Dentre as 18 espécies do gênero Yersinia, as espécies Y. enterocolitica, Y. pseudotuberculosis e Y. pestis foram extensivamente caracterizadas em diversos aspectos como ecologia, epidemiologia e mecanismos de patogenicidade. Sete das 15 espécies restantes (Y. aldovae, Y. bercovieri, Y. frederiksenii, Y. intermedia, Y. kristensenii, Y. mollaretii e Y. rohdei), usualmente conhecidas como Y. enterocolitica-like, até o momento, não tiveram seu potencial patogênico caracterizado e são, geralmente, consideradas não-patogênicas. Entretanto, dados da literatura sugerem que algumas dessas espécies possam causar doença. Esses dados estimularam o surgimento de questões sobre os mecanismos pelos quais as espécies de Y. enterocolitica-like possam interagir com as células do hospedeiros e causar doenças. Esse projeto teve como principal objetivo caracterizar o potencial patogênico de linhagens de Y. enterocolitica-like, especificamente das espécies Y. frederiksenii, Y. kristensenii e Y. intermedia. No presente trabalho, o potencial patogênico de 118 linhagens de Y. enterocolitica-like (50 Y. frederiksenii, 55 Y. intermedia e 13 Y. kristensenii) foi avaliado pela pesquisa da presença dos genes relacionados à virulência ail, fepA, fepD, fes, hreP, myfA, tccC, ystA, ystB e virF por PCR. Além disso, a habilidade de algumas linhagens de Yersinia de aderir e invadir células Caco-2 e HEp-2 após diferentes períodos de incubação, bem como, de sobreviver no interior de macrófagos humanos U937 foi testada. Aspectos morfológicos da adesão bacteriana foram visualizados por microscopia eletrônica. Finalmente, a presença de possíveis novos mecanismos de virulência foi avaliada a partir do sequenciamento de RNA de uma linhagem de Y. enterocolitica-like. As linhagens estudadas apresentaram os seguintes genes: Y. frederiksenii, fepA (44%), fes (44%) e ystB (18%); Y. intermedia, ail (53%), fepA (35%), fepD (2%), fes (97%), hreP (2%), ystB (2%) e tccC (35%); e Y. kristensenii, ail (62%), ystB (23%), fepA (77%), fepD (54%), fes (54%) e hreP (54%). De modo geral, as linhagens de Y. enterocolitica-like tiveram a habilidade de aderir e invadir células Caco-2 e HEp-2 inferior à da linhagem altamente patogênica Y. enterocolitica 8081. Contudo, Y. kristensenii FCF 410 e Y. frederiksenii FCF 461 apresentaram elevado potencial de invasão a células Caco-2 após cinco dias de pré-incubação, os quais foram 45 e 7,2 vezes maiores do que o controle Y. enterocolitica 8081, respectivamente, porém, o gene ail não foi detectado nessas linhagens. O ensaio de sobrevivência em macrófagos humanos U937 ii mostrou que as linhagens de Y. frederiksenii FCF 461 (40,0%) e Y. frederiksenii FCF 379 (24,6%) tiveram porcentagens de sobrevivência superior à de Y. enterocolitica 8081 (13,4%). Todavia, linhagens de Y. intermedia e Y. kristensenii apresentaram uma capacidade reduzida de sobreviver em macrófagos. A microscopia eletrônica de varredura mostrou as bactérias em contato com a filipódia celular. As bactérias foram distribuídas tanto individualmente quanto em pequenos aglomerados. Portanto, podemos concluir que a presença dos genes relacionados à virulência encontrados nas Y. enterocolitica-like estudadas indicou o possível potencial patogênico de algumas dessas linhagens. Os ensaios de adesão e invasão a células de mamíferos sugerem que a patogenicidade de Y. kristensenii e Y. frederiksenii possa ser linhagem-dependente. O ensaio de sobrevivência em macrófagos humanos U937 evidenciou o potencial patogênico de algumas linhagens de Y. frederiksenii. Em conjunto, os resultados obtidos sugerem a existência de mecanismos de virulência alternativos aos mecanismos clássicos descritos para Y. enterocolitica patogênica. Contudo, a presença de possíveis novos mecanismos de virulência não pode ser verificada, uma vez que a plataforma 454 GS Junior (Roche) não se mostrou adequada para a realização de sequenciamento de RNA de amostras provenientes de interações com células devido à baixa cobertura obtida. / Among the 18 species of the Yersinia genus, Y. enterocolitica, Y. pseudotuberculosis and Y. pestis were extensively characterized in different subjects as ecology, epidemiology and pathogenicity mechanisms. Seven among the remaining 15 species (Y. aldovae, Y. bercovieri, Y. frederiksenii, Y. intermedia, Y. kristensenii, Y. mollaretii and Y. rohdei), often called Y. enterocolitica-like have not their pathogenic potential characterized and are usually considered to be nonpathogenic. However, literature data suggest that some of these species can cause diseases. These data stimulate the upsurge of questions about the mechanisms of which Y. enterocolitica-like species may interact with host cells and cause diseases. The main objective of this preject was to characterize the pathogenic potential of Y. enterocolitica-like strains, specifically of the species Y. frederiksenii, Y. kristensenii and Y. intermedia. This work evaluated the pathogenic potential of 118 Y. enterocolitica-like strains (50 Y. frederiksenii, 55 Y. intermedia and 13 Y. kristensenii) searching for the presence of the virulence-related genes ail, fepA, fepD, fes, hreP, myfA, tccC, ystA, ystB and virF by PCR. Besides, Yersinia strains ability of adhesion and invasion to Caco-2 and HEp-2 cells after different pre-incubation periods, and its survival within human macrophages U937 were tested. Morphologic aspects of bacterial adhesion were observed by scanning electronic microscopy. Finally, the presence of new possible virulence mechanisms was evaluated through RNA sequencing of one Y. enterocolitica-like strain. The studied strains showed the following genes: Y. frederiksenii, fepA (44%), fes (44%) and ystB (18%); Y. intermedia, ail (53%), fepA (35%), fepD (2%), fes (97%), hreP (2%), ystB (2%) and tccC (35%); and Y. kristensenii, ail (62%), ystB (23%), fepA (77%), fepD (54%), fes (54%) and hreP (54%). Usually Y. enterocolitica-like strains presented less ability of adhere and invade Caco-2 and HEp-2 cells than the highly pathogenic strain Y. enterocolitica 8081. On the other hand, Y. kristensenii FCF 410 and Y. frederiksenii FCF 461 showed high potential of invasion in Caco-2 cells after 5 days of pre-incubation, which were 45 and 7.2 times higher than the control Y. enterocolitica 8081 respectively, but ail gene was not found in these strains. Survival assay in human macrophages U937 showed that Y. frederiksenii FCF 461 (40.0%) and Y. frederiksenii FCF 379 (24.6%) strains presented survival percentages higher than Y. enterocolitica 8081 (13.4%). However, Y. intermedia and Y. iv kristensenii strains showed a reduced capability of surviving in macrophages. Scanning electron microscopy showed bacteria at the surface in contact with the cellular filopodia. The bacteria were distributed either individually or in small clumps. Therefore, it may be concluded that the presence of virulence-related genes in some of the Y. enterocolitica-like strains indicated their possible pathogenic potential. Mammal cells adhesion and invasion assays suggest that the pathogenicity of Y. kristensenii and Y. frederiksenii may be strain-dependent. Human macrophages U937 surviving assay highlighted the pathogenic potential of some Y. frederiksenii strains. Together, the results suggest the existence of alternative virulence mechanisms other than the classical mechanisms described for pathogenic Y. enterocolitica. However, we could not verify the presence of possible new virulence mechanisms because 454 GS Junior (Roche) platform was not suitable for RNA sequencing of strains from cells interaction due its low coverage obtained.

O câncer de mama é o tipo de câncer mais comumente detectado em mulheres de todo o mundo. Na maioria das pacientes, a causa de morte se deve, principalmente, à doença metastática que pode se desenvolver a partir do tumor primário. O processo metastático envolve uma complexa cascata de eventos, incluindo a quebra organizada dos componentes da matriz extracelular por metaloproteinases de matriz (MMPs). A atividade das MMPs é precisamente regulada por inibidores específicos, os inibidores teciduais das MMPs (TIMPs). Dado seu papel na progressão tumoral, níveis elevados de MMPs têm sido associados com prognóstico desfavorável para pacientes com câncer. Por outro lado, sendo os TIMPs proteínas multifuncionais, níveis elevados de TlMP-1 e de TIMP-2 correlacionam com agressividade do tumor e prognóstico ruim em diferentes tipos de câncer, incluindo o câncer de mama. O gene supressor de metástase RECK codifica uma glicoproteína de membrana capaz de inibir a invasão e a metástase tumoral através da regulação negativa da atividade de MMPs envolvidas em carcinogênese: MMP-2, MMP-9 e MMP-14 (MT1-MMP). A fim de analisar o papel das MMPs e de seus inibidores (TIMPs e RECK) na progressão tumoral do câncer de mama, o perfil de expressão destes genes foi detectado, através de ensaios de Real-Time PCR, em um painel de cinco linhagens celulares de carcinoma de mama humano com diferentes potenciais invasivos e metastáticos e em 72 amostras teciduais de tumores primários de mama e 30 amostras teciduais de borda normal adjacente ao tumor. O perfil de expressão protéica de RECK foi avaliado em 236 amostras de tumores primários de mama através de ensaios de Tissue Microarray. Além disso, a atividade proteolítica das MMPs foi detectada em ensaios de Zimografia. Os resultados obtidos indicam que a progressão do câncer de mama humano está relacionada com um aumento dos níveis de expressão das MMPs e de seus inibidores específicos. O aumento dos níveis de expressão dos TIMPs parece estar relacionado ao seu papel como proteína multifuncional que pode estar funcionando de maneira a promover, mais do que suprimir, a progressão tumoral. Níveis elevados da expressão protéica de RECK estão associados com pior prognóstico. No entanto, para pacientes em estádios clínicos avançados, altos níveis de expressão de RECK podem estar correlacionados com melhor prognóstico, dependendo do balanço MMP/inibidor. Os níveis de expressão das MMPs apresentaram correlação positiva em relação aos níveis de expressão de seus inibidores específicos, sugerindo a existência de fatores e vias de sinalização comuns envolvidas na regulação coordenada destes genes. Além disso, a síntese do inibidor pode estar relacionada a uma resposta celular ao aumento da expressão e atividade de proteases. O balanço transcricional enzima/inibidor favorece a enzima nas amostras tumorais e, de modo contrário, o inibidor específico nas amostras de borda normal, sugerindo o balanço como o principal fator na determinação da degradação da MEC em processos invasivos e metastáticos. Os resultados obtidos podem contribuir para um melhor entendimento da complexidade dos mecanismos envolvidos na metástase do câncer de mama. / Breast cancer is among the most common tumors affecting women. Like most solid tumors, metastatic disease rather than the primary tumor itself is responsible for death. The metastatic process involves a complex cascade of events, including the organized breakdown of the extracellular matrix by matrix metalloproteinases (MMPs). The activity of these proteases is tightly regulated by specific inhibitors, known as tissue inhibitors of MMPs (TIMPs). Consistent with their role in tumor progression, high levels of a number of MMPs have been shown to correlate with poor prognosis in human cancers. On the other hand, TIMPs are multifunctional molecules with high levels of TIMP-1 and TIMP-2 having been shown to predict adverse prognosis and correlate with tumor aggressiveness in several different human cancers, including breast cancer. The RECK metastasis suppressor gene encodes a membrane-associated MMP regulator protein that is able to suppress tumor invasion and metastasis by negatively regulating MMPs involved in carcinogenesis, namely: MMP-2, MMP-9 and MMP-14 (MT1-MMP). In order to analyse the role of these genes in breast cancer progression, the expression levels of MMPs and theirs inhibitors were detected by Real Time PCR in a panel of five human breast cancer cell lines displaying different degrees of invasiveness and metastatic potential and in 72 primary breast cancer and 30 adjacent normal tissue specimens. The RECK protein expression profile was also examined in 236 primary breast cancer tissue specimens by Tissue Microarray technology. The proteolytic activity of MMPs was examined by Zymography. The results suggest that high expression levels of MMPs and their inhibitors are correlated with breast cancer progression. High levels of TIMP transcript may be involved in tumor-promoting activity as a result of their multifunctional role. Increased levels of the RECK protein are correlated with poor prognosis for the patient. However, high levels of RECK would be expected to confer a favorable prognosis to patients with advanced disease. The expression levels of MMPs significantly correlated with the levels of TIMPs and may be explained by coordinate correlation of these molecules or, alternatively, the synthesis of an inhibitor may be a cellular reaction to the presence of the protease. The enzyme/inhibitor balance at the transcriptional level favors the enzyme in tumor tissue and the inhibitor in adjacent normal tissue. It is probably the parameter that will determine the matrix degradation at invasion and metastatic process. Our results are likely to contribute for better understanding of the complex mechanisms involved in breast cancer metastasis.

Abstract Between October 1999 and October 2003, 30 adult and 48 young (< 1 year) raccoon dogs (Nyctereutes procyonoides) were monitored using radio-telemetry in an area of Germany which has been occupied by this invasive alien species since the early 1990s. Additionally, three pairs of raccoon dogs were observed by continuous radio-tracking during the first six weeks after parturition in 2003. Furthermore 136 raccoon dog pubs were ear-tagged between June 1999 and August 2006. No adult animals dispersed from the area during the study period and home ranges tended to be used for several years, probably for life. The average annual home range size, calculated using 95% fixed kernel, was 382.2 ha ± 297.4 SD for females (n = 30 seasonal home ranges) and 352.4 ha ± 313.3 SD for males (n = 32 seasonal home ranges). Paired raccoon dogs had home ranges of similar size, with pair sharing the same area all year round. Raccoon dogs occupied large core areas (85% kernel) covering 81.2% of their home ranges. The home ranges were at their smallest during the mating season. The slightly larger size of home ranges in winter suggests that, due to the temperate climate, raccoon dogs do not hibernate in Germany. Males and females formed a long-term (probably lifelong) pair bond. Same-sex neighbours ignored each other and even adjacent males/females showed neither preference nor avoidance. Thus, it can be assumed that the raccoon dog in Central Europe is monogamous without exclusive territories, based on the results of home range overlap analysis and interaction estimations. Habitat composition within home ranges and within the whole study area was almost equal. Although, percentage shares of farmland and meadow was 16.35% smaller and 12.06% higher within the home ranges, respectively. All nine habitat types (farmland, forest, settlement, water, meadows, maize fields, small woods, reeds and hedges) were used opportunistically by raccoon dogs. No significant, recognisable difference for habitat preferences between seasons was detected. Male and female raccoon dog showed equal habitat preference pattern. A comparison of active and inactive locations in different habitats found no remarkable differences. Habitat composition of individual home ranges was used to classify animals. If the percentage of forest within a home range exceeded 50% the individual was classified as a ‘forest type’ raccoon dog. If the percentage of forest habitats within a home range was less than 5%, the share of pastureland was mean 81.82% ± 16.92 SD. Consequently the individual was classified as a ‘agrarian type’ raccoon dog. Neither habitat preference nor habitat selection process differed between the two ‘types’. Habitat use and preference is discussed with relation to the ability of the raccoon dog to expand its range towards Western Europe. Males spent noticeably more time (40.5% of the time ±11.7 SD) alone with the pups than females (16.4% of the time ±8.5 SD). Females had noticeably larger 95% kernel home ranges (98.24 ha ±51.71 SD) than males (14.73 ha ±8.16 SD) and moved much longer daily distances (7,368 m ±2,015 SD) than males (4,094 m ±2,886 SD) in six weeks postpartum. The raccoon dogs being studied left the breeding den in the 6th week after the birth of the pups. In situ video observation showed that the male carried prey to the den to provide the female and the litter with food. A clear division of labour took place among parents during the period in which the pups were nursed: males guarded the litter in the den or in close vicinity of it, while the females foraged to satisfy their increased energy requirements. There were relocations of 59 (43.4%) ear-tagged young racoon dogs and mean distance from marking point was 13.5 km ±20.1 SD. Dispersal mortality rate was 69.5% among young raccoon dogs. Most animals (55.9%) were recovered nearer than 5 km from the marking point, whereas only 8.5% relocations were recorded further than 50 km from the marking point. There was no difference in the distances of relocations between sexes. Most (53.7%) relocations of ear-tagged young raccoon dogs were in August and September and, only 34.1% were recorded from October to April. Hunting (55 %) and traffic (27 %) were the major mortality factors. Radio-collared young raccoon dogs generally dispersed between July and September. The mean natal home range size (MCP 100%) with and without excursions was 502.6 ha ±66.4 SD (n = 9) and 92.1 ha ±66.4 SD (n = 17), respectively. There were no differences between sexes in the month of dispersal. The direction of travel for dispersing animals appeared to be random, with distances from 0.5 km to 91.2 km. A highly flexible dispersing behaviour is certainly one of the reasons which contribute to the high expansion success of the species.

Between October 1999 and October 2003, 30 adult and 48 young (< 1 year) raccoon dogs (Nyctereutes procyonoides) were monitored using radio-telemetry in an area of North-East Germany which has been occupied by this invasive alien species since the early 1990s. Additionally, three pairs of raccoon dogs were observed by continuous radio-tracking during the first six weeks after parturition in 2003. Furthermore 136 raccoon dog pubs were ear-tagged between June 1999 and August 2006. No adult animals dispersed from the area during the study period and home ranges tended to be used for several years, probably for life. The average annual home range size, calculated using 95% fixed kernel, was 382.2 ha ± 297.4 SD for females (n = 30 seasonal home ranges) and 352.4 ha ± 313.3 SD for males (n = 32 seasonal home ranges). Paired raccoon dogs had home ranges of similar size, with pair mates sharing the same area all year round. Raccoon dogs occupied large core areas (85% kernel) covering 81.2% of their home ranges. The home ranges were at their smallest during the mating season. The slightly larger size of home ranges in winter suggests that, due to the temperate climate, raccoon dogs do not hibernate in Germany. Males and females formed a long-term (probably lifelong) pair bond. Same-sex neighbours ignored each other and even adjacent males/females showed neither preference nor avoidance. Thus, it can be assumed that the raccoon dog in Central Europe is monogamous without exclusive territories, based on the results of home range overlap analysis and interaction estimations. Habitat composition within home ranges and within the whole study area was almost equal. Although, percentage shares of farmland and meadow was 16.35% smaller and 12.06% higher within the home ranges, respectively. All nine habitat types (farmland, forest, settlement, water, meadows, maize fields, small woods, reeds and hedges) were used opportunistically by raccoon dogs. No significant, recognisable difference for habitat preferences between seasons was detected. Male and female raccoon dog showed equal habitat preference pattern. A comparison of active and inactive locations in different habitats found no remarkable differences. Habitat composition of individual home ranges was used to classify animals. If the percentage of forest within a home range exceeded 50% the individual was classified as a ‘forest type’ raccoon dog. If the percentage of forest habitats within a home range was less than 5%, the share of pastureland was mean 81.82% ± 16.92 SD. Consequently the individual was classified as a ‘agrarian type’ raccoon dog. Neither habitat preference nor habitat selection process differed between the two ‘types’. Habitat use and preference is discussed with relation to the ability of the raccoon dog to expand its range towards Western Europe. Males spent noticeably more time (40.5% of the time ±11.7 SD) alone with the pups than females (16.4% of the time ±8.5 SD). Females had noticeably larger 95% kernel home ranges (98.24 ha ±51.71 SD) than males (14.73 ha ±8.16 SD) and moved much longer daily distances (7,368 m ±2,015 SD) than males (4,094 m ±2,886 SD) in six weeks postpartum. The raccoon dogs being studied left the breeding den in the 6th week after the birth of the pups. In situ video observation showed that the male carried prey to the den to provide the female and the litter with food. A clear division of labour took place among parents during the period in which the pups were nursed: males guarded the litter in the den or in close vicinity of it, while the females foraged to satisfy their increased energy requirements. There were relocations of 59 (43.4%) ear-tagged young raccoon dogs and mean distance from marking point was 13.5 km ±20.1 SD. Dispersal mortality rate was 69.5% among young raccoon dogs. Most animals (55.9%) were recovered nearer than 5 km from the marking point, whereas only 8.5% relocations were recorded further than 50 km from the marking point. There was no difference in the distances of relocations between sexes. Most (53.7%) relocations of ear-tagged young raccoon dogs were in August and September and, only 34.1% were recorded from October to April. Hunting (55 %) and traffic (27 %) were the major mortality factors. Radiocollared young raccoon dogs generally dispersed between July and September. The mean natal home range size (MCP 100%) with and without excursions was 502.6 ha ±66.4 SD (n = 9) and 92.1 ha ±66.4 SD (n = 17), respectively. There were no differences between sexes in the month of dispersal. The direction of travel for dispersing animals appeared to be random, with distances from 0.5 km to 91.2 km. A highly flexible dispersing behaviour is certainly one of the reasons which contribute to the high expansion success of the species.

The diploma thesis focuses on changes of the life of the German minority that was living in the Czechoslovak Socialist Republic at the turn of the 1960s and 1970s, in connection with the then press. The thesis deals with an influence of the reviving process, which is called Prague Spring and then the period of normalization on the progress of status of the German minority in the Czechoslovakia. The thesis presents events of the year 1968, that are related to the national minorities policy which led to reception of the Constitutional Act No. 144/1968 Coll., about the status of the national groups in the state. I also analyse the activisation of the members of the German minority for the purpose of formation of their cultural association named "Kulturní sdružení občanů ČSSR německé národnosti" (the Cultural association of the citizen of the German nationality in the Czechoslovak Socialist Republic), which was enabled on the ground of the aforesaid constitutional law. The text also deals with the start of the period of normalization and its consequences for the German minority. The thesis also analyses the Czechoslovak media at the turn of the 1960s and 1970s with special regard to the only periodical press in the German language called the "Prager Volkszeitung". A very important part of the text is...

This film is an autobiographical documentary which tells the story of the process of documenting the filmmaker's trip to his land of heritage. As his plans for his journey and film begin to go awry, he begins to question the entire process of trying to connect with nation and place.

Human related activities have influenced the rivers of the southern Western Cape since as early as the 1700’s. As there is no detailed information available on ecological status of the Holsloot River, a tributary of the Breede River, this study aimed to gain insight into the effect of impacts associated with human activities on the habitat integrity of this river. The study intended to understand how seasonal changes, catchment characteristics and events are reflected in the ecological status of habitats along the river by applying bio-monitoring and river health measurements at selected sites in the upper, middle and lower reaches of the Holsloot River and compare the results to that of an undisturbed reference site. Results obtained in this study are compared with data gathered in 2008/2009 to determine if the ecological status of the river had changed in the period between the two sampling times. The study included assessment of the ecological status of the river based on standard bio-monitoring protocol (SASS5, IHI, IHAS and VEGRAI) as well as in situ water quality analysis (pH, dissolved oxygen, electrical conductivity and total dissolved solids). The construction of the instream Stettynskloof Dam changed the configuration of the riparian zone and river channel in the upper catchment area. Agricultural- and other human related activities, with consequent water abstraction, non-point-source pollution, loss of riparian vegetation, as well as dense stands of alien invader plants influence flow patterns and affects river ecology, especially in the dry summer months. Providing sufficient stream flow and adequate water levels, human related activities can create a larger variety of habitat types available that can support larger biodiversity and higher productivity. The level of inundation and stream flow, influenced by water abstraction as well as irrigation return-flow from extensive drainage systems especially in the dry months, contribute to the loss of biodiversity in the middle and lower reaches of the river. Where the upper reaches of the river are largely natural with few modifications, the habitat integrity deteriorates in the middle reaches so much so that ecosystem functioning are collectively impaired in lower reaches due to human related impacts. Sensitive macro-invertebrates found at lower seriously impacted parts of the river however, were in all probability washed down from lower impacted upstream habitats and may expectedly be able to again occupy habitats downstream if water quality and habitat availability improves. / Environmental Sciences / M. Sc. (Environmental Science)

This project investigates Russian nationalist organisations during the invasion of Ukraine, to study their collective identity and practises. It features an ethnography of 26 organisations, their websites and social media. Their content is coded via a semiotic framework, categorising their symbols, images, content posted and other identity tropes. It is then analysed by applying political, sociological and communication theories. The study’s background is the 2022 Russian invasion of Ukraine, with its dynamics of war and cultural struggle. The results are that Russian collective identities simplified their communication already by the end of March 2022, abandoning many of the tropes they initially supported, as well as their symbols, limiting themselves to attribute the label of Nazism to Ukraine and its government, avoiding the promotion of frontline volunteer activities and considering Russia’s struggle as global. Finally, most continued to evaluate the invasion of Ukraine as proceeding positively, and at the time half the organisations referred to it as special military operation.

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No description available.