La protéine Damaged DNA-Binding 2 (DDB2) est connue pour son rôle dans la réparation de l'ADN lésé par les UV. Cependant, le laboratoire a montré que cette protéine est surexprimée naturellement dans les cellules tumorales mammaires non métastatiques et active leur prolifération, en favorisant leur entrée en phase de transition G1/S du cycle cellulaire. Il a été montré que cette nouvelle activité biologique de DDB2 dépend de sa capacité à intervenir dans la transcription de gènes cibles, comme celui codant l'enzyme anti-oxydante, la superoxyde dismutase à manganèse (SOD Mn). Sur la base que DDB2 est peu ou pas exprimée dans les cellules tumorales mammaires métastatiques, ce travail a consisté à étudier le rôle de cette protéine dans les capacités invasives de ces cellules. Dans un 1er temps, nous avons montré que les cellules tumorales mammaires hautement métastatiques (MDA-MB231 et SKBR3), lorsqu'elles surexpriment DDB2 après introduction de son gène, ont des capacités migratrices et invasives in vitro, ainsi que des propriétés in vivo à développer des métastases pulmonaires, fortement réduites, en association avec une diminution importante de l'expression de la métalloprotéase matricielle 9 (MMP-9). De même, lors d'une analyse rétrospective sur 92 échantillons cliniques provenant de patientes, une corrélation inverse entre l'expression de DDB2 et le haut grade (SBR>ou =3) des tumeurs mammaires est observée. Dans un 2ème temps, nous avons identifié le mécanisme moléculaire par lequel DDB2 agit négativement sur les capacités invasives des cellules tumorales mammaires. Nous avons montré que DDB2 intervient positivement sur l'expression du gène codant I kappa B alpha (IkBa), en se fixant sur une séquence d'ADN localisée dans la région proximale du promoteur, qui entraîne en conséquence une forte diminution de l'activité du facteur de transcription NF-kB. Ce dernier est connu pour son rôle dans les capacités invasives et migratrices des cellules tumorales mammaires métastatiques, en régulant de nombreux gènes cibles comme celui codant la MMP-9. Nous avons montré, que l?inhibition de l'expression d'IkBa, par ARN interférence restaure en partie les propriétés invasives des cellules tumorales mammaires métastatiques surexprimant DDB2, en association avec une réexpression de MMP-9. Dans un 3ème temps, nous avons également montré dans les cellules tumorales mammaires métastatiques, que l?expression constitutivement élevée de la SOD Mn, en l'absence de DDB2, dépend de l'activité conjointe des facteurs de transcription NF-kB et Sp1, révélant ainsi un autre mécanisme moléculaire impliqué dans les propriétés invasives de ces cellules. L'ensemble de ce travail contribue ainsi à mieux comprendre comment les cellules tumorales mammaires progressent vers un statut invasif et renforce également l'idée que DDB2 présente un intérêt clinique potentiel, comme marqueur prédictif de la progression métastatique des tumeurs mammaires. Enfin, la relation entre la DDB2, NF-kB et la SOD Mn représente une voie intéressante pour le développement de nouvelles thérapies anticancéreuses.

Agronomijos studijų programos magistro baigiamasis darbas 40 puslapių, 10 paveikslų, 2 lentelės, lietuvių kalba. Tyrimo objektas: paprastojo miežio (Hordeum vulgare L.) vasarinės veislės ,,Simba'' agrocenozė. Tyrimo tikslas: Įvertinti ilgalaikio skirtingo intensyvumo žemės dirbimo poveikį miežių agrocenozei, taikant intensyvias technologijas. Tyrimo uždaviniai: 1. Apžvelgti mokslinę literatūrą ir atlikti sisteminę analizę tiriamuoju klausimu. 2. Aprašyti atlikto eksperimento vykdymo sąlygas ir metodus. 3. Tyrimais nustatyti ir įvertinti ilgalaikio skirtingo intensyvumo žemės dirbimo poveikį struktūros patvarumui, miežių sudygimui, pasėlių piktžolėtumui ir miežių produktyvumui. Tyrimo metodai: mokslinės literatūros apžvalga, loginė analizė ir sintezė, statistinė analizė, palyginamoji analizė, grafinio modeliavimo technikos. Tyrimo rezultatai: • Pirmoji dalis pristato Lietuvos ir užsienio autorių mokslinės literatūros analizę. Mokslinė medžiaga atrinkta, susisteminta ir analizuota. • Antroje dalyje aprašyti koncentruojamasi ties bandymo schema ir parametrais, mokslinio eksperimento variantais ir duomenų analizės stebėjimais ir metodais. • Trečioji dalis identifikuoja dirvožemio struktūros patvarumą, miežių sudygimą ir stiebų produktyvumą, miežių derlių, pasėlių piktžolėtumą bei miežių produktyvumą. Nustatyta, kad ilgalaikis skirtingo intensyvumo žemės dirbimas neturėjo esminės įtakos dirvožemio struktūros patvarumui. Nenustatyta esminės įtakos ir miežių produktyvumui... [toliau žr. visą tekstą] / Final work of University Undergraduate/Master Studies 47 pages, 9 figures, 2 tables, the Lithuanian language. Object of the research: barley (Hordeum vulgare L.) spring variety Simba'' agrocenoses. Aim of the research: to evaluate the long-term effect of different soil tillage intensity on barley agrocenosis applying intensive technologies. Objectives of the research: 1. Review of the literature and perform a systematic analysis of the relevant issue. 2. Describe an experiment carried out with the conditions and methods. 3. Studies to identify and evaluate the long-term differences in the intensity of tillage effects on structural stability, barley germination, weed infestation and barley productivity. Research methods: review of scientific literature logical analysis and synthesis, statistical analysis, comparative analysis, graphical modelling techniques. Research results: • Part One presents Lithuanian and foreign scientific literature. Scientific material selected, structured and analysed. • Part Two describe the focus is on testing scheme and the parameters of the scientific variants of the experiment and data analysis methods and observations. • Part Three identifies the soil structure stability, barley germination and stem productivity of barley harvest, weed and barley crop productivity. It was established, that tillage intensity had no significant effect on soil aggregate stability and on barley productivity parameters except the number of germinating barley... [to full text]

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Biodiversity rich regions worldwide face threats from various global change agents. This research quantifies environmental influences on vegetation, and the impacts of exotic woody plant invasion and anthropogenic nitrogen (N) deposition in a global biodiversity hotspot. The study was conducted in the montane grasslands of the Nilgiris, Western Ghats, and outlines potential management options for this region. Specifically, I examined (1) the role of environmental factors in influencing native plant distribution and ecosystem properties, (2) the status and impact of exotic shrub (Scotch broom, henceforth broom) invasion, (3) the role of disturbances in the success of broom, (4) the role of fire in restoring invaded grasslands, and (5) the impacts of terrestrial N loading on the grassland ecosystem. I used experiments and surveys to assess these. Distributions of several key species were explained by a few complex environmental gradients. In invaded-grasslands, broom populations consisted mainly of intermediate size and age classes, with no clear indication of population decline. Invasion negatively impacted plant community structure and drastically changed composition, favoring shade-tolerant and weedy species. However, invasion did not greatly alter ecosystem function. Fire successfully eliminated mature broom stands, but resulted in a short-term increase in broom seedling recruitment. At the end of 18 months, the fire effects on uninvaded-grasslands were not apparent, but there was no conclusive evidence of the formerly invaded patches attaining the composition of uninvaded-grasslands following burning. N fertilization strongly influenced soil N dynamics, and shoot N concentrations, but effects on aboveground production were weak. Surprisingly, N enrichment had positive effects on diversity in the short-term. It is clear that these grasslands need immediate management intervention to forestall degradation from invasion. Fire could be used to eliminate mature broom stands and deplete persistent seedbanks, which will facilitate colonization by shade-intolerant grassland plants. Active restoration should be mindful of environmental preferences of framework species. Long-term studies of the impacts of N deposition in the context of disturbances will help determine realistic critical thresholds and utilize disturbances to buffer the potential adverse effects of increasing N loading.

Analyses of the large-scale population structure of pathogens enable the identification of migration patterns, diversity reservoirs or longevity of populations, the understanding of current evolutionary trajectories and the anticipation of future ones. A detailed analysis of populations in centre of diversity should enable to infer the adaptive capacity of the pathogen and identify potential sources for new invasions. Puccinia striiformis f.sp. tritici (PST) is the causal agent of wheat yellow/stripe rust, and despite a worldwide distribution, this fungus remains a model species for invasion studies, due to its long-distance migration capacity and recurrent local emergence of new strains. Little is known about the ancestral relationship of the worldwide PST population with unknown center of origin. We used multilocus microsatellite genotyping to infer the worldwide population structure of PST and the origin of new invasions, analysing a set of isolates representative of sampling performed over six continents. Bayesian and multivariate clustering methods partitioned the isolates into six distinct genetic groups, corresponding to distinct geographic areas. The assignment analysis confirmed the Middle East-Red Sea Area as the most likely source of newly spreading, high-temperature-adapted strains; Europe as the source of South American, North American and Australian populations; and Mediterranean-Central Asian populations as the origin of South African populations. The existence of strong population subdivision at worldwide level shows that major genetic groups are not markedly affected by recent dispersal events. However, the sources for recent invasions and the migration routes identified emphasize the importance of human activities on the recent long-distance spread of the disease. The analyses of linkage disequilibrium and genotypic diversity indicated a strong regional heterogeneity in levels of recombination, with clear signatures of recombination in the Himalayan (Nepal and Pakistan) and near-Himalayan (China) regions and a predominant clonal population structure in other regions. To explain the variability in diversity and recombination of worldwide PST populations, we assessed their sex ability in terms of telial production, the sex-specific structures that are obligatory for PST sexual cycle, in a set of 56 isolates representative of these worldwide geographical origins. We confirmed that the variability in genotypic diversity/ recombination was linked with the sex ability, pinpointing the Himalayan region as the possible center of origin of PST, from where it then spread worldwide. The reduced sex ability in clonal populations certainly reflects a loss of sexual function, associated to migration in areas where sexual alternate host is lacking, or not necessary for the completion of epidemic cycle. Approximate Bayesian computation analyses confirmed an out of Himalaya spread of PST, with Pakistan and China being the most ancestral population. A detailed analysis of Pakistani population at regional level revealed the existence of a strong population subdivision, a high genotypic diversity and the existence of recombination signature at each location reflecting the role of sexual recombination in the temporal maintenance at local level. A time spaced sampling of PST in the valley of Tianshui (China) inspired the development of a new estimator, allowing to quantify the relative contribution of sexual reproduction and effective population size on the basis of clonal resampling within and between years. A sexual reproduction rate of 74% (95% confidence interval [CI]: 38-95%) and effective population size of 1735 (95% CI: 675-2800) was quantified in Chinese PST population. The description of the origin and migration routes of PST populations worldwide and at its centre of diversity contributes to our understanding of PST evolutionary potential, and is helpful to build disease management strategies.

Salmonella enterica serovar Typhimurium is a facultative intracellular pathogen that has evolved to take advantage of the eukaryotic host cells it inhabits during infection. It uses bacterial effectors translocated into the host cell cytosol to manipulate host cell machinery and establish a replicative niche. In this thesis I study the function of two of these effectors, SopD and SopB, which have been shown to act cooperatively to induce phenotypes associated with gastroenteritis (fluid secretion and neutrophil influx into the intestinal lumen). In addition to promoting gastroenteritis, SopD has also been implicated in systemic and persistent infection of mice. Although recently implicated in invasion, the precise function of SopD has remained elusive. Here I show that SopD affects membrane dynamics during S. Typhimurium invasion of epithelial cells. SopD promotes membrane sealing and macropinosome formation, events that may have important consequences for efficiency of bacterial cell entry in vivo. Furthermore, we demonstrate that SopD is recruited to the invasion site membranes through the phosphatase activity of SopB, suggesting a mechanism for their cooperative action during induction of gastroenteritis. Unlike SopD, SopB has been a focus of intense research efforts and its role in invasion as a phosphoinositide phosphatase is well documented. However, we have observed that SopB also inhibits fusion of lysosomes with Salmonella-containing vacuoles (SCVs) following invasion. This ability depends on SopB-mediated reduction of negative membrane charge of the SCV during invasion by hydrolysis of the phosphoinositide PI(4,5)P2. Membrane charge alterations driven by SopB result in removal of Rab GTPases from the SCV that depend on electrostatic interactions for their targeting. Two of these Rabs, Rab23 and Rab35 were previously shown to promote phagosome-lysosome fusion. Therefore their removal from the SCV may promote SCV trafficking away from the degradative endocytic pathway of host cells. This represents a new mechanism by which an invasion associated effector controls SCV maturation. Together, this work advances our knowledge of the interaction between S. Typhimurium and its host. This research also suggests a new mechanism by which pathogens other than S. Typhimurium could promote their intracellular survival.

Coccidiosis in chicken causes great economic losses. Increasing resistance of Eimeria species to anticoccidials has forced the search for alternative methods of control. The present study evaluates the anticoccidial activity of some anti-Eimeria tenella antibody fragments expressed in pea plants. Both in vitro and in vivo infection assays including indirect immunofluorescence, in vivo evaluation of antibody neutralization and cell culture invasion-inhibition assays were used to study the inhibitory effect of these antibody fragments on E. tenella sporozoites. Seven of nine antibody fragments (Ab1, Ab4, Ab5, Ab6, Ab7, Ab8 and Ab9) showed binding to sporozoites of E. tenella in an indirect immunofluorescence test. Only two antibodies (Ab4 and Ab5) cross reacted with sporozoites of E. maxima, E. acervulina and E. brunetti. The localization of specific fluorescence differed between species. Ab binding with sporozoites was seen in the area of both anterior and posterior refractile bodies in case of E. tenella, E. brunetti, and E. maxima but was only observed in the posterior refractile body in case of E. acervulina. No antibody binding was observed on merozoites. The suitability of antibody fragments to alter the infectivity of E. tenella sporozoites to Madin Darby Bovine Kidney cells (MDBK) was examined in vitro and the invasion-inhibition rates were quantified by flow cytometry. To assess the inhibitory effect on parasite reproduction, the in vivo antibody neutralization assay was done by retrograde infection of chicken with sporozoites previously incubated with antibody fragments. In vitro invasion rates were reduced by incubation with antibody fragments by approximately 24 to 45 %, with Ab6 and Ab7 showing the most distinct effect. However, proliferation rates (PR) of the respective MDBK cultures were also clearly reduced by 15 to 26 %. PR of MDBK cells treated with 1:1000, 1:100, 1:10 and undiluted mixed antibody fragments were reduced by 1%, 10%, 16%, and 26% with a reduction of invasion rates by 0%, 9%, 15% and 18%, respectively. Immune sera reduced the invasion rates by 16% to 70% and increased PR of the host cells. It appeared that the preparations of the antibody fragments contained compounds cytotoxic to MDBK cells and thus invasion inhibition could not be unequivocally evaluated in vitro. However, after incubation with antibody fragments sporozoites displayed a reduced ability to reproduce after intracloacal application to chicken (especially Ab1, Ab3, Ab5 and Ab9). Other antibody fragments (Ab2, Ab4, Ab6, Ab7 and Ab8) were less capable to reduce sporozoite infectivity and reproduction. More investigations are still required to study the possible use of antibody fragments and their application to infected chicken exposed to coccidiosis.

Made available in DSpace on 2016-06-02T19:32:14Z (GMT). No. of bitstreams: 1 6709.pdf: 4195083 bytes, checksum: 80992e3623fd375eb0fcff062a4c0f5d (MD5) Previous issue date: 2015-02-05 / Financiadora de Estudos e Projetos / Biological invasion is a threat to biodiversity, due to competition for resources between invaders and native organisms, decreasing the overall biodiversity. So, studies about the invasiveness of the ecossystems and invasion potential of exotic plants several environments must be taken to control this species. Hedychium coronarium, from the Himalayas, is an example of a high invasivisiness plant, taken to different places in the world, mainly due to ornamental uses. This study evaluated the development (height, diameter, birth and ramets mortality and internal rhyzome anatomy) of plants under different moisture contents, in in situ and in vitro experiments. We found the existence of plasticity for both vegetative ramets growth, and the internal rhyzome anatomy, which may contribute to the invasive potential of this plant. / A invasão biológica é uma ameaça à biodiversidade, já que ocorre competição por recursos entre organismos invasores e nativos, diminuindo a biodiversidade global. Assim, estudos sobre a invasão dos ecossistemas e potencial invasão de plantas exóticas em ambientes diversos devem ser realizados para que haja controle das espécies. Hedychium coronarium, proveniente do Himalaia, é um exemplo de planta com alta invasivisidade, levada a diferentes lugares do mundo, sobretudo devido ao potencial ornamental que fornece. O presente estudo avaliou o desenvolvimento (altura, diâmetro, nascimento, mortalidade dos rametas e anatomia interna do rizoma) da planta em ambientes com diferentes umidades, com experimentos in situ e in vitro. Foi encontrado indício de plasticidade, tanto em crescimento vegetativo dos rametas, quanto na anatomia interna dos rizomas, o que pode contribuir para a invasivisidade dessa planta.

Le récepteur-canal P2X7 est fortement exprimé et est fonctionnel dans la lignée de cellules cancéreuses mammaires humaines hautement invasives MDA-MB-435s. L’activation de P2X7 par l’ATP extracellulaire est responsable de l'émission des prolongements cellulaires et l'augmentation de la migration cellulaire. En outre, l’activation de P2X7 augmente l’invasion cellulaire à travers la matrice extracellulaire et fait intervenir la libération de forme mature de cathepsines à cystéine dans le milieu extracellulaire. L’inhibition pharmacologique de P2X7 diminue l’invasivité des cellules cancéreuses dans un modèle de micrométastases chez le poisson zèbre. Nous avons également montré que l’émodine (1,3,8-trihydroxy-6-méthylanthraquinone) une anthraquinone isolée de Rheum officinale Baill (Rhubarbe chinoise) inhibe l’invasivité des cellules cancéreuses humaines via l’antagonisme de P2X7 et n’as pas d’effet sur les autres récepteurs P2X. Nos résultats démontrent un nouveau mécanisme entre la fonctionnalité de P2X7 dans les cellules cancéreuses et l’invasivité cellulaire, un paramètre clé dans la croissance tumorale et le développement des métastases. Ceci suggère également un rôle thérapeutique potentiel pour les antagonistes des P2X7. / P2X7 receptor channel is highly expressed and fully functional in the highly invasive human breast cancer cell line MDA-MB-435s. Its activation by extracellular ATP is responsible for the extension of neurite-like cellular prolongations, and the increase in cell migration. Furthermore, P2X7 activation enhanced invasion through the extracellular matrix and was related to the increase of mature forms of cysteine cathepsins in the extracellular medium. Pharmacological targeting of P2X7 decreases cancer cell invasiveness in a zebrafish model of micrometastases. We also showed that emodin (1,3,8-trihydroxy-6-methylanthraquinone) an anthraquinone derivative originally isolated from Rheum officinale Baill (Chinese Rhubarb) inhibits human cancer cell invasiveness by specifically antagonizing the P2X7 and not the other members of the P2X family. Our results demonstrate a novel mechanistic link between P2X7 functionality in cancer cells and invasiveness, a key parameter in tumour growth and in the development of metastases. These results also suggest a potential therapeutic role for the newly developed P2X7 antagonists.

Submitted by Noeme Timbo (noeme.timbo@metodista.br) on 2017-05-30T18:28:32Z No. of bitstreams: 1 Aline Silva Ferreira.pdf: 1292757 bytes, checksum: 95de701640eb65c840c7627a1e07ed1a (MD5) / Made available in DSpace on 2017-05-30T18:28:32Z (GMT). No. of bitstreams: 1 Aline Silva Ferreira.pdf: 1292757 bytes, checksum: 95de701640eb65c840c7627a1e07ed1a (MD5) Previous issue date: 2017-02-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Due to competitivity, companies adhered to Information and Communication Technologies (ICT) for increasing productivity and results. In this sense, technology improved quality and productivity in workers’ routine, but excessive use of ICT can lead to the technological stress known as technostress. Some aspects of creating technological stress relates to information overload and constant connectivity, denominated as techno-overload and techno-invasion. This study analyzed reflexes of the technostress in the organizational behavior. Using a quantitative approach, data was collected through a research instrument with 331 questionnaires answered. It concludes that techno-overload and techno-invasion influence positively work-family conflict, so excessive use of ITC can generate discomfort in the professional’s life. Results point that it was not possible to indicate influence of techno-overload and techno-invasion relating to affective organizational commitment and turnover intention. It verifies that there is low influence of the Techno-invasion relating to job satisfaction, and it was not possible to confirm influence on job satisfaction by techno-overload. / Devido à competitividade de mercado, as empresas aderiram as Tecnologias da Informação e Comunicação (TIC) buscando cada vez mais elevar sua produtividade e os resultados da organização. Nesse sentido, a tecnologia melhorou a qualidade e a produtividade na rotina dos trabalhadores, mas o uso excessivo das TIC pode levar ao estresse tecnológico conhecido como Tecnoestresse. Alguns aspectos de criação de estresse tecnológico estão relacionados à sobrecarga de informações e conectividade constante, denominados como Tecnosobrecarga e Tecnoinvasão. Este estudo analisou os reflexos do Tecnoestresse no Comportamento Organizacional. Através de uma abordagem quantitativa, os dados foram coletados por meio de um instrumento de pesquisa com obtenção das respostas de 331 questionários. Conclui-se que a Tecnosobrecarga e a Tecnoinvasão influenciam positivamente o Conflito Trabalho – Família, desta forma o excesso do uso da TIC pode gerar dissabores na vida do profissional. De acordo com os resultados deste estudo não foi possível afirmar que há influência da Tecnosobrecarga e Tecnoinvasão em relação ao Comprometimento Organizacional Afetivo e a Intenção de Rotatividade. Verificou-se que existe uma baixa influência da Tecnoinvasão em relação à satisfação no trabalho, e não foi possível confirmar a influência referente à satisfação no trabalho pela Tecnosobrecarga.