Yes / Virus-encoded miRNAs are emerging as key regulators of persistent infection and host-cell immune evasion. Merkel cell polyomavirus (MCPyV), the predominant aetiological agent of Merkel cell carcinoma (MCC), encodes a single miRNA, MCV-miR-M1, which targets the oncogenic MCPyV large T antigen (LT). MCV-miR-M1 has previously been shown to play an important role in establishment of long-term infection, however, the underlying mechanism is not fully understood. A key unanswered question is whether, in addition to auto-regulating LT, MCV-miR-M1 also targets cellular transcripts to orchestrate an environment conducive for persistent infection. To address this, we adopted an RNA-Seq-based approach to identify cellular targets of MCV-miR-M1. Intriguingly, bioinformatics analysis of transcripts that are differentially expressed in cells expressing MCV-miR-M1 revealed several genes implicated in immune evasion. Subsequent target validation led to the identification of the innate immunity protein, SP100, as a direct target of MCV-miR-M1. Moreover, MCV-miR-M1-mediated modulation of SP100 was associated with a significant decrease in CXCL8 secretion, resulting in the attenuation of neutrophil chemotaxis towards Merkel cells harbouring synthetic MCPyV. Based on these observations we propose that MCV-miR-M1 targets key immune response regulators to help facilitate persistent infection, which is a pre-requisite for cellular transformation in MCC. / Funded in part by a University of Bradford studentship to PA and a Royal Society research award to JRB.

Os poliomavírus compõem uma grande família de vírus que causam infecções primárias geralmente na infância, e se mantem em condições subclínicas. Em situações de imunossupressão podem causar algumas doenças. Os indivíduos com HIV/AIDS frequentemente apresentam deficiência imunológica e por isso podem exibir maior risco de doenças causadas pelos poliomavírus. A utilização da saliva no diagnóstico e acompanhamento de doenças infecciosas tem sido explorado na literatura. As vantagens de usar a saliva para rastreio se pautam especialmente na coleta não invasiva e segurança no manuseio. O presente estudo teve como objetivo, detectar e quantificar o DNA dos poliomavírus BKV, JCV, de células Merkel e TSV, em fluídos orais (saliva, lavado bucal e fluído gengival crevicular) e comparar com a detecção em soro e urina, meios usualmente utilizados para detecção. Foram coletadas 299 amostras de 42 indivíduos, sendo 22 HIV positivos (GE) e 20 pacientes controle (GC). No GE, 63,6% dos pacientes apresentaram positividade para JCV em pelo menos uma amostra analisada, 54,5% foram positivos para BKV, 18,2% para células Merkel e não houve amostra positiva para TSV. No GC, 45% exibiu positividade para o JCV em pelo menos uma amostra analisada, 80% para BKV e nenhuma participante controle exibiu positividade para células Merkel e TSV. Não houve diferença de frequência de detecção viral entre os grupos estudados em relação às amostras coletadas, ou ainda em relação à idade ou sexo. Entretanto, nas amostras de fluídos orais houve maior prevalência de detecção para o BKV e para células Merkel. Concluímos que fluídos orais, especialmente saliva e lavado bucal, podem ser usados para o rastreamento do BK e JC; e que os indivíduos HIV positivos, sob tratamento antirretroviral não exibem frequências maior de poliomavírus, comparativamente a indivíduos controle. / Polyomavirus is one of the large family of viruses that cause primary infections usually in childhood, and can remain subclinical. In immunosuppression may cause some diseases. Individuals with HIV/AIDS often have immune deficiencies and may be at increased risk for diseases caused by polyomaviruses. The use of saliva in the diagnosis and follow-up of infectious diseases has been explored in the literature. The advantages of using saliva for screening are based on non-invasive collection and handling safety. The aim of present study was to detect and quantify the DNA from BKV, JCV, Merkel cell and TSV polyomaviruses in oral fluids (saliva, mouthwash and gingival crevicular fluid) and to compare it with serum and urine detection, the means usually used for detection. A total of 299 samples were collected from 42 individuals, 22 HIV positive (GE) and 20 control patients (GC). In GE, 63,6% of the patients presented positive for JCV in at least one sample analyzed, 54,5% were positive for BKV, 18,2% for Merkel cell and there was no positive sample for TSV. In GC, 45% showed JCV positivity in at least one analyzed sample, 80% in BKV, and no control participant exhibited positivity for Merkel cell and TSV. There was no difference in the frequency of viral detection among the groups studied in relation to the samples collected, or in relation to age or gender. However, in oral fluid samples there was a higher prevalence of detection for BKV and Merkel cell. We conclude that oral fluids, especially saliva and mouthwash, can be used for the screening of BK e JC; and that HIV positive individuals under antiretroviral treatment do not exhibit higher frequencies of polyomavirus compared to healthy control subjects.

Os poliomavírus compõem uma grande família de vírus que causam infecções primárias geralmente na infância, e se mantem em condições subclínicas. Em situações de imunossupressão podem causar algumas doenças. Os indivíduos com HIV/AIDS frequentemente apresentam deficiência imunológica e por isso podem exibir maior risco de doenças causadas pelos poliomavírus. A utilização da saliva no diagnóstico e acompanhamento de doenças infecciosas tem sido explorado na literatura. As vantagens de usar a saliva para rastreio se pautam especialmente na coleta não invasiva e segurança no manuseio. O presente estudo teve como objetivo, detectar e quantificar o DNA dos poliomavírus BKV, JCV, de células Merkel e TSV, em fluídos orais (saliva, lavado bucal e fluído gengival crevicular) e comparar com a detecção em soro e urina, meios usualmente utilizados para detecção. Foram coletadas 299 amostras de 42 indivíduos, sendo 22 HIV positivos (GE) e 20 pacientes controle (GC). No GE, 63,6% dos pacientes apresentaram positividade para JCV em pelo menos uma amostra analisada, 54,5% foram positivos para BKV, 18,2% para células Merkel e não houve amostra positiva para TSV. No GC, 45% exibiu positividade para o JCV em pelo menos uma amostra analisada, 80% para BKV e nenhuma participante controle exibiu positividade para células Merkel e TSV. Não houve diferença de frequência de detecção viral entre os grupos estudados em relação às amostras coletadas, ou ainda em relação à idade ou sexo. Entretanto, nas amostras de fluídos orais houve maior prevalência de detecção para o BKV e para células Merkel. Concluímos que fluídos orais, especialmente saliva e lavado bucal, podem ser usados para o rastreamento do BK e JC; e que os indivíduos HIV positivos, sob tratamento antirretroviral não exibem frequências maior de poliomavírus, comparativamente a indivíduos controle. / Polyomavirus is one of the large family of viruses that cause primary infections usually in childhood, and can remain subclinical. In immunosuppression may cause some diseases. Individuals with HIV/AIDS often have immune deficiencies and may be at increased risk for diseases caused by polyomaviruses. The use of saliva in the diagnosis and follow-up of infectious diseases has been explored in the literature. The advantages of using saliva for screening are based on non-invasive collection and handling safety. The aim of present study was to detect and quantify the DNA from BKV, JCV, Merkel cell and TSV polyomaviruses in oral fluids (saliva, mouthwash and gingival crevicular fluid) and to compare it with serum and urine detection, the means usually used for detection. A total of 299 samples were collected from 42 individuals, 22 HIV positive (GE) and 20 control patients (GC). In GE, 63,6% of the patients presented positive for JCV in at least one sample analyzed, 54,5% were positive for BKV, 18,2% for Merkel cell and there was no positive sample for TSV. In GC, 45% showed JCV positivity in at least one analyzed sample, 80% in BKV, and no control participant exhibited positivity for Merkel cell and TSV. There was no difference in the frequency of viral detection among the groups studied in relation to the samples collected, or in relation to age or gender. However, in oral fluid samples there was a higher prevalence of detection for BKV and Merkel cell. We conclude that oral fluids, especially saliva and mouthwash, can be used for the screening of BK e JC; and that HIV positive individuals under antiretroviral treatment do not exhibit higher frequencies of polyomavirus compared to healthy control subjects.

No description available.

This thesis describes basis of blockchain technology implementation for SAP Cloud platform with emphasis to security and safety of critical data which are stored in blockchain. This diploma thesis implements letter of credit to see and control business process administration. It also compares all the possible technology modification. Thesis describes all elementary parts of software which are necessary to implement while storing data and secure integrity. This thesis also leverages ideal configuration of each programable block in implementation. Alternative configurations of possible solutions are described with pros and cons as well. Another part of diploma thesis is actual working implementation as a proof of concept to cover letter of credit. All parts of code are design to be stand alone to provide working concept for possible implementation and can source as a help to write productive code. User using this concept will be able to see whole process and create new statutes for whole letter of credit business process.

Aim of this diploma thesis was to prepare two protein antigens and two monoclonal antibodies, all based on VP2 minor protein of human polyomaviruses BK virus and Merkel Cell Polyomavirus. One monoclonal antibody was being prepared against unique part of VP2 protein (N-terminal epitope, not present in VP3 protein). A cell line producing such monoclonal antibody has never been established before due to low immunogenicity of the epitope. Our approach was successful in terms of mouse immunization, however, serious problems with hybridoma line stability appeared later during the preparation process. Preparation of antibody targeted to the sequence of VP2 protein of Merkel Cell Polyomavirus was another aim of this thesis. Mouse immunization and hybridoma fusion were performed successfully. After four rounds of cloning in order to purify an established clone, nine clones were cultivated in larger scale. This cultivation probably led to diminished antibody specificity and loss of production ability in most of the hybridoma cells. One more cloning should give rise to an established clone with sufficient production. Two preparations of protein antigens were performed in two expression systems. DNA encoding C-terminally truncated protein VP2 of BK virus fused with His-tag was cloned into a vector suitable for...

Polyomaviruses are small non-enveloped DNA viruses infecting birds and mammals, including human. Their capsid consists of the major capsid protein, VP1, and two minor capsid proteins, VP2 and VP3. The VP2 and VP3 proteins are supposed to have an important function in the transport of viral genome into the cell nucleus, which is a key step to facilitate viral replication. VP2 and VP3 proteins of mouse polyomavirus and SV40 have an ability to bind and disrupt cellular membranes. This feature is believed to be involved in the transport of viral genome into the nucleus. Plasmids carrying genes of the minor capsid proteins of Merkel cell polyomavirus were prepared in order to produce and visualize these proteins in mammalian cells. These proteins are known to have very unusual sequences compared to other human polyomaviruses or related mouse polyomavirus. When produced alone, the minor capsid proteins of Merkel cell polyomavirus did not significantly interact with cellular membranes, unlike the minor proteins of the mouse polyomavirus. The second goal of this work was to prepare mouse polyomavirus mutants with deletion in hydrophobic domains of VP2 and VP3 proteins. These domains are likely responsible for the mentioned membrane interactions. Prepared mutants were non-infectious. The loss of infectivity was not...

Immer wieder fallen im Verlauf von Wahlkämpfen Schlagworte wie Personalisierung, Negativismus und Sensationalismus. Während diese Eigenschaften auf modern geführte Wahl-Kampagnen in den USA durchaus zutreffen mögen, so bescheinigen Studien deutscher Wahlforschungsinstitute für die Bundesrepublik doch recht durchwachsene Befunde. Im Zuge der Bundestagswahl 2005 haben wir uns mittels einer Inhaltsanalyse mit der Kandidatendarstellung Angela Merkels in der deutschen Qualitätspresse beschäftigt. Dabei ging es uns besonders um die Feststellung eines Wandels in der Darstellung Angela Merkels, welcher die Wähler in ihrer Wahlentscheidung beeinflusst haben könnte und um erkennbare Indizien für einen generellen Trend zur Personalisierung. / During electoral campaigns one can always hear key words like personalization, negativism and sensationalism. While these terms may represent the harsh reality of modern campaigns in the US, the results for Germany are rather ambiguous, according to prominent scientific studies. Following the chronology of the German 2005 election of the Bundestag, we tried to evaluate the presentation of candidate Angela Merkel, who ran for chancellor’s office, in the German quality press. Our main goal was the verification of a change in the way the papers presented Angela Merkel, hence possibly influencing the electorate. Furthermore, we tried to find proof of a general trend of personalization in Germany’s elections of 2005.

This master thesis is concerned with the role of the chancellor in the foreign policy with respect to the institutional possibilities that the German political system provides him, and also with respect to the personality of the chancellor - it strives to answer the question, if it is possible to identify the personality impact in the chancellor foreign policy positions. As the case example the current chancellor Angela Merkel has been chosen, who serves for a period long enough to carry out the research and who is at the same time well-respected personality abroad. The thesis comes to the conclusion that the chancellor has the rights to intervene in the foreign policy and uses those rights. By means of the discourse and content analysis of chancellor's speeches it has been possible to identify foreign policy preferences of the chancellor which can be associated with her personal life experience.

This thesis concerns German foreign policy towards Russia during the governments of G. Schroeder and A. Merkel with a view to analyzing this policy from a number of perspectives. The first objective is to introduce an analysis of German-Russian relations in the areas of international politics, security, economy and energy policy from 1998 to the present. The second objective is to compare the approaches of both chancellors in foreign policy and identify whether a certain continuity in German foreign policy vis-á-vis Russia has been preserved or whether the policy has been modified, and if so, in what areas.