e carvacrol por meio da liberação de ácaros no centro de uma arena com metade pulverizada e metade não pulverizada. A análise da composição química dos óleos essenciais de L. gracilis identificou o timol (52,41%) e o carvacrol (49,36%) como compostos majoritários dos genótipos LGRA-106 e LGRA-109, respectivamente. O genótipo LGRA-106 apresentou maior toxicidade (CL50 = 4,73 μL/mL) ao ácaro-da-necrose em comparação com LGRA-109 (CL50 = 30,99 μL/mL). A toxicidade do LGRA-106 não diferiu do seu composto majoritário timol (CL50 = 5,34 μL/mL). O óleo essencial do LGRA-106 foi 2,3 vezes mais tóxico que do seu composto majoritário timol para CL90. A CL50 e a CL99 dos genótipos LGRA-106 (4,73 e 13,18 μL/mL-1), LGRA-109 (30,99 e 54,33 μL/mL-1) e de carvacrol (6,84 e 45,97 μL/mL-1), repeliram o ácaro-da-necrose após 24 h de exposição. A CL50 e CL99 de timol (5,34 e 56,35 μL/mL) apresentaram efeito neutro sobre o ácaro-da-necrose após 24 h de exposição. Conclui-se que o óleo essencial do genótipo LGRA-106 apresenta potencial para desenvolvimento de um produto para o controle do ácaro-da-necrose.

This dissertation analyses the effects of current fisheries practices and management regulations of the green spiny lobster (Panulirus gracilis) in the Eastern Tropical Pacific Region (ETP). P. gracilis has reached a critical state in the ETP. Country-based studies report that between 60 and 98% of lobsters caught in the wild are under the minimum landing size (MLS). This means that spiny lobsters are being extracted before reproducing and contributing to the replenishment of interconnected populations. The recovery of green spiny lobster populations in the ETP and the future maintenance of a sustainable fishery will depend on effective management decisions and on taking in account environmental factors that influence the population dynamics of the lobsters. In the first study (Chapter 2), the B52 Spiny Lobster individual based simulation model was used for conducting a population viability analysis to quantify the effect of current fishing practices and the effect of varying management regulations on minimum landing size (MLS) and fishing effort. The best suit of regulations to maintain the highest abundance, production of offspring and catch is to protect juveniles and egged females, and to establish a MLS that assures the reproduction of individuals before being extracted. This study revealed regional variations, however the patterns and the causes for variation were not yet clear. This led to the next chapters in this dissertation. In Chapter 3, I used a meta-analysis to explore regional lobster variability by comparing published studies from the ETP. The objective was to identify patterns of variation related to geographic and environmental factors of the region that can inform the establishment and evaluation of coordinated regulations. Morphological relationships showed to be more variable at northern latitudes, where the mean annual sea surface temperatures are higher than at lower temperatures at the Equator. In terms of management, MLS regulations should be adapted accounting for the effect of sea surface temperature and its variation. Additionally, it was observed that monitoring methodologies are not standardized within the region and even in some cases, neither within countries. Furthermore, in most places monitoring of the spiny lobster fishery happens sporadically, only in Galapagos takes place every year. Identifying patterns of variations can improve the accuracy of prediction models which can help to explore, design, and apply more effective management measures, as well as promote regional coordination to support the recovery and maintenance of spiny lobsters. In Chapter 4, I contrast current Ecuadorian minimum landing size (MLS) regulation to lobster empirical measurements within Ecuador in order to recognize potential pitfalls for management enforcement. I used linear regression and multiple regression models with the objective of identifying potential relative size variations of the individuals caught in the wild over time and in the different fishing areas in Ecuador, as well as to analyse the effect of locality, sex, age and mean SST on the tail length/total length ratio. Morphological relations were significantly different among sexes, in time and by all sites. Most importantly, this study shows that current minimum size regulations are not applicable to all sites. Additionally, I found that water temperature has a significant effect on morphological relationship variations. However, it was not the main site-specific variable responsible for explaining such variations. In general, this work emphasizes the need for length data collection standardization and the consideration of temporal and spatial variation implications in national and regional fishery management planning, enforcement and evaluation.