A research report submitted by the Wits School of Arts, Film and Television Department, University of the Witwatersrand, Johannesburg, in partial fulfilment of the requirements for the degree of Masters in Film and Television. Johannesburg 2015 / This research report looks at the politics of memory, commemoration and representation in the new South Africa with a focus on 32-Battalion. The research draws on interviews and testimonies of ex-SADF soldiers who were members of the unit and unpacks how when considering memory and remembering, a multitude of viewpoints emerge. Factors such as the impact of the ideological transition from apartheid to democracy and the resultant impact on ex-SADF soldiers as well as the reshaping of the country’s official history which has been shaped to suit the current political climate, are considered. The research reflects on how these political processes which include exclusions of unwanted histories have affected nation building in South Africa post 1994.

O carcinoma epidermóide representa mais de 90% das neoplasias malignas de cabeça e pescoço, apresentando taxas elevadas de morbi-mortalidade, porém pouco se sabe sobre as vias de sinalização que estão envolvidas na progressão tumoral. Têm sido relatado na literatura, que alguns estímulos podem ativar a holoenzima PI3K que, por sua vez, desencadeia um processo que induz a fosforilação da proteína Akt (pAkt) que leva a ativação e a translocação do NF-B do citoplasma para o núcleo, onde ocorre a transcrição de genes envolvidos na proliferação e invasão celular. Assim, esse estudo analisou, através dos métodos de Imunofluorescência e Western Blot, a expressão das proteínas pAkt, NF-B e Ciclina D1 em três linhagens de células de carcinoma epidermóide de cabeça e pescoço (HN6, HN30 e HN31) submetidas a cultivo tridimensional e ensaio de invasão (gerando clones invasivos HN6.1, HN30.1 e HN31.1), ambos realizados com Matrigel®. O pAkt apresentou marcação citoplasmática e nuclear nas linhagens celulares HN6, HN30, HN6.1 e na HN30 submetida ao cultivo tridimensional, todavia, as linhagens celulares HN31, HN30.1, HN31.1 e HN6/HN31 cultivadas tridimensionalmente, apresentaram positividade predominantemente nuclear. No caso do NF-B, a localização foi marcadamente citoplasmática em todas as linhagens celulares cultivadas bi ou tridimensionalmente, porém, com exceção da HN31.1, o padrão de marcação foi citoplasmático e nuclear nos clones invasivos. Por fim, a Ciclina D1 exibiu imunopositividade apenas nuclear. A técnica de Western Blot mostrou diminuição nos níveis de expressão das proteínas analisadas quando as células foram cultivadas em Matrigel®, sendo, na maioria das vezes, essa redução estatisticamente significante, exceto no caso da linhagem celular HN6, que apresentou um aumento significativo de Ciclina D1. Os clones invasivos HN6.1 e HN30.1 exibiram elevação significativa dos níveis de expressão de pAkt e NF-B porém, a HN31.1 apresentou aumento de pAkt e discreta diminuição de NF-B, mas ambos os valores não estatisticamente significantes. Em relação à Ciclina D1, houve um aumento dos seus níveis em todas as linhagens invasivas, sendo que apenas na HN6.1 foi estatisticamente significante. Esses resultados mostraram que, nas linhagens celulares avaliadas, quando cultivadas em ambiente tridimensional, a significativa redução dos níveis de expressão de proteínas da via de sinalização PI3K ocorreu devido a uma possível fase adaptativa dessas células ao Matrigel® que seria seguida provavelmente por uma etapa de aumento do potencial proliferativo e posterior transdiferenciação de algumas células para ganho de fenótipo mais agressivo. Além disso, este estudo mostrou a participação da via de sinalização Akt/NF-B/Ciclina D1 no processo de invasão das células de carcinoma epidermóide de cabeça e pescoço (HN6.1 e HN30.1). / Squamous cell carcinoma represents more than 90% of the head and neck malignant tumors, with high mortality rates. Nevertheless, the knowledge about signaling pathways involved in tumor progression remains unclear. The relationship between pAkt and NF-B is well established in carcinogenesis. It is known that the activation of PI3K can be induced by some factors what leads to Akt phosphorilation (pAkt). This further event starts an activation cascade that induces NF-B translocation from the cytoplasm into the nucleus where the transcription of genes enrolled in cellular proliferation and invasion will be done. Therefore, this study analyzed the status of pAkt, NF-B and Cyclin D1 proteins by Immunofluorescence and Western Blot methods in three head and neck squamous cell carcinoma cell lines (HN6, HN30 and HN31) submitted to three-dimensional culture model and an in vitro invasion assay (invasive clones HN6.1, HN30.1 e HN31.1), both with Matrigel®. pAkt expression was detected in cytoplasm and nucleus in HN6, HN30, HN6.1 and HN30 cultured with Matrigel®, however, HN31, HN30.1, HN31.1 cell lines and HN6/HN31 submitted to three-dimensional culture model showed nuclear expression. NF-B localization was strongly cytoplasmatic in all cell lines cultured with or without Matrigel®, but, regarding HN31.1, NF-B protein expression was nuclear and cytoplasmatic in the invasive clones. Cyclin D1 was nuclear in all cell lines analyzed. Western blot assays showed a decrease in pAkt, NF-B and Cyclin D1 expression levels in all cells lines in three-dimensional culture model, most of them statistically significant, but It was detected a considerable increase in Cyclin D1 expression in HN6 cultured in threedimensional model. Moreover, HN6.1 and HN30.1 showed a significant enhance in pAkt and NF-B levels but, HN31.1 demonstrated a raise in pAkt and a slight decline in NF-B, both were not statistically significant. Finally, an increase in Cyclin D1 expression levels was illustrated in all cell lines but, only HN6.1 showed a statistic signal. These results suggest that, in the cell lines studied, when cultured in threedimensional model, a noteworthy reduction in expression levels of PI3K signaling pathway proteins happened for the reason that a possibly adaptation phase of these cells to Matrigel® was occurring, in the first moment, but it would be probably followed by an increase proliferative potential stage and subsequent transdifferentiation of some cells that could show, consequently, a more aggressive phenotype. At last, this study revealed the participation of Akt/NF-B/Cyclin D1 signaling pathway in invasion process of head and neck squamous cell carcinoma (HN6.1 and HN30.1).

O presente trabalho teve como objetivo o estudo da variabilidade genética e dos fatores de virulência de isolados de U. diversum. As cepas foram submetidas a sequenciamento dos genes da urease e 16S rRNA e a testes para verificar os fatores de virulência: cápsula, fosfolipase C, IgAse e adesão e invasão. A análise do sequênciamento parcial do gene 16S rRNA resultou na presença de polimorfismos em 44 posições da seqüência, que diferenciou as amostras em sete grupos. Em relação aos fatores de virulência, os dados mostraram que as cepas estudadas apresentaram uma camada densa ao redor da membrana celular dos microrganismos e atividade de fosfolipase C. No entanto, não foi observado a atividade de IgAse nas cepas. Em relação a atividade de invasão, observou-se que os ureaplasma estudados puderam ser visualizados no interior de células Hep-2 com apenas um minutos de infecção, sendo observados em uma região perinuclear, mas não no interior do núcleo. Além disto, pode verificar que entre 1% a 10% dos ureaplasmas estudos penetraram na célula pelo teste da gentamicina. / The aim of the present study was the study of genetic variability and virulence factors of U. diversum clinical isolates. The strains were submitted to sequencing for 16S rRNA and urease genes. Moreover, the strains were analyzed to the virulence factors: capsule, phospholipase C, IgA protease and adhesion and invasion into Hep-2 cells. The sequencing of parcial 16S rRNA gene showed polymorphic patterns into 44 positions. These polymorphisms clustered the strains in seven groups. For the virulence factors, ureaplasma cells showed a dense-stained external capsule-like structure surrounding the cell membrane. A high level of phospholipase C activity was also detected in 31 studied ureaplasma. However, no strains showed IgA protease activity. For the invasion assay, the isolates and strains used were detected inside the cells after infection of one minute. The invasions of the ureaplasmas surrounded the nuclear region but were not observed inside the nuclei. The gentamicin invasion assay detected that 1% to 10% of studied ureaplasmas were inside the infected cells.

No Brasil, foram introduzidas diversas espécies de Pinus e a espécie P. elliottii Engelm apresenta o comportamento invasor em unidades de conservação de Cerrado, no estado de São Paulo. O objetivo deste estudo foi comparar P. elliottii com outras duas espécies do mesmo gênero, P. caribaea Moret e P. oocarpa Shiede ex Schltdl, através de: viabilidade e germinabilidade das sementes, em laboratório; sobrevivência das plântulas logo após a germinação e acompanhamento de seu crescimento em estufa; germinabilidade em fisionomias campestres; e chuva de sementes dentro de talhões. Para P. elliottii, também foi verificada a distância de dispersão das sementes. As sementes utilizadas nos testes em laboratório foram coletadas em Águas de Santa Bárbara (Estação Ecológica de Santa Bárbara - EEcSB; Floresta de Águas de Santa Bárbara - FASB) e Itirapina (Estação Experimental de Itirapina - EExI). Primeiramente, as sementes foram colocadas em água para separação das sementes cheias das vazias (flutuabilidade). Em seguida, as sementes cheias foram testadas quanto à viabilidade por meio de sal de tetrazólio e quanto à germinabilidade. As sementes germinadas foram acompanhadas até os cotilédones ficarem visíveis e uma parte destas teve o crescimento acompanhado até a 24ª semana. A germinabilidade em fisionomias campestres foi realizada apenas na EEcSB, onde as sementes das três espécies foram semeadas em diferentes graus de umidade do solo. Coletores de sementes foram instalados dentro dos talhões de Pinus spp., tanto na EEcSB-FASB como na EExI, para quantificar a chuva de sementes. A distância de dispersão das sementes de P. elliottii foi avaliada apenas na EEcSB, durante o período de maior dispersão de sementes (março a maio), e teve como classes de distâncias em relação ao talhão: 10, 30, 60, 100, 150 e 250 m. Os resultados dos testes em laboratório mostraram que P. caribaea e P. elliottii apresentam as maiores proporções de sementes cheias (>70%) e P. oocarpa e P. elliottii, as maiores taxas de viabilidade (>90%) e germinabilidade (>90%). Dentre as sementes germinadas das três espécies, mais de 90% desenvolveram-se até o aparecimento dos cotilédones. Quanto ao crescimento, P. caribaea e P. oocarpa apresentaram maior vigor em relação a P. elliottii. Em campo, as três espécies apresentaram, de maneira similar, baixíssima porcentagem de germinação (<1,5% do total semeado), preferencialmente em solos mais úmidos. A chuva de sementes de P. elliottii dentro de talhões foi muito maior (pelo menos o dobro) em comparação com as outras duas espécies. A distância de dispersão das sementes de P. elliottii foi maior nos primeiros 30 m, mas chegou até os 150 m. Portanto, P. caribaea e P. oocarpa apresentaram condições de se tornarem invasores pela viabilidade de suas sementes, vigor de crescimento e germinação em campo, mas o fator determinante para o sucesso na invasão P. elliottii é, provavelmente, a pressão de propágulos, maior do que as outras duas espécies, causada pela sua alta produção de sementes e intenso plantio. / Several pine species of the genus Pinus have been introduced in Brazil, and P. elliottii Engelm is presently considered an invasive species in the Brazilian savanna (Cerrado) in São Paulo State. The aim of this study was to compare P. elliottii with two other Pinus species, P. caribaea Moret and P. oocarpa Shiede ex Schltdl according to: seed viability and germinability, in laboratory; survival of seedlings soon after germination and their growth, in greenhouse; germination at field conditions; and seed rain inside the plantation. The distance of seed distance dispersal was also verified for P. elliottii. The seeds for the laboratory tests were collected in Águas de Santa Bárbara (Santa Barbara Ecological Station - EEcSB; Águas de Santa Bárbara Forest - FASB) and Itirapina (Itirapina Experimental Station - EExI). First, the seeds were placed in water to separate the full from the empty seeds (flutuability). Then full seeds were tested by viability (tetrazolium) and germinability. Germinated seeds were monitored until the cotyledons were visible and some of those were monitored up the 24th week to verify their growth rate. In the EEcSB, seeds of the three species were sown at different levels of soil moisture to observe the germination in the field. In order to quantify the species seed rain seed collectors were installed inside the Pinus spp. stands, in both EEcSB-FASB and EExI. The distance of P. elliottii seed spread was evaluated only at the EEcSB, during the greater period of seed dispersion (March to May), at the distances of 10, 30, 60, 100, 150 and 250 m from the planted areas. The results of laboratory tests showed that P. caribaea and P. elliottii have the highest proportions of full seeds (> 70%) and P. oocarpa and P. elliottii, the highest viability (> 90%) and germination rates (> 90%). Among the germinated seeds of the three species more than 90% developed to seedling stage. Concerning seedling growth, P. caribaea and P. oocarpa showed greater vigor than P. elliottii. In the field, the three species showed similar and very low germination rates (<1.5% of the total sown), preferably in more humid soils. The seed rain of P. elliottii inside the plantation stand was much higher (at least twice) compared to the other two species. The dispersal distance of P. elliottii seeds was higher in the first 30 m, but reached 150 m. The results show that P. caribaea and P. oocarpa are able to become invaders due to their high values of seed viability and germination, and vigor growth, however, the determining factor for the invasive success of P. elliottii is probably its higher propagule pressure, caused by several human mediated introductions, and high production of seeds.

Neste trabalho, esclarecemos tópicos da patogenicidade de EIEC que sustentam a sua menor virulência quando comparada à S. flexneri, e mostramos a importância das células dendríticas (CD) nesse processo. Estudou-se o comportamento de EIEC e S. flexneri quando em contato com células Caco-2, avaliando-se uma cinética de expressão dos genes envolvidos na invasão e disseminação bacteriana. Em geral, todos os genes foram menos expressos em EIEC, fato corroborado pelo fenótipo de disseminação bacteriana, onde EIEC foi menos eficiente do que Shigella. Também foi avaliada a modulação da resposta inflamatória de células dendríticas intestinais murinas pela produção de citocinas, expressão de moléculas co-estimulatórias e apresentação de antígenos, após desafio das células com as bactérias. Os resultados sugerem que EIEC induz a uma resposta protetora ao hospedeiro, enquanto que Shigella estaria \"driblando\" o sistema imune, além de provavelmente super-estimular o sistema imune adaptativo, fato que poderia levar a um agravamento da doença. As ações integradas das células Caco-2, células dendríticas e estímulos bacterianos foram estudadas em co-cultura celular. Observou-se que EIEC e suas proteínas secretadas induzem a migração das CDs ao compartimento apical da co-cultura; nada foi observado quando o desafio se deu com Shigella. Também foram avaliadas as concentrações de citocinas inflamatórias no microambiente infeccioso formado. A citocina TNF-&#945;, bem como CCL20 e MCP-1 foram mais proeminentes após estímulo com EIEC, fato que poderia explicar parcialmente a migração das CDs ao lado apical da co-cultura após estímulo com EIEC e suas proteínas secretadas. Nossas evidências experimentais indicam que a doença desencadeada por EIEC é mais restrita a um determinado local da infecção, ou seja, não é capaz de se disseminar a ponto de estender a lesão tecidual de forma mais drástica, como Shigella. Esse fenômeno pode estar associado à menor expressão de seus dos fatores de virulência e à resposta imune inata induzida no sítio de infecção, o que levaria, fatalmente, à resolução da doença. / In this study, we clarify topics of pathogenicity from EIEC that support its lower level of virulence when it is compared to S. flexneri, and we have shown the importance of dendritic cells (DC) in this process. We studied the conduct of EIEC and S. flexneri when they were in contact with Caco-2 cells and we analyzed the kinetics of the genes expression that was involved in the spread and invasion of the bacteria. In general, all genes were expressed less in EIEC, as demonstrated by the phenotype of the bacterial spread, where EIEC was less efficient than Shigella. We also analyzed the modulation of the inflammatory response by the murine intestinal dendritic cells by the production of cytokine, expression of co-stimulators molecules and antigens presentation, after the interaction of the cells with the bacteria. The results showed that EIEC induces a response that protects the host while Shigella manipulate the host intestinal innate and adaptive immune system and it probably over-stimulates the adaptive immune system which could let the disease worse. The integrated actions of Caco-2 cells, dendritic cells and bacterial stimulus, were studied in a co-culture cell. We observed that EIEC and its secreted proteins induce the migration of the DCs to the apical compartment of the co-culture; nothing was observed related to Shigella. We also evaluated the concentrations of the inflammatory cytokines at the infective micro environment that was formed. The cytokine TNF-&#945;, as CCL20 and MCP-1 were more prominent after been stimulated with EIEC, a fact that could partially explain the migration of DCs to the apical side of the co-culture after the stimulus with EIEC and its secreted proteins. Our experimental evidence shows that the disease triggered by the EIEC is more restricted at a definite infection place, which means that it is not capable of disseminating beyond a certain point to extend the tissue\'s injury and let it worsen, as Shigella do. This phenomenon can be associated with the lower level of expression of its virulence factors and to the immune response induced in the infection site, what could finally lead to the eradication of the disease.

Neospora caninum (filo Apicomplexa) é um parasita obrigatório intracelular como todos os membros deste filo, alguns reconhecidos por causarem doenças com impacto relevante na saúde humana (Plasmodium e Toxoplasma) e veterinária (Babesia, Eimeria e Cryptosporidium). Causador da neosporose, N. caninum vem emergindo como um dos maiores causadores de abortos infecciosos em bovinos, levando a consideráveis perdas econômicas na bovinocultura mundial. Devido à sua recente descoberta, o conhecimento sobre diversos processos bioquímicos de N.caninum ainda é limitado, demandando novas pesquisas para a compreensão de seus mecanismos de sobrevivência e consequente identificação de alvos para intervenção terapêutica. O processo de invasão celular é bastante investigado em pesquisas envolvendo apicomplexas, uma vez que a sobrevivência desses parasitas depende do sucesso de sua entrada na célula hospedeira. Proteínas secretadas de organelas filo-específicas (micronemas, roptrias e grânulos densos) estão intimamente envolvidas com a invasão celular. Elas são responsáveis pela interação inicial com a célula hospedeira, participam da junção de movimento formada no momento da invasão, e contribuem para a estabilização do vacúolo parasitóforo. Neste trabalho as proteínas secretadas por taquizoítas de N. caninum foram investigadas de duas formas: (1) por caracterização molecular de proteínas com domínio Apple; e (2) por estudo do secretoma do parasita. Os domínios proteicos do tipo Apple são caracterizados pela capacidade de interação proteína-proteína e proteína-carboidrato, e estão presentes em algumas proteínas micronêmicas com propriedades adesivas. Neste trabalho três proteínas de N. caninum contendo domínios Apple foram caracterizadas: MIC17A, MIC17B e MIC17C. A análise das sequências proteicas e das estruturas dos domínios Apple, obtidas por modelagem molecular, mostraram alta identidade sequencial e estrutural entre MIC17A e MIC17C. Apesar de ser paráloga às outras duas, MIC17B apresenta diferenças importantes em sua sequência e estrutura. Para MIC17B e MIC17C foram realizados experimentos de detecção das proteínas nativas nos extratos total e secretado do taquizoíta que sugerem diferentes formas de processamento entre essas proteínas no parasita. Para MIC17B foi confirmada a localização em micronemas, num padrão diferente do observado para MIC17C. Os ensaios de invasão combinados aos de localização indicam que estas proteínas estejam relacionadas ao processo de invasão celular, porém, suas funções permanecem desconhecidas. O secretoma é o conjunto de proteínas secretadas pelo parasita e, para explorar a composição deste extrato (ESA) no taquizoíta de N. caninum, duas abordagens complementares foram utilizadas. Na primeira abordagem foram identificadas as proteínas presentes no ESA por espectrometria de massas. Na segunda abordagem realizou-se uma ii quantificação relativa das proteínas, marcadas por dois isótopos, nos extratos totais de taquizoítas submetidos ou não ao estímulo secretório. O resultado esperado seria com as proteínas secretadas diminuídas no parasita estimulado. Em ambas as abordagens foram utilizadas técnicas de espectrometria de massas de alta resolução (nanoLC-MS/MS), o que resultou num alto número de identificações; 615 proteínas no ESA e 2011 proteínas quantificadas. A comparação das duas abordagens permitiu o reconhecimento de proteínas com maior probabilidade de secreção. Uma rede de interação entre as proteínas diferencialmente expressas foi predita, gerando resultados que, associados às informações sobre as proteínas aumentadas, permitiram uma investigação sobre proteínas potencialmente envolvidas com a regulação do metabolismo relacionado à secreção. Os resultados obtidos por ambos os estudos aqui demonstrados somam conhecimento acerca do parasita N. caninum e demonstram ser úteis para guiar a busca e seleção de alvos a serem investigados para o desenvolvimento de terapêutica contra a neosporose. / Neospora caninum (Apicomplexa phylum) is an obligatory intracellular parasite like all members from this phylum, some causing diseases with relevant impact on human (Plasmodium and Toxoplasma) and veterinary (Babesia, Eimeria and Cryptosporidium) health. Causative agent of neosporosis, N. caninum has emerged as one of the leading causes of infectious abortion in cattle, generating huge economical losses in worldwide livestock. Due to its recent discovery, knowledge of N. caninum biochemical processes remains scarce, demanding new research for comprehending its survival mechanisms and, consequently, identifying new targets for therapeutic intervention. The invasion process has often been investigated in apicomplexans since their survival depends on the success of their entry into the host cell. Proteins secreted from phylum-specific organelles (micronemes, rhoptries and dense granules) are deeply involved with invasion. They are responsible for the initial interaction with the host cell; participate of the moving junction formed in the moment of invasion; and contribute for the stabilization of the parasitophorus vacuole. In this study, the proteins secreted by N. caninum tachyzoites were investigated in two ways: (1) the molecular characterization of Apple domaincontaining proteins; and (2) exploring the parasite secretome. The Apple protein domains are characterized by the ability to interact as protein-protein and proteincarbohydrate, and are present in some microneme proteins with adhesive properties. Here three N. caninum proteins containing Apple domains were characterized: MIC17A, MIC17B and MIC17C. Analyses of the Apple domains sequences and structures, obtained by molecular modeling, revealed high sequential and structural identities between MIC17A and MIC17C. Although being a paralog of the other two proteins, MIC17B presents significant differences in its sequence and structure. Experiments were performed for native MIC17B and MIC17C detection in the total and secreted tachyzoite extracts, suggesting different processing forms for these proteins in the parasite. For MIC17B, the microneme localization was confirmed, differently from the pattern observed for MIC17C. Invasion and localization assays indicated that these proteins are related to the cell invasion process; nevertheless, their functions remain unknown. The secretome is the set of proteins secreted by the parasite and, to explore this extract (ESA) composition in N. caninum, two complementary approaches were used. Firstly proteins present in ESA were identified by mass spectrometry. In the second approach, a relative quantification was performed on the proteomes of ethanol stimulated/non stimulated tachyzoites, expecting that the secreted proteins would be down regulated at the stimulated parasite. Both approaches were performed with high resolution mass spectrometry techniques (nanoLC-MS/MS), reaching a high number of identifications: 615 proteins iv in ESA and 2011 quantified proteins. The comparison between both approaches allowed the recognition of the most likely secreted proteins. An interaction network was predicted, involving the differentially expressed proteins. These results, associated with the information of up regulated proteins, allowed the investigation of proteins potentially involved with the secretion metabolism regulation. The findings from our two studies add up knowledge about N. caninum and demonstrate to be useful in guiding the search and selection for new targets for therapeutic development against neosporosis.

O carcinoma epidermóide representa mais de 90% das neoplasias malignas de cabeça e pescoço, apresentando taxas elevadas de morbi-mortalidade, porém pouco se sabe sobre as vias de sinalização que estão envolvidas na progressão tumoral. Têm sido relatado na literatura, que alguns estímulos podem ativar a holoenzima PI3K que, por sua vez, desencadeia um processo que induz a fosforilação da proteína Akt (pAkt) que leva a ativação e a translocação do NF-B do citoplasma para o núcleo, onde ocorre a transcrição de genes envolvidos na proliferação e invasão celular. Assim, esse estudo analisou, através dos métodos de Imunofluorescência e Western Blot, a expressão das proteínas pAkt, NF-B e Ciclina D1 em três linhagens de células de carcinoma epidermóide de cabeça e pescoço (HN6, HN30 e HN31) submetidas a cultivo tridimensional e ensaio de invasão (gerando clones invasivos HN6.1, HN30.1 e HN31.1), ambos realizados com Matrigel®. O pAkt apresentou marcação citoplasmática e nuclear nas linhagens celulares HN6, HN30, HN6.1 e na HN30 submetida ao cultivo tridimensional, todavia, as linhagens celulares HN31, HN30.1, HN31.1 e HN6/HN31 cultivadas tridimensionalmente, apresentaram positividade predominantemente nuclear. No caso do NF-B, a localização foi marcadamente citoplasmática em todas as linhagens celulares cultivadas bi ou tridimensionalmente, porém, com exceção da HN31.1, o padrão de marcação foi citoplasmático e nuclear nos clones invasivos. Por fim, a Ciclina D1 exibiu imunopositividade apenas nuclear. A técnica de Western Blot mostrou diminuição nos níveis de expressão das proteínas analisadas quando as células foram cultivadas em Matrigel®, sendo, na maioria das vezes, essa redução estatisticamente significante, exceto no caso da linhagem celular HN6, que apresentou um aumento significativo de Ciclina D1. Os clones invasivos HN6.1 e HN30.1 exibiram elevação significativa dos níveis de expressão de pAkt e NF-B porém, a HN31.1 apresentou aumento de pAkt e discreta diminuição de NF-B, mas ambos os valores não estatisticamente significantes. Em relação à Ciclina D1, houve um aumento dos seus níveis em todas as linhagens invasivas, sendo que apenas na HN6.1 foi estatisticamente significante. Esses resultados mostraram que, nas linhagens celulares avaliadas, quando cultivadas em ambiente tridimensional, a significativa redução dos níveis de expressão de proteínas da via de sinalização PI3K ocorreu devido a uma possível fase adaptativa dessas células ao Matrigel® que seria seguida provavelmente por uma etapa de aumento do potencial proliferativo e posterior transdiferenciação de algumas células para ganho de fenótipo mais agressivo. Além disso, este estudo mostrou a participação da via de sinalização Akt/NF-B/Ciclina D1 no processo de invasão das células de carcinoma epidermóide de cabeça e pescoço (HN6.1 e HN30.1). / Squamous cell carcinoma represents more than 90% of the head and neck malignant tumors, with high mortality rates. Nevertheless, the knowledge about signaling pathways involved in tumor progression remains unclear. The relationship between pAkt and NF-B is well established in carcinogenesis. It is known that the activation of PI3K can be induced by some factors what leads to Akt phosphorilation (pAkt). This further event starts an activation cascade that induces NF-B translocation from the cytoplasm into the nucleus where the transcription of genes enrolled in cellular proliferation and invasion will be done. Therefore, this study analyzed the status of pAkt, NF-B and Cyclin D1 proteins by Immunofluorescence and Western Blot methods in three head and neck squamous cell carcinoma cell lines (HN6, HN30 and HN31) submitted to three-dimensional culture model and an in vitro invasion assay (invasive clones HN6.1, HN30.1 e HN31.1), both with Matrigel®. pAkt expression was detected in cytoplasm and nucleus in HN6, HN30, HN6.1 and HN30 cultured with Matrigel®, however, HN31, HN30.1, HN31.1 cell lines and HN6/HN31 submitted to three-dimensional culture model showed nuclear expression. NF-B localization was strongly cytoplasmatic in all cell lines cultured with or without Matrigel®, but, regarding HN31.1, NF-B protein expression was nuclear and cytoplasmatic in the invasive clones. Cyclin D1 was nuclear in all cell lines analyzed. Western blot assays showed a decrease in pAkt, NF-B and Cyclin D1 expression levels in all cells lines in three-dimensional culture model, most of them statistically significant, but It was detected a considerable increase in Cyclin D1 expression in HN6 cultured in threedimensional model. Moreover, HN6.1 and HN30.1 showed a significant enhance in pAkt and NF-B levels but, HN31.1 demonstrated a raise in pAkt and a slight decline in NF-B, both were not statistically significant. Finally, an increase in Cyclin D1 expression levels was illustrated in all cell lines but, only HN6.1 showed a statistic signal. These results suggest that, in the cell lines studied, when cultured in threedimensional model, a noteworthy reduction in expression levels of PI3K signaling pathway proteins happened for the reason that a possibly adaptation phase of these cells to Matrigel® was occurring, in the first moment, but it would be probably followed by an increase proliferative potential stage and subsequent transdifferentiation of some cells that could show, consequently, a more aggressive phenotype. At last, this study revealed the participation of Akt/NF-B/Cyclin D1 signaling pathway in invasion process of head and neck squamous cell carcinoma (HN6.1 and HN30.1).

O presente trabalho teve como objetivo o estudo da variabilidade genética e dos fatores de virulência de isolados de U. diversum. As cepas foram submetidas a sequenciamento dos genes da urease e 16S rRNA e a testes para verificar os fatores de virulência: cápsula, fosfolipase C, IgAse e adesão e invasão. A análise do sequênciamento parcial do gene 16S rRNA resultou na presença de polimorfismos em 44 posições da seqüência, que diferenciou as amostras em sete grupos. Em relação aos fatores de virulência, os dados mostraram que as cepas estudadas apresentaram uma camada densa ao redor da membrana celular dos microrganismos e atividade de fosfolipase C. No entanto, não foi observado a atividade de IgAse nas cepas. Em relação a atividade de invasão, observou-se que os ureaplasma estudados puderam ser visualizados no interior de células Hep-2 com apenas um minutos de infecção, sendo observados em uma região perinuclear, mas não no interior do núcleo. Além disto, pode verificar que entre 1% a 10% dos ureaplasmas estudos penetraram na célula pelo teste da gentamicina. / The aim of the present study was the study of genetic variability and virulence factors of U. diversum clinical isolates. The strains were submitted to sequencing for 16S rRNA and urease genes. Moreover, the strains were analyzed to the virulence factors: capsule, phospholipase C, IgA protease and adhesion and invasion into Hep-2 cells. The sequencing of parcial 16S rRNA gene showed polymorphic patterns into 44 positions. These polymorphisms clustered the strains in seven groups. For the virulence factors, ureaplasma cells showed a dense-stained external capsule-like structure surrounding the cell membrane. A high level of phospholipase C activity was also detected in 31 studied ureaplasma. However, no strains showed IgA protease activity. For the invasion assay, the isolates and strains used were detected inside the cells after infection of one minute. The invasions of the ureaplasmas surrounded the nuclear region but were not observed inside the nuclei. The gentamicin invasion assay detected that 1% to 10% of studied ureaplasmas were inside the infected cells.

<p>Имајући&nbsp; у&nbsp; виду&nbsp; значај&nbsp; површинских&nbsp; вода&nbsp; у продору&nbsp; и&nbsp; ширењу&nbsp; страних&nbsp; инвазивних врста,&nbsp; циљеви&nbsp; овог&nbsp; рада&nbsp; били&nbsp; су&nbsp; да&nbsp; се представи&nbsp; актуелно&nbsp; стање&nbsp; присуства&nbsp; и дистрибуције&nbsp; акватичних&nbsp; инвазивних биљних&nbsp; врста&nbsp; у&nbsp; површинским&nbsp; копненим водама,&nbsp; анализира&nbsp; степен&nbsp; инвазије рипаријалних&nbsp; зона&nbsp; на&nbsp; подручју&nbsp; Србије, одреди&nbsp; везаност&nbsp; анализираних&nbsp; инвазивних<br />врста&nbsp; за&nbsp; поједине&nbsp; типове&nbsp; станишта&nbsp; у рипаријалу река и канала и анализира утицај различитих&nbsp; станишних&nbsp; и&nbsp; антропогених фактора&nbsp; на&nbsp; присуство&nbsp; и&nbsp; абунданцу&nbsp; 26 одабраних&nbsp; инвазивних&nbsp; таксона.&nbsp; Теренска истраживања&nbsp; вршена&nbsp; су&nbsp; у&nbsp; периоду&nbsp; од&nbsp; 2013. до&nbsp; 2016.&nbsp; године.&nbsp; За&nbsp; потребе&nbsp; овог&nbsp; рада анализирано је 250 локалитета, од чега 217 у рипаријалу&nbsp; 39&nbsp; река&nbsp; и&nbsp; 33&nbsp; у&nbsp; рипаријалу&nbsp; шест<br />деоница&nbsp; канала&nbsp; хидросистема&nbsp; Дунав-Тиса-Дунав.&nbsp; Уз&nbsp; податке&nbsp; о&nbsp; бројности&nbsp; и<br />покровности&nbsp; заступљених&nbsp; биљних&nbsp; врста, сакупљани&nbsp; су&nbsp; и&nbsp; подаци&nbsp; о&nbsp; типу&nbsp; станишта&nbsp; (у складу&nbsp; са&nbsp; EUNIS&nbsp; класификацијом), релевантним&nbsp; физичким&nbsp; и<br />хидроморфолошким&nbsp; карактеристикама истраживаних&nbsp; водотокова&nbsp; и&nbsp; доминантним антропогеним&nbsp; утицајима&nbsp; на&nbsp; истраживаном локалитету&nbsp; (у&nbsp; складу&nbsp; са&nbsp; стандардним протоколом&nbsp; RHS&nbsp; методе).&nbsp; Статистичка обрада&nbsp; података&nbsp; вршена&nbsp; је&nbsp; у&nbsp; софтверу CANOCO&nbsp; 5.0,&nbsp; применом&nbsp; анализе&nbsp; главних компоненти&nbsp; (РСА),&nbsp; канонијскe коресподентнe&nbsp; анализе&nbsp; (CCA)&nbsp; и&nbsp; анализе редундантности&nbsp; (RDA).&nbsp; На&nbsp; територији Србије&nbsp; забележено&nbsp; је&nbsp; присуство&nbsp; осам акватичних&nbsp; инвазивних&nbsp; биљних&nbsp; врстаПрема&nbsp; броју&nbsp; налаза&nbsp; истичу&nbsp; се&nbsp; врсте Vallisneria spiralis, Azolla filiculoides и Elodea&nbsp; nuttallii.&nbsp; Примарно су заступљене у текућим,<br />у односу на стајаће воде, при чему се мрежа&nbsp; канала&nbsp; ХС&nbsp; ДТД&nbsp; и&nbsp; речни&nbsp; токови&nbsp; који&nbsp; припадају&nbsp; сливу&nbsp; Дунава&nbsp; могу&nbsp; сматрати&nbsp; основним&nbsp; акватичним&nbsp; коридорима&nbsp; њиховог&nbsp; ширења.&nbsp; Од&nbsp; 26&nbsp; таксона&nbsp; инвазивних&nbsp; биљака&nbsp; чије&nbsp; је&nbsp; присуство&nbsp; праћено&nbsp; и&nbsp; анализирано&nbsp; у&nbsp; рипаријалним&nbsp; подручјима,&nbsp; таксони&nbsp; са<br />највећим&nbsp; бројем&nbsp; налаза&nbsp; су&nbsp;<em> Xanthium&nbsp; strumarium</em>&nbsp; subsp.&nbsp; <em>italicum,&nbsp; Amorpha</em> <em>fruticosa, Erigeron&nbsp; canadensis,&nbsp; Robinia&nbsp; pseudoacacia&nbsp; и&nbsp; Echinochloa&nbsp; crus-galli.</em>&nbsp; Речни&nbsp; сливови Дунава,&nbsp; Јужне&nbsp; Мораве,&nbsp; Западне&nbsp; Мораве&nbsp; и&nbsp; Тимока&nbsp; истичу&nbsp; се&nbsp; по&nbsp; заступљености&nbsp; истраживаних таксона. На основу података о дистрибуцији,&nbsp; бројности&nbsp; и&nbsp; покровности&nbsp; анализираних&nbsp; таксона&nbsp; сливови&nbsp; Дунава, Колубаре и Западне Мораве издвојили су се&nbsp; као&nbsp; коридори&nbsp; ширења&nbsp; највећег&nbsp; броја<br />истраживаних&nbsp; таксона.&nbsp; Четири&nbsp; типа&nbsp; станишта&nbsp; издвојила&nbsp; су&nbsp; се&nbsp; према<br />заступљености&nbsp; инвазивних&nbsp; биљних&nbsp; врста.&nbsp; Таксони који су забележени у највећем броју&nbsp; типова&nbsp; станишта&nbsp; су:&nbsp; <em>Amorpha&nbsp; fruticosa, Erigeron&nbsp; canadensis,&nbsp;</em> <em>Robinia&nbsp; pseudoacacia, Echinochloa crus-galli&nbsp;</em> и&nbsp; <em>Xanthium strumarium subsp</em>.&nbsp; <em>italicum.</em>&nbsp; Као&nbsp; резултат&nbsp; нумеричких&nbsp; анализа&nbsp; издвојени&nbsp; су&nbsp; физички&nbsp; и<br />хидроморфолошки параметри истраживаних&nbsp; водотокова&nbsp; и&nbsp; доминантни&nbsp; антропогени&nbsp; утицаји који на истраживаном локалитетима утичу&nbsp; на&nbsp; присуство&nbsp; и&nbsp; абунданцу&nbsp; анализираних&nbsp; таксона.&nbsp; У&nbsp; условима&nbsp; глобалних&nbsp; климатских&nbsp; промена&nbsp; може&nbsp; се&nbsp; очекивати да ће се&nbsp; у наредном периоду под снажним&nbsp; притиском&nbsp; ширења&nbsp; и&nbsp; доминације&nbsp; инвазивних врста наћи сливови Саве, Велике Мораве&nbsp; и&nbsp; Јужне&nbsp; Мораве.&nbsp; У&nbsp; рипаријалним областима на југозападу наше земље (долине Лима,&nbsp; Ибра&nbsp; и&nbsp; њихових&nbsp; притока)&nbsp; као&nbsp; и&nbsp; рипаријалним&nbsp; зонама&nbsp; планинских&nbsp; и високопланинских&nbsp; области&nbsp; такође&nbsp; је очекивано&nbsp; да&nbsp; ће&nbsp; доћи&nbsp; до&nbsp; повећања&nbsp; броја присутних&nbsp; инвазивних&nbsp; врста&nbsp; и&nbsp; њиховог даљег ширења.</p> / <p>Imajući&nbsp; u&nbsp; vidu&nbsp; značaj&nbsp; površinskih&nbsp; voda&nbsp; u prodoru&nbsp; i&nbsp; širenju&nbsp; stranih&nbsp; invazivnih vrsta,&nbsp; ciljevi&nbsp; ovog&nbsp; rada&nbsp; bili&nbsp; su&nbsp; da&nbsp; se predstavi&nbsp; aktuelno&nbsp; stanje&nbsp; prisustva&nbsp; i distribucije&nbsp; akvatičnih&nbsp; invazivnih biljnih&nbsp; vrsta&nbsp; u&nbsp; površinskim&nbsp; kopnenim vodama,&nbsp; analizira&nbsp; stepen&nbsp; invazije riparijalnih&nbsp; zona&nbsp; na&nbsp; području&nbsp; Srbije, odredi&nbsp; vezanost&nbsp; analiziranih&nbsp; invazivnih<br />vrsta&nbsp; za&nbsp; pojedine&nbsp; tipove&nbsp; staništa&nbsp; u riparijalu reka i kanala i analizira uticaj različitih&nbsp; stanišnih&nbsp; i&nbsp; antropogenih faktora&nbsp; na&nbsp; prisustvo&nbsp; i&nbsp; abundancu&nbsp; 26 odabranih&nbsp; invazivnih&nbsp; taksona.&nbsp; Terenska istraživanja&nbsp; vršena&nbsp; su&nbsp; u&nbsp; periodu&nbsp; od&nbsp; 2013. do&nbsp; 2016.&nbsp; godine.&nbsp; Za&nbsp; potrebe&nbsp; ovog&nbsp; rada analizirano je 250 lokaliteta, od čega 217 u riparijalu&nbsp; 39&nbsp; reka&nbsp; i&nbsp; 33&nbsp; u&nbsp; riparijalu&nbsp; šest<br />deonica&nbsp; kanala&nbsp; hidrosistema&nbsp; Dunav-Tisa-Dunav.&nbsp; Uz&nbsp; podatke&nbsp; o&nbsp; brojnosti&nbsp; i<br />pokrovnosti&nbsp; zastupljenih&nbsp; biljnih&nbsp; vrsta, sakupljani&nbsp; su&nbsp; i&nbsp; podaci&nbsp; o&nbsp; tipu&nbsp; staništa&nbsp; (u skladu&nbsp; sa&nbsp; EUNIS&nbsp; klasifikacijom), relevantnim&nbsp; fizičkim&nbsp; i<br />hidromorfološkim&nbsp; karakteristikama istraživanih&nbsp; vodotokova&nbsp; i&nbsp; dominantnim antropogenim&nbsp; uticajima&nbsp; na&nbsp; istraživanom lokalitetu&nbsp; (u&nbsp; skladu&nbsp; sa&nbsp; standardnim protokolom&nbsp; RHS&nbsp; metode).&nbsp; Statistička obrada&nbsp; podataka&nbsp; vršena&nbsp; je&nbsp; u&nbsp; softveru CANOCO&nbsp; 5.0,&nbsp; primenom&nbsp; analize&nbsp; glavnih komponenti&nbsp; (RSA),&nbsp; kanonijske korespodentne&nbsp; analize&nbsp; (CCA)&nbsp; i&nbsp; analize redundantnosti&nbsp; (RDA).&nbsp; Na&nbsp; teritoriji Srbije&nbsp; zabeleženo&nbsp; je&nbsp; prisustvo&nbsp; osam akvatičnih&nbsp; invazivnih&nbsp; biljnih&nbsp; vrstaPrema&nbsp; broju&nbsp; nalaza&nbsp; ističu&nbsp; se&nbsp; vrste Vallisneria spiralis, Azolla filiculoides i Elodea&nbsp; nuttallii.&nbsp; Primarno su zastupljene u tekućim,<br />u odnosu na stajaće vode, pri čemu se mreža&nbsp; kanala&nbsp; HS&nbsp; DTD&nbsp; i&nbsp; rečni&nbsp; tokovi&nbsp; koji&nbsp; pripadaju&nbsp; slivu&nbsp; Dunava&nbsp; mogu&nbsp; smatrati&nbsp; osnovnim&nbsp; akvatičnim&nbsp; koridorima&nbsp; njihovog&nbsp; širenja.&nbsp; Od&nbsp; 26&nbsp; taksona&nbsp; invazivnih&nbsp; biljaka&nbsp; čije&nbsp; je&nbsp; prisustvo&nbsp; praćeno&nbsp; i&nbsp; analizirano&nbsp; u&nbsp; riparijalnim&nbsp; područjima,&nbsp; taksoni&nbsp; sa<br />najvećim&nbsp; brojem&nbsp; nalaza&nbsp; su&nbsp;<em> Xanthium&nbsp; strumarium</em>&nbsp; subsp.&nbsp; <em>italicum,&nbsp; Amorpha</em> <em>fruticosa, Erigeron&nbsp; canadensis,&nbsp; Robinia&nbsp; pseudoacacia&nbsp; i&nbsp; Echinochloa&nbsp; crus-galli.</em>&nbsp; Rečni&nbsp; slivovi Dunava,&nbsp; Južne&nbsp; Morave,&nbsp; Zapadne&nbsp; Morave&nbsp; i&nbsp; Timoka&nbsp; ističu&nbsp; se&nbsp; po&nbsp; zastupljenosti&nbsp; istraživanih taksona. Na osnovu podataka o distribuciji,&nbsp; brojnosti&nbsp; i&nbsp; pokrovnosti&nbsp; analiziranih&nbsp; taksona&nbsp; slivovi&nbsp; Dunava, Kolubare i Zapadne Morave izdvojili su se&nbsp; kao&nbsp; koridori&nbsp; širenja&nbsp; najvećeg&nbsp; broja<br />istraživanih&nbsp; taksona.&nbsp; Četiri&nbsp; tipa&nbsp; staništa&nbsp; izdvojila&nbsp; su&nbsp; se&nbsp; prema<br />zastupljenosti&nbsp; invazivnih&nbsp; biljnih&nbsp; vrsta.&nbsp; Taksoni koji su zabeleženi u najvećem broju&nbsp; tipova&nbsp; staništa&nbsp; su:&nbsp; <em>Amorpha&nbsp; fruticosa, Erigeron&nbsp; canadensis,&nbsp;</em> <em>Robinia&nbsp; pseudoacacia, Echinochloa crus-galli&nbsp;</em> i&nbsp; <em>Xanthium strumarium subsp</em>.&nbsp; <em>italicum.</em>&nbsp; Kao&nbsp; rezultat&nbsp; numeričkih&nbsp; analiza&nbsp; izdvojeni&nbsp; su&nbsp; fizički&nbsp; i<br />hidromorfološki parametri istraživanih&nbsp; vodotokova&nbsp; i&nbsp; dominantni&nbsp; antropogeni&nbsp; uticaji koji na istraživanom lokalitetima utiču&nbsp; na&nbsp; prisustvo&nbsp; i&nbsp; abundancu&nbsp; analiziranih&nbsp; taksona.&nbsp; U&nbsp; uslovima&nbsp; globalnih&nbsp; klimatskih&nbsp; promena&nbsp; može&nbsp; se&nbsp; očekivati da će se&nbsp; u narednom periodu pod snažnim&nbsp; pritiskom&nbsp; širenja&nbsp; i&nbsp; dominacije&nbsp; invazivnih vrsta naći slivovi Save, Velike Morave&nbsp; i&nbsp; Južne&nbsp; Morave.&nbsp; U&nbsp; riparijalnim oblastima na jugozapadu naše zemlje (doline Lima,&nbsp; Ibra&nbsp; i&nbsp; njihovih&nbsp; pritoka)&nbsp; kao&nbsp; i&nbsp; riparijalnim&nbsp; zonama&nbsp; planinskih&nbsp; i visokoplaninskih&nbsp; oblasti&nbsp; takođe&nbsp; je očekivano&nbsp; da&nbsp; će&nbsp; doći&nbsp; do&nbsp; povećanja&nbsp; broja prisutnih&nbsp; invazivnih&nbsp; vrsta&nbsp; i&nbsp; njihovog daljeg širenja.</p> / <p>Bearing&nbsp; in&nbsp; mind&nbsp; the&nbsp; significance&nbsp; of&nbsp; inland surface waters for the introduction and spread of invasive&nbsp; alien&nbsp; species&nbsp; (IAS),&nbsp; the&nbsp; aims&nbsp; of&nbsp; this research&nbsp; were&nbsp; to&nbsp; present&nbsp; the&nbsp; contemporary records&nbsp; of&nbsp; aquatic&nbsp; invasive&nbsp; plant&nbsp; species&nbsp; in surface&nbsp; waters,&nbsp; analyse&nbsp; the&nbsp; level&nbsp; of&nbsp; invasion&nbsp; of riparian&nbsp; areas&nbsp; in&nbsp; Serbia,&nbsp; determine&nbsp; the&nbsp; habitat preferences&nbsp; of&nbsp; the&nbsp; analysed&nbsp; IAS&nbsp; and&nbsp; determine the effects of different habitat characteristics and anthropogenic&nbsp; influences&nbsp; on&nbsp; the&nbsp; presence&nbsp; and bundance&nbsp; of&nbsp; the&nbsp; 26&nbsp; analysed&nbsp; invasive&nbsp; plants. Field&nbsp; research&nbsp; was&nbsp; conducted&nbsp; during&nbsp; the&nbsp; 2013-2016&nbsp; period.&nbsp; A&nbsp; total&nbsp; of&nbsp; 250&nbsp; field&nbsp; sites&nbsp; were analysed&nbsp; as&nbsp; part&nbsp; of&nbsp; this&nbsp; research,&nbsp; 217&nbsp; in&nbsp; the riparian zones of 39 rivers and 33 in the riparian zones&nbsp;&nbsp;&nbsp; of&nbsp; six&nbsp; canal&nbsp; sections&nbsp; of&nbsp; the&nbsp; hydrosystem Danube-Tisa-Danube&nbsp; (HS&nbsp; DTD).&nbsp; In&nbsp; addition&nbsp; to data on the cover and abundance of the analysed plant species, data on habitat type (following the EUNIS&nbsp; classification),&nbsp; relevant&nbsp; physical&nbsp; and hydromorhological&nbsp; parameters&nbsp; of&nbsp; the&nbsp; studied watercourses&nbsp; and&nbsp; dominant&nbsp; anthropogenic influences an analysed field sites were recorded (following&nbsp; the&nbsp; standard&nbsp; RHS&nbsp; protocol). Statistical&nbsp; data&nbsp; analysis&nbsp; was&nbsp; done&nbsp; in&nbsp; CANOCO 5.0&nbsp;&nbsp; software,&nbsp; using&nbsp; the&nbsp; principal&nbsp; component analysis&nbsp; (PCA),&nbsp; canonical&nbsp; correspondence analysis (CCA) and redundancy analysis (RDA). Eight&nbsp; aquatic&nbsp; invasive&nbsp; plant&nbsp; species&nbsp; were recorded for the territory of Serbia. Species with the highest number of records were:&nbsp; Vallisneria spiralis,&nbsp; Azolla filiculoides&nbsp; and&nbsp; Elodea nuttallii. They&nbsp; are&nbsp; primarily&nbsp; present&nbsp; in&nbsp; running,&nbsp; in comparison&nbsp; with&nbsp; standing&nbsp; waters,&nbsp; and&nbsp; the&nbsp; canal network&nbsp; of&nbsp; the&nbsp; HS&nbsp; DTD&nbsp; and&nbsp; rivers&nbsp; of&nbsp; the Danube catchment area are considered to be their main&nbsp; aquatic&nbsp; corridor&nbsp; of&nbsp; spread.&nbsp; Of&nbsp; the&nbsp; 26 invasive plant taxa whose presence&nbsp; was recorded and&nbsp; analysed&nbsp; in&nbsp; the&nbsp; riparian&nbsp; areas&nbsp; of&nbsp; Serbia,Xanthium strumarium&nbsp; subsp.&nbsp; italicum,&nbsp; Amorpha fruticosa,&nbsp; Erigeron&nbsp; canadensis,&nbsp; Robinia pseudoacacia&nbsp; and&nbsp; Echinochloa&nbsp; crus-&nbsp; galli&nbsp; were taxa&nbsp; with&nbsp; the&nbsp; highest&nbsp; number&nbsp; of&nbsp; records.Catchment&nbsp; areas&nbsp; of&nbsp; the&nbsp; Danube,&nbsp; Južna&nbsp; Morava, Zapadna&nbsp; Morava&nbsp; and&nbsp; Timok&nbsp; rivers&nbsp; were characterised&nbsp; by&nbsp; the&nbsp; highest&nbsp; levels&nbsp; of&nbsp; invasion. Based&nbsp; on&nbsp; their&nbsp; distribution&nbsp; data,&nbsp; cover&nbsp; and abundance of the analysed taxa, catchment areas of&nbsp; the&nbsp; Danube,&nbsp; Kolubara&nbsp; and&nbsp; Zapadna&nbsp; Morava rivers&nbsp; are&nbsp; highlighted&nbsp; as&nbsp; the&nbsp; main&nbsp; corridors&nbsp; of spread for the majority of the studied IAS. Four habitat&nbsp; types&nbsp; stand&nbsp; out&nbsp; by&nbsp; the&nbsp; number&nbsp; of analysed IAS. Taxa which were recorded in the highest&nbsp; number&nbsp; of&nbsp; habitat&nbsp; types&nbsp; are:&nbsp; Amorpha fruticosa,&nbsp; Erigeron&nbsp; canadensis,&nbsp; Robinia pseudoacacia,&nbsp; Echinochloa&nbsp; crus-galli&nbsp; and Xanthium strumarium subsp. italicum.&nbsp; A number of&nbsp; physical&nbsp; and&nbsp; hydromorphological&nbsp; parameters and&nbsp; dominant&nbsp; anthropogenic&nbsp; influences&nbsp; of&nbsp; the studied&nbsp; field&nbsp; sites&nbsp; were&nbsp; shown&nbsp; to&nbsp; affect&nbsp; the presence&nbsp; and&nbsp; abundance&nbsp; of&nbsp; the&nbsp; studied&nbsp; invasive plants.&nbsp; The catchment areas of the Sava, Velika Morava&nbsp; and&nbsp; Južna&nbsp; Morava&nbsp; rivers&nbsp; are&nbsp; excpected to&nbsp; be&nbsp; under&nbsp; a&nbsp; strong&nbsp; pressure&nbsp; of&nbsp; the&nbsp; spread&nbsp; and dominance&nbsp; of&nbsp; IAS&nbsp; unnder&nbsp; the&nbsp; conditions&nbsp; of global&nbsp; climate&nbsp; changes.&nbsp; Riparian&nbsp; areas&nbsp; in&nbsp; the southwestern parts of Serbia (valleys of&nbsp; Lim and Ibar&nbsp; rivers&nbsp; and&nbsp; their&nbsp; tributaries)&nbsp; and&nbsp; riparian zones&nbsp; in&nbsp; the&nbsp; mountain&nbsp; and&nbsp; high- mountain&nbsp; areas are also expected to experience an increase in the number&nbsp; of&nbsp; invasive&nbsp; species&nbsp; and&nbsp; their&nbsp; spread rates.</p>

This thesis is a broadly chronological study of representations of Japan and the Japanese in Australian novels, stories and memoirs from the late nineteenth century to the twenty-first century. Adopting Edward Said???s Orientalist notion of the `Other???, it attempts to elaborate patterns in which Australian authors describe and evaluate the Japanese. As well as examining these patterns of representation, this thesis outlines the course of their development and change over the years, how they relate to the context in which they occur, and how they contribute to the formation of wider Australian views on Japan and the Japanese. The thesis considers the role of certain Australian authors in formulating images and ideas of the Japanese ???Other???. These authors, ranging from fiction writers to journalists, scholars and war memoirists, act as observers, interpreters, translators, and sometimes ???traitors??? in their cross-cultural interactions. The thesis includes work from within and outside ???mainstream??? writings, thus expanding the contexts of Australian literary history. The major ???periods??? of Australian literature discussed in this thesis include: the 1880s to World War II; the Pacific War; the post-war period; and the multicultural period (1980s to 2000). While a comprehensive examination of available literature reveals the powerful and continuing influence of the Pacific War, images of ???the stranger???, ???the enemy??? and later ???the ally??? or ???partner??? are shown to vary according to authors, situations and wider international relations. This thesis also examines gender issues, which are often brought into sharp relief in cross-cultural representations. While typical East-West power-relationships are reflected in gender relations, more complex approaches are also taken by some authors. This thesis argues that, while certain patterns recur, such as versions of the ???Cho-Cho-San??? or ???Madame Butterfly??? story, Japan-related works have given some Australian authors, especially women, opportunities to reveal more ???liberated??? viewpoints than seemed possible in their own cultural context. As the first extensive study of Japan in Australian literary consciousness, this thesis brings to the surface many neglected texts. It shows a pattern of changing interests and interactions between two nations whose economic interactions have usually been explored more deeply than their literary and cultural relations.