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Western blot analysis of oral Selenomonas speciesKuhner, Matthias K. January 1989 (has links)
Thesis (M.S.)--University of Michigan, Ann Arbor, 1989. / Typescript (photocopy). Includes bibliographical references (leaves 78-86). Also issued in print.
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Western blot analysis of oral Selenomonas speciesKuhner, Matthias K. January 1989 (has links)
Thesis (M.S.)--University of Michigan, Ann Arbor, 1989. / Typescript (photocopy). eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 78-86).
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Molecular mechanisms and therapies in metastatic retinoblastoma and other malignanciesTarlton, John Francis January 1998 (has links)
No description available.
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Western blot analysis of Bacteroides gingivalis antigen reactivity in human seraLee, Seok-Woo. January 1988 (has links)
Thesis (M.S.)--University of Michigan, Ann Arbor, 1988. / Typescript (photocopy). Includes bibliographical references (leaves 127-142). Also issued in print.
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Western blot analysis of Bacteroides gingivalis antigen reactivity in human seraLee, Seok-Woo. January 1988 (has links)
Thesis (M.S.)--University of Michigan, Ann Arbor, 1988. / Typescript (photocopy). eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 127-142).
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Caracterización y detección de tipos de canales de potasio presentes en membranas apicales de sinciciotrofoblasto placentario humanoDe Gregorio, Nicole January 2011 (has links)
Memoria para optar al Título Profesional de Médico Veterinario / La gestación, en todas las especies, es apoyada por una estructura que permite que el nuevo ser pueda desarrollarse y crecer adecuadamente dentro del vientre materno: la placenta. El transporte de solutos a través de ella se realiza, específicamente, a través de una célula epitelial multinucleada: el sinciciotrofoblasto (hSTB). Al ser un epitelio continuo, todo soluto debe atravesar, obligatoriamente, tanto su membrana apical como su membrana basal (BM). La membrana apical del hSTB se divide en dos subdominios diferentes entre sí: membrana apical pesada (MVM) y membrana apical liviana (LMVM).
Para desarrollar el modelo de transporte transplacentario, es necesario identificar las vías conductivas existentes en las membranas del hSTB. Existen numerosos estudios acerca de canales iónicos y transportadores en ellas, sin embargo, en particular, los canales de K+ (principal ión intracelular) han sido pobremente descritos. Un estudio preliminar demostró la existencia funcional de corrientes de K+ en LMVM de hSTB.
El objetivo del presente estudio, fue caracterizar funcional y estructuralmente que tipo de canales de K+ son los que subyacen las corrientes identificadas previamente en LMVM de hSTB de placentas humanas normales.
Experimentos de Inmunohistoquímica revelaron la presencia de TREK-1 en membrana apical del hSTB y de BKCa en el citoplasma del sinciciotrofoblasto. Adicionalmente, mediante Immunoblotting, se detectó la presencia de KIR2.1, KV2.1, TASK-1 y TREK-1 en ambos subdominios de membrana apical, mientras que la expresión de BKCa fue, principalmente, en BM.
La detección funcional de los canales de K+ que demostraron estar presentes en LMVM (KIR2.1, KV2.1, TASK-1 y TREK-1) por Immunoblotting, fue realizada utilizando la técnica de Patch-Clamp en configuración parche escindido (116 sellos de 16 placentas independientes). Se utilizaron bloqueos diferenciales para poder disecar corrientes con diferentes características. Se obtuvieron corrientes de K+ muy sensibles al bloqueo por BaCl2 (69,82%) de manera reversible. Adicionalmente, se obtuvieron registros de corrientes con sensibilidad a MgCl2 y CsCl2 (29% y 25%, respectivamente). Estas corrientes son características del canal KIR2.1. Por otro lado, se obtuvieron corrientes que demostraron tener un gran sensibilidad al bloqueador TEA a concentraciones milimolares (79,86%), las que corresponderían al canal KV2.1. En contraste, se obtuvieron corrientes que no presentaron efectos a los bloqueadores mencionados y que sin embargo, mostraron diferentes respuestas a la acidificación del medio. Es así como, un canal que pudiese corresponder a TASK-1 se obtuvo al registrar corrientes bloqueadas a pH 6,1 (bloqueo de un 28,92%) y una corriente que mostró una fuerte activación a pH 6,6 (63,08% de aumento sobre la corriente inicial), cuyo candidato más probable es TREK-1.
Estos experimentos demostraron, en base a su farmacología y respuesta a cambios del medio, la presencia de cuatro canales de K+ (KIR2.1, KV2.1, TASK-1 y TREK-1) en LMVM del hSTB lo que se correlaciona directamente con lo observado mediante Western blots
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Identifying immunogenic pneumococcal proteins with roles in colonization of human epitheliumCassibry, Abigail 10 May 2024 (has links) (PDF)
Streptococcus pneumoniae (pneumococcus) is a Gram positive opportunistic bacterium that is a primary cause of pneumonia in young children and immunocompromised individuals. This microorganism colonizes the nasopharynx of all age groups and health levels and is easily transmitted through respiratory droplets or aerosols. While capsule-based vaccines are available, these are becoming more obsolete as S. pneumoniae strains are undergoing serotype replacement, thus evading detection, and antibiotic resistant strains are increasing in prevalence. This study has taken a different approach by identifying immunogenic proteins that elicit antibodies which block attachment of bacteria to host epithelial cells. Proteins with the most potential were assessed by analyzing the reactivities of human sera with varying degrees of colonization blockage. Proteins which reacted with the primary protective antibody in mucosal sites, IgA, were prioritized. Identification of those proteins through mass spectrometry will be crucial in creating more effective, long-term protection against S. pneumoniae infections.
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Studium interakce vybraných chemopreventivních sloučenin a potravních karcinogenů s cytochromy P450 / Study on the interaction of chemopreventive compounds and food born carcinogens with cytochrome P450 enzymesBrabencová, Eliška January 2013 (has links)
The use of food supplements containing natural chemopreventive compounds increased in recent years. Some of the most popular chemopreventive compounds are flavonoids. Due to their natural origin, flavonoids are generally accepted as safe compounds. They exert antioxidant, anti-cancer and anti-inflammatory properties. However, flavonoids should be considered as foreign compounds (xenobiotics). Flavonoids interact with many enzymes, among the most important belong cytochromes P450 (CYPs), key enzymes of the first phase of biotransformation of xenobiotics (e.g. drugs, carcinogens). CYPs catalyze reactions leading mainly to detoxification of xenobiotics. However, some CYPs are involved in the activation of carcinogens, particularly CYP1A1 and CYP1A2 activate e.g. heterocyclic amines. Flavonoids might enhance the activation of carcinogens via induction of these CYPs or stimulation of their activities and hence, increase the risk of a cancer development. The thesis is focused on the influence of flavonoids and food carcinogens on the induction and activity of CYP1A1 and CYP1A2 in liver and small intestine of rats. For the purpose of this study, the small intestine was dissected into three parts: proximal (nearest to stomach), middle and distal. Western blotting was used for the evaluation of CYP...
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Estudo da resposta imunológica de anticorpos IgG, IgM e IgA, subclasses de IgG (IgG1 e IgG3) e avidez de IgG, por Western Blotting, em amostras de soros de pacientes com tuberculose pulmonar e comparação de resultados com métodos microbiológicos e dosagem de interferon-gama / Study of the immune response of IgG, IgM and IgA, IgG subclasses (IgG1 and IgG3) and IgG avidity by Western blotting in serum samples collected from patients with pulmonary tuberculosis and comparing results with microbiological methods and gamma interferon determinationFelix, Alvina Clara 29 April 2011 (has links)
Apesar das recomendações da Organização Mundial da Saúde, na reunião ministerial realizada em Amsterdam, Holanda em 2000, a tuberculose continua em ritmo crescente, atingindo principalmente os países em desenvolvimento e pessoas com o sistema imunológico comprometido, principalmente as infectadas pelo vírus da imunodeficiência adquirida. Os métodos utilizados no diagnóstico continuam os mesmos utilizados por muitos anos, cujas limitações impedem a ação rápida dos programas de saúde que buscam interromper a cadeia de transmissão da doença. Continuando uma linha de pesquisa do Laboratório de Soroepidemiologia e Imunobiologia do IMTSP, utilizando o método do Western Blotting, procuramos neste trabalho ampliar os conhecimentos da resposta imunológica de anticorpos em pacientes com tuberculose pulmonar, clinica e laboratorialmente definida, avaliando a participação das imunoglobulinas IgG, IgA e IgM, subclasses de IgG (IgG1 e IgG3) e a avidez da imunoglobulina IgG em amostras de soros colhidas no início e no final do tratamento. Nossos resultados mostraram que o melhor marcador imunológico foi a imunoglobulina IgG por apresentar melhor desempenho diagnóstico quando comparada com os resultados dos métodos microbiológicos e de dosagem de interferon gama, Quantiferon TB Gold. As frações protéicas que apresentaram melhor desempenho diagnóstico foram as de 38 e 30 KDa / Despite the recommendations of the World Health Organization, during the Ministerial meeting held in Amsterdam, Holland in 2000, tuberculosis continues at an important increasing rate, affecting mostly developing countries and people with severe compromised immune systems, especially those infected with human acquired immunodeficiency virus. The methods used for diagnosis are the same used for many years, whose limitations prevent the rapid action of countries health programs that seek to stop the chain of disease transmission. Continuing a line of research of the Laboratory of Seroepidemiology and Immunobiology of IMTSP using the method of Western blotting, in this study we tried to broaden the knowledge of the immune response of antibodies in patients with pulmonary tuberculosis, clinical and laboratory defined by evaluating the participation of IgG, IgA and IgM, IgG subclasses (IgG1 and IgG3) and immunoglobulin IgG avidity in serum samples collected in the beginning and end of treatment. Our results showed that the immunoglobulin IgG was best immunological marker when compared with the results of microbiological methods and determination of interferon gamma, QuantiFERON TB Gold. The proteins fractions that showed better diagnosis performance were 38 and 30 KDa
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pH final e suas alterações na concentração de HSP27 e HSP70 e no perfil proteômico do músculo esquelético de bovinos cruzados / Ultimate pH and its alterations in HSP27 and HSP70 concentrations and crossbred cattle skeletal muscle proteomic profilePereira, Mikaele Alexandre 02 March 2018 (has links)
Este estudo teve como objetivo investigar incialmente o efeito do pHf na qualidade da carne, nas concentrações de HSP27 e HSP70, bem como no perfil proteômico da carne (Longissimus thoracis) de bovinos cruzados - 1/2 (Simental Sul Africano x Nelore) de forma a contribuir para a predição de biomarcadores que poderão ser agregados em programas de melhoramento genético. Foram avaliados 415 bovinos (180 fêmeas e 235 machos imunocastrados) terminados em confinamento. As carcaças foram classificadas de acordo com pHf, mensurados 48 horas após o abate, em dois grupos: normal (pHf ≤ 5,80) e intermediário (pHf = 5,81 - 6,19). Os parâmetros físico-químicos pH, cor (L*, a* e b*), perda de água por cocção (PAC, %) e força de cisalhamento (FC, kg), foram mensurados para avaliar a qualidade da carne em dois tempos de maturação (1 e 14 dias). Para a quantificação de HSP27 e HSP70 foram selecionadas 38 amostras em função dos grupos de pHf, dentro de cada tempo de maturação, totalizando 76 amostras. Para a realização das análises proteômicas foram selecionadas três amostras de cada grupo de pHf nos dois tempos de maturação, totalizando doze amostras. Observou-se interação (p<0,05) entre os grupos de pHf e os tempos de maturação para a cor e FC. A concentração de HSP27 foi maior (p<0,05) em pHf intermediário e com 1 de maturação, para a HSP70 só foi possível observar maior concentração (p<0,05) aos 14 dias de maturação, as correlações indicam que as HSPs modulam a qualidade da carne. Foram identificadas 13 proteínas diferencialmente expressas (p<0,05) envolvidas com funções metabólicas (triosefosfato isomerase - TPI1; fosfoglicerato mutase 2 - PGAM2; malato desidrogenase - MDH1; piruvato quinase - PKM2; e adenalato quinase isoenzima 1 - AK1), estrutural (troponina I tipo 2 - TTNI2; tropomiosina cadeia beta - TPM2; troponina C tipo 2 - TNNC2; e cofilina 2 - CFL2) e defesa em resposta ao estresse (proteína deglicase DJ-1 - PARK7; αβ-cristalina - CRYAB; proteína do choque térmico 27 - HSPB1; e peroxirredoxina 1 - PRDX1). As correlações demonstraram que essas proteínas estão envolvidas na determinação do pHf e dos demais atributos da qualidade da carne. A bioinformática permitiu observar que essas proteínas estão envolvidas nas três categorias de ontologia gênica, sendo 18 termos de componentes celulares, 1 termo de função molecular e 61 termos dos processos biológicos, foram identificadas também 6 vias metabólicas. A proteína MDH1, esteve associada a mecanismos de produção de energia, confirmando o potencial desta como um biomarcador para a determinação do pHf assim como para a cor da carne, enquanto que a TPI1 foi relacionada somente no estabelecimento do pHf. Outras proteínas como a PKM2, com função metabólica, e a PARK7, com função protetora, estiveram associadas aos atributos maciez e cor da carne, respectivamente. Os resultados obtidos são de grande relevância no âmbito das ciências da carne e do melhoramento genético animal, por permitir a identificação de possíveis biomarcadores que poderão ser utilizados na seleção animal. / The objective of this study was to investigate the effect of pHf on meat quality, on the concentrations of HSP27 and HSP70, as well as on the proteomic profile in beef (Longissimus thoracis) of crossbreed cattle - 1/2 (South African Simmental x Nellore) - in order to explore the contribute to the prediction of biomarkers that can be added to the process of animal selection in breeding programs. Were evaluated 415 cattle (180 females and 235 immunocastrated males) feedlot-finished. Carcasses were classified according to pHu, measured 48 hours after slaughter, in two groups: normal (pHu ≤ 5.80) and intermediate (pHu = 5.81 - 6.19). The physical-chemical parameters pH, color values (L *, a * and b *), cooking loss (CL,%) and shear force (SF, kg) were measured to evaluate the meat quality in two aging times (1 and 14 days). For quantification of HSP27 and HSP70, 38 samples were selected according to the pHu groups, and aging time, totaling 76 samples. To perform the proteomic analysis were selected three samples from each group and aging time, totaling twelve samples. Color and SF meat quality attributes showed interaction (p<0.05) between pHu groups and aging times. The concentration of HSP27 was higher (p<0.05) at intermediate pHu and at 1 day of aging. It was only possible to observe a higher concentration (p<0.05) for HSP70 at 14 days of aging. Correlations analysis indicate that HSPs modulate the quality of meat. Thirteen differentially expressed proteins (p<0.05) were identified as involved with metabolic (triosephosphate isomerase - TPI, phosphoglycerate mutase 2 - PGAM2; malate dehydrogenase, cytoplasmic - MDH1; pyruvate kinase - PKM2; and adenylate kinase isoenzyme 1- AK1), structural (troponin I type 2 - TTNI2; tropomyosin beta chain - TPM2; troponin C type 2 - TNNC2; and cofilin 2 - CFL2) and defense in response to stress functions (protein deglycase DJ-1- PARK7; αβ-cristallin - CRYAB; heat shock protein 27 - HSPB1; and peroxiredoxin 1 - PRDX1). Correlations analysis showed that these proteins are involved in the determination of pHu and other attributes of meat quality. Using bioinformatics it was possible to observe that these proteins are involved in three gene ontology categories, which were 18 cellular components terms, 1 molecular function term and 61 biological processes terms. Other six metabolic pathways have also been identified. The MDH1 protein was associated with energy production mechanisms, confirming its potential as a biomarker for the determination of pHu as well as meat color, while TPI was related to the establishment of pHu only. Other proteins such as PKM2, with metabolic function, and PARK7, with protective function, were associated with the attributes of meat tenderness and color, respectively. The obtained results are of great relevance in the field of meat science and animal breeding genetic, it allows the identification of biomarkers that can be used in animal selection.
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