Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2018-07-25T13:41:21Z No. of bitstreams: 2 Tese - Aline de Arvelos Salgado - 2018.pdf: 8292746 bytes, checksum: b28ed506e404b8c0b4373382d667712a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-07-26T10:41:59Z (GMT) No. of bitstreams: 2 Tese - Aline de Arvelos Salgado - 2018.pdf: 8292746 bytes, checksum: b28ed506e404b8c0b4373382d667712a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-07-26T10:41:59Z (GMT). No. of bitstreams: 2 Tese - Aline de Arvelos Salgado - 2018.pdf: 8292746 bytes, checksum: b28ed506e404b8c0b4373382d667712a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-06-22 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Shallow reservoirs, with large surface area and long hydraulic retention times, present conditions conducive to the development of cyanobacteria. Due to the potential risk that cyanobacteria can cause to public health, the importance of the management of water bodies to control the toxic blooms, especially in reservoirs destined to the public supply, is evident. Thus, there is a clear need to develop techniques that can help existing monitoring, with the possibility of extending the area of ​​coverage in monitoring programs. Thus, the present study aimed to combine several methods of data acquisition, mainly obtaining continuously, such as the use of sensors for in situ evaluation of phytoplankton, especially cyanobacteria, through the detection of chlorophyll-a and phycocyanin, as well as the use of remote sensing for phytoplankton monitoring and with this information to build water quality prediction scenarios through mathematical modeling. The procedure aimed to improve data acquisition, especially the spatialization of data. For this purpose, the dynamics and water quality of the João Leite reservoir was characterised over 30 months using physical, chemical and biological parameters and trophic status index, to know the seasonality of the quality of the of the water body and present the main parameters and factors of water quality change. A continuous and spatial phytoplankton monitoring method was proposed for mapping concentrations of chlorophyll- a and cyanobacteria from Sentinel-2 satellite images with in situ and inferential observations to evaluate the performance of these to provide information on cyanobacterial blooms. The CE-QUAL-W2 model was calibrated from field data to evaluate the water quality dynamics and hydrodynamics of the reservoir. As a result, João Leite reservoir presented characteristic and similar patterns of the variation of the parameters in hydrological periods, and the results showed that the use and the occupation of the soil reflect in the characteristics of the quality of the water. In the majority of the evaluated points and times, good oxygenation of the waters and low concentrations of nutrients were demonstrated, being proved by the trophic state indexes. Both chlorophyll-a and cyanobacteria showed correlation between data measured in situ and estimated. The results can considered satisfactory since with the data of reflectance it was possible to validate the atmospheric correction, and the maps of estimate of chlorophyll- a and cyanobacteria presented the places and the months more prone to the greater proliferation of cyanobacteria. Thus, these results demonstrate the importance of the use of remote sensing for the monitoring of water quality, since with the use of this tool there is greater possibility of point coverage, being able to have a synoptic visualisation of the studied environment, identifying the most subject areas to the proliferation of cyanobacteria. The results of the modeling indicate that the reservoir presents periodic stratification, and in the warmer months the stratification is more evident and that in the colder months a mixture of the water column is noticed, possibly due to in those months there are more incidental winds in the region. In general, the highest values ​​of algae and cyanobacteria occur in the warmer months, where there is a higher solar incidence and larger periods of thermal stratification occur. It was also presented that the amount of algae and cyanobacteria on the surface changes the amount of oxygen dissolved in the surface region while the demand for sediment oxygen may be associated with both the oxygen consumption in the sediments and can also be used as a way to simulate the consumption of oxygen by the submerged organic matter, which is visible in the shallower areas of the reservoir. The results of the CE-QUAL-W2 model provided a comprehensive understanding of the spatial and temporal dynamics of the parameters analyzed during the modeling periods. The application of this model with the integration of fluorometers and remote sensing for the evaluation of water quality, should be better suited to water resource managers, to help them better understand the dynamics of physicochemical and biological processes and to be used as a warning system in cases of cyanobacteria blooms and to propose the best mitigation strategies. / Reservatório de água lênticos, rasos, com grande área de superfície e longos tempos de retenção hidráulica apresentam condições propícias para o desenvolvimento de cianobactérias. Devido o potencial risco que cianobactérias podem causar a saúde pública, evidencia-se a importância do gerenciamento dos corpos hídricos a fim de controlar as florações tóxicas, principalmente em reservatórios destinados ao abastecimento público. Assim, há uma clara necessidade de desenvolver técnicas que possam auxiliar os monitoramentos existentes, com a possibilidade de estender a área de abrangência em programas de monitoramento. O objetivo geral dessa pesquisa é avaliar um método de monitoramento contínuo e espacial de fitoplâncton, especialmente quanto à presença de cianobactérias, por meio da ficocianina e clorofila “a”, de modo a se ter informações de qualidade da água, para efetuar a previsão de cenários da dinâmica de cianobactérias, empregando modelagem matemática. Para tanto, foi caracterizada a dinâmica e a qualidade das águas do reservatório João Leite, ao longo de 30 meses, por meio de análises dos parâmetros físicos, químicos e biológicos, e índice de estado trófico, a fim de conhecer a sazonalidade da qualidade da água do corpo hídrico e apresentar os principais parâmetros e fatores de alteração da qualidade das águas. Foi proposto um método de monitoramento contínuo e espacial de fitoplâncton, para confecção de mapas de concentrações de clorofila “a” e cianobactérias a partir de imagens do satélite Sentinel-2, com observações in situ e a partir de inferências, para avaliar o desempenho desses em fornecer informações sobre proliferações de cianobactérias. Por último, o modelo CE-QUAL-W2 foi calibrado a partir de dados obtidos em campo, para avaliar a dinâmica da qualidade da água e hidrodinâmica do reservatório. Como resultado, o reservatório João Leite apresentou padrões característicos e semelhantes da variação dos parâmetros em períodos hidrológicos e os resultados demonstraram que o uso e a ocupação do solo refletem nas características da qualidade da água. Na maior parte dos pontos e tempos avaliados, foram apresentadas boa oxigenação das águas e baixas concentrações de nutrientes, sendo comprovados pelos índices de estado trófico. Tanto a clorofila “a” quanto cianobactérias demonstraram correlações entre os dados medidos in situ e os estimados. Os resultados podem ser considerados satisfatórios, uma vez que com os dados de refletância foi possível validar a correção atmosférica e os mapas de estimativa de clorofila “a” e cianobactérias apresentaram os locais e os meses mais propensos à maior proliferação de cianobactérias. Assim, esses resultados demonstram a importância do uso do sensoriamento remoto para o monitoramento da qualidade das águas, uma vez que com o uso dessa ferramenta há maior possibilidade de abrangência de pontos, podendo ter uma visualização sinóptica do ambiente estudado, identificando as áreas mais sujeitas à proliferação de cianobactérias. Os resultados da modelagem indicam que o reservatório tem a tendência a se comportar com estratificação periódicas, sendo que nos meses mais quentes a estratificação é mais evidente e que nos meses mais frios uma mistura da coluna d’água é notada, possivelmente em função de que nesses meses há mais ventos incidentes na região. De um modo geral, os maiores valores de algas e cianobactérias ocorrem nos meses mais quentes, em que há maior incidência solar e ocorre maiores períodos de estratificação térmica. Foi ainda apresentado que a quantidade de algas e cianobactérias na superfície altera a quantidade de oxigênio dissolvido na região da superfície enquanto a demanda por oxigênio sedimentar, pode estar associado tanto ao consumo de oxigênio nos sedimentos quanto pode ser também usada como forma de simular o consumo de oxigênio pela matéria orgânica submersa, que é visível nas áreas mais rasas do reservatório. Dessa forma, foi apresentado que os resultados do modelo CE-QUAL-W2 forneceram uma compreensão abrangente da dinâmica espacial e temporal dos parâmetros analisados durante os períodos modelados. A aplicação deste modelo com a integração do uso de fluorômetros e sensoriamento remoto para a avaliação da qualidade da água, devem ser mais indicados aos gestores de recursos hídricos, para auxiliá-los a entender melhor a dinâmica dos processos físico- químicos e biológicos e principalmente para serem utilizados como um sistema de alertas em casos de florações de cianobactérias, para que possam propor as melhores estratégias de mitigação.

C-ficocianina (C-PC) é um pigmento solúvel em água e está presente em Arthrospira platensis. Este pigmento é constituído por duas subunidades, &#945 e &#946, com massas moleculares de 16 e 17 kDa, respectivamente. Recentemente foi demonstrado que a subunidade &#946 da C-PC de Anabaena tem atividade antitumoral. Os principais objetivos deste estudo foram clonar e expressar a subunidade &#946 de C-PC de A. platensis em amostras de Escherichia coli e verificar a capacidade da proteína recombinante e da proteína C-PC em induzir ou não apoptose em células tumorais (HEp-2) e não tumorais (fibroblastos). As células foram tratadas com 100 µg das proteínas recombinante e C-PC por 6 h. Após tratamento, as células foram coradas com azul de toluidina (Metódo: Concentração Crítica de Eletrólitos - CCE). As células em apoptose foram detectadas pela coloração e por alterações morfológicas. Os resultados obtidos demonstram que ambas as proteínas, &#946-PC recombinante e C-PC, são capazes de induzir apotose em células HEp-2 e não induzi-la em células fibroblásticas. / C-phycocyanin (C-PC) is a water soluble pigment and is present in Arthrospira platensis. It is consisted of two subunits, &#945 and &#946, with molecular masses of 16 and 17 kDa, respectively. Recently it was shown that the &#946 subunit of C-PC Anabaena has antitumor activity. The main objectives of this study were cloning and expression the &#946 subunit of C-PC (&#946-PC) of A. platensis in Escherichia coli and check the ability of the C-PC and recombinant proteins in apoptotic induction in cancer (HEp-2) and non-cancer (fibroblasts) cells. The fibroblasts and HEp-2 cells were treated with 100 µg of recombinant and C-PC, proteins respectively for 6 h. After treatment, the cells were stained with toluidine blue (Critical Electrolyte Concentration Method - CEC). Apoptosis cells were detected by staining and morphological changes. The results show that the recombinant &#946-PC and C-PC proteins are able to induce apotosis in HEp-2 cells and dont induce it in fibroblasts cells.

Indiana University-Purdue University Indianapolis (IUPUI) / This thesis presents the results of an analysis of predicting phytoplankton pigment concentrations (chlorophyll a and phycocyanin) from remotely sensed imagery. Hyperspectral airborne and hand-held reflectance spectra were acquired on three reservoirs (Geist, Morse and Eagle Creek) in Central Indiana, USA. Concurrent with the reflectance acquisition, in situ samples were collected and analyzed in laboratories to quantify the pigment concentration and other water quality parameters. The resultant concentration was then linked to Airborne Imaging Spectrometer for Applications (AISA) reflectance spectra for the sampling stations to develop predictive models. AISA reflectance spectra were extracted from the imagery which had been processed for radiometric calibration and geometric correction. Several previously published algorithms were examined for the estimation of pigment concentration from the spectra. High coefficients of determination were achieved for predicting chlorophyll a in two of the three reservoirs (Geist R2 = 0.712, Morse R2 = 0.895 and Eagle Creek Reservoir R2 = 0.392). This situation was similar for PC prediction, where two of the three reservoirs had high coefficients of determination between pigment concentration and reflectance (Geist R2 = 0.805, Morse R2 = 0.878 and Eagle Creek Reservoir R2 = 0.316). The results of this study show that reflectance spectra collected with an airborne hyperspectral imager are statistically significant, p < 0.03, in predicting chlorophyll a and phycocyanin pigment concentration in all three reservoirs in this study without the consideration of other parameters. The algorithms were then applied to the AISA image to generate high spatial resolution (1 m2) maps of Chlorophyll a and Phycocyanin distribution for each reservoir.

Indiana University-Purdue University Indianapolis (IUPUI) / This thesis applies several different remote sensing techniques to data collected from 2005 to 2007 on central Indiana reservoirs to determine the best performing algorithms in estimating the cyanobacterial pigments chlorophyll a and phycocyanin. This thesis is a set of three scientific papers either in press or review at the time this thesis is published. The first paper describes using a curve fitting model as a novel approach to estimating cyanobacterial pigments from field spectra. The second paper compares the previous method with additional methods, band ratio and semi-empirical algorithms, commonly used in remote sensing. The third paper describes using a partial least squares (PLS) method as a novel approach to estimate cyanobacterial pigments from field spectra. While the three papers had different methodologies and cannot be directly compared, the results from all three studies suggest that no type of algorithm greatly outperformed another in estimating chlorophyll a on central Indiana reservoirs. However, algorithms that account for increased complexity, such as the stepwise regression band ratio (also known as 3-band tuning), curve fitting, and PLS, were able to predict phycocyanin with greater confidence.

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Additionally, cyclosporin A, an inhibitor of the mitochondrial permeability transition (MPT) pore, failed to prevent hIAPP-induced DeltaPsim collapse, cytochrome c and AIF release and caspase-3 activation, indicating that the MPT pore was not involved in hIAPP-induced apoptosis. On the other hand, potential crosstalk between the extrinsic and intrinsic apoptotic pathways was demonstrated by cleavage of Bid by caspase-8 in the apoptotic process triggered by hIAPP. / It is widely accepted that human islet amyloid polypeptide (hIAPP) aggregation plays an important role in the loss of insulin-producing pancreatic beta cells. Insulin secretion impairment and cell apoptosis can be due to mitochondrial dysfunction in pancreatic beta cells. hIAPP-induced cytotoxicity is mediated by the generation of reactive oxygen species (ROS). Phycocyanin (PC) is a natural compound from blue-green algae that is widely used as food supplement. Currently, little information is available about the effect of hIAPP on mitochondrial function of beta cells and protection of PC against hIAPP-induced cytotoxicity. In this thesis, I hypothesize that hIAPP may impair beta cell function with the involvement of mitochrondrial dysfunction, and this effects could be attenuated by PC. Therefore, the aim of this study was to investigate the role of mitochondria in hIAPP-induced apoptosis, the in vitro protective effects of PC and explore the underlying mechanisms. / It was found that hIAPP induced apoptosis in INS-1E cells with the disruption of mitochondrial function, as evidenced by ATP depletion, mitochondrial mass reduction, mitochondrial fragmentation and loss of mitochondrial membrane potential (DeltaPsim). Further molecular analysis showed that hIAPP induced changes in the expression of Bcl-2 family members, release of cytochrome c and apoptosis-inducing factor (AIF) from mitochondria into cytosol, activation of caspases and cleavage of poly (ADP-ribose) polymerase. Interestingly, the hIAPP-induced mitochondrial dysfunction in INS1-E cells was effectively restored by co-treatment with PC. / Our results showed that hIAPP inhibited the INS-1E cell growth in a dose-dependent manner. However, cytotoxicity of hIAPP was significantly attenuated by co-incubation of the cells with PC. hIAPP induced DNA fragmentation and chromatin condensation, which were key characteristics of cell apoptosis. These changes were inhibited by PC as examined by TUNEL assay and DAPI staining. Moreover, PC significantly prevented the hIAPP-induced overproduction of intracellular ROS and malonaldehyde (MDA), as well as changes of activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) enzymes. Furthermore, hIAPP triggered the activation of mitogen-activated protein kinases (MAPKs) such as c-Jun N-terminal kinase (JNK) and p38 kinase, and these effects were effectively suppressed by PC. / Taken together, I have demonstrated for the first time the involvement of mitochondrial dysfunction in hIAPP-induced INS-1E cell apoptosis, which was attenuated by PC through attenuating oxidative stress, modulating JNK and p38 pathways and reducing mitochondrial dysfunction. / Li, Xiaoling. / Adviser: Juliana Chung Ngor Chan. / Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 150-159). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Le stress oxydatif joue un rôle prépondérant en tant que messager secondaire dans la régulation de nombreux processus cellulaires tels que l’apoptose, la survie et la prolifération et serait impliqué dans l’ensemble des étapes de la carcinogenèse pulmonaire. L’activité physique et la nutrition sont deux facteurs pouvant moduler le stress oxydatif et les mécanismes associés. La bétaïne et la C-Phycocyanine sont deux micronutriments reconnus pour avoir des effets antioxydants, anti-inflammatoires et antiprolifératifs. Récemment notre équipe a montré in vitro qu’un traitement en bétaïne et/ou la C-phycocyanine diminuait la viabilité des cellules A549 (carcinome pulmonaire). L’objectif général de ce travail de thèse était d’évaluer l’effet de facteurs nutritionnels (bétaïne, C-PC ou activité physique) sur la croissance tumorale de cellules A549 implantées chez le rat Nude et de déterminer les mécanismes sous-jacents. Dans un premier temps, nous avons étudié l’effet d’une supplémentation nutritionnelle (bétaïne ou C-phycocyanine) associée ou non à la pratique d’une activité physique volontaire (roue d’activité) sur l’équilibre redox et l’inflammation, chez des rats sains. Nous avons montré que la bétaïne et la C-phycocyanine augmentaient les défenses antioxydantes tandis que l’activité physique volontaire n’avait pas d’effet si elle n’était pas couplée à une supplémentation. Nous avons également mis en évidence que C-phycocyanine inhibait l’augmentation de Cox-2 musculaire induite par l’activité physique. Dans un second temps, nous avons étudié l’effet de la bétaïne et/ou la C-Phycocyanine sur la croissance des cellules A549 implantées chez des rats Nude. Nous avons montré que ces deux micronutriments associés ou non ralentissaient la croissance des tumeurs pulmonaires, au travers de mécanismes communs (activation de NF-kappaB, augmentation de la peroxydation lipidique et de l’expression de cytokine pro-inflammatoire (IL-1-beta, Cox-2 et TNF-alpha) au sein de la tumeur) et de mécanismes propres à chaque micronutriment. La C-phycocyanine a induit une diminution du ratio AKT phosphorylé / AKT total et une augmentation du ratio p38 phosphorylé / p38 total, mécanisme en faveur de l’apoptose et de l’autophagie. La bétaïne associée à la C-phycocyanine a augmenté le ratio caspase-3 / pro-caspase-3. Dans un dernier temps, nous avons évalué l’effet de l’activité physique volontaire sur la croissance tumorale des cellules A549 implantées chez des rats Nude. Nous avons mis en évidence que l’activité physique volontaire ralentissait la croissance des tumeurs pulmonaires induites, sans différence significative avec la bétaïne et/ou la C-phycocyanine. Il apparait que l’augmentation de la peroxydation lipidique, l’activation de la MAPK p38 et de NF-kappaB, et l’inhibition d’AKT, favorisant la mort cellulaire soient impliquées dans cette diminution tumorale. Un régime enrichi en bétaïne et/ou C-phycocyanine ralentit la croissance cellulaire d’adénocarcinome pulmonaire implanté chez le rat, suggérant leur intérêt dans l’action anti-carcinogène pulmonaire. L’activité physique semble jouer sur les mêmes mécanismes. Nos résultats méritent d’être confirmés par des protocoles à plus large échelle et suggèrent de possibles applications chez des patients porteurs de tumeurs pulmonaires. / Oxidative stress seems to play a crucial role as a secondary messenger in the regulation of several cellular processes such as apoptosis, survival and proliferation, and could be involved in all steps of the lung carcinogenesis (i.e. initiation, promotion and progression). Physical activity and nutrition are two factors able to modulate oxidative stress and associated mechanisms. Betaine and C-phycocyanin are two known micronutrients having antioxidant, anti-inflammatory and anti-proliferative effects. Previously, our team showed that betaine and/or C-phycocyanin treatment decreased the viability of A549 cells in vitro (pulmonary adenocarcinoma cell line). The main objective of this work was to evaluate the effect of nutritional factors (betaine, C-phycocyanin) or physical activity on growth of implanted A549 cells in Nude rats and to determine underlying mechanisms.Firstly, we studied the effect of nutritional supplementation (betaine or C-phycocyanin) combined or not with voluntary physical activity (wheel running) on redox balance and inflammation in healthy rats. We showed that betaine and C-phycocyanin increased antioxidant defenses, whereas voluntary physical activity did not have an effect when it was not associated with micronutrient supplementation. We also observed that C-phycocyanin inhibited physical activity-induced muscle Cox-2 activity increase.Secondly, we studied the effect of betaine and/or C-phycocyanin on growth of implanted A549 cells in Nude rats. We showed that these two micronutrients, whether associated or supplied separately, slowed down the lung tumour growth through similar mechanisms (NF-kappaB activation and increase of lipid peroxidation and expression of pro-inflammatory cytokines (IL-1beta, Cox-2 et TNF-alpha) in tumour). Also, some mechanisms were specific for each micronutrient or their combination. C-phycocyanin induced a decrease of phosphorylated AKT / total AKT ratio, and an increase of phosphorylated p38 / total p38 ratio, both mechanisms promoting apoptosis and autophagy. On the other hand, betaine associated with C-phycocyanin increased caspase-3 / pro-caspase-3 ratio.Finally, we studied the effect of voluntary physical activity on growth of implanted A549 cells in Nude rats. We showed that voluntary physical activity slowed down the lung tumour growth, without significant difference if animals were supplied with betaine or/and C-phycocyanin. It seems that the increase of lipid peroxidation, NF-kappaB and p38 activation, and AKT inhibition, all having a role in promotion of a cell death, are responsible for the tumour growth slowdown following the physical activity. In conclusion, diet enriched with betaine or/and C-phycocyanin slows down the growth of pulmonary adenocarcinoma cells implanted in rats, suggesting their interest in anti-cancer activity. Physical activity seems to act on similar mechanisms as these micronutrients. Our results have to be confirmed with further studies, but are already suggesting a potential application in lung cancer patients.

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