Claritromicina era considerada um antibiótico de escolha para infecções causadas pelo complexo Mycobacterium abscessus, entretando, recentemente falhas no tratamento com este antibiótico têm sido reportadas. A resistência adquirida a claritromicina está relacionada à mutações pontuais que acarretam substituição da adenina na posição 2058 ou na posição 2059 na região do gene rrl que codifica o domínio peptidil transferase do rRNA 23S. Um mecanismo secundário de resistência à claritromicina tem sido descrito como resistência induzida, que é conferida pelo polimorfismo T/C no nucleotídeo 28 do gene erm(41). A resistência adquirida pode ser detectada em até 3 dias de incubação do M. abscessus com a claritromicina enquanto que a resistência induzida requer mais do que 5 dias de incubação. Por outro lado, o uso de marcadores moleculares para detecção de resistência adquirida e induzida no complexo M. abscessus têm sido propostos. O objetivo desse estudo foi avaliar o perfil de suscetibilidade e os marcadores moleculares de resistência à claritromicina no complexo M. abscessus. Um total de 42 isolados de um estudo prévio de vigilância, entre os anos 2007 e 2013 foram utilizados. O perfil de suscetibilidade para a claritromicina foi determinado por microdiluição em caldo com leituras em 3, 5, 7 e 14 dias. Mutações nos genes rrl e erm(41) foram avaliados por PCR com primers específicos e posterior sequenciamento. Resistência à claritromicina, em até 3 dias de incubação, foi observada em 31 dos 42 (73,8%) isolados. Resistência induzida à claritromicina foi observada em 6 de 11 (54,5%) isolados que apresentaram resistência após 5 ou 7 dias de incubação. Todos os isolados com resistência induzida foram M. abscessus subsp. massiliense. Além disso, todos os 28 isolados de M. abscessus subsp. massiliense apresentaram deleção em erm(41). Apenas cinco isolados foram sensíveis à claritromicina após 14 dias de incubação. Nenhum dos 42 isolados apresentaram mutação pontual na região de peptidil transferase do rRNA 23S e todos os isolados apresentaram o polimorfismo T/C no nucleotídeo 28 do gene erm(41). Os dados deste estudo indicam a falta de correlação dos marcadores moleculares com a expressão de resistência à claritromicina. / Infections due to Mycobacterium abscessus complex used to respond to clarithromycin treatment but more recently treatment failure with this antibiotic has been reported. Acquired resistance to clarithromycin is related to substitutions at the adenine either at position 2058 or at position 2059 in a region of the rrl gene encoding to the peptidyltransferase domain of the 23S rRNA. A secondary mechanism related to clarithromycin resistance has been described as an inducible resistance, conferred by T/C polymorphism at the 28th nucleotide in erm(41) gene. Acquired resistance can be detected up to 3 days of incubation of the M. abscessus with the clarithromycin while inducible resistance requires more than 5 days of incubation. Molecular markers to detect acquired and inducible resistance in M. abscessus complex isolates were proposed. This study evaluated the profile of susceptibility and the molecular markers of clarithromycin resistance in M. abscessus complex. A total of 42 isolates from a previous surveillance study (2007 to 2013) were used in this study. The susceptibility profile for clarithromycin was determined by broth microdilution with reads at 3, 5, 7 and 14 days. Mutations in rrl and erm(41) genes were evaluated by PCR with specific primers followed by sequencing. Clarithromycin resistance, up to 3 days of incubation, was observed in 31 of 42 (73.8%) isolates. Inducible clarithromycin resistance was observed in 6 of 11 (54.5%) isolates which presented resistance only after 5 or 7 days of incubation. All isolates with inducible resistance were identified as M. abscessus subsp. massiliense. Moreover, all 28 M. abscessus subsp. massiliense had a deletion in erm(41). Only five isolates proved to be susceptible to clarithromycin after 14 days of incubation. None of the 42 isolates presented a point mutation in the peptidyltransferase region of the 23S rRNA (rrl) and all isolates presented the T/C polymorphism at the 28th nucleotide of the erm(41) gene. The data of this study indicates a lack of correlation of molecular markers of clarithromycin resistance for both acquired and inducible resistance to clarithromycin.

Diplomová práce se zabývá problematikou zapojení České republiky do systému ERM II. Teoretická část práce stručně popisuje proces evropské integrace, a to nejen v obecné rovině, ale hlavně v rovině hospodářské a měnové. Tato část se věnuje také zapojení ČR do evropských struktur, představení mechanismu ERM II a dále vymezuje některé důležité pojmy z této oblasti. Praktická část práce je věnována podrobné analýze plnění jednotlivých maastrichtských konvergenčních kritérií Českou republikou, kde u kritéria stability měnového kurzu je také velmi komplexně popsán stav ČR v souvislosti se zapojením do systému ERM II. Analýza jednotlivých kritérií je poté stručně shrnuta.

Práce se zabývá otázkou připravenosti České republiky na přijetí jednotné měny euro, respektive otázkou připravenosti na vstup do mechanismu směnných kurzů ERM II (Exchange rate mechanism). Zabývá se historií evropské měnové integrace a ustanovením Maastrichtských konvergenčních kritérií a jejich dodržováním. V rámci konvergenčních kritérií hodnotí na základě Konvergenční zprávy dosavadní výsledky plnění kritérií jak Českou republikou tak ostatními členskými státy, které usilují o vstup do ERM II. Dále rozebírá mechanismu ERM II jak z historického hlediska, tak z procedurálního hlediska a také z pohledu zemí, které se tohoto mechanismu již zúčastnily a nebo jsou právě jeho součástí. Snaží se nastínit možná rizika, která by mohla vyplynout z pobytu koruny v tomto mechanismu, jakožto mechanismu fixování zúčastněné měny na euro v rámci tzv.fluktuačního pásma

<p>Proteins of the band 4.1 superfamily are characterized by their sequence similarity to the ERM proteins ezrin, radixin and moesin, which are involved in cell motility, adhesion of cells, and signal transduction events. Little is however known of the function of ERM proteins in the nervous system, though an essential role for radixin and moesin in neuronal growth cone motility has been suggested. </p><p> This thesis is focused on the cloning, functional characterization and description of the tissue distribution in rat brain of MIR, a novel member of the band 4.1 superfamily. </p><p> The cDNA of MIR encods a protein of 445 amino acids which is composed of an ERM-homology domain and a RING finger, separated by an interregion. To reveal the cellular function of MIR, PC12 cell lines overexpressing MIR was generated and observed to inhibit NGF stimulated neurite outgrowth. </p><p> To elucidate the signal transduction of MIR by which it exerts its physiological activity, the yeast two-hybrid system was employed to screen for proteins that interact with MIR. A number of interactors known to regulate the cytoskeleton was obtained - among them myosin regulatory light chain-B which controls the actomyosin complex - and a novel type 2 membrane protein denoted NSAP for its similarity to saposin A-D. Overexpressed NSAP induced neurite outgrowth in PC12 cells and enhanced cell adhesion in fibroblasts. </p><p> The tissue distribution of MIR in rat brain, as determined by immunohistochemistry studies, showed that MIR is localized especially to neurons in hippocampus and cerebellum. The chromosomal localization of the MIR gene was assessed to 6p22.3-23, a region lost in the 6p23 deletion syndrome.</p><p> These results suggests that MIR is expressed in neurons in discrete regions of rat brain where it may regulate neurite outgrowth by modulating the cytoskeleton.</p>

Proteins of the band 4.1 superfamily are characterized by their sequence similarity to the ERM proteins ezrin, radixin and moesin, which are involved in cell motility, adhesion of cells, and signal transduction events. Little is however known of the function of ERM proteins in the nervous system, though an essential role for radixin and moesin in neuronal growth cone motility has been suggested. This thesis is focused on the cloning, functional characterization and description of the tissue distribution in rat brain of MIR, a novel member of the band 4.1 superfamily. The cDNA of MIR encods a protein of 445 amino acids which is composed of an ERM-homology domain and a RING finger, separated by an interregion. To reveal the cellular function of MIR, PC12 cell lines overexpressing MIR was generated and observed to inhibit NGF stimulated neurite outgrowth. To elucidate the signal transduction of MIR by which it exerts its physiological activity, the yeast two-hybrid system was employed to screen for proteins that interact with MIR. A number of interactors known to regulate the cytoskeleton was obtained - among them myosin regulatory light chain-B which controls the actomyosin complex - and a novel type 2 membrane protein denoted NSAP for its similarity to saposin A-D. Overexpressed NSAP induced neurite outgrowth in PC12 cells and enhanced cell adhesion in fibroblasts. The tissue distribution of MIR in rat brain, as determined by immunohistochemistry studies, showed that MIR is localized especially to neurons in hippocampus and cerebellum. The chromosomal localization of the MIR gene was assessed to 6p22.3-23, a region lost in the 6p23 deletion syndrome. These results suggests that MIR is expressed in neurons in discrete regions of rat brain where it may regulate neurite outgrowth by modulating the cytoskeleton.

The ability to spatially restrict specific activities across the cell cortex functionally defines individual cells and tissues. This is achieved, in part, via the assembly of protein complexes that link the plasma membrane to the underlying cortical actin cytoskeleton. The neurofibromatosis type 2 (NF2) tumor suppressor Merlin and closely related ERM proteins (Ezrin, Radixin and Moesin) are a special class of such membrane:cytoskeleton associated proteins that function to organize specialized cortical domains. In addition to their high degree of similarity, mounting evidence suggests that Merlin/ERMs share a functional relationship, which is largely unexplored. Unlike Merlin, the ERMs are not known to inhibit cell proliferation; in fact, Ezrin is thought to promote tumor metastasis. Defining the relationship between Merlin and the ERMs is essential to appreciating their respective roles in cancer development. Here I demonstrate a novel role for Merlin and the ERMs in generating cortical asymmetry in the absence of external cues. Our data reveal that Merlin functions to restrict the cortical distribution of Ezrin, which in turn positions the interphase centrosome in single epithelial cells and 3D organotypic cultures. In the absence of Merlin, ectopic cortical Ezrin yields mispositioned centrosomes, misoriented spindles and aberrant epithelial architecture. Furthermore, in tumor cells with centrosome amplification, the failure to restrict cortical Ezrin abolishes centrosome clustering, yielding multipolar mitoses. Consistent with a functional relationship, I observe a strong genetic interaction between Nf2 and Ezrin in the mouse intestine in vivo. Finally, I begin to address the basis of their functional interaction by testing whether they are coordinately regulated by the Ste-20 like kinase SLK. Altogether, these data uncover fundamental roles for Merlin/ERM proteins in spatiotemporally organizing the cell cortex in vitro and in vivo and suggest that Merlin’s role in promoting cortical heterogeneity may contribute to tumorigenesis by disrupting cell polarity, spindle orientation and potentially genome stability.

Enterprise is often described as risk for reward, but it may be possible to reduce the risk while improving returns. According to SEDA, failure rates of SMMEs in South Africa range from 70 to 80 percent. The need for this study arose when it was found that most SMMEs did not have a formal system in place to mitigate their risks right from the outset in the feasibility study, the business plan design and the start–up of the business. This lack of mitigation controls could be a result of a lack of understanding of the enterprise risk management (ERM) methodology or an inappropriate ERM decision–making model to assist them in a way that would mitigate their risk and minimise financial losses. The ERM approach can anticipate unplanned occurrences and is a systematic way of foreseeing the future. Entrepreneurs and business owners take on risks to pursue new business objectives within their respective risk appetites. This study also evaluated several models of risk identification and the ERM methodology. In this study an ERM model, ISO 31000, was applied in a business case and a comparison was made between the risks identified in the business plan and the ERM approach. / Thesis (M.B.A.)--North-West University, Potchefstroom Campus, 2011.

Enterprise is often described as risk for reward, but it may be possible to reduce the risk while improving returns. According to SEDA, failure rates of SMMEs in South Africa range from 70 to 80 percent. The need for this study arose when it was found that most SMMEs did not have a formal system in place to mitigate their risks right from the outset in the feasibility study, the business plan design and the start–up of the business. This lack of mitigation controls could be a result of a lack of understanding of the enterprise risk management (ERM) methodology or an inappropriate ERM decision–making model to assist them in a way that would mitigate their risk and minimise financial losses. The ERM approach can anticipate unplanned occurrences and is a systematic way of foreseeing the future. Entrepreneurs and business owners take on risks to pursue new business objectives within their respective risk appetites. This study also evaluated several models of risk identification and the ERM methodology. In this study an ERM model, ISO 31000, was applied in a business case and a comparison was made between the risks identified in the business plan and the ERM approach. / Thesis (M.B.A.)--North-West University, Potchefstroom Campus, 2011.

La Espectroscopia por Resonancia Magnética de Protón (ERM 1H) es una técnica no invasiva de diagnóstico que, utilizando el mismo material que los estudios convencionales de imagen por RM, estudia el contenido bioquímico de los tejidos. De esta manera permite valorar el estado metabólico de los tejidos.En esta tesis se aborda el tema de la aplicación clínica de la espectroscopia por resonancia magnética de protón (ERM 1H) en el diagnóstico de los tumores cerebrales.Contiene tres artículos publicados en European Radiology, Radiology y Neuroradiology en los años 2002 y 2003. En ellos se propone un método para clasificar los tumores cerebrales en base a sus características en ERM 1H. Para ello se utilizan los percentiles de 10 y 90% de determinadas resonancias, dependiendo de los casos, como puntos de corte en diversos algoritmos. Los algoritmos se adaptan en cada trabajo dependiendo de la pregunta a responder.En el primer trabajo (Majós et al, European Radiology 2003) se trabaja la clasificación de los tumores más frecuentes en cuatro grupos (1-meningioma, 2-astrocitoma de bajo grado, 3-astrocitoma anaplásico, y 4-glioblastoma+metástasis). Los resultados, validados en un grupo de test, son de 82% clasificaciones correctas.En el segundo trabajo (Majós et al, Radiology 2002) se estudia un grupo tumoral menos prevalente: los Tumores Neuroectodérmicos Primitivos (TNEP) en adultos. Se estudian sus características espectroscópicas y los resultados que se obtienen al aplicar estas características en el diagnóstico diferencial con los tumores cerebrales más frecuentes en el adulto (meningioma, astrocitoma de bajo grado, astrocitoma anaplásico, glioblastoma y metástasis). Los resultados obtenidos, validados también en un grupo test, son de 78% clasificaciones correctas, 13% clasificaciones incorrectas y 9% casos inclasificables. En este artículo se concluye que la ERM 1H proporciona información útil para la diferenciación clínica entre TNEP y los tumores cerebrales más frecuentes en el adulto.En el tercer artículo (Majós et al, Neuroradiology 2003) se estudian los meningiomas de aspecto radiológico atípico. Los autores estudian en profundidad las características espectroscópicas de los meningiomas y los aspectos que pueden ser de mayor utilidad para diferenciarlos de los tumores cerebrales más frecuentes. Estos aspectos se aplican en un algoritmo similar al elaborado en los otros dos trabajos. En un segundo tiempo se identifican los meningiomas que por su aspecto radiológico pueden ser considerados atípicos. Estos meningiomas constituyen un 14% del total de meningiomas (5/37). Se les aplica el algoritmo elaborado con el global de meningiomas para realizar los diagnósticos diferenciales surgidos a partir de la valoración de las imágenes, obteniendo una clasificación correcta en 4 de 5 casos (80%) y 10 de 12 procedimientos de diagnóstico diferencial (83%). La conclusión de este trabajo es que la ERM 1H puede ser aplicada con éxito en tumores en que el diagnóstico de meningioma no está suficientemente claro con los estudios de imagen.En global estos trabajos aportan una valoración de la utilidad clínica de la espectroscopia, concluyendo que su aplicación puede ser útil en determinados casos. Se sugiere la ERM 1H como una técnica adicional a ser utilizada para mejorar el diagnóstico prequirúrgico de los tumores cerebrales, máxime teniendo en cuenta que se trata de un método no invasivo y que puede realizarse en el mismo procedimiento que el estudio por RM convencional.

Claritromicina era considerada um antibiótico de escolha para infecções causadas pelo complexo Mycobacterium abscessus, entretando, recentemente falhas no tratamento com este antibiótico têm sido reportadas. A resistência adquirida a claritromicina está relacionada à mutações pontuais que acarretam substituição da adenina na posição 2058 ou na posição 2059 na região do gene rrl que codifica o domínio peptidil transferase do rRNA 23S. Um mecanismo secundário de resistência à claritromicina tem sido descrito como resistência induzida, que é conferida pelo polimorfismo T/C no nucleotídeo 28 do gene erm(41). A resistência adquirida pode ser detectada em até 3 dias de incubação do M. abscessus com a claritromicina enquanto que a resistência induzida requer mais do que 5 dias de incubação. Por outro lado, o uso de marcadores moleculares para detecção de resistência adquirida e induzida no complexo M. abscessus têm sido propostos. O objetivo desse estudo foi avaliar o perfil de suscetibilidade e os marcadores moleculares de resistência à claritromicina no complexo M. abscessus. Um total de 42 isolados de um estudo prévio de vigilância, entre os anos 2007 e 2013 foram utilizados. O perfil de suscetibilidade para a claritromicina foi determinado por microdiluição em caldo com leituras em 3, 5, 7 e 14 dias. Mutações nos genes rrl e erm(41) foram avaliados por PCR com primers específicos e posterior sequenciamento. Resistência à claritromicina, em até 3 dias de incubação, foi observada em 31 dos 42 (73,8%) isolados. Resistência induzida à claritromicina foi observada em 6 de 11 (54,5%) isolados que apresentaram resistência após 5 ou 7 dias de incubação. Todos os isolados com resistência induzida foram M. abscessus subsp. massiliense. Além disso, todos os 28 isolados de M. abscessus subsp. massiliense apresentaram deleção em erm(41). Apenas cinco isolados foram sensíveis à claritromicina após 14 dias de incubação. Nenhum dos 42 isolados apresentaram mutação pontual na região de peptidil transferase do rRNA 23S e todos os isolados apresentaram o polimorfismo T/C no nucleotídeo 28 do gene erm(41). Os dados deste estudo indicam a falta de correlação dos marcadores moleculares com a expressão de resistência à claritromicina. / Infections due to Mycobacterium abscessus complex used to respond to clarithromycin treatment but more recently treatment failure with this antibiotic has been reported. Acquired resistance to clarithromycin is related to substitutions at the adenine either at position 2058 or at position 2059 in a region of the rrl gene encoding to the peptidyltransferase domain of the 23S rRNA. A secondary mechanism related to clarithromycin resistance has been described as an inducible resistance, conferred by T/C polymorphism at the 28th nucleotide in erm(41) gene. Acquired resistance can be detected up to 3 days of incubation of the M. abscessus with the clarithromycin while inducible resistance requires more than 5 days of incubation. Molecular markers to detect acquired and inducible resistance in M. abscessus complex isolates were proposed. This study evaluated the profile of susceptibility and the molecular markers of clarithromycin resistance in M. abscessus complex. A total of 42 isolates from a previous surveillance study (2007 to 2013) were used in this study. The susceptibility profile for clarithromycin was determined by broth microdilution with reads at 3, 5, 7 and 14 days. Mutations in rrl and erm(41) genes were evaluated by PCR with specific primers followed by sequencing. Clarithromycin resistance, up to 3 days of incubation, was observed in 31 of 42 (73.8%) isolates. Inducible clarithromycin resistance was observed in 6 of 11 (54.5%) isolates which presented resistance only after 5 or 7 days of incubation. All isolates with inducible resistance were identified as M. abscessus subsp. massiliense. Moreover, all 28 M. abscessus subsp. massiliense had a deletion in erm(41). Only five isolates proved to be susceptible to clarithromycin after 14 days of incubation. None of the 42 isolates presented a point mutation in the peptidyltransferase region of the 23S rRNA (rrl) and all isolates presented the T/C polymorphism at the 28th nucleotide of the erm(41) gene. The data of this study indicates a lack of correlation of molecular markers of clarithromycin resistance for both acquired and inducible resistance to clarithromycin.