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Nachweis Proteinkinase C abhängig exprimierter Gene in AstrozytomenSchulz, Timm 19 September 2003 (has links)
Die Proteinkinase C (PKC) ist eine wichtige Signaltransduktionskomponente, deren Aktivierung die Expression zahlreicher Gene induziert und zur Zelldifferenzierung und Zellproliferation führt. Ein besonders hohes Expressionsniveau der PKC findet man in vielen Tumoren. So korreliert in malignen Gliazellen das Expressionsniveau der PKC mit deren Wachstumsgeschwindigkeit. Es wird angenommen, daß die aktivierte PKC eine wichtige Rolle in der Tumorpromotion hat. In der vorliegenden Arbeit wurde untersucht, ob in Astrozytomzellinien Gene zu finden sind, die nach PKC-Aktivierung durch den Phorbol-Ester TPA differentiell exprimiert werden. Zunächst wurden kultivierte Zellen der Astrozytomzellinie LN-405 mit TPA respektive dem PKC-Inhibitor Chelerythrin behandelt. Nach Gewinnung der mRNA aus der zuvor isolierten RNA wurden in einem mehrstufigem PCR-Verfahren (SSH) cDNA-Abschnitte gewonnen, die zu vermeintlich differentiell exprimierten Genen gehören. Diese cDNA-Abschnitte wurden in Plasmid-Vektoren eingefügt, kloniert und zur Bestimmung sequenziert. Um falsch positive Sequenzen zu erkennen, wurden die zuvor radioaktiv markierten cDNA-Abschnitte mit Northernblots hybridisiert. Gleichzeitig ließ sich so ein zeitabhängiger Anstieg der Expression nach PKC-Stimulation untersuchen. Durch den PCR-Select-Assay (SSH) konnten insgesamt 11 Gene gefunden werden, die sich in der radioaktiven Northernblot-Hybridisierung, als nach PKC-Aktivierung differentiell exprimiert, darstellen ließen. Dabei bestätigt der gefundene Zusammenhang zwischen PKC-Aktivierung und differentieller Exprimierung bei fünf der 11 Gene (IL-8, Calpain, Interferon-gamma Rezeptor 2, Methionin Adenosyltransferase, beta-2 adrenerger Rezeptor) Ergebnisse anderer Autoren, wobei dieser Zusammenhang nur bei zwei Genen (IL-8 und Calpain) auch in Astrozytom- bzw. Gliom-Zellen schon früher gezeigt werden konnte. Sechs Gene (M-Phase Phosphoprotein-1, ect2-Onkogen, ERM-Gen, Ornithin-Decarboxylase-Antizym 2, MHC-bindendes Protein 2, Sequenz aus Cosmid F0811) wurden in der vorliegenden Arbeit erstmalig als PKC-abhängig exprimiert beschrieben. Die gefundenen Gene haben auf verschiedene Funktionen der Zellen Einfluß. So beeinflussen sie die Regulation des Zellzyklus (MPP1, ect2-Oncogen), die Immunregulation (MBP-2, IL-8, Interferon-gamma Rezeptor 2), die Signaltransduktion (beta-2 AR), die Transkription (ERM-Gen), die Proteinsynthese (ODC-Antizym, MAT), die Wachstumskontrolle (ODC-Antizym) und die Regulation der PKC selbst (Calpain). Für fünf Gene läßt sich ein eindeutiger Zusammenhang mit der Tumorpromotion herstellen: IL-8 (Angioneogenese), MBP-2 (Immunsuppression), ERM-Gen (Transkriptionspromotion), MAT (allgemein fördernder Einfluß auf den Metabolismus) und ect2-Oncogen (Oncogen). / The protein kinase C (PKC) is one of the major signal transduction systems and its activation leads to the induction of the expression of several genes, to cell differentiation and cell proliferation. Very high expressed PKC are found in many tumors. In malignant glia cells the expression of PKC correlates with their proliferation rate. The PKC activity has an important role for the tumor promotion. The object of this paper, was to investigated, if there are genes differentialy expressed after activation of PKC through the phorbol-ester TPA in astrocytoma cell lines. The astrocytoma cell line LN-405 was incubated with TPA and the PKC-inhibitor chelerythrine respectively. After isolation of RNA and mRNA the suppression subtractive hybridization (SSH) was used to isolate differentially expressed cDNA fragments. These cDNA fragments were inserted into the T/A cloning vector, cloned and sequenced. To detect false positives the cDNA fragments were analysed with northern blot technique. Examined was also a time-dependent acceleration of expression after TPA treatment. 11 genes were detected by suppression subtractive hybridization, showing differentially expressed in the northern blot hybridization. Five of the genes were found differentially expressed after PKC activation before (IL-8, calpain, interferon gamma receptor 2, beta-2-adrenergic receptor, methionine adenosyltransferase alpha), two of these genes (IL-8, calpain) also in astrocytoma- and glioma-cells respectively. Six genes (M-phase phosphoprotein 1, ect2-onkogene, erm gene, ornithine decarboxylase antizyme 2, MHC binding protein 2, sequence from Cosmid F0811) were described as PKC dependent expressed for the first time. The genes detected influence several cell functions. They are involved in cell-cycle regulation (MPP1, ect2-oncogene), immuneregulation (MBP-2, IL-8, interferon gamma receptor 2), signal transduction (beta 2 adrenergic receptor), transcription (erm-gene), synthesis of proteins (ODC-antizyme 2, MAT), growth control (ODC-antizyme) and regulation of PKC (Calpain). Five genes show a clear connection to tumor promotion: IL-8 (angioneogenesis), MPB-2 (immunesuppression), erm gene (promotion of transcription), MAT (promotion of metabolism) and ect2-oncogene (oncogene).
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Caracterização da elevação do Rio Grande a partir de elementos do campo de gravidade terrestre / Characterization of the Rio Grande Rise from elements of the terrestrial gravity fieldMarília Takaguti Dicezare 05 February 2018 (has links)
Dados de altimetria por satélite contêm informações importantes para o mapeamento de estruturas tectônicas em regiões oceânicas, como falhas, zonas de fratura e montes submarinos. A grande disponibilidade e densidade desses dados permitem a identificação de feições do assoalho oceânico com boa precisão. Este trabalho tem como objetivo investigar as características estruturais da Elevação do Rio Grande, no Atlântico Sul, através de elementos do campo de gravidade terrestre. Para isso, são utilizados dados de altura da superfície do mar (SSH) provenientes dos satélites das missões ERS1-GM, Geosat-GM e Seasat. Com o cálculo da derivada direcional da SSH ao longo das trilhas ascendentes e descendentes dos satélites, foram obtidos os gradientes de superfície do mar (SSG), que ressaltam os curtos comprimentos de ondas associados às importantes feições oceânicas estudadas. Também foram calculados os gradientes da altura geoidal (desvio da vertical) para auxiliar na interpretação do sinal da SSG. Através da SSH foi possível identificar estruturas de maior porte, como o rift da elevação, e algumas fraturas e montes submarinos maiores. Por outro lado, a SSG forneceu maiores detalhes sobre as feições já caracterizadas pela SSH e de toda a região, revelando também diversas outras estruturas de menor dimensão. O posicionamento das feições identificadas por ambas as grandezas, SSH e SSG, é bastante preciso. Entretanto, fatores como a direção e a orientação das trilhas dos satélites e a presença de estruturas adjacentes podem influenciar a resposta da SSG para uma determinada feição tectônica, por isso, as trilhas ascendentes e descendentes dos três satélites podem apresentar respostas diferenciadas. Sendo assim, recomenda-se analisar os dois conjuntos de trilhas de várias missões altimétricas para obter maiores informações das características estruturais das feições investigadas. O estudo também permitiu identificar possíveis estruturas com uma resposta característica de montes submarinos, nas trilhas descendentes de SSH, que não foram caracterizados anteriormente na literatura e não possuem correspondente nos modelos topográficos/batimétricos. / Satellite altimetry data contain important information for mapping tectonic structures in oceanic regions, such as faults, fracture zones and seamounts. The great availability and spatial density of these data allow one to identify ocean floor features with good accuracy. This work aims to investigate structural characteristics of the Rio Grande Rise, in South Atlantic, through elements of the terrestrial gravity field. We used sea surface height (SSH) data from satellite missions ERS1-GM, Geosat-GM and Seasat to calculate sea surface gradients (SSG), which are the SSH directional derivative along the ascending and descending satellite tracks. SSG emphasize the short wavelengths associated with the important oceanic features studied. Geoid gradients (deflection of the vertical) were also calculated to assist in the interpretation of the SSG signal. By analyzing sea surface heights, it is possible to identify larger structures, such as the rift of the rise, some fractures and large seamounts. In contrast, sea surface gradients provide greater details of the features characterized by the SSH and the entire area, also revealing several smaller seamounts. The positioning of the structures identified by both SSH and SSG is fairly accurate. However, factors such as direction and orientation of the satellite tracks and the presence of adjacent structures may influence the SSG response to a given tectonic feature. For this reason, the ascending and descending tracks of the three satellites may have different responses. Therefore, it is recommended that one analyzes the two sets of tracks from the several altimetric missions to obtain more information on the structural characteristics of the features. The study also allowed us to identify possible structures with a characteristic response of seamounts on SSH descending tracks, which were not previously characterized in the literature and do not have a similar correspondent in topographic/bathymetric models.
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以Scrum觀念及SSH框架為基礎進行系統開發之研究 / A study on system development based on scrum concept and SSH framework宋洪炘, Sung, Hung Hsin Unknown Date (has links)
由於現今商業環境時常變動,使得使用者需求在系統開發過程中頻繁改變。傳統系統開發方式無法迅速回應使用者需求變更,強調彈性開發流程的敏捷開發方式逐漸廣為接受。敏捷開發並未探討系統設計議題而系統設計不佳會導致系統品質低落,使用軟體框架可以快速建立良好的分層結構,有助於穩定產品品質並提高軟體再用性。本研究以Scrum為基礎,結合Struts、Spring、Hibernate整合框架,建立一套系統開發方法,包含敏捷開發方式的彈性流程,兼顧系統擴充性及可維護性,研究方法為概念形成法與系統展示法,並以Java Web-Based雛型系統為例,說明相關作業內容。 / Due to the quick variation in the business environment, user requirements change frequently during the system development process. This induces the rising popularity of agile development method which emphasizes the development flexibility during the process. However, agile development method does not address the design issues and poor system design can easily cause poor system quality. Software framework helps to quickly build a good hierarchy structure and thus improve the product quality and software reusability. This study proposes a system development method based on Scrum with the integration of Struts、Spring and Hibernate frameworks. The method is embedded with the development flexibility from the agile method, as well as the system extensibility and maintainability from the software frameworks. The study is conducted with the research method of concept formulation, and would be continued with a Java Web-Based prototype development as an example to illustrate the relevant activities.
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Analise da expressão diferencial de genes de citros em resposta a infecção por Xanthomonas axonopodis pv. citri / Differential gene expression of citrus in response to infection by Xanthomonas axonopodis pv. citriCamillo, Luciana Rodrigues 13 July 2006 (has links)
Orientador: Celso Eduardo Benedetti / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-07T07:02:38Z (GMT). No. of bitstreams: 1
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Previous issue date: 2006 / Resumo: A doença cancro cítrico, causada pela bactéria Xanthomonas axonopodis pv. citri (Xac), emergiu como uma das principais ameaças à citricultura brasileira pois afeta todas as variedades comerciais de citros, diminuindo a produção e qualidade dos frutos e podendo se dispersar rapidamente em áreas de cultivo de citros. Entre os gêneros de Xanthomonas, foram encontrados muitos genes associados com patogenicidade e virulência, entretanto, pouco se conhece sobre os mecanismos envolvidos nas interações entre Xac e laranjeira e os genes envolvidos no desenvolvimento dos sintomas do cancro cítrico em folhas de citros (Citrus sinensis) em resposta à infecção por Xac. Neste trabalho, foi analizada a expressão diferencial de genes envolvidos no desenvolvimento do cancro cítrico em folhas de laranja, em resposta à infecção por Xac. Para tanto foram construídas duas bibliotecas de Hibridização Subtrativa Suprimida (SSH) com mRNAs de folhas infiltradas com Xac ou H20 após 10 dias de inoculação. O "screnning" das bibliotecas foi feito por dot blot e 17 genes diferencialmente expressos foram sequenciados e identificados por homologia no banco de dados ncbi-BLAST contra o banco de dados de EST de citros. A expressão gênica diferencial foi analizada por Northern Blot e PCR em tempo real (qPCR). Para complementar nossos dados, a expressão dos 17 genes diferenciais foi aI).alisada a partir de cDNAs de folhas de citros que foram infiltradas com Xac, H20 ou Xanthomonas axonopodis pv. aurantifolii (Xaa), que não é patogênica à laranja, mas causa cancro no limão galego. Nossos resultados demonstraram que Xac e Xaa induzem e reprimem o mesmo grupo de genes, porém em diferentes níveis. Nós identificamos que ambos patóg;enos alteram as vias de transdução de sinal de auxina, tráfico e fusão de vesículas, resposta de defesa e doença, síntese de proteínas e ciclo celular, metabolismo de carbono-nitrogênio e metabolismo secundário. A análise desses genes poderá ser de grande importância para o entendimento dos eventos que levam ao desenvolvimento dos sintomas do cancro / Abstract: The citrus canker disease, caused by Xanthomonas axonopodis pv. citri (Xac), has emerged as one of the major threats to the Brazilian citriculture because it affects all commercial citrus varieties, decreases the production and quality of the fruits and can spread rapidly in the citrus growing areas. The symptoms include canker lesions, leading to abscission of fruits and leaves and general tree decline. Within the genus Xanthomonas, several genes have been found associated with pathogenicity and virulence. However, little is known about the mechanisms involved in the Xac- citrus interaction and the development of the canker disease. In this work we analyzed the differential expression of genes involved in the development of the canker disease in sweet orange (Citrus sinensis) leaves in response to Xac infection. Therefore we constructed two Supression Subtracted Hybridization (SSH) libraries with mRNA from leaves infiltrated with Xac or water after 10 days of inoculation. The libraries were screnned by dot blot and the 17 differentially expressed genes were sequenced and identified through BLAST searches against the Citrus EST and protein databases. The differential gene expression was evaluated by Northem blot and Real Time PCR (qPCR). To complement our analysis, the expression of 17 genes was compared in cDNA samples from citrus leaves that had been infilt:r:ated with Xac, water or X axonopodis pv. aurantifolii (Xaa), wich is not pathogenic to orange but causes canker in key lime. Our results show that Xac and Xaa induce and repress a similar set of genes, however at different leveis. We found that both pathogens altered the pathways of auxin transport and signaling, vesicle trafficking and transport, cell cycle and protein synthesis, photorespiration and disease response. Further analysis of these genes will be of great importance to understand the events triggering the development of the canker symptoms / Mestrado / Genetica de Microorganismos / Mestre em Genética e Biologia Molecular
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Caracterização molecular de genes preferencialmente expressos na fase leveduriforme patogênica de ´Paracoccidioides brasiliensis´ através das técnicas de ´Macroarray´ e de SSH (Suppression Substractive Hybridization) / Molecular characterization of preferentially expressed genes in the yeast pathogenic phase of Paracoccidioides brasiliensis through the techniques of Macroarray and SSH (Suppression Subtraction Hybridization)Everaldo dos Reis Marques 22 December 2005 (has links)
Paracoccidioides brasiliensis, um fungo termodimórfico, é o agente causador da paracoccidioidomicose (PCM), a micose sistêmica prevalente da América Latina. A patogenicidade aparenta estar intimamente relacionada com a transição dimórfica da forma de micélio para a de levedura, que é induzida pela mudança da temperatura do ambiente pela temperatura do hospedeiro mamífero. Há poucas informações disponíveis sobre genes de P. brasiliensis que são necessários durante a fase patogênica. Nós, então, realizamos as técnicas de SSH (Suppression Subtraction Hybridization) e de Macroarray com o objetivo de identificar genes que sejam preferencialmente expressos na fase leveduriforme do isolado Pb18. Genes identificados em ambos os procedimentos estão mais expressos na fase leveduriforme e estão envolvidos em metabolismo básico, transdução de sinal, crescimento e morfogênese e metabolismo do enxofre. Para testar se as mudanças observadas na expressão gênica refletem as diferenças entre as condições de crescimento usadas para obter as duas formas morfológicas preferivelmente às diferenças intrínsecas dos tipos celulares, nós realizamos experimentos com RT-PCR em tempo real utilizando preparações de RNA derivadas de ambas as fases, micélio e levedura, crescidas a 26°C e 37°C nos meios de cultura completos (YPD e Sabouraud) e meio mínimo. Vinte genes, incluindo AGS1 ( -1,3-glucan synthase) e TSA1 (thiol-specific antioxidant), foram mostrados como mais expressos na levedura patogênica em relação ao micélio. Embora a expressão de RNA mensageiro foi bastante diferente em relação aos meios completos e meio mínimo, mostramos uma tendência geral para que esses genes serem mais expressos nas células leveduriformes patogênicas de P.x brasiliensis. Além disso, mostramos a complementação dos genes METR e SCONC de P. brasiliensis e uma cepa com estes genes deletados de Aspergillus nidulans, sugerindo uma possível homologia entre eles. Mostramos também a análise de genes da via do metabolismo do enxofre foram mais expressos na levedura patogênica de P. brasiliensis em relação ao micélio saprofítico. / Paracoccidioides brasiliensis, a thermodimorphic fungus, is the causative agent of paracoccidioidomycosis (PCM), a prevalent systemic mycosis in Latin America. Pathogenicity appears to be intimately related to the dimorphic transition from the hyphal to the yeast form, which is induced by a shift from environmental temperature to the temperature of the mammalian host. Little information is available on the P. brasiliensis genes necessary during the pathogenic phase. We have therefore undertaken Suppression Subtraction Hybridization (SSH) and macroarray analyses with the aim of identifying genes that are preferentially expressed in the yeast phase. Genes identified by both procedures as being more highly expressed in the yeast phase are involved in basic metabolism, signal transduction, growth and morphogenesis, and sulfur metabolism. In order to test whether the observed changes in gene expression reflect the differences between the growth conditions used to obtain the two morphological forms rather than differences intrinsic to the cell types, we performed real-time RT-PCR experiments using RNA derived from both yeast cells and mycelia that had been cultured at 37 and 26°C in either complete medium (YPD or Sabouraud) or minimal medium. Twenty genes, including AGS1 ( 1,3-glucan synthase) and TSA1 (thiol-specific antioxidant), were shown to be more highly expressed in the yeast cells than in the hyphae. Although their levels of expression could be different in rich and minimal media, there was a general tendency for these genes to be more highly expressed in the yeast cells. Moreover, complementation of P. brasiliensis METR and SCONC genes in strains of Aspergillus nidulans with these genes deleted suggested a possible homology between them. We show the analyses of genes involved in the xii sulphur metabolism pathway and these genes were more expressed in the pathogenic yeast than saprophytic mycelia of P. brasiliensis.
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Virus manipulateurs du comportement des insectes : prévalence et influence sur la structure des communautés hôtes : exemple de l’association Leptopilina boulardi / LbFV / Viruses manipulating insect behaviour : prevalence and impact on the structure of host community : example of the association Leptopilina boulardi / LbFVPatot, Sabine 10 July 2009 (has links)
Les symbioses eucaryotes/micro-organismes constituent une importante source d’innovation évolutive et de diversité écologique. Ces associations sont très répandues chez les insectes, en particulier chez les insectes parasitoïdes (insectes parasites d’autres insectes) qui hébergent en particulier une grande diversité de virus transmis verticalement. Leurs effets directs sur les parasitoïdes ainsi que les effets indirects sur la structure des communautés sont à l’heure actuelle mal compris. Nous avons abordé ces questions au travers l’étude d’un virus héritable (LbFV) ayant la particularité de manipuler le comportement de superparasitisme de son hôte, l’hyménoptère parasitoïde de drosophiles Leptopilina boulardi. La mise au point d’un outil moléculaire diagnostic de l’infection nous a permis de montrer que ce virus, spécifique à L. boulardi, peut atteindre de fortes prévalences dans les populations d’hôtes. Nous avons également mis en évidence un effet de la présence du virus sur les interactions compétitives interspécifiques au sein de la communauté des parasitoïdes de drosophiles. L’approche intégrée de ce travail constitue une étape importante dans la connaissance du rôle des virus héritables sur l’écologie et l’évolution de leurs hôtes / Eukaryots/microorganisms symbiosis is an important source of evolutionary novelty and ecological diversification. These associations are widespread in insects, particularly in parasitoids (insects that parasitize other insects) where a broad diversity of vertically transmitted viruses are found. However, their direct and indirect effects on host community are poorly understood. In this thesis, we used a system involving a Drosophila parasitoid, Leptopilina boulardi and a heritable virus LbFV that manipulates the behaviour of the parasitoid by increasing its tendency to lay eggs in a host that is already parasitized (superparasitism). Using a viral molecular marker developed in this work, we showed very high prevalences of the virus in L. boulardi populations. Additionally, we found a strong effect of the virus on interspecific competition in the Drosophila parasitoid community. The integrative approach of this work is an important step in understanding the role of heritable viruses in parasitoid ecology and evolution
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Efeitos da infecção por Rickettsia rickettsii sobre o perfil de expressão gênica do carrapato vetor Amblyomma cajennense. / Effects of infection with Rickettsia rickettsii on the gene expression profile of the tick vector Amblyomma cajennense.Larissa Almeida Martins 06 May 2014 (has links)
O agente etiológico da Febre Maculosa das Montanhas Rochosas (RMSF), conhecida no Brasil como Febre Maculosa Brasileira, é a bactéria Rickettsia rickettsii. Essa bactéria é transmitida ao homem pela picada de diferentes espécies de carrapatos ixodídeos. No Brasil, os vetores são Amblyomma cajennense e A. aureolatum. As taxas de prevalência de R. rickettsii nas populações de carrapatos de áreas endêmicas para RMSF são baixas, em geral abaixo de 1%. Essa baixa prevalência parece estar associada a menores taxas reprodutivas e de sobrevivência de linhagens infectadas, sugerindo que R. rickettsii seja patogênica também para os seus vetores. Infecções experimentais demonstraram que 80-100% dos indivíduos de uma colônia de A. aureolatum mantida em laboratório são infectados por R. rickettsii, enquanto apenas 10-60% de A. cajennense adquirem a bactéria. Esses dados indicam que as respostas dessas duas espécies de carrapatos à infecção sejam diferentes, resultando em diferentes taxas de prevalência da bactéria. Dessa maneira, a caracterização molecular das interações entre carrapatos do gênero Amblyomma e a bactéria R. rickettsii é importante, podendo gerar informações não somente para o esclarecimento acerca dos mecanismos de patogenicidade de R. rickettsii para os carrapatos, mas também para um melhor entendimento dos mecanismos responsáveis pela aparente restringência de A. cajennense à infecção. Assim, os objetivos do presente estudo foram: (i) analisar os efeitos da infecção por R. rickettsii sobre o perfil de expressão gênica de carrapatos A. cajennense por hibridação subtrativa por supressão (SSH), (ii) validar os dados de SSH por reação em cadeia de polimerase quantitativa precedida por transcrição reversa (RT-qPCR) e (iii) caracterizar funcionalmente dois genes com expressão induzida pela infecção por RNA de interferência (RNAi). Após a análise bioinformática dos dados de SSH, 44 sequências únicas foram obtidas, das quais 36 representam genes com expressão induzida e 8 genes com expressão reprimida pela infecção. A indução dos genes codificadores da subunidade I da citocromo c oxidase (COX1), da subunidade IV da NADH desidrogenase, de uma proteína com domínio de inibidor de serina-proteases Kunitz-type (papilina-like), identificados por SSH, e de um peptídeo antimicrobiano (hebraeína), foi confirmada por RT-qPCR. O silenciamento gênico da hebraeína e da papilina-like não teve nenhum efeito na aquisição de R. rickettsii pelo vetor, indicando que, isoladamente, não são responsáveis pela proteção de A. cajennense contra a infecção. Os dados gerados pelo presente estudo abrem perspectivas para que outros genes sejam avaliados quanto ao seu papel na aquisição de R. rickettsii, os quais, no futuro, podem ser considerados como alvos para o desenvolvimento de vacinas. / The etiologic agent of the Rocky Mountain Spotted Fever (RMSF), also known as Brazilian Spotted Fever in Brazil, is the bacterium Rickettsia rickettsii. This rickettsia is transmitted to humans by the bite of various tick species. In Brazil, Amblyomma cajennense and A. aureolatum are known as vectors. The prevalence rates of R. rickettsii infected ticks in RMSF endemic areas are low, oscillating around 1%. These low prevalence rates seems to be associated with lower reproductive and survival rates of infected ticks, suggesting that R. rickettsii is also pathogenic to its vectors. Experimental infections with R. rickettsii have demonstrated that 80 to 100% of A. aureolatum ticks from a laboratory colony acquire this bacterium, whereas only 10 to 60% of A. cajennense ticks become infected. These results indicate that the responses of these two tick species against infection are different, resulting in different prevalence rates of the bacterium. Therefore, the elucidation of the interactions between ticks of the genera Amblyomma and the bacterium R. rickettsii at a molecular level is important to provide information to better understand the mechanisms of pathogenicity of R. rickettsii against ticks as well as for the elucidation of the mechanisms responsible for the apparent refractoriness of A. cajennense against infection. Therefore, the objectives of the current study were: (i) analyze the effets of the infection with R. rickettsii on the gene expression of ticks A. cajennense by suppression subtractive hybridization (SSH), (ii) validate SSH data by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and (iii) functionally characterize two genes induced by infection using RNA interference (RNAi). After bioinformatics analysis of SSH data, 44 unique sequences were obtained, among which 36 represent genes with expression induced and 8 repressed genes by infection. The induction of genes encoding subunit I of cytochrome c oxidase (COX1), the NADH dehydrogenase subunit IV, a protein containing Kunitz-type inhibitor domain (papilin-like), identified by SSH, and an antimicrobial peptide (hebraein), was confirmed by RT-qPCR. The effects of knockdown of hebraein and papilin-like encoding genes had no effect on the acquisition of R. rickettsii by the vector. Data of the current study may be used to evaluate the role of other genes in acquisition of R. rickettsii, which, in the future, may be considered as target for vaccine development.
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Automatisk kontroll av status för switch-portar / Automatic control of switchport statusDan, Östergren January 2020 (has links)
I stora nätverksmiljöer kan det idag vara svårt att få en komplett sammanställning av hur många switch-portar som är i bruk. Den lösning som vanligtvis används är manuell inloggning till aktuell switch för kontroll, ett moment med tidsåtgång och som ofta utförs vid enstaka tillfällen. Information riskerar därför att upptäckas för sent med en åtgärd som blir reaktiv. Metoder för att tillgängliggöra informationen på ett sådant sätt att den till stor del kan bli proaktiv kan istället möjliggöra åtgärder i tid men även tillhandahålla aktuell information mer lättillgängligt och i slutändan spara både tid och utgifter för ett företag. Arbetet inriktar sig på framtagandet av en lösning med specifika krav med en inriktning för att förenkla nuvarande moment; att på ett enkelt sätt kunna ta del av dagsaktuell information om status för switch-portar, att kunna få en övergripande bild av fördelningen av switch-portar för en anläggning och möjlighet för notifiering vid gränsöverskridelser. Inledande görs en undersökning av aktuella lösningar som finns att tillgå inom området idag, där flera visar sig ha brister men även saknar nödvändiga funktioner. Då de lösningar som finns tillgängliga idag inte stämmer överens med kraven finns en motivering till framtagandet av en lösning med egna funktioner. Den lösning som tas fram utvecklas med separata funktioner för att inhämta information, tolka, presentera och notifiera information för administratör. För att garantera en stabil lösning med kontinuerlig drift installerades en server i en extern datahall där en linuxdistribution användes. Två olika protokoll för inhämtning av information jämförs med praktiska tester, inloggning via SSH, samt SNMP-poll, där den valda lösningen bygger på SSH som metod på grund av säkerhetsaspekter, men där SNMP visar mer kompatibilitet mellan tillverkare och modeller av hårdvara. Vidare beskrivs tillvägagångssätt hos de funktioner som utvecklats för tolkning av inhämtad information samt de svårigheter som uppstått i samband med detta, för undvikandet av feltolkningar. Olika alternativ för presentation av information till administratör jämförs, där den valda lösningen blev åtkomst via webbsida, detta på grund av det grundläggande stöd oberoende av plattform. Något som exempelvis en applikation inte kan ge i samma omfattning. Bland de olika notifieringsmetoder som undersöktes föll valet även i detta fall på en plattformsoberoende metod, där notifiering via e-post ansågs både enkel att implementera och med ett brett stöd hos klienter. Varningar vid förangivna kriterier av antal lediga switch-portar kunde därmed tas emot av administratör. Samtliga funktioner fungerade vid utförandet som planerat och lösningen används av kunden. Det finns några förslag på förbättringsåtgärder där SNMP istället med fördel kan användas med bredare stöd hos andra tillverkare och modeller, samt även skyddsfunktioner vid tolkning av information. / In large network environments today, it can be difficult to get a complete summary of how many switchports are in use. The solution that is usually used is manual login to the current switch for control, a step with time consumption and which is often performed on occasion. Information therefore risks being discovered too late with a measure that becomes reactive. Methods for making the information available in such a way that it can largely become proactive can instead enable timely action but also provide up-to-date information more easily accessible and ultimately save both time and expenses for a company. The work focuses on the development of a solution with specific requirements with a focus on simplifying current steps; to be able to easily access up-to-date information on the status of switchports, to be able to get an overall picture of the distribution of switchports for a facility and the possibility of notification in the event of border crossings. Initially, an investigation is made of current solutions that are available in the area today, where several turn out to have shortcomings but also lack the necessary functions. As the solutions available today do not comply with the requirements, there is a motivation for developing a solution with its own functions. The solution that is put together is developed with separate functions for collecting information, interpreting, presenting and notifying information to the administrator. To ensure a stable solution with continuous operation, a server was installed in an external datacenter where a Linux distribution was used. Two different protocols for obtaining information are compared with practical tests, login via SSH, and SNMP-poll, where the chosen solution is based on SSH as a method due to security reasons, but where SNMP shows more compatibility between manufacturers and models of hardware. Furthermore, the procedures of the functions that have been developed for the interpretation of collected information and the difficulties that have arisen in connection with this, for the avoidance of misinterpretations are described. Different options for presenting information to the administrator are compared, where the chosen solution was accessed via a website, this is due to the basic support regardless of platform. Something that, for example, an application cannot provide to the same extent. Among the various notification methods examined, the choice also fell in this case on a platform-independent method, where notification via e-mail was considered both easy to implement and with broad support among clients. Warnings for the specified criteria of the number of available switchports could thus be received by the administrator. All functions worked during the execution as planned and the solution is used by the customer. There are some suggestions for improvement measures where SNMP can instead be used to advantage with broader support from other manufacturers and models, as well as protection functions when interpreting information.
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HoneyRAN : A Medium-Interaction Honeypot for Radio Access Networks Mimicking a Command-Line Interface in a Baseband Unit / HoneyRAN : En medelinteraktion honungskruka för radioaccessnät som imiterar en kommandoradsgränssnitt i en basbandsenhetCho, Daniel January 2021 (has links)
There is a lack of understanding of the threat vectors and attacks in Radio Access Network (RAN) systems. In addition, there is a lack of knowledge in adversary behaviour and tactics in RAN. To gain an increased understanding of the threat landscape in RANs and potentially increase the overall security for RANs, this thesis project designed, implemented, and evaluated a honeypot for Ericsson’s RAN. Developing a suitable honeypot required an initial literature study of honeypots, RANs, and Long-Term Evolution (LTE) networks. In addition, previous research and work related to this topic were included in the literature study. The open-source Secure Shell Protocol (SSH) honeypot project, Cowrie, was used as a fundamental building block for the honeypot. Moreover, a high-level architecture of the honeypot system (HoneyRAN) was designed and used as a basis for the implementation process. For evaluating the honeypot, a penetration test was conducted in which an internal Ericsson penetration tester evaluated the honeypot. The Elasticsearch, Logstash, and Kibana (ELK) stack was utilised to facilitate log analysis. The results from the penetration test were better than expected and are promising as HoneyRAN achieved the three claimed goals/objectives: understanding adversarial behaviour and tactics, stalling the adversaries, and early detection of attacks. The outcome of the evaluation suggests that HoneyRAN is a good initial approach for increasing the overall security of RANs and that honeypots are an effective deception technique for stalling attackers. The conclusions from the evaluation of HoneyRAN are that HoneyRAN’s implementation using Cowrie achieved an essential desired outcome, namely realism, deception strategies are essential to implement deception techniques (such as honeypots) effectively, and the design and implementation choices of HoneyRAN provides a foundation for other solutions. / Det finns begränsad förståelse för hotvektorer och attacker i RAN system. Dessutom finns det även begränsad kunskap om fientlig beteende och taktiker som används i RAN. För att få en ökad förståelse för de hotbilderna som finns i RAN så designade, implementerade och utvärderade detta avhandlingsprojekt en så kallad honungskruka för Ericssons RAN. Att utveckla en lämplig honungskruka krävde en inledande litteraturstudie på honungskrukor, RAN, och LTE nätverk. I litteraturstudien krävdes det även att man utförde en forskning om tidigare forskning och arbeten relaterat till detta ämne. En översikt på de komponenter som behövdes för att implementera honungskrukan (HoneyRAN) skapades och användes som grund för implementeringsprocessen. För att utvärdera systemet genomfördes ett penetrationstest som utfördes av en intern penetrationstestare på Ericsson. ELK stack användes för att underlätta logganalys. Resultaten från penetrationstestet var bättre än förväntat och är lovande eftersom honungskrukan uppnådde de tre påstådda målen: att få en förståelse av fientlig beteende och taktik, att uppehålla motståndarna, och att upptäcka tidiga faser av attacker. Dessutom tyder resultatet på att HoneyRAN är ett bra första steg till att öka den övergripande säkerheten i RAN och att honungskrukor är en effektiv vilseledningsteknik för att uppehålla attackerare. Slutsatserna från utvärderingen av HoneyRAN är att användingen av Cowrie uppnådde en väsentlig faktor, nämligen att HoneyRAN ser realistisk ut, vilseledningsstrategier är viktiga för att kunna effektivt implementera en honungskruka, och designvalen som togs för implementeringen av Honey- RAN kan utgöra en grund för framtida lösningar att bygga vidare på.
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Localized electronic states of a centrosymmetric SSH solitonBédard, Maude 12 1900 (has links)
La matière condensée moderne porte un intérêt particulier pour la classe de matériaux formée par les isolants topologiques. Ils sont différents des isolants typiques par leurs intéressantes propriétés quantiques; ils se comportent comme des isolants dans leur intérieur, mais contiennent des états conducteurs sur leur surface. On peut mieux comprendre le comportement de certains systèmes en matière condensée, tel que les chaînes de polyacétylène, en étudiant un système unidimensionnel simple : le modèle de Su-Schrieffer-Heeger (SSH). Le modèle SSH décrit des fermions sans spin sautant sur un réseau unidimensionnel où les amplitudes de saut alternent d’un site à l’autre. Ce modèle, bien que simpliste, expose les propriétés clés des isolants topologiques tel que les états délocalisés dans tout le réseau ainsi que les états exponentiellement localisés aux frontières du réseau. Dans ce projet, nous étudions le modèle SSH, mais en ajoutant un défaut central dans le réseau qu’on appelle un soliton. Dans notre cas, le soliton consiste en un site central donc les amplitudes de saut sont les mêmes d’un côté et de l’autre. Nous trouvons un ensemble de solutions complet incluant des états de basse énergie localisés aux frontières ainsi que des états de haute énergie localisés au soliton. / Topological insulators are a class of materials that have attracted much attention in modern condensed matter. They are different from typical insulators as they exhibit interesting quantum properties; they behave as insulators in their interior but have conducting states on their surface. We can better understand the properties of low dimensional condensed matter systems (like poly-acetylene chains) by studying a toy model known as the Su-Schrieffer-Heeger (SSH) Model. The SSH model describes spinless fermions hopping on a one-dimensional lattice with staggered hopping amplitudes. Such a toy model exhibits key properties of topological insulators, such as bulk states (delocalized states across the lattice) and edge states (exponentially localized states at the boundaries of the lattice). In this project, we study the SSH model with an added central defect to the chain, which we call a soliton. In our case, the soliton consists of a central site with the same hopping amplitude on either side. We study the impact of such a defect on the properties of the system; we find a complete set of solutions including near-zero-energy edge states as well as high-energy states localized at the soliton.
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