As formas de defesa encontradas na natureza são incrivelmente diversas e envolvem estratégias que minimizam as chances de encontro com o predador ou as chances de escape diante de um ataque. A liberação de substâncias químicas é uma forma de defesa amplamente difundida entre artrópodes e vários estudos já demonstraram sua eficiência contra o ataque de predadores. Apesar de trazerem benefícios óbvios em termos de defesa, a produção de vários desses compostos químicos deve ser custosa para as presas. Esta tese teve como objetivo geral explorar os custos da produção de defesas químicas em um grupo particular de artrópodes, os opiliões. No capítulo 1, investigamos como a produção de ovos pode comprometer a produção de defesas químicas (benzoquinonas) e quais as conseqüências da redução do volume de secreção liberada sobre a capacidade de defesa das fêmeas de Acutisoma longipes. Nossos resultados apontam que a quantidade de secreção produzida por fêmeas ovígeras é quase 50% inferior à das fêmeas não-ovígeras. Como conseqüência, a secreção liberada por fêmeas ovígeras é menos eficiente em deter formigas e aranhas do que a secreção liberada por fêmeas não-ovígeras. No capítulo 2, investigamos como a quantidade e a qualidade da dieta influenciam a produção de defesas químicas (benzoquinonas) em Magnispina neptunus. Demonstramos que a produção de defesas químicas é condição dependente, pois indivíduos bem alimentados produziram mais secreção que indivíduos mal alimentados. Entretanto, indivíduos que receberam aporte extra dos precursores da secreção não incrementaram nem a quantidade total nem a concentração da secreção. Por fim, o capítulo 3 teve como objetivo investigar possíveis demandas conflitantes (trade-offs) entre o esforço de acasalamento (produção de armamentos) e o esforço somático (defesas químicas) de machos e fêmeas em um clado de opiliões neotropicais. Usando uma abordagem comparativa, mostramos que fêmeas produzem mais defesas químicas (benzoquinonas) que machos, mas não há uma relação negativa entre o dimorfismo sexual morfológico e o dimorfismo na quantidade de defesas químicas produzidas por machos e fêmeas. Coletivamente, os resultados obtidos aqui apontam que a produção de defesas químicas em opiliões é custosa e que está sujeita a demandas conflitantes com outros componentes de aptidão. / The forms of animal defenses found in nature are incredibly diverse and comprise many strategies that minimize the changes of a prey being detected or attacked by a predator. The emission of chemical secretions is a widespread defense among arthropods, and several studies have already demonstrated the efficiency of chemical defenses against predation. Although chemical defenses confer obvious survival benefits, the production of deterrent compounds may be costly for the individuals. The main goal of this thesis was to evaluate the costs of producing chemical defenses in a particular arthropod group, the harvestmen. In chapter 1 we investigated how egg production may compromise the production of chemical defenses (benzoquinones) by females of Acutisoma longipes, and whether a reduction in the amount of secretions released by ovigerous females make then more vulnerable to predation. Our results indicate that ovigerous females produce almost 50% less secretion than non-ovigerous females. Moreover, the low amount of secretion released by ovigerous females is less efficient than the amount released by non−ovigerous females in deterring ants and spiders. In chapter 2 we investigated how the quantity and quality of food influence the production of chemical defenses (benzoquinones) in Magnispina neptunus. We demonstrate that the production of chemical defenses is clearly condition dependent because well fed individuals produced more secretion than poorly fed individuals. However, individuals that received in the diet an input of the benzoquinones\' precursor did not show an increase in the amount and concentration of secretion released. Finally, in chapter 3 we investigated possible trade-offs between mating effort (i.e., investment in weaponry) and somatic effort (i.e., investment in chemical secretions) in males and females of several harvestman species belonging to the family Gonyleptidae. Using a comparative approach, we showed that females consistently produce more secretion than males, but there is no negative relationship between morphological sexual dimorphism and sexual dimorphism in the amount of secretion released by males and females. Taken together, our findings indicate that the production of chemical defenses in harvestmen is costly and is under allocation trade-offs with other fitness components.

Este trabalho apresenta o primeiro estudo dos estados aniônicos de baixa energia da para-Benzoquinona (p-BQ) por meio de cálculos de espalhamento elástico de elétrons pela molécula neutra. Foi utilizado o método multicanal de Schwinger implementado com pseudopotenciais de Bachelet, Hamann e Schlüter, na aproximação de núcleos fixos, e nas aproximações estático-troca e estático-troca mais polarização. As seções de choque integrais foram calculadas em duas geometrias do alvo, otimizadas para o estado fundamental da molécula neutra e do ânion. Os estados aniônicos encontrados foram aracterizados e comparados com resultados experimentais, realizados com a técnica de espectroscopia de transferência eletrônica (ETS). Três estados * foram observados, sendo um ligado e dois metaestáveis, em acordo com os experimentos. Uma ressonância de Feshbach foi encontrada nos cálculos, tal qual é observada experimentalmente e sugerida em diversos estudos teóricos de estado ligado. Acredita-se que a aproximação utilizada para descrever estados excitados do alvo poderá ser reconsiderada, em que é sugerida uma proximação mais sofisticada, a ser utilizada em cálculos futuros. Com este estudo, é possível uma melhor compreensão dos resultados experimentais ETS, assim como é permitida uma interpretação de estudos teóricos sobre os estados aniônicos da p-BQ. Ademais, este trabalho possibilita próximos estudos de espalhamento envolvendo quinonas mais complexas. / This work present the first study of low-energy anionic states in para-benzoquinone (p-BQ) by electron scattering calculations. The elastic scattering calculations were done in the fixed-nuclei approximation, with the Schwinger multichannel method implemented with pseudopotentials of Bachelet, Hamann and Schlüter, in the static-exchange and in the static-exchange plus polarization approximations. The integral cross sections were performed in two target geometries optimized for the neutral and anion ground states. The anionic states found were characterized and compared with experimental results obtained by electron transfer spectroscopy (ETS). Three * states were observed, a bound state and two metastable, in good agreement with experimental results. A Feshbach resonance was found in calculations, also it is observed experimentally and it was suggested in several theoretical studies of bound state. It is believed that the approach used to describe the target excited states can be reconsidered, as it is suggested a more sophisticated approach to be used in future calculations. With this study, a better understanding of experimental results ETS is possible, as it allowed an interpretation of theoretical studies of the anionic states of p-BQ. Moreover, this work enables further scattering studies involving more complex quinones.

No description available.

Este trabalho apresenta o primeiro estudo dos estados aniônicos de baixa energia da para-Benzoquinona (p-BQ) por meio de cálculos de espalhamento elástico de elétrons pela molécula neutra. Foi utilizado o método multicanal de Schwinger implementado com pseudopotenciais de Bachelet, Hamann e Schlüter, na aproximação de núcleos fixos, e nas aproximações estático-troca e estático-troca mais polarização. As seções de choque integrais foram calculadas em duas geometrias do alvo, otimizadas para o estado fundamental da molécula neutra e do ânion. Os estados aniônicos encontrados foram aracterizados e comparados com resultados experimentais, realizados com a técnica de espectroscopia de transferência eletrônica (ETS). Três estados * foram observados, sendo um ligado e dois metaestáveis, em acordo com os experimentos. Uma ressonância de Feshbach foi encontrada nos cálculos, tal qual é observada experimentalmente e sugerida em diversos estudos teóricos de estado ligado. Acredita-se que a aproximação utilizada para descrever estados excitados do alvo poderá ser reconsiderada, em que é sugerida uma proximação mais sofisticada, a ser utilizada em cálculos futuros. Com este estudo, é possível uma melhor compreensão dos resultados experimentais ETS, assim como é permitida uma interpretação de estudos teóricos sobre os estados aniônicos da p-BQ. Ademais, este trabalho possibilita próximos estudos de espalhamento envolvendo quinonas mais complexas. / This work present the first study of low-energy anionic states in para-benzoquinone (p-BQ) by electron scattering calculations. The elastic scattering calculations were done in the fixed-nuclei approximation, with the Schwinger multichannel method implemented with pseudopotentials of Bachelet, Hamann and Schlüter, in the static-exchange and in the static-exchange plus polarization approximations. The integral cross sections were performed in two target geometries optimized for the neutral and anion ground states. The anionic states found were characterized and compared with experimental results obtained by electron transfer spectroscopy (ETS). Three * states were observed, a bound state and two metastable, in good agreement with experimental results. A Feshbach resonance was found in calculations, also it is observed experimentally and it was suggested in several theoretical studies of bound state. It is believed that the approach used to describe the target excited states can be reconsidered, as it is suggested a more sophisticated approach to be used in future calculations. With this study, a better understanding of experimental results ETS is possible, as it allowed an interpretation of theoretical studies of the anionic states of p-BQ. Moreover, this work enables further scattering studies involving more complex quinones.

As formas de defesa encontradas na natureza são incrivelmente diversas e envolvem estratégias que minimizam as chances de encontro com o predador ou as chances de escape diante de um ataque. A liberação de substâncias químicas é uma forma de defesa amplamente difundida entre artrópodes e vários estudos já demonstraram sua eficiência contra o ataque de predadores. Apesar de trazerem benefícios óbvios em termos de defesa, a produção de vários desses compostos químicos deve ser custosa para as presas. Esta tese teve como objetivo geral explorar os custos da produção de defesas químicas em um grupo particular de artrópodes, os opiliões. No capítulo 1, investigamos como a produção de ovos pode comprometer a produção de defesas químicas (benzoquinonas) e quais as conseqüências da redução do volume de secreção liberada sobre a capacidade de defesa das fêmeas de Acutisoma longipes. Nossos resultados apontam que a quantidade de secreção produzida por fêmeas ovígeras é quase 50% inferior à das fêmeas não-ovígeras. Como conseqüência, a secreção liberada por fêmeas ovígeras é menos eficiente em deter formigas e aranhas do que a secreção liberada por fêmeas não-ovígeras. No capítulo 2, investigamos como a quantidade e a qualidade da dieta influenciam a produção de defesas químicas (benzoquinonas) em Magnispina neptunus. Demonstramos que a produção de defesas químicas é condição dependente, pois indivíduos bem alimentados produziram mais secreção que indivíduos mal alimentados. Entretanto, indivíduos que receberam aporte extra dos precursores da secreção não incrementaram nem a quantidade total nem a concentração da secreção. Por fim, o capítulo 3 teve como objetivo investigar possíveis demandas conflitantes (trade-offs) entre o esforço de acasalamento (produção de armamentos) e o esforço somático (defesas químicas) de machos e fêmeas em um clado de opiliões neotropicais. Usando uma abordagem comparativa, mostramos que fêmeas produzem mais defesas químicas (benzoquinonas) que machos, mas não há uma relação negativa entre o dimorfismo sexual morfológico e o dimorfismo na quantidade de defesas químicas produzidas por machos e fêmeas. Coletivamente, os resultados obtidos aqui apontam que a produção de defesas químicas em opiliões é custosa e que está sujeita a demandas conflitantes com outros componentes de aptidão. / The forms of animal defenses found in nature are incredibly diverse and comprise many strategies that minimize the changes of a prey being detected or attacked by a predator. The emission of chemical secretions is a widespread defense among arthropods, and several studies have already demonstrated the efficiency of chemical defenses against predation. Although chemical defenses confer obvious survival benefits, the production of deterrent compounds may be costly for the individuals. The main goal of this thesis was to evaluate the costs of producing chemical defenses in a particular arthropod group, the harvestmen. In chapter 1 we investigated how egg production may compromise the production of chemical defenses (benzoquinones) by females of Acutisoma longipes, and whether a reduction in the amount of secretions released by ovigerous females make then more vulnerable to predation. Our results indicate that ovigerous females produce almost 50% less secretion than non-ovigerous females. Moreover, the low amount of secretion released by ovigerous females is less efficient than the amount released by non−ovigerous females in deterring ants and spiders. In chapter 2 we investigated how the quantity and quality of food influence the production of chemical defenses (benzoquinones) in Magnispina neptunus. We demonstrate that the production of chemical defenses is clearly condition dependent because well fed individuals produced more secretion than poorly fed individuals. However, individuals that received in the diet an input of the benzoquinones\' precursor did not show an increase in the amount and concentration of secretion released. Finally, in chapter 3 we investigated possible trade-offs between mating effort (i.e., investment in weaponry) and somatic effort (i.e., investment in chemical secretions) in males and females of several harvestman species belonging to the family Gonyleptidae. Using a comparative approach, we showed that females consistently produce more secretion than males, but there is no negative relationship between morphological sexual dimorphism and sexual dimorphism in the amount of secretion released by males and females. Taken together, our findings indicate that the production of chemical defenses in harvestmen is costly and is under allocation trade-offs with other fitness components.

Pentachlorophenol, (PCP) is an organochlorine compound which was first developed in the 1930’s. PCP is said to be the most toxic of the chlorophenols and is classified as a hazardous substance and a probable human carcinogen. PCP has proven to be cytotoxic to a number of cell lines translating to its effect on various organs. The aim of the study was to assess organochlorine-induced hepatotoxicity in a mechanistic manner using an in-house developed procedure. Also, the possible hepatoprotective effect of methanolic extracts of the bark of two medicinal plants, Burkea africana (BA) and Syzygium cordatum (SC), as well as the known hepatoprotective agent, N-acetyl cysteine (NAC), were investigated. In addition to PCP, two of its major metabolites, tetrachloro-1,2-hydroquinone (TCHQ) and tetrachloro-1,4-benzoquinone (TCBQ) were also evaluated. A hepatocarcinoma cell line (HepG2) was used to investigate the effect of these compounds on different parameters of cellular function. Cytotoxicity was assessed using the neutral red uptake assay. Cytochrome P4501A1 (CYP1A1) activity was determined using ethoxy-resorufin-O-deethylation as surrogate. Generation of reactive oxygen species (ROS) was investigated by measuring dichlorofluorescein diacetate cleavage. Effects on mitochondrial membrane potential were determined using JC-1 staining, whilst necrosis was investigated by assessing plasma membrane integrity using propidium iodide (PI)staining. The degree of apoptotic death was determined by quantifying caspase-3 activity. Assays were repeated with an additional 1 h pre-treatment of the cells with either NAC, SC or BA in order to investigate whether these compounds were able to protect against the toxicity induced by PCP and its metabolites. The IC50 values of PCP, TCHQ and TCBQ were 68.0, 144.0 and 129.4 μM, respectively. All three test compounds induced CYP1A1 activity with PCP being the most potent. TCBQ produced extensive ROS generation. TCHQ also induced ROS generation, whilst PCP appeared to have no significant effect on ROS generation. All test compounds caused mitochondrial depolarization. None of the test compounds caused an increase in necrotic cell death. PCP, TCHQ and TCBQ had negligible effects on apoptosis. Both SC and BA alleviated the toxic effects observed in cells treated with PCP. Minor increases in viability occurred in cells pre-treated with plant extracts prior to exposure to both metabolites. NAC, as well as both plant extracts, greatly reduced CYP 1A1 activity induced by PCP. NAC, SC and BA exacerbated CYP1A1 induction in cells exposed to concentrations of TCBQ and TCHQ that initially produced little or no effect on CYP1A1 activity. Contrarily, decreased CYP1A1 activity was observed in cells exposed to concentrations of TCBQ and TCHQ where extensive induction of CYP1A1 occurred. NAC, as well as both plant extracts, suppressed ROS generation in cells exposed to all test compounds. In cells exposed to PCP and TCBQ more extensive mitochondrial depolarization was seen when pre-treated with NAC and plant extracts than when exposed to the compounds alone. Negligible effects were seen in pre-treated cells exposed to TCHQ. BA and SC caused increases in necrotic death in cells exposed to the test compounds. NAC, BA and SC had negligible effects on the changes in caspase-3 activity induced by the test compounds. From the results it is proposed that PCP induces its own metabolism by increasing CYP1A1 activity. It also causes mitochondrial insult which could lead to the opening of the mitochondrial permeability transition pore and subsequent release of cytochrome C, activation of caspases and eventually apoptotic cell death. With regard to TCHQ and TCBQ, results suggest that extensive ROS generation caused damage to various cellular macromolecules and that this could be the main cause of their toxicity. NAC, SC and BA appeared to alleviate toxicity in certain instances. Further investigation is required in order to assess them as possible hepatoprotective agents. Copyright / Dissertation (MSc)--University of Pretoria, 2011. / Pharmacology / unrestricted

Methionine sulfoxide reductase A (MSRA) and MSRB are responsible for the repairing of methionine-R-sulfoxide (Met-S-SO) and methionine-S-sulfoxide (Met-R-SO) back to me-thionine, respectively. Five MSRA isoforms and four MSRB isoforms are discovered in the unicellular green alga Chlamydomonas reinhardtii. Whether high light regulates CrMSR ex-pression via photosynthetic electron transport (PET) was examined. By checking the se-quence of PCR product of each isoform, quantitative real-time primers were designed for discrimination of isoform expression. Light ≥ 300 £gE m-2 s-1 and PET inhibitors inhibited PSII activity (Fv/Fm, Fv´/Fm´) and photosynthetic O2 evolution rate, particularly 1,000 £gE m-2 s-1, in which it did not recover after 3 h. A transfer to dark decreased CrMSRA2, CrMSRA3, CrMSRB1.1, CrMSRB1.2, CrMSRB2.1 mRNA levels but increased CrMSRA4 mRNA levels. When exposed to 50, 300, 600, or 1,000 £gE m-2 s-1, CrMSRA2, CrMSRA3, CrMSRA5, CrMSRB1.1, CrMSRB2.1 and CrMSRB2.2 mRNA levels increased as light ≥ 300 £gE m-2 s-1, and concomitantly CrMSRA4 mRNA level decreased. Changes in mRNA levels increased as light intensity increased. The treatment of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) in 1,000 £gE m-2 s-1 inhibited high light effect, and the treatment of 2,5-dibromo-3-methyl-6- isopropyl-p- benzoquinone (DBMIB) in 50 £gE m-2 s-1 increased CrMSRA3, CrMSRA5 and CrMSRB2.2 mRNA levels but decreased CrMSRA4 mRNA level. The application of phena-zine methosulfate (PMS), an electron donor to P700+ that promotes cyclic electron transport, in 300 £gE m-2 s-1 enhanced the increase of CrMSRA3 and CrMSRA5 mRNA levels by high light but inhibited the decrease of CrMSRA4 mRNA level, reflecting a role of cyclic PET. The above results let us to draw a conclusion that plastoquinone as reduced status mediates the expression of CrMSRA3, CrMSRA4, CrMSRA5 and CrMSRB2.2 by high light. The im-plication of linear electron transport and cyclic electron transport on the regulation of CrMSR gene expression will be discussed.We speculated that the high light up-regulation of CrMSR mRNA expression offers the resistance of Chlamydomonas to photooxidative stress.