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Besarani, Dler. "Anti-Vimentin Antibodies in Renal Transplantation." Thesis, Imperial College London, 2010. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.526360.
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Wong, Kai-lun, and 黃棨麟. "Nanomechanical studies of vimentin intermediate filaments." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2012. http://hub.hku.hk/bib/B49799617.
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Intermediate filaments, microtubules and microfilaments are the major components of the cytoskeleton. Though it is known that intermediate filaments play an important role in the mechanical behaviour of cells, it is surprising that their mechanical properties are far from being fully understood.
The morphology and assembly process of the vimentin intermediate filaments (IFs) were studied using transmission electron microscopy (TEM) and atomic force spectroscopy (AFM). The width of the vimentin was found to change as the assembly proceeded. This finding agrees with the literature about the compaction process of vimentin IFs. The width of the IFs decreased gradually, while the range of width increased within the first few minutes after assembly initiation, and then decreased at last and became stable at 12.80±2.20nm. The average length of the IFs increased with decreasing rate. The length attained 485.60±162.23nm at 120 minutes. The range of length increased which revealed the assembly process was randomly occurring between filaments in the solution. The height of the IFs obtained with AFM did not show the periodicity in contrary to the literature. It may be due to the flattening of IFs on the functionalized mica(AP-mica) surface, or the periodicity was not prominent to be observed morphologically.
In the force spectroscopy study, the nanomechanical properties of individual vimentin intermediate filaments were studied using AFM. Fresh vimentin intermediate filaments and samples fixed with glutaraldehydewere examined, and force-displacement curves with nano-scale resolutions of different vimentin intermediate filament samples were analysed. The use of glutaraldehydefixative provided cross-linking of the IF, and the structural change will result in differences in their force-displacement curves which helped to provide comparison with the non-fixed samples in order to identify the structure-mechanical property relationship. Statistical studies of these curves revealed that tearing off of protofilaments from the mature intermediate filaments (with and without glutaraldehyde) occurred inthe low force regime below 100pN, and successive tearing off events were observed readily below 25 nm separations, which were comparable with the lengths of domains of around 20 nm. Different features of sawtooth indicated the possibility of sliding mechanism in vimentin IF, and the sliding was found to occur at 30.44±13.41pN. Helical domain unfoldings were observed only in the non-fixed samples to start at 10.19±5.63pN on average witha mean increase of 42.12±26.74nm. This force agreed with the prediction of the extended Bell model described in the literature and the length increase was around double of the domain length, which indicated the uncoiling of the coiled-coils. The force-displacement curves also reveal different modes of failure of the vimentin intermediate filaments including protofilaments slippage/sliding and entropic elasticity. A new tearing off model was hence proposed based on different modes of failure and a previous model developed for desmin filaments reported in literature.published_or_final_version
Orthopaedics and Traumatology
Doctoral
Doctor of Philosophy
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Carter, D. Vaughan. "The role of vimentin antibodies in transplantation." Thesis, University of Newcastle Upon Tyne, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.424154.
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Vechio, Aluana Maria da Costa Dal. "Expressão da vimentina em cultivo tridimensional de linhagens celulares derivadas de carcinoma epidermóide de boca." Universidade de São Paulo, 2008. http://www.teses.usp.br/teses/disponiveis/23/23141/tde-08042009-160336/.
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O carcinoma epidermóide representa mais de 90% das neoplasias malignas de cabeça e de pescoço, apresentando taxas elevadas de morbi-mortalidade. Proteínas relacionadas à invasão e proliferação celular estão em evidência devido ao seu envolvimento na carcinogênese, a exemplo da vimentina, encontrada em células de origem mesodérmica. Sua presença em células epiteliais neoplásicas contribui na transição epitélio mesenquimal e está associada à tumorigênese, à invasão celular e à metástase. O propósito deste estudo foi analisar através de métodos qualitativos (imunofluorescência e imunoistoquímica) e quantitativos (Western Blot) a expressão da vimentina em linhagens celulares de carcinoma epidermóide de cabeça e de pescoço (CECP) e em uma linhagem de queratinócitos imortalizados (HaCat) submetidas ao cultivo tridimensional em Matrigel®. O grupo controle foi representado pelas mesmas linhagens cultivadas na ausência de Matrigel®. A Vimentina apresentou marcação citoplasmática em algumas células das linhagens estudadas, exceto na HaCat, com evidente diminuição da sua expressão quando submetida ao cultivo com Matrigel®. Esses resultados foram confirmados por Western Blot. A expressão da Vimentina em diferentes linhagens de CECP pode variar dependendo da linhagem analisada, da reação de suas células aos componentes da matriz extracelular e da técnica utilizada para avaliação da expressão da proteína.Head and neck squamous cell carcinoma (HNSCC) represents more than 90% of all head and neck malignancies, causing more deaths than any other oral disease. Proteins related to cancer growth, invasion and metastasis are in evidence due to their involvement in carcinogens, such as vimentin. This protein is observed in mesenquimal cells, however, it is considered a common finding in cervix, breast and bladder tumours. Thus its presencence in epithelial neoplasic cells contributes to epithelial-mesenchymal transition associated with tumorigenesis and tumor progression. The aim of this study was to analyse through Western Blot, Immunohistochemistry and Immunofluorescence methods, the expression of Vimentin in three different HNSCC cell lines and HaCat cell line (immortalized keratinocytes) submitted to a 3D assay into Matrigel®. The control group was represented by the same cell lines, without any treatment. Results showed that Vimentin had citoplasmatic staining in some cell of lines studied, except for HaCat cells, with evident decrease in its expression when submitted to cultive into Matrigel®. These findings were confirmed by Western Blot. Taking these results together, we conclude that in squamous cell carcinoma, the Vimentin is related to the process of tumour invasion and metastasis. This fact was showed by the reduction of its expression after treatment with Matrigel®. Therefore, the expression of Vimentin in different cell lines of HNSCC may vary according to the stimulus and, fundamentally, the localization of the tumor and the individual characteristics of neoplasic cells.
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Gianelo, Maikol Carlos Simões. "Estudo da resposta regenerativa do músculo sóleo de ratas bebês após procedimento de imobilização e reabilitação pelo alongamento." Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/17/17152/tde-02072015-122858/.
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Modelos de desusos do músculo esquelético como imobilização gessada, suspensão são frequentemente utilizados em grupos de pesquisas experimentais. Esse tempo de desuso por período prolongado pode determinar alterações significativas na citoarquitetura muscular. Este estudo teve como objetivo avaliar os aspectos morfológicos do músculo sóleo de ratas em desenvolvimento pós-natal que tiveram seus membros posteriores direitos imobilizados, e posteriormente foram submetidas ao protocolo passivo de alongamento (alongamento manual passivo intermitente), por um período de sete dias. Utilizou-se 20 ratas da raça Wistar (Rattus Norvegicus Albinus) com 21 dias de idade, dividas em cinco grupos: Grupo Controle 21 dias (GC21- Animais com 21 dias), Grupo Imobilizado (GI- Animais de 21 dias que foram imobilizados por 7 dias), Grupo Imobilizado e Alongado (GIA- Animais de 21 dias que foram imobilizados por 7 e reabilitados pelo alongamento durante 3 dias) e Grupo Alongado (GA- Animais de 21 dias não imobilizados por 7 dias e posteriormente alongamentos durante 3 dias), Grupo Controle 30 (GC30 - Animais com 30 dias). Fragmentos dos músculo sóleo foi processado sob diferentes métodos histoquímicos, coloração hematoxilina-eosina e picro-sirius. As variáveis foram avaliadas inter- e intra-grupos através de técnicas estatísticas como: Teste de Kruskall Wallis e pós teste de Dunn. Conclusão: Os resultados indicaram que o músculo sóleo de ratas bebês sofreram modificações citoarquiteturais significativas quando o alongamento manual intermitente foi usado como recurso terapêutico após 7 dias de desuso do segmento posterior direito (imobilização em flexão plantar). Nos músculos imobilizados, ambas as proteínas (desmina e vimentina) tiveram seus conteúdos reduzidos em relação aos valores controle (21 ou 30 dias), indicando balanço negativo para o tecido pós-desuso. A quantidade dessas proteínas não foi modificada nos animais submetidos somente ao procedimento de alongamento intermitente. Os animais que sofreram imobilização e que foram reabilitados, a quantidade de desmina não aumentou significativamente, não atingindo valores similares ao grupo controle 30 dias. Esses dados sugerem que os filamentos de desmina necessitam de tempo superior a 3 dias de reabilitação por alongamento intermitente para restabelecerem a arquitetura intersticial das fibras e consequentemente favorecerem a transdução mecânica de sinais entre os meios intra e extracelular. Porém, o efeito citoarquitetural do alongamento sobre os filamentos intermediários deve ser acompanhado longitudinalmente e confirmados em adicionais estudos bioquímicos e moleculares.Disuses models of skeletal muscle as immobilization , suspension are often used in experimental research groups. This time of disuse for a prolonged period can cause significant changes in muscle cytoarchitecture .This study aimed to evaluate the morphology of the soleus muscle of rats in postnatal development that had its members later immobilized rights, and subsequently were subjected to passive stretching protocol (passive manual stretching flashes), for a period of seven days. We used 20 Wistar rats (Rattus Norvegicus Albinos) race with 21 days of age , divided into five groups: Control Group (CG21- Animal 21 days ), Immobilized Group (IG- Animal 21 days that were immobilized for 7 days ) , Immobilized and Stretched Group (ISG - Animal 21 days that were immobilized for 7 and rehabilitated by stretching for 3 days) and Stretched Group (SG -Animal 21 days not immobilized for 7 days and subsequently stretching for 3 days), Control Group (CG30 - Animals 30 days).Fragments of the soleus muscle was processed under different histochemical methods, hematoxylin - eosin staining and picro-sirius. Variables were evaluated inter - and intra - groups through statistical techniques such as Kruskall Wallis and Dunn\'s post test . Conclusion: The results indicated that the soleus muscle of rats were babies citoarquiteturais significant changes when the manual stretching intermittently been used as a therapeutic resource after 7 days of disuse of the right posterior segment ( immobilization in plantar flexion). The immobilized muscles , both proteins (desmin and vimentin ) content had decreased compared to control values (21 or 30 days), indicating negative after- tissue balance disuse. The amount of these proteins was not modified in animals subjected only to intermittent stretching procedure. The animals underwent immobilization have been rehabilitated and that the amount of desmin was not significantly increased, not reaching values similar to the control group 30 days. These data suggest that desmin filaments need to 3 days longer than the time for rehabilitation to restore the intermittent stretching interstitial fiber architecture and hence favor the mechanical transduction of signals between intra-and extracellular media. However, the effect of stretching on cytoarchitectural intermediate filaments should be followed longitudinally and confirmed in additional biochemical and molecular studies .
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Pattabiraman, Sundararaghavan [Verfasser]. "Vimentin protects differentiating stem cells from stress / Sundararaghavan Pattabiraman." Göttingen : Niedersächsische Staats- und Universitätsbibliothek Göttingen, 2020. http://d-nb.info/1223171582/34.
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McGinn, Mary Catherine. "Interplay Between Keratin and Vimentin Expression in Oral Cancer." VCU Scholars Compass, 2010. http://scholarscompass.vcu.edu/etd/49.
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Previous research in our laboratory found that inhibiting expression of vimentin, a marker of epithelial-to mesenchymal transition, inhibited cell growth and motility in vitro and in vivo. Tumors derived from vimentin knockdown cells showed features of epithelial redifferentiation and increased expression of differentiation-specific keratins. It is unknown what causes re-expression of keratins when vimentin is inhibited. Although, canonical Wnt signaling may activate NF-κB and repress of keratin and/or induce vimentin expression through β-catenin. We hypothesize that downregulation of differentiation-specific keratins contributes to tumor progression, mediated directly or indirectly by expression of vimentin. Vimentin-negative HN4 cells were transfected with plasmids encoding wild-type, PKCε-phosphomimetic, or unphosphorylatable versions of vimentin. Expression of vimentin was confirmed by western blot and immunofluorescence. Effects on cell growth and motility were determined using MTT, cell proliferation, and wound-closure assays. These results indicate that mutation of vimentin PKCε-phosphorylation sites cause changes in proliferation and filament assembly. Treatment of cells with an NF-κB inhibitor or 5-Aza-C, which allows re-expression of the Wnt inhibitor DKK3, led to a decrease in proliferation. These results suggest that inhibiting Wnt signaling removes the inhibition on GSK-3β and prevents activation of NF-κB, which decreases proliferation.
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Kirmse, Robert. "Studium der Wachstumskinetik von Intermediärfilamenten mit Hilfe von Vimentin." [S.l. : s.n.], 2007. http://nbn-resolving.de/urn:nbn:de:bsz:16-opus-83413.
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Fay, Nikta. "Parvoviral interactions with the cytoskeleton : exposing vimentin – the forgotten player." Thesis, University of British Columbia, 2014. http://hdl.handle.net/2429/50672.
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There are three structurally and functionally distinct cytoskeleton components: actin filaments, microtubules, and intermediate filaments (IFs). Among the three cytoskeleton networks IFs are understudied; consequently, there is a lack of information about the role of IFs during early viral infection. IFs have long been known to serve structural functions within the cell, and recently, additional functions have been elucidated, including novel roles during infection by many viruses. During early infection with the parvovirus minute virus of mice (MVM), prior to viral replication, I have found that the virus induces dramatic morphological changes in mouse fibroblast cells. This observed change in the shape of infected cells may be a result of the virus using the host cytoskeleton to aid in the mechanism of intracellular trafficking. Thus, this thesis focuses on MVM and its effects on the cytoskeleton components, especially IFs, during infection. Using fluorescence microscopy techniques, I found that during early infection with MVM, after endosomal escape, the vimentin IF network was considerably altered, yielding collapsed immunofluorescence staining near the nuclear periphery. Furthermore, I found that vimentin plays an important role in the infection cycle of MVM. The number of cells successfully replicating MVM was reduced in infected cells in which the vimentin network was pharmacologically modified or in cells lacking a vimentin network; viral endocytosis, however, remained unaltered. Perinuclear accumulation of MVM-containing vesicles and progression of MVM through the endocytic pathway was reduced in cells lacking vimentin. Cells lacking vimentin, accumulated virions in early endosomes up to 2 h post-infection compared to wild type cells. Thus, my data supports a model where vimentin facilitates a productive MVM infection, presenting possibly a dual role: (1) during progression of MVM through the endocytic pathway and (2) following MVM escape from endosomes.Science, Faculty of
Zoology, Department of
Graduate
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Rato, Leila Sofia Coelho. "Vimentin interacts with the Akt/mTOR pathway mediating cell growth." Master's thesis, Universidade de Aveiro, 2017. http://hdl.handle.net/10773/22372.
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Mestrado em Bioquímica - Bioquímica ClínicaA vimentina é uma proteína da classe III dos filamentos intermédios que promove processos tais como proliferação, migração e invasão celular através da interação com diferentes vias de sinalização. No entanto, o papel da vimentina no crescimento celular é ainda pouco conhecido. Neste estudo, observamos que fibroblastos isolados de embriões de ratinhos sem vimentina (Vim -/- MEFs) eram mais pequenos que o tipo normal (WT). Assim, o objetivo deste estudo era entender de que forma a vimentina regula o crescimento celular. Com recurso a modelos in vitro, técnicas de microscopia e técnicas bioquímicas descobrimos que Vim -/- MEFs tinham menor volume e concentração de proteínas quando comparadas com WT MEFs. Adicionalmente, a síntese proteica e ativação de mTORC1 estavam significativamente reduzidas em Vim -/- MEFs. Através de co-imunoprecipitação, descobrimos que a vimentina interage com os complexos mTORC2 e TSC. Assim, postulamos que a vimentina regula o crescimento celular por interação com proteínas da via de sinalização AKT/mTO
Vimentin is a type III intermediate filament protein that takes part in cell proliferation, migration and invasion, by acting as a signalling scaffold. The role of vimentin in cell growth, however, is poorly understood. We observed that vimentin knockout mouse embryonic fibroblasts (Vim -/- MEFs) were smaller than the wild type (WT). Therefore, this work aimed to understanding how vimentin regulates cell growth. Using in vitro models, imaging techniques and biochemical approaches, we have found that the volume and protein concentration of Vim -/- MEFs is lower when compared to WT MEFs. Further, protein synthesis and mammalian target of rapamycin complex 1 (mTORC1) activation was attenuated in Vim -/- MEFs. By co-immunoprecipitation we found that vimentin interacts with mammalian target of rapamycin complex 2 (mTORC2) and tuberous sclerosis protein complex (TSC) after insulin stimulation. Consequently, we postulate that vimentin regulates cell growth by interacting with proteins of the AKT/mTOR pathway
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Botelho, Tessa de Lucena. "Expressão imunoistoquímica das proteínas c-erbB-2 e vimentina em carcinomas epidermóides bucais em correlação com características clínicas e prognóstico." Universidade de São Paulo, 2009. http://www.teses.usp.br/teses/disponiveis/23/23141/tde-29092009-081641/.
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Das neoplasias malignas que ocorrem na boca, 95% são representados pelo Carcinoma Epidermóide Bucal. Esta é uma doença usualmente agressiva, com comportamento biológico imprevisível e prognóstico desfavorável sendo a infiltração local e consequente emissão de metástases a principal causa de morte dos pacientes. A identificação de marcadores moleculares que possam predizer o curso clínico da doença, orientar a determinação do tratamento, bem como conduzir o desenvolvimento de novas terapias que melhorem os índices de sobrevida, tem sido o objetivo de inúmeras pesquisas. O presente estudo procurou determinar a correlação da expressão imunoistoquímica da c-erbB-2 e vimentina com características clínicas da neoplasia e o prognóstico dos pacientes a partir da análise retrospectiva de 65 casos de Carcinoma Epidermóide Bucal. c-erbB-2 e vimentina estavam expressos em 61,54% e 70,8% das amostras analisadas, respectivamente. Não foi observada correlação estatisticamente significante entre a expressão destes marcadores e as características clínicas avaliadas, porém houve uma tendência à expressão imunoistoquímica da vimentina em relação ao índice N. Quanto à sobrevida dos pacientes, esta foi influenciada pelo gênero dos pacientes, hábito de fumar cigarro, estádio clínico da doença, índice N e modalidade de tratamento submetido, sendo o gênero o único fator prognóstico independente detectado. A expressão imunoistoquimica da c-erbB-2 e vimentina não se demonstraram preditivos de sobrevida em carcinomas epidermóides de boca.About of malignancies in oral mucosa, 95% are represented by oral squamous cell carcinoma (OSCC). This disease is usually aggressive, with unpredictable biological behavior and poor prognosis and the local infiltration and consequent metastases the main cause of death of patients. The identification of molecular markers that may predict the clinical course of disease, guide the treatment as well as lead the development of new therapies that improve the rates of survival, has been the goal of many studies. This study examined the correlation of immunohistochemical expression of c-erbB-2 and vimentin with clinical features and prognosis of OSCC patients from the retrospective analysis of 65 cases. c-erbB-2 and vimentin were expressed in 61.54% and 70.8% of samples, respectively. There was no statistically significant correlation between the expression of these markers and clinical characteristics, but a tendency to vimentin expression in the lymph node status. The patients survival, was influenced by their gender, smoking habits the, clinical stage of disease, N index and modalities of treatment. The gender was the only independent prognostic factor detected. The immunohistochemical expression of c-erbB-2 and vimentin did not show up predictive of survival in oral cavity squamous cell carcinoma.
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Block, Johanna Lena [Verfasser]. "Stress-Strain Behavior of Single Vimentin Intermediate Filament / Johanna Lena Block." Göttingen : Universitätsverlag Göttingen, 2018. http://d-nb.info/1170672000/34.
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Mahesh, Balakrishnan. "Role of the anti-vimentin response in rejection of murine cardiac allografts." Thesis, Imperial College London, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.498312.
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Zehetmayer, Barbara. "Molekulargenetische Analyse der Kandidatengene p97/VCP, Myotilin und Vimentin bei hereditären Myopathien." Diss., Ludwig-Maximilians-Universität München, 2015. http://nbn-resolving.de/urn:nbn:de:bvb:19-180438.
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Die Gruppe der degenerativen Myopathien mit Proteinaggregaten und myofibrillären Veränderungen ist eine genetisch und klinisch heterogene Gruppe der erblichen Muskelerkrankungen. In der folgenden Arbeit wurde das Hauptaugenmerk auf zwei spezielle Erkrankungen gelegt. Dabei handelte es sich um die IBMPFD (Inclusion body myopathy, Paget‘s disease of bone and frontotemporal dementia – Einschlusskörpermyopathie mit Morbus Paget und frontotemporaler Demenz) und eine Form der Gliedergürteldystrophie (LGMD1A-Limb Girdle Muscular Dystrophy 1A). Beides sind Erkrankungen des Erwachsenenalters und eher durch eine langsame Progredienz gekennzeichnet.
Ziel der Arbeit war eine klinische Charakterisierung von betroffenen Patienten mit genauer molekulargenetischer Analyse der drei Kandidatengene VCP/p97 (VCP), Myotilin (MYOT) und Vimentin (VIM). Hierzu wurde bei der Verdachtsdiagnose einer degenerativen Myopthie oder Einschlusskörpermyopathie die Patienten-DNA auf mögliche krankheitsauslösende Mutationen untersucht. Des Weiteren folgten klinische Vergleiche des Phänotyps bestimmter Mutationen und spezifischere zellbiologische Untersuchungen zum besseren Verständnis der Pathogenese der Erkrankungen.
Im Rahmen dieser Arbeit wurde die genomische DNA von insgesamt 42 Patienten molekulargenetisch untersucht. Bei zwei Patienten wurde dabei die seltene Mutation R159C im VCP-Gen identifiziert, die klinisch zum Phänotyp einer IBMPFD passte, welche weiters in funktionellen Analysen im Zellkulturmodell untersucht wurde. Hierbei zeigte sich eine Störung der Myofibrillenbildung und Zelldifferenzierung.
Bei der dominanten Gliedergürteldystrophie LGMD1A konnte ein krankheitsassoziierter Basenaustausch im MYOT-Gen neu identifiziert werden, der mit der Erkrankung innerhalb der betroffenen Familie segregierte, In Kontrollpopulationen ohne neuromuskuläre Erkrankungen wurde diese Genvariante nicht nachgewiesen. Zusätzlich wurde die Pathogenität der Mutation durch die phylogenetische Konservierung der entsprechenden Aminosäureposition und bioinformatische Vorhersage-Algorithmen gestützt.
Zur Identifizierung weiterer genetischer Ursachen hereditärer Einschlusskörpermyopathien wurde das Kandidatengen Vimentin mittels direkter Sequenzierung bei einem Patientenkollektiv von 28 Patienten mit einer Aggregatmyopathie unklarer Genese untersucht. Das Protein Vimentin ist ein Typ-3-Intermediärfilament aus der Gruppe der Desmine. Sein Vorkommen in pathologischen Proteinablagerungen bei myofibrillären Myopathien legte einen Zusammenhang mit anderen Proteinaggregatmyopathien nahe, so dass es als interessantes neues Kandidatengen erschien. Hier fand sich jedoch kein Hinweis darauf, dass Mutationen im Vimentin-Gen eine häufige Ursache für Aggregatmyopathien in dem untersuchten Patientenkollektiv sind.
Für Patienten mit einer erblichen Aggregatmyopathie ist eine genaue genetische Diagnostik eine wichtige Information, da sie bei der Klassifikation der Erkrankung hilfreich ist, Aussagen über die Prognose vereinfacht und auch weiterführende, interdisziplinäre Diagnostik begründet. Zusätzlich hat eine genetische Erkrankung auch Auswirkungen auf die Familienplanung, sodass der sorgfältigen Diagnosestellung eine professionelle humangenetische Beratung folgen sollte.
Derzeit beschränkt sich die Therapie der in dieser Arbeit behandelten Erkrankungen überwiegend auf supportive Maßnahmen. In Zukunft sind aber auch spezifische Gentherapien vorstellbar, die eine präzise genetische Diagnostik als Basis benötigen. Bei einer Vielzahl von bisher unheilbaren, degenerativen neuromuskulären Erkrankungen bildet eine konsequente intensive Forschung auf molekular- und zellbiologischer Ebene die Grundlage neuer translationaler Forschungsansätze für künftige kausale Therapieoptionen.
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Denz, Manuela [Verfasser]. "Influence of Ions on the Assembly of Vimentin Intermediate Filaments / Manuela Denz." Göttingen : Niedersächsische Staats- und Universitätsbibliothek Göttingen, 2020. http://d-nb.info/1227039670/34.
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Kraxner, Julia [Verfasser]. "Vimentin Intermediate Filament Softening - Recovery Behavior and Post-Translational Modifications / Julia Kraxner." Göttingen : Niedersächsische Staats- und Universitätsbibliothek Göttingen, 2021. http://nbn-resolving.de/urn:nbn:de:gbv:7-21.11130/00-1735-0000-0008-590D-7-0.
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Ben, Azzouz Eya. "Étude physiopathologique des infections à Tropheryma whipplei." Thesis, Aix-Marseille, 2019. http://www.theses.fr/2019AIXM0243.
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La maladie de Whipple est une maladie systémique rare, causée par l’agent bactérien Tropheryma whipplei.T. whipplei présente un tropisme particulier pour les macrophages. Elle induit un profil d’activation macrophagique atypique, caractérisé par l’expression de molécules anti-inflammatoires, la sécrétion d’IL-16 et l’induction de l’apoptose. Au cours de cette étude, nous nous étions fixé comme objectif d’étudier l’interaction entre T. whipplei et les cellules myéloïdes (macrophages et monocytes). Dans un premier temps, nous nous sommes intéressés à une molécule immuno-régulatrice HLA-G impliquée dans les mécanismes de tolérance. Nous avons montré que HLA-G est fortement exprimée et secrétée in-vivo chez les patients atteints de la maladie de Whipple classique. Par ailleurs, nous avons constaté qu’in-vitro l’infection des monocytes par T. whipplei induisait l’expression de HLA-G accompagnée d’une faible sécrétion de TNF. D’autre part, nous avons montré que le blocage de HLA-G restaure la sécrétion de TNF par les monocytes et inversement, le blocage du TNF favorise la réplication de T. whipplei dans les macrophages.Dans un second temps, nous nous sommes intéressés aux évènements précoces de l’interaction entre T. whipplei et les cellules immunitaires dans le but d’identifier un partenaire cellulaire susceptible d’interagir avec T. whipplei. Nous avons pu montrer que la vimentine, une protéine du cytosquelette appartenant aux filaments intermédiaires est impliquée de manière spécifique dans l’adhésion et l’internalisation de T. whipplei. En définitive, ces travaux ont permis d’apporter des éléments de réponse pour mieux comprendre les infections à T. whippleiWhipple disease is a rare systemic disease caused by the bacterial agent Tropheryma whipplei. T. whipplei presents a particular tropism for macrophages. It induces an atypical macrophage activation program, characterized by the expression of anti-inflammatory molecules, the secretion of IL-16 and the induction of apoptosis. During this thesis, we analyzed the interaction between T. whipplei and myeloid cells (macrophages and monocytes). First, we focused on an immunomodulatory molecule, HLA-G which has previously been associated with immunotolerance mechanisms. We showed that HLA-G is strongly expressed and secreted in vivo in patients with classical Whipple disease. In addition, we found that in vitro, infection of monocytes by T.whipplei induced HLA-G expression accompanied with low TNF secretion. On the other hand, we have shown that a neutralizing antibody against HLA-G increased TNF secretion by monocytes in response to T whipplei, while a TNF inhibitor promoted bacterial replication. Thus, we were able to highlight the involvement of HLA-G in the persistence of T. whipplei within immune cells, then providing a proper environment for its replication.Second, we looked at the early events of T. whipplei interaction with immune cells in order to identify a cell partner who could interact with T. whipplei. We showed that vimentin, a cytoskeletal protein belonging to the intermediate filaments, is specifically involved in the adhesion and internalization of T. whipplei. In conclusion, this work has provided some answers to a better understanding of T. whipplei infections
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Paulin-Levasseur, Micheline. "Cellular dynamics of vimentin filaments and their spatial relationship to microtubules in lymphocytes." Thesis, University of Ottawa (Canada), 1987. http://hdl.handle.net/10393/5396.
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Tezcan, Okan. "Metastatic Behaviour Of Doxorubicin Resistant Mcf-7 Breast Cancer Cells After Vimentin Silencing." Master's thesis, METU, 2013. http://etd.lib.metu.edu.tr/upload/12615553/index.pdf.
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Chemotherapy is one of the common treatments in cancer therapy. The effectiveness of chemotherapy is limited by several factors one of which is the emergence of multidrug resistance (MDR). MDR is caused by the activity of diverse ATP binding cassette (ABC) transporters that pump drugs out of the cells. There are several drugs which have been used in treatment of cancer. One of them is doxorubicin that intercalates and inhibits DNA replication. However, doxorubicin has been found to cause development of MDR in tumors. It has been reported that there is a correlation between multidrug resistance and invasiveness of cancer cells. Vimentin is a type III intermediate filament protein that is expressed frequently in epithelial carcinomas correlating with invasiveness and also poor prognosis of cancer. There are several studies that have shown the connection between expression level of vimentin and invasiveness. In this study, MCF-7 cell line (MCF-7/S), which is a model cell line for human mammary carcinoma, and doxorubicin resistant MCF-7 cell line (MCF-7/Dox) were used. The resistant cell line was previously obtained by stepwise selection in our laboratory. The main purpose of this study was to investigate changes of metastatic behaviour in MCF-7/Dox cell line, after transient silencing of vimentin gene by siRNA. In conclusion, down-regulation of vimentin gene expression in MCF-7/Dox cell lines was expected to change the characteristics in migration and invasiveness shown by migration and invasion assays.
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Bird, Danielle N. "Regulatory Analysis of Vimentin Expression in Metastatic Versus Nonmetastatic Breast Cancer Cell Lines." VCU Scholars Compass, 1994. http://scholarscompass.vcu.edu/etd/4366.
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The intermediate filament gene family, composed of six classes, shows both tissue and development-specific expression. Vimentin is a unique member of the intermediate filament protein family: although it is expressed in cells of mesenchymal origin, vimentin may also be expressed with other intermediate filament proteins during early stages of development. In some cases, as differentiation continues, vimentin is normally down-regulated whereas other intermediate filament proteins, like desmin, glial fibrillary acidic protein, or neurofilaments are turned on in muscle, glial cells, or neurons, respectively. In some metastatic cancers, including breast and prostate cancers, vimentin is aberrantly expressed, despite the embryonic origin of the metastatic cell. From a Northern analysis (Thompson et al., 1992; Stover et al., 1994), it was determined that vimentin mRNA is highly abundant in the metastatic breast carcinoma cell line, MDA-MB-231, while in the nonmetastatic breast carcinoma cell line, MCF-7, no vimentin mRNA is present.
A central question to metastasis is, how is the vimentin gene expressed in the metastatic but not the nonmetastatic cancer cell? Several cis-acting elements localized to the 5'-end of the chicken vimentin gene and trans-acting factors important in the regulation of vimentin gene expression have been identified. By sequence homology, comparable elements can be found in the human vimentin gene. Most notable of these is a unique silencer element and an overriding element, referred to as an antisilencer. These elements bind a 90 kDa and 120 kDa protein, respectively, in chicken, mouse, and human nuclear extracts. Previously, the silencer factor was found to be missing in nuclear extracts from a metastatic breast cancer cell line MDA-MB-231 but abundant in the nonmetastatic counterpart MCF-7 (Stover et al., 1994). The antisilencer exhibits the opposite pattern. Here, various combinations of these elements have been fused to the bacterial chloramphenicol acetyltransferase reporter gene, CAT. Transcriptional activity was then compared in the different breast cancer cell lines in order to try and understand how vimentin is expressed in the metastatic but not the nonmetastatic cancer cell.
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Leong, Hon Sing. "The PAI-1-vitronectin-vimentin ternary complex : mechanism of extracellular assembly and role in transplant vasculopathy." Thesis, University of British Columbia, 2008. http://hdl.handle.net/2429/2509.
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The active state of plasminogen activator inhibitor type-1 (PAI-1) is prolonged when it forms a complex with vitronectin (VN), a major serum protein. Active PAI-1 in the PAI-1:VN complex serves many functions related to fibrinolysis and cell migration but key to these effects is its extracellular distribution. PAI-1:VN complexes can bind to exposed vimentin (VIM) on activated platelet and platelet microparticles, resulting in the assembly of PAI-1:VN:VIM ternary complexes. However, the manner in which the vimentin cytoskeleton is presented extracellularlyi s not well understood.
I hypothesized that PAI-1:VN:VIM ternary complex assembly occurs on cell surfaces when microparticle release leads to exposure of vimentin cytoskeleton which can lead to either assembly of the ternary complex or become involved in an autoimmune response specific for vimentin.
To follow the intracellular and extracellular fate of PAI-1, I constructed an expression vector encoding PAI-1-dsRed, a fluorescent form of PAI-1, which would permit live cell tracking of PAI-1 in megakaryocytes and endothelial cells. Secondly, to study how vimentin is expressed on platelets and platelet microparticles, flow cytometry was used to isolate vimentin positive platelets or PMP's and atomic force microscopy was performed to image platelets or PMP's at nanoscale resolution. From these studies, I propose a model of vimentin expression in which the junction of microparticle release results in the exposure of cytoskeletal vimentin on both the cell and the microparticle. This exposed vimentin could potentially induce VN multimerization on the same cell surface leading to incorporation of multiple PAI-1:VN complexes.
Finally, I investigated how anti-vimentin antibodies can induce platelet:leukocyte conjugate formation. To achieve this, in vitro tests were performed to determine the binding site of anti-vimentin antibodies (AVA's) and how they induce blood cell activation. Overall, my results suggest that vimentin exposure in our model of microparticle release can lead to ternary complex assembly if suitable quantities of PAI-1 are released during platelet activation. In the setting of transplant vasculopathy with high titres of AVA's, vimentin-positive granulocytes can bind these autoantibodies, which then leads to platelet activation and the formation of platelet:leukocyte conjugates.
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Kim, Ryung Rae. "Investigation of the action of new pentapeptides entities and the interaction between human group IIA phospholipase A2 and vimentin." Thesis, The University of Sydney, 2018. http://hdl.handle.net/2123/19853.
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The cyclic pentapeptide c2 is a promising anti-cancer agent. The thesis investigates the mechanism of action of c2, focusing on its ability to prevent the interaction between hGIIA and vimentin. Computational studies using molecular dynamics and docking predicted that hGIIA would be stable as a monomer in physiological environments and provided possible binding conformations of c2 compound in hGIIA and vimentin. Using recombinant hGIIA and vimentin, assays were conducted to investigate the binding of hGIIA to vimentin and the role of c2 in inhibiting the interaction. Surface plasmon resonance and enzyme immunoassay studies showed evidence of hGIIA binding at the coil 2 domain of vimentin, and that c2 can inhibit this binding. Radiolabelled c2 displayed high affinity towards the coil 2 domain of vimentin relative to the coil 1 domain, suggesting that c2 prevents the interaction between hGIIA and vimentin possibly by binding to the hGIIA binding site on vimentin. From these results combined with the computational docking work, it is predicted that the most probable binding site of c2 on vimentin is located between Met347 and Asn357. In the advanced prostate cancer cell line PC-3, exogenously added hGIIA did not significantly increase the proliferation rate of the cells, and nor was this affected by vimentin silencing. The prostaglandin E2 level in PC-3 cells were reduced by the exogenously added hGIIA, and c2 had an opposing effect to hGIIA. Such results are a contrast to data that has come from early stage prostate cancer cells. The role of hGIIA potentially has changing roles depending on the stage and type of the cancer, and the mechanism of action of hGIIA in the prostate cancer cells has the potential to be highly complex. It was also demonstrated that c2 can bind to prostate cancer cells without the expression of hGIIA. It suggests the incorporation of c2 may be dependent on cell surface proteins such as vimentin to which it has affinity rather than hGIIA.
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Lazaruk, O. V. "Vimentin expression of parenchymal cells and stromal cells of ductal breast carcinoma: comparative characteristics." Thesis, Sumy State University, 2016. http://essuir.sumdu.edu.ua/handle/123456789/44936.
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Ductal breast carcinoma ranks first among all malignant tumours of the reproductive system in women. To study the processes that occur when changing the normal structures gland tumor should be considered in the processes that occur in the tumour site and area around the tumour. To study the transformation in the fabric around the tumour is widely used immunohistochemical detection methods, including vimentin. Vimentin - a protein that is expressed in normal cells of mesenchymal origin. Increased expression of vimentin is observed in a variety of epithelial tumours. This, in turn, shows the epithelial-mesenchymal transformation, by which the tumour becomes different characteristics: fast growth, capacity for invasion, metastasis, tumour resistance to treatment and prognosis.
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Paccione, Rachel J. "Vimentin Overexpression Contributes To the Biological Properties of Metastatic Head and Neck Cancer Cells." VCU Scholars Compass, 2005. http://scholarscompass.vcu.edu/etd/1084.
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Epithelial to mesenchymal transition occurs in the later stages of epithelial tumor progression, with cells expressing mesenchymal markers. Of these, the intermediate filament protein vimentin is frequently upregulated in metastatic carcinomas. Previously, microarray studies showed that the gene encoding vimentin is highly upregulated in metastatic HN12 cells compared to a related primary tumor cell line. In this study, we confirmed this difference using real-time quantitative PCR, western blot analysis, and immunostaining. Furthermore, EGF and TGF-β, growth factors that induce migration and invasion of HN12 cells, produced synergistic increases in vimentin expression. To assess the contribution of vimentin to the biological properties, HN12 cells were stably transfected with a plasmid that directs synthesis of vimentin shRNA. Clones expressing decreased amounts of vimentin were isolated and characterized. These cells showed significantly reduced proliferation compared to non-targeting controls. Moreover, downregulation of vimentin led to a decrease in cell motility, as well as reducing their ability to invade through a basement membrane substitute. Using transient transfection assays, vimentin promoter activity was determined in HN12 cells to define regulatory elements important for controlling vimentin upregulation in the absence or presence of EGF and TGF-β. Taken together, the data indicate that overexpression of vimentin is important for proliferation and invasion of metastatic HN12 cells, and suggest that EGF- dependent pathways target binding elements in the proximal vimentin promoter, while TGF-β is likely to act in an AP1-dependent manner. Furthermore, both growth factors appear to synergize by stimulating promoter activation through the ASE site, suggesting involvement of Stat-dependent pathways in regulation of vimentin expression in HN12 cells.
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Lardier, Nathan. "Mechanics of intermediate filaments and microtubules in living cells." Electronic Thesis or Diss., Université Paris sciences et lettres, 2021. http://www.theses.fr/2021UPSLS058.
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Bien que largement étudiée in vitro, la mécanique du cytosquelette reste encore peu explorée dans les cellules vivantes. Nous utilisons une technique de micromanipulation intracellulaire basée sur des pinces optiques pour appliquer des forces directement sur les filaments du cytosquelette afin de sonder la mécanique des microtubules et des filaments intermédiaires et de nous intéresser à leur interaction mécanique. En mesurant simultanément la force appliquée aux filaments et leur déflexion, c'est-à-dire la déformation des filaments perpendiculairement à leur axe longitudinal, en fonction du temps, nous pouvons déduire les courbes force-déflexion des filaments et caractériser la rigidité des filaments intermédiaires de vimentine et des microtubules. Par un ajustement linéaire des courbes force-déflexion dans le régime des faibles forces, nous montrons que les microtubules ont une rigidité effective plus faible que la vimentine lors de la déflexion. Nous appliquons ensuite des forces deux fois sur le même faisceau de cytosquelette pour montrer que les filaments de vimentine, mais pas les microtubules, se rigidifient d'un facteur supérieur à trois lors de déflexions répétées. Nous caractérisons plus en détail le couplage mécanique entre les filaments de vimentine et les microtubules en utilisant des agents déstabilisateurs et stabilisateurs des microtubules et en augmentant l'acétylation des microtubules. De façon intéressante, nous constatons que ces modifications n'affectent pas la rigidité effective des filaments de vimentine alors que la déstabilisation ou l'acétylation des microtubules réduit significativement la rigidité des filaments de vimentine lors de déflexions répétées. Dans l'ensemble, ces résultats suggèrent que les microtubules favorisent la rigidification des faisceaux de vimentine lors de contraintes mécaniques répétées. En revanche, dans les cellules où la vimentine est inactivée, les propriétés mécaniques des microtubules sont inchangées. Nos résultats soulignent l'importance des interactions entre les microtubules et les filaments intermédiaires dans la mécanique cellulaire et suggèrent que les filaments intermédiaires de vimentine sont des structures mécano-sensibles qui présentent des réponses dépendantes des contraintes mécaniques passéesAlthough extensively studied in vitro, the mechanics of the cytoskeleton is still largely unexplored in living cells. We use an intracellular optical tweezers-based micromanipulation technique to apply forces directly on cytoskeletal filaments in order to probe microtubules and intermediate filament mechanics and focus on how they interact mechanically. Measuring simultaneously the force applied to the filaments and their deflection, i.e. the deformation of the filaments perpendicular to their axis, as a function of time, allows us to deduce the force-deflection curves of the filaments and to characterize the rigidity of vimentin intermediate filaments and microtubules. By fitting the force-deflection curves at small forces, we show that microtubules have a lower effective stiffness than vimentin upon deflection. We then apply forces twice on the same cytoskeletal bundle to show that vimentin filaments, but not microtubules, stiffen more than three times upon repeated deflections. We further characterize the mechanical coupling between vimentin filaments and microtubules by using microtubule destabilizing and stabilizing drugs and by increasing microtubule acetylation. Interestingly, we find that these modifications do not affect the effective stiffness of vimentin filaments while destabilizing or acetylating microtubules significantly reduces vimentin filament stiffening upon repeated deflection. Altogether, these results suggest that microtubules promote stiffening of vimentin bundles under repeated mechanical stress. In sharp contrast, in cells knockout for vimentin, the mechanical properties of microtubules are unchanged. Our findings highlight the importance of the interactions between microtubules and intermediate filaments in cell mechanics and suggest that vimentin intermediate filaments are mechanosensitive structures which exhibit history-dependent mechanoresponses
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Espejel, del Moral María del Carmen. "Histomorphologische und immunhistologische Charakterisierung der Endometrose beim Rind." Doctoral thesis, Universitätsbibliothek Leipzig, 2012. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-99530.
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In Anlehnung an die Definition beim Pferd (SCHOON et al. 1992, 1997) wird die bovine Endometrose als endometriale periglanduläre und/oder stromale Fibrose mit Alteration der betroffenen Drüsen definiert (RODENBUSCH 2011). Eine eingehende histomorphologische und immunhistologische Charakterisierung der Endometrose existiert bisher bei der Stute (KENNEY u. DOIG 1986, SCHOON et al. 1992, HOFFMANN et al. 2009), jedoch nicht beim Rind. Das Ziel dieser Arbeit ist daher die histomorphologische und immunhistologische Charakterisierung der verschiedenen Endometroseformen beim Rind.
Die Auswertung der Proben erfolgt am Institut für Veterinär-Pathologie der Universität Leipzig. In Abhängigkeit von den klinisch-gynäkologischen Befunden werden diese Proben in drei Gruppen unterteilt. Gruppe A1: Endometriumbioptate (n=12) von vier klinisch-gynäkologisch gesunden fertilen Rindern in definierten Zyklusphasen; Gruppe A2: Endometriumbioptate (n=36) von 36 klinisch genitalgesunden Rindern mit mindestens einer Abkalbung und Gruppe B: Uterusquerschnitte (n=69) von 69 sub-/ infertilen Rindern.
Die Proben werden anhand der von HOFFMANN (2006) definierten Kriterien auf histopathologische Veränderungen hin untersucht und charakterisiert. Das histomorphologische Erscheinungsbild der involvierten periglandulären Stromazellen erlaubt die Einteilung der Endometrose in eine aktive, inaktive und gemischte Fibrose, die, je nach der Integrität des Drüsenepithels, einen destruierenden oder nicht destruierenden Charakter aufweist. Zur Charakterisierung der Endometroseformen werden neben histomorphologischen Kriterien die Intermediärfilamente Desmin, Vimentin und Zytokeratin sowie die Expression von α-Aktin und Laminin berücksichtigt. Die deskriptive statistische sowie die Inferenzstatistik-Auswertung erfolgen unter Zuhilfenahme der Software SPSS 18.
Eine aktive Fibrose tritt bei 96,2 % der Rinder auf; 1,9 % weisen eine inaktive und 1,9 % eine gemischte Endometrose auf. Ein nicht destruierendes Erscheinungsbild der Endometrose kann bei 88,6 % der untersuchten Rinder nachgewiesen werden. Bei 11,4 % der untersuchten Rinder liegt eine Endometrose mit destruierendem Charakter vor. Die Endometrose betrifft bei 81 % der Rinder Einzeldrüsen und bei 19 % Drüsennester. Drüsennester sind häufiger bei mittel- und hochgradigen Endometrosen nachweisbar. 20 % der Proben mit nicht destruierender Endometrose und 58 % der Proben mit destruierender Endometrose weisen eine entzündliche Infiltration der Drüsen auf, wobei ein Zusammenhang zwischen der entzündlichen Infiltration der endometrotischen Drüsen und dem destruierenden Charakter der Endometrose festgestellt werden kann.
Zusätzlich zur Endometrose zeigen 61 % der Rinder eine Endometritis, 12,4 % eine Perivaskulitis und 66 % eine interkarunkuläre und/oder intrakarunkuläre Angiosklerose. Insgesamt findet sich nur bei 24 % der Fälle ausschließlich eine Endometrose, bei 10,5 % eine Endometrose und zugleich eine Endometritis, bei 16 % liegt eine Endometrose zusammen mit einer Angiosklerose vor. Eine Kombination von Endometrose, Angiosklerose und Endometritis ist bei 49,5 % der Proben nachweisbar. Insgesamt bestehen jedoch keine erkennbaren statistischen Zusammenhänge zwischen den einzelnen Befundkombinationen.
Aufgrund der immunhistologischen Untersuchung kann konstatiert werden, dass die periglandulären Stromazellen innerhalb der Endometrose eine stromale Koexpression von Desmin, Vimentin und α-Aktin aufweisen, welche ein für Myofibroblasten charakteristisches Merkmal ist. Ein kleiner Prozentsatz der Drüsenepithelzellen in der destruierenden Endometrose reagiert multifokal positiv mit dem Vimentinantikörper. Dies ist möglicherweise Ausdruck einer Fehldifferenzierung zur Stabilisierung der Zelle oder Anzeichen einer intensivierten (pathologischen) Proliferation. Die Lamininexpression der Basallamina der endometrotisch veränderten Drüsen ist, insbesondere bei der destruierenden Endometrose, diskontinuierlich und geht mit einer Auffaserung der Basallamina einher. Vermutlich lassen sich die umfangreichen Basallaminaalterationen auf von Myofibroblasten sezernierte Enzyme zurückführen.
Bei Rindern kann kein Zusammenhang zwischen der Endometrose und dem Alter der Rinder oder der Anzahl der Kalbungen festgestellt werden. In der vorliegenden Arbeit dominiert die geringgradig aktive nicht destruierende Endometrose gegenüber den anderen bovinen Endometroseformen. Die sub-/ infertilen Kühe (Gruppe B) zeigen häufiger eine schwerere und destruierende Endometrose als die klinisch gesunden Rinder (Gruppe A1, A2). Die klinisch-gynäkologisch gesunden Rinder in definierten Zyklusphasen weisen variable Endometroseformen oder Endometrosegrade auf. Die sub-/ infertilen Rinder zeigen eine höhere Güstzeit (252,82 ± 163,83 Tage) als die klinisch-gynäkologisch gesunden Tiere mit mindestens einer Abkalbung (94 ± 28,4 Tage). Die längere Güstzeit bei den sub- und infertilen Rindern könnte somit eine Folge des insgesamt in Charakter und Grad stärker geschädigten Endometriums bei dieser Gruppe sein. Somit kann unter Berücksichtigung der vorliegenden Ergebnisse angenommen werden, dass die aufgeführten Alterationen die Fertilität des Rindes negativ beeinflussen.
Die Ergebnisse ermöglichen eine histomorphologische und immunhistologische Charakterisierung der Endometrose beim Rind. Anhand der Ergebnisse der hier durchgeführten detaillierten Untersuchungen ist es möglich, eine präzisere Deskription degenerativer endometrialer Befunde vorzunehmen. In Hinblick auf die Fertilität bei Vorliegen einer bovinen Endometrose wird somit die Grundlage für zukünftige prognostische Bewertungen gelegt.
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Thiagarajan, Praveena S. "VIMENTIN IS A PHOSPHORYLATED TARGET OF MCP-1-INDUCED PKCβ ACTIVATION AND AN ENDOGENOUS LIGAND FOR THE INNATE IMMUNE RECEPTOR DECTIN-1." Kent State University / OhioLINK, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=kent1399566096.
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井川, 敬介. "Polo-like kinase1はvimentinのリン酸化を介して分裂期において初期エンドソームの膜融合を阻害する." 京都大学 (Kyoto University), 2014. http://hdl.handle.net/2433/188844.
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Wu, Wei. "Unveiling the neglected roles of nucleoprotein NLS2 and cellular vimentin during Influenza A virus infection." Thesis, University of British Columbia, 2014. http://hdl.handle.net/2429/51474.
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Influenza A virus exploits the cellular transport machinery during the early stages of infection. It enters cells by endocytosis and takes advantage of the endocytic trafficking to move towards the perinuclear region with the assistance of actin filaments and microtubules. A recent proteomic study identified vimentin as a putative interacting protein of influenza viral components. However, the role of vimentin during influenza A infection has not yet been determined. After endocytosis, the viral ribonucleopotein complexes (vRNPs), containing the RNA viral genome, the viral polymerases, and several copies of nucleoprotein, are released from late endosomes and enter the nucleus for replication. Two nuclear localization sequences (NLSs), NLS1 and NLS2, on nucleoprotein mediate the nuclear import of vRNPs. The function of NLS1 has been well studied, however, the role of NLS2 remains to be defined.
This thesis has two major aims: to characterize the function of NLS2 and to determine the role of vimentin during influenza infection. For the first aim, I use a chimeric protein (5GFP) fused to NLS2, in combination with RNAi of several importin α isoforms and biochemical assays, and found that NLS2 is able to mediate the nuclear import of 5GFP by interacting with importin α1, α3, α5, and α7. NLS2 contains only a single amino acid difference at position 17 between different strains of influenza A virus, which could be either lysine (K) or arginine (R). I found that NLS2K induces more nuclear accumulation of 5GFP than NLS2R. Using site-directed mutagenesis I demonstrated that NLS2K contains two independent functional basic clusters, while NLS2R only has one. Moreover, my study also revealed that inhibiting the function of NLS1 and NLS2 impairs the nuclear import of vRNPs and further inhibits viral infection.
For the second aim I followed influenza A virus infection in both vimentin null cells and vimentin RNAi knock-down cells, and found that vimentin plays a role in releasing vRNPs from endosomes. In summary, my work dissects the basic mechanisms involved in influenza A virus endocytic trafficking and nuclear import, which provide us with better insights into the viral-host interaction during influenza A virus infection.Science, Faculty of
Zoology, Department of
Graduate
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Gumprecht, Annett. "Analyse morphometrischer Messungen an Astrozyten des Hippokampus von Wildtypmäusen im Vergleich zu GFAP-/- - Vimentin-/- - Mäusen." Doctoral thesis, Universitätsbibliothek Leipzig, 2013. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-129946.
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Die Forschungsergebnisse der letzten Jahre beweisen, dass die Informationsverarbeitung im ZNS auf eine ausgewogene Interaktion zwischen den Neuronen und den Astrozyten im Sinne eines funktionellen Netzwerkes angewiesen ist. Allerdings liegen nur unzureichende Erkenntnisse über den strukturellen Charakter dieser symbiotischen Beziehung vor.
Zu den grundlegenden Aufgaben der Astrozyten gehört die Modulation der synaptischen Aktivität von Neuronen, Aufrechterhaltung der extrazellulären Homoöstase, Ausbildung der Blut-Hirn-Schranke, Pufferung der lokalen Kaliumkonzentration und die Synthese von Zytokinen, Wachstumsfaktoren sowie Neurotransmittern.
Ein Hauptbestandteil des Zytoskelettes der Astrozyten stellen die Intermediärfilamente (z.B. GFAP, Vimentin) dar. Funktionell dienen sie dem Aufbau von Zell-Zell- Kontakten, der Stabilisierung des Zytoskelettes und der Verankerung der Zellorganellen im Zytoplasma.
Charakteristisch für den strukturellen Aufbau der Astrozyten ist das GFAP, welches hauptsächlich in den Zellfortsätzen lokalisiert ist. Aktuelle Forschungen legen nahe, dass es sowohl unter physiologischen Alterungsprozessen als auch im Rahmen von pathologischen Vorgängen im ZNS (beispielsweise chronischer Alkoholabusus, Alexanderkrankheit, Ischämien und Epilepsie) zu einem prozentualen Anstieg der GFAP-Expression kommt, wobei der Einfluss dieser erhöhten GFAP-Synthese auf die Funktionsfähigkeit der Astrozyten noch nicht umfassend geklärt werden konnte.
Im Zentrum dieser Dissertation steht deshalb die Fragestellung, haben Alter und Intermediärfilamente (GFAP, Vimentin) Einfluss auf Morphologie und Zellzahl der Astrozyten im Hippokampus einer Maus?
Initial erfolgte die Volumenbestimmung der Astrozytendomänen mittels LSM- Aufnahmen im Reflexionsmodus sowie Vermessung nach Bearbeitung der Präparate mit der Silber Imprägnationstechnik. Die Domäne eines Astrozyten stellt das von einem Astrozyten mit Soma und allen Fortsätzen okkupierte Volumen dar.
Nach der Auswahl von S100ß als immunhistochemischen Astrozytenmarker wurde die Zellzählung in entsprechenden Hippokampusarealen durchgeführt.
Aus den ermittelten Daten wurde rechnerisch der Überlappungsgrad der hippokampalen Astrozytendomänen bestimmt. Von entscheidendem Interesse war dabei die Möglichkeit einer Überschneidung benachbarter Astrozytendomänen.
Aus den Messungen resultiert ein ca 1,6 fach grösseres Volumen der Wildtyp-Tiere im Vergleich zu den Doppel-knock-out-Mäusen. Im Gegensatz dazu zeigten die Ergebnisse der Zellzählung sowohl in der Kontrollgruppe als auch in der Gruppe der Versuchstiere eine vergleichbare Astrozytenanzahl pro mm3.
Entsprechend lag der Überlappungsfaktor bei den Doppel-Knock-out-Tieren (0,49) unter dem der Wildtyp-Tiere (0,7). Die abschließende Auswertung erbrachte in allen Untersuchungsgruppen einen Überlappungsfaktor < 1.
Ausgehend von der essentiellen Bedeutung der Astrozyten für die Funktionsfähigkeit der Nervenzellen käme es z.B. im Rahmen pathologischer Prozesse, welche mit Gliazellschäden einhergehen, bei einem Überlappungsfaktor < 1 zu erheblichen Engpässen in der neuronalen Versorgung, sowie in der Kompensation äußerer Einflüsse. Die Wahrscheinlichkeit irreversibler Schädigung der Nervenzellen steigt.
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Knecht, Sharmon M. "PI3K mediates S. aureus invasion leading to peri-nuclear vimentin collapse in human endothelial cells." Virtual Press, 2005. http://liblink.bsu.edu/uhtbin/catkey/1319545.
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Staphylococcus aureus (S. aureus) is a medically important bacterial pathogen associated with many diseases and infections of the respiratory system, wound sites, surgical incisions, and other portals of entry and exit. S. aureus is able to invade cells via mechanisms that have yet to be fully characterized. Vimentin, a protein filament of the animal cell cytoskeleton, and phosphoinositide 3-kinase (PI3K), a family of kinases responsible for initiating several cell signaling events, were found to be associated with S. aureus invasion. Confocal microscopy revealed that the vimentin network in human umbilical vein endothelial cells (HUVECs) undergoes dynamic rearrangement in steady state under control conditions. However, cells infected with S. aureus demonstrated peri-nuclear collapse of the vimentin network. Pre-treatment with LY294002, a drug that inhibits PI3K activity, decreased invasion of S. aureus and paralyzed the vimentin network. These data suggest that PI3K mediates S. aureus infection and vimentin rearrangement.Department of Biology
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Wiseman, J. "Relocalisation of β-globin coding sequences by myosin heavy chain and vimentin 3' untranslated regions." Thesis, University of Aberdeen, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.318982.
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The presence of a localisation signal within the 3' untranslated regions of myosin heavy chain and vimentin messenger RNAs was investigated by studying mRNA distribution in myoblasts and fibroblasts which were transfected with β-globin and chimaeric β-globin-myosin and β-globin-vimentin gene constructs. In cells transfected with constructs containing either a fragment of the β-globin gene containing both coding and 3' untranslated sequences or the β-globin coding sequences alone, in situ hybridisation revealed that the mRNAs transcribed from these constructs were distributed throughout the cytoplasm without any evident localisation. In contrast, within cells transfected with constructs containing β-globin coding sequences linked to the myosin heavy chain or vimentin 3' untranslated sequences there was a strong perinuclear localisation of the chimaeric mRNAs. This indicated that loss of its 3' untranslated region does not affect distribution of β-globin mRNA whereas the 3' untranslated sequences of both myosin heavy chain and vimentin cause a relocalisation of β-globin mRNA. The ability of the myosin heavy chain 3' untranslated region to relocalise β-globin coding sequences was investigated in the more complex cellular environment of skeletal muscle by intra-muscular injection of β-globin and chimaeric β-globin-myosin constructs. In muscle injected with β-globin constructs in situ hybridisation revealed that mRNAs transcribed from these constructs were distributed throughout the muscle section with no apparent localisation.
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Mostajeran, Zahra [Verfasser]. "The influence of vimentin on actin dynamics and force generation in RPE1 cells / Zahra Mostajeran." Saarbrücken : Saarländische Universitäts- und Landesbibliothek, 2021. http://d-nb.info/1237268761/34.
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Jungert, Kerstin. "Untersuchungen zur Bedeutung des Transkriptionsfaktors Sp1 in der TGF-beta-1-vermittelten Progression des Pankreaskarzinoms." [S.l. : s.n.], 2006. http://nbn-resolving.de/urn:nbn:de:bsz:289-vts-55615.
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Nair, K. Sukumaran. "The role of anti-vimentin antibodies in graft dysfunction after cardiac transplantation in a mouse model." Thesis, Imperial College London, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.436124.
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Dune, Ana Cláudia [UNESP]. "Expressão de recptor de estrógeno, vimentina, TGFbeta, e marcador de macrófagos em tumor ósseo de células gigantes em gatos domésticos." Universidade Estadual Paulista (UNESP), 2008. http://hdl.handle.net/11449/95976.
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O tumor ósseo de células gigantes apresenta 3 diferentes tipos celulares, sendo duas estromais: fibroblastos neoplásicos e células mononucleares; e o terceiro tipo, células gigantes. Propôs-se que este tumor é de linhagem monócito-macrofágica e acredita-se que as células gigantes se formam por fusão de células mononucleares. Aparentemente os fibroblastos neoplásicos que expressam o fator transformador de crescimento TGFbeta1 estão envolvidos no recrutamento das células gigantes para o tumor. Com o intuito de compreender melhor a histogênese, o envolvimento do estrógeno e a expressão de receptores TGFbeta1, foi realizado este estudo em casos deste tumor em gatos domésticos. Para tanto foi utilizado o método imuno-enzimático Streptoavidina-biotina utilizando-se o anticorpo primário anti-vimentina, clone 3B4 (Dako A/s, Denmark); o anticorpo marcador de macrófago, MAC387 (Dako A/s, Denmark); o anticorpo para receptores de estrógeno, clone 15D (Dako A/s, Denmark) e o anticorpo marcador para TGFBeta1 (Santa Cruz Biotechnology). Os resultados foram analisados pela porcentagem e desvio padrão de células marcadas para cada anticorpo e permitiram concluir que: o TOCG de gatos domésticos, assim como em humanos, tem origem mesenquimal e expressa receptores de estrógeno e de TGF 1 e as células gigantes do tumor não reagem com o clone 387, marcador de células de linhagem mielomonocítica.
Giant cell tumor of bone are composed of 3 different cell types: round mononuclear stromal cells, spindle-shaped mononuclear stromal cells, and giant cells. Some authors assert giant cell could arise by fusion of mononuclear cell mielomonocytic. Aparently neoplasic fibroblast that expression TGF is involved in the recruitment of giant cells from tumor. For better understand of histogenesis, the involved of receptors estrogen and of expression of TGF receptors, achieved this study in this cases tumor in domestics cats. By using the immune-enzymatic Streptoavidin- biotin using the primary antibody anti-vimentin, clone VIM 3B4 (Dako A/s, Denmark); antibody myeloid/histocyte clone MAC 387 (Dako A/s, Denmark); antibody estrogen receptor clone 1D5 (Dako A/s, Denmark) and the antibody TGFb1 (Santa Cruz Biotechnology). The results analyzed for percent and standara deviation of marks cells for each antibody and permissive to come to a conclusion: the GCT of bone in domestic cats, like humans, has mesenchymal origin and has expression of estrogen receptors and of TGFbeta, and giants cells this tumor not react with the clone 387, mark the cells myeloidmonocyte lineage.
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TUVERI, ROSSANA. "Identification & characterization of natural and synthetic compounds as new anticancer agents." Doctoral thesis, Università degli Studi di Cagliari, 2016. http://hdl.handle.net/11584/266770.
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This thesis collects the work I have done during the three-year PhD Course. During my first
year I have started a path that has allowed me to acquire different techniques devoted to set up
and maintain primary cell cultures and cancer derived cell lines as well as to evaluate the
cytotoxicity of potential novel synthetic inhibitors of human cancer cells. Part of the second and
all the third year was spent at the University of Cape Town, South Africa, in the laboratory of
prof. Ariel Katz under the supervision of proff. Roger Hunter and Catherine H. Kaschula
investigating the anticancer activity of Z-ajoene, a garlic compound.
Overall, the main aim of all the my research project was to identify and characterize natural or
synthetic compounds as new antineoplastic agents. The results obtained are divided according
to the research topics addressed:
Anticancer activity of new Phenanthroline compounds (Part I);
The garlic compound Z-ajoene as anticancer agents (Part II).
Studies referring to the Part have been carried out at the University of Cagliari and were
focalized on the evaluation of new Cu(II) -phenanthroline complexes as a potent antineoplastic
agents against various solid and suspension tumours. The [Cu(1,10-phenanthrolin
-5,6-diol)2(OH2)](ClO4)2 complex appears to be the most potent compound against human
leukemia, prostate and lung cancer cell lines. The results obtained on the biological activity of
this class of compounds, providing valuable information for the design of new anticancer drugs,
have been published in the Journal of Inorganic Biochemistry (2014).
As for the Part II of my research, I focused on the mechanisms underlying the anti-tumoral
activity of garlic compound Z-ajoene on human triple –negative breast cancer cells. The results
indicate that Z-ajoene localizes in the ER of MDA-MB-231 cells where it activates the unfolded
protein response (UPR) and ER stress. These findings have been published in the Molecular
Carcinogenesis journal (2015)
Moreover, immunofluorescence studies support the concept that the Z-ajoene main target is a
ER-resident chaperon protein (PDI), whose functional alteration may well be the cause of the
cytotoxic effect. Another molecular target of Z-ajoene is the cytoskeleton protein Vimentin.
Z-ajoene interacts with Vimentin through a S–thiolation causing the disruption of Vimentin
filaments and therefore an alteration of the cell morphology. Given that Vimentin is known to
participate to the early stage of the metastatic process, I also investigated the potential effect of
Z-ajoene at non-cytotoxic concentrations in a specific cell assay and found that it effectively
inhibits cell migration, both in the absence and presence of a chemotactic agent. The metastatic
inhibition induced by Z-ajoene seems caused by modification of several signaling pathways as
expression of Axl and Src proteins, and phosphorylation of β–catenin were changed. Although
following inhibition of cell migration, a reduction of Vimentin expression was to be expected,
Z-ajoene treatment surprisingly induced an upregulation of Vimentin. We interpreted this result
as a consequence of Z-ajoene binding to Vimentin which unable this protein to perform its
physiologic functions (manuscripts in preparation).
Altogether, the data of my in vitro study indicate that Z-ajoene is a promising chemotherapeutic
agent simultaneously acting on different molecular targets, also able to affect the metastatic
process in cells derived from highly invasive breast tumors. Due to its potential use in the clinic,
preclinical evaluation in xenograft mouse models of cancer are ongoing.
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Zehetmayer, Barbara [Verfasser], and Sabine [Akademischer Betreuer] Krause. "Molekulargenetische Analyse der Kandidatengene p97/VCP, Myotilin und Vimentin bei hereditären Myopathien / Barbara Zehetmayer. Betreuer: Sabine Krause." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2015. http://d-nb.info/1069743380/34.
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Carse, Sinead. "Characterisation of human surfactant protein A and recombinant human vimentin in their modulation of HPV16 pseudovirus infection." Master's thesis, Faculty of Health Sciences, 2019. https://hdl.handle.net/11427/31719.
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Infection by oncogenic human papillomavirus (HPV) is the primary cause of cervical cancer, where low-and middle-income countries (LMIC) have the highest incidence. Prophylactic HPV vaccines exist but LMIC have limited access. Therefore, alternative preventative measures against HPV infection and cervical cancer progression are needed. Two human proteins have been identified in our laboratory that modulate HPV16 pseudovirus (HPV16-PsVs) infection in vitro, namely surfactant protein A (SP-A) and recombinant human vimentin (rhVim). Previous work suggested SP-A mediated immune recognition of HPV since SP-A-coated HPV16- PsVs enhanced viral uptake by RAW264.7 murine macrophages. These initial observations were confirmed using a murine C57BL/6 cervicovaginal challenge model: pre-incubation of HPV16- PsVs with purified human SP-A significantly reduced the level of HPV16-PsV infection in vivo. Moreover, when isolated cells from female reproductive tracts of naïve C57BL/6 mice were incubated with HPV16-PsVs and stained for selected innate immune cell populations by flow cytometry, significant increases in viral uptake by eosinophils, neutrophils, monocytes and macrophages were observed over time using SP-A-pre-coated virions compared to control particles. Compared to SP-A mediated modulation of HPV infection through activation of innate immune responses, rhVim was suggested to directly interfere with HPV entry into host cells. Indeed, supplementation with non-filamentous rhVim resulted in decreased viral uptake by NIKS cells which was confirmed in vivo using the murine C57BL/6 cervicovaginal HPV16-PsVs challenge model. Co-localisation analysis employing confocal imaging, revealed that rhVim-coated HPV16- PsVs co-localised, to a lesser degree, with surface-expressed heparan sulphate proteoglycans (HSPGs) than control particles. Removal of surface HSPGs on NIKS cells decreased HPV16-PsVs cell surface binding and internalisation, while pre-incubation of HPV16-PsVs with rhVim decreased viral particle binding and internalisation to a greater extent. This indicates that rhVim may modulate HPV16 infection by interfering with its attachment to HSPGs as well as viral engagement with the yet unknown entry receptor(s). In summary, both SP-A and vimentin modulate HPV16-PsVs infection by different mechanisms. These in vivo studies strongly confirm previous in vitro observations, rendering both proteins potentially suitable for further development into possible candidates for use in topical microbicides, which may provide protection against new HPV infections.
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Ishigooka, Nozomi. "Predicting factors for disappearance of anti-mutated citrullinated vimentin antibodies in sera of patients with rheumatoid arthritis." Doctoral thesis, Kyoto University, 2020. http://hdl.handle.net/2433/245829.
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京都大学0048
新制・課程博士
博士(医学)
甲第22144号
医博第4535号
新制||医||1039(附属図書館)
京都大学大学院医学研究科医学専攻
(主査)教授 松田 秀一, 教授 生田 宏一, 教授 杉田 昌彦
学位規則第4条第1項該当
Doctor of Medical Science
Kyoto University
DFAM
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Schorling, Jamie J. "Biochemical and Immunocytochemical Characterization of Canine Corneal Cells Cultured in Two Different Media." Thesis, Virginia Tech, 2007. http://hdl.handle.net/10919/32443.
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The study purpose was to determine whether canine corneal cultures demonstrate superior growth when cultured in a fully defined epithelial selective medium, Epilife®, compared to Dulbecco's modification of Eagle's medium (DMEM) with fetal bovine serum (FBS), and to characterize cultured canine corneal cells. Superficial keratectomies were performed on three dogs. Samples were trypsinized to separate cell layers. Post-trypsinization, immunohistochemistry confirmed that epithelial cells had been released from the stroma. Both cell populations (presumed epithelial cells and stromal tissues) were cultured in DMEM with FBS or Epilife®. First passage cells were fixed for immunocytochemistry and prepared for PCR. Immunocytochemical staining for pancytokeratin, vimentin, and E-cadherin was evaluated, and immunofluorescence for zonula occludens-1 was attempted. Amplification of cytokeratin 5 (CK5) mRNA was assessed by PCR. Primary presumed epithelial cells grew faster when cultured in DMEM with FBS compared to Epilife®. Stromal tissue segments in Epilife® medium failed to adhere to culture plates, indicating that this medium may inhibit attachment and growth of non-epithelial tissues. Staining of corneal tissue segments confirmed that epithelial layers were pancytokeratin and E-cadherin positive, while stromal cells were vimentin positive. Immunocytochemistry of cultured cells revealed that epithelial cells stained positively for pancytokeratin, vimentin, and E-cadherin, while stromal cells remained only vimentin positive. Greater amplification of CK5 mRNA occurred from epithelial cells grown in Epilife® compared to epithelial cells in DMEM with FBS or the stromal cells. Based on PCR results, Epilife® medium may support retention of the epithelial characteristic of CK5 mRNA expression better than DMEM with FBS.Master of Science
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Colakoglu, Gulsen. "Assembly Dynamics of Intermediate Filaments." The Ohio State University, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=osu1247691189.
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Pritchard, Adaleigh Elizabeth. "Modeling End-to-End Annealing of Intermediate Filaments." The Ohio State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=osu1397743583.
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Paula, Fernanda Oliveira de. "Avaliação da imunoexpressão de vimentina e de osteopontina no reparo ósseo precoce de defeitos preenchidos com enxerto bovino associado à terapia laser de baixa intensidade." Universidade Federal de Juiz de Fora (UFJF), 2010. https://repositorio.ufjf.br/jspui/handle/ufjf/2638.
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O objetivo deste estudo foi avaliar, pelo método de imunoistoquímica, a expressão de vimentina e osteopontina durante as fases iniciais de reparo de defeitos ósseos criados em fêmures de ratos Wistar albinus tratados com enxerto ósseo bovino orgânico Gen-ox® em associação com terapia laser de baixa intensidade. Foram selecionadas de forma aleatória 24 amostras emblocadas de arquivo provenientes de trabalho experimental desenvolvido anteriormente, no qual foram efetuadas análises histológicas e histomorfométricas de fêmures de cinquenta ratos. Obtiveram-se lâminas histológicas dos blocos de diferentes animais, os quais estavam previamente divididos, de acordo com o tratamento realizado, da seguinte forma: grupo I (controle), grupo II (Gen-Ox®) e grupo III (LLLT e Gen-Ox®). Foram elaboradas quatro lâminas para cada um dos tempos experimentais de 3, 5, 7 e 15 dias para cada grupo. Duas lâminas foram utilizadas para analisar a expressão da osteopontina e duas para vimentina, totalizando 48 lâminas. Em cada lâmina considerou-se dois campos para análise, sendo um campo na região da interface osso-defeito e o outro próximo ao periósteo, em um total de 96 campos. Para realização da reação imunoistoquímica anti-vimentina e anti-osteopontina utilizou-se o método clássico do complexo avidina-biotina peroxidase anti-peroxidase. A marcação positiva foi determinada pela identificação de coloração castanha intracitoplasmática nas reações com ambos os anticorpos. Os cortes foram analisados em microscópio Zeiss em aumentos de 200x, 400x e 1000x, em toda extensão, por dois diferentes observadores. Determinou-se, por método de contagem semiquantitativo, a média de intensidade de células positivas marcadas nos campos dos tratamentos propostos em cada período, o qual foi classificado pelo sistema de escore de acordo com os seguintes parâmetros: 0 = ausência de marcação; + = marcação leve (até 1/3 de células positivas); ++ = marcação moderada (até 2/3 de células positivas); e +++ = marcação intensa (acima de 2/3 de células marcadas). Os resultados mostraram que todos os grupos apresentaram marcação para vimentina e para osteopontina em todos os períodos do experimento. Observou-se marcação celular mais acentuada para vimentina no período cicatricial inicial no grupo III. Não foram verificadas diferenças na marcação para osteopontina nos animais submetidos à terapia laser de baixa intensidade associada ao enxerto quando comparado aos outros grupos.
The aim of this study was to evaluate, by immunohistochemistry, the expression of vimentin and osteopontin during the early stages of repair of bone defects created in femurs of Wistar albinus rats treated with organic bovine bone graft Gen-ox® and associated with low level laser therapy. Twenty four embedded samples were randomly selected from the file of a previous experimental work, in which histological analysis and histomorphometry of the femurs of fifty rats was performed. Histological slides were obtained from blocks of different animals which were divided in accordance to previous treatment as follow: group I (control), group II (Gen-Ox ® ) and group III (LLLT and Gen-Ox ®). Four slides were prepared for each of the experimental time of 3, 5, 7 and 15 days for each group. Of the four slides, two were assessed for the expression of osteopontin and two of vimentin, in a total of 48 slides. On each slide two fields were considered for analysis: one in the bone-defect interface and the other near the periosteum, in a total of 96 fields. To perform the immunohistochemistry anti-vimentin and anti-osteopontin, we used the classical avidin-biotin peroxidase anti-peroxidase method. The positive staining was determined by identification of intracytoplasmic brown color in the reactions with both antibodies. The sections were analyzed in Zeiss microscope at a magnification of 200x, 400x and 1000x by two different observers. The average intensity of positive cells stained in the fields in each period was determined by a semiquantitative counting method which was classified by a scoring system as follow: 0 = no marking; + = mild labeling (up to one third of positive cells); + + = moderate labeling (up to two thirds of positive cells) and + + + = intense labeling (over two thirds of labeled cells). The results showed that all groups had marked for vimentin and osteopontin in all periods of the experiment. We observed a stronger cell labeling for vimentin in the initial healing period in group III. There were no differences in cell labeling for osteopontin in animals subjected to low level laser therapy associated with graft as compared to other groups.
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Block, Johanna Lena [Verfasser], Sarah [Akademischer Betreuer] Köster, Sarah [Gutachter] Köster, and Andreas [Gutachter] Janshoff. "Stress-Strain Behavior of Single Vimentin Intermediate Filaments / Johanna Lena Block ; Gutachter: Sarah, Köster; Andreas Janshoff ; Betreuer: Sarah, Köster." Göttingen : Niedersächsische Staats- und Universitätsbibliothek Göttingen, 2018. http://d-nb.info/1164231197/34.
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Rapatoni, Liane. "Expressão de CK5 e vimentina/E-caderina nos diferentes subtipos de carcinomas ductais mamários." Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/17/17145/tde-27102013-000552/.
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Introdução: Os marcadores moleculares têm sido utilizados para identificar subgrupos de tumores com comportamento clínico distinto, inclusive quanto ao padrão de recidiva. Objetivo: Caracterizar a expressão imunoistoquímica de vimentina (VIM) e E-caderina (CDH1) em carcinomas ductais invasivos (CDI) de mama e sua associação com a expressão de citoqueratina 5 (CK5), e características clínico-patológicas. Métodos: Microarranjos teciduais (TMA) foram construídos a partir de 82 amostras de CDI de mama. Imunoistoquímica (IHQ) foi realizada para determinar os receptores hormonais (RH; receptores de estrógeno e progesterona) e receptor do fator de crescimento epidérmico humano 2 (HER2), VIM, CDH1, CK5 e Ki-67. Os tumores foram classificados como luminal A (RH+, HER2-), luminal B (RH+, HER2+ ou Ki-67 alto), HER2 hiperexpresso (RH-, HER2+) e triplo negativo (TN; RH-, HER2-). Resultados: O fenótipo VIM+/CDH1-/low não foi observado nos tumores luminal A, B e HER2 hiperexpresso, enquanto que este fenótipo estava presente em 61,9% dos TN (p= 0,0001). A mediana de Ki-67 em tumores VIM+/CDH1-/low foi 13,6 (variação, 17,8-45,4), em comparação com 9,8 (variação de 4,1-38,1) nos não- VIM+/CDH1-/low (p= 0,0007). O acometimento linfonodal foi menos freqüente em pacientes com VIM+/CDH1-/low do que nos não- VIM+/CDH1-/low (23% X 61%, teste de 2, p = 0,01). A sobrevida livre de doença em 5 anos foi de 61,5% e 83,7% (teste de log-rank, p= 0,02) e a sobrevida global em 5 anos foi de 51,2% e 83,5% (teste de log-rank, p= 0,03) em pacientes com tumores com fenótipo VIM+/CDH1-/low e não VIM+/CDH1-/low, respectivamente. Conclusão: A expressão de VIM e CDH1 identificaram um subconjunto de CDI de mama com fenótipo mesenquimal com alto grau histológico e alto índice mitótico.Introduction: Molecular markers have been used to identify subgroups of tumors with distinct clinical behavior, including recurrence pattern. Objective: To characterize the immunohistochemical expression of vimentin (VIM) and E-cadherin (CDH1) in invasive ductal carcinoma (IDC) of the breast and its association with cytokeratin 5 (CK5) expression and clinicopathological features. Methods: A tissue microarray was constructed from 82 IDC breast cancer specimens. Immunohistochemistry (IHC) was used to determine hormone receptor (HR; estrogen and progesterone receptors) status and human epidermal growth factor receptor 2 (HER2), VIM, CDH1, CK5, and Ki-67 expression. Tumors were classified as luminal A (HR+, HER2-), luminal B (HR+, HER2+ or high Ki-67), HER2 enriched (HR-, HER2+), and triple negative (TNBC; HR-, HER2-). Results: The VIM+/CDH1-/low phenotype was not observed in luminal A, luminal B and HER2 enriched tumors, whereas this phenotype was present in 61.9% of triple negative (TNBC) tumors (p = 0.0001). The median Ki-67 index in VIM+/CDH1-/low tumors was 13.6 (range, 17.8-45.4) compared with 9.8 (range, 4.1-38.1) in non-VIM+/CDH1-/low tumors (p= 0.0007). The presence of lymph node metastasis was less frequent in patients with VIM+/CDH1-/low tumors than in those with non-VIM+/CDH1-/low tumors (23% vs. 61%; 2 test, p= 0.01). The 5-years disease-free survival was 61.5% and 83.7% (log-rank test; p= 0.02) and the 5-year overall survival was 51.2% and 83.5% (log-rank test; p= 0.03) in patients with VIM+/CDH1-/low phenotype and non-VIM+/CDH1-/low phenotype tumors, respectively. Conclusion: The expression of VIM and CDH1 identified a subset of IDC breast cancer of the mesenchymal phenotype with high histological grade and high mitotic index.
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Suasnavas, Edison A. "Characterization and Potential Utility of Porcine Trophoblast-Derived Stem-Like Cells." DigitalCommons@USU, 2013. http://digitalcommons.usu.edu/etd/1540.
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In mammals, the trophoblast lineage of the embryo is specified before implantation. It is restricted to become the fetal portion of the placenta. We have isolated and cultured trophoblast-derived cells from day 10 and day 13 porcine embryos. These cells demonstrate morphological and biological characteristics that make them unique. We have demonstrated that these cells can grow in vitro in a defined, serum-replacement medium for over a year without showing any signs of senescence. Trophoblast-derived cells placed into serum-containing medium, however, rapidly senesce and fail to proliferate. Gene expression analysis by RT-PCR and Fluidigm analysis of cells in culture from 0-30 days confirmed expression of genes involved in trophoblast function (CDX2, TEAD4, CYP17A1, HSD17B1, FGFR2, PLET, HAND1) as well as some genes known to mediate pluripotency (POU5F1, KLF4, CMYC). These experiments revealed changes in gene expression over time and in response to serum-containing medium. We have demonstrated that these trophoblast-derived cells are easily stably transfected with an exogenous transgene (eGFP) by a variety of methods, and show the ability to survive and to be passaged repeatedly after transfection. Also, immunofluorescence analysis results demonstrated that these cells do not only demonstrate epithelial characteristics by the expression of KRT18, but also they show expression of VIMENTIN which is a protein found in mesenchymal cells. These findings contradict studies done by Ramsoondar in 1993 and Flechon in 1995 which reported the negative expression of VIMENTIN in similar cells. In summary, early embryonic porcine trophoblast-derived cells have demonstrated unique characteristics which have taken us to the conclusion that they could be used as valuable tools for laboratory work. Anticipated applications include the study of trophoblast physiology as well as possible solutions for improving efficiency of transgenesis by somatic cell nuclear transfer and for pluripotency reprogramming of cells.
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Brennich, Martha. "Cation induced self-assembly of intermediate filaments." Doctoral thesis, Niedersächsische Staats- und Universitätsbibliothek Göttingen, 2012. http://hdl.handle.net/11858/00-1735-0000-001A-6BD9-E.
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Tuleneva, O. А., and I. S. Davydenko. "Immunohistochemical vimentin concentration in the endothelium of the terminal chorionic villi in the aspect of various forms of placental insufficiency." Thesis, БДМУ, 2017. http://dspace.bsmu.edu.ua:8080/xmlui/handle/123456789/16789.
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Bowers, Hannah Elizabeth, and Jennifer Hall. "THE EFFECTS OF ESTROGEN-INDUCED STROMAL CELL EFFECTORS, OSTEOPONTIN AND VIMENTIN, ON CHLAMYDIA INFECTIONS IN A NON-POLARIZED CELL CULTURE MODEL." Digital Commons @ East Tennessee State University, 2018. https://dc.etsu.edu/asrf/2018/schedule/98.
Full textAbstract:
Chlamydia is the most reported sexually transmitted infection in the US and is caused by the obligate intracellular bacterium Chlamydia trachomatis. Typically, this presents as a lower genital tract infection (cervicitis or urethritis), but can ascend to the upper genital tract, causing pelvic inflammatory disease, tubal infertility, epididymitis, or ectopic pregnancy. While chlamydia infections can be cured with a single-dose oral antibiotic, repeat infections are common and having multiple chlamydial infections increases a woman’s risk of developing serious chronic conditions. Previous research has shown that estrogen has a positive effect on C. trachomatis infections—an important finding, connecting fluctuating estrogen levels in females to variance in pathogenesis.The mechanism behind this hormonal influence remains unknown; however, previous work in our laboratory indicates that estrogen-stimulated stromal cell effectors play a role in enhancing C. trachomatis infections in a polarized endometrial epithelial Ishikawa (IK)/stromal (SHT-290) cell co-culture model. Specifically, our data indicate that estrogen exposure stimulates osteopontin and vimentin release from stromal cells in co-culture with endometrial epithelial cells. Furthermore, we noted that Chlamydia-infected, polarized Ishikawa cells exposed to a combination of recombinant osteopontin and estrogen released significantly more infectious chlamydia than cultures exposed to estrogen alone. Most tissue culture models being used today employee non-polarized cells. Given the fact that epithelial cell polarization is known to impact C. trachomatis serovar E development, in the current study we sought to determine if the estrogen-induced stromal cell effectors, osteopontin and vimentin, affect C. trachomatis viability and infectivity in non-polarized Ishikawa cells. Non-polarized Ishikawa cells were exposed to osteopontin or vimentin in the presence or absence of estrogen, infected with C. trachomatis serovar E, and collected for examination of chlamydial infectivity and progeny production. Our initial data show that osteopontin and vimentin impact chlamydial progeny production in a concentration dependent fashion, with higher concentrations of recombinant effectors +/- estrogen significantly decreasing progeny production. These data suggest that polarization of host cells influences the way hormone-stimulated effectors interact with the cell to impact on chlamydial infection. Future research goals are to explore other stromal effectors such as fibronectin with estrogen and to study the cell signaling mechanism osteopontin and vimentin use to affect chlamydial infections in polarized epithelial cell cultures.