RESUMO
This study explored the evolutionary mechanism by which the clinical isolate PA110514 yields the imipenemresistant derivative PA116136. Both isolates were examined by PFGE and SDS-PAGE, which led to the identification of a new insertion sequence, ISPa133. This element was shown to have distinct chromosomal locations in each of the original isolates that appeared to explain the differences in imipenem susceptibilty. In strain PA110514, ISPa133 is located 56 nucleotides upstream of the translational start codon, which has no effect on expression of the porin OprD. However, in strain PA116136 ISPa133 it is located in front of nucleotide 696 and, by interrupting the coding region, causes a loss of OprD expression, thus conferring imipenem resistance. In vitro experiments mimicking the natural conditions of selective pressure yielded imipenem-resistant strains in which ISPa133 similarly interrupted oprD. A mechanism is proposed whereby ISPa133 acts as a mobile switch, with its position in oprD depending on the degree of selective pressure exerted by imipenem.
Assuntos
Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Elementos de DNA Transponíveis , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Resistência beta-Lactâmica , Códon de Iniciação , DNA Bacteriano/química , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Eletroforese em Gel de Poliacrilamida , Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Técnicas de Inativação de Genes , Humanos , Dados de Sequência Molecular , Mutagênese Insercional , Proteoma/análise , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/isolamento & purificação , Análise de Sequência de DNARESUMO
This study explored the evolutionary mechanism by which the clinical isolate PA110514 yields the imipenemresistant derivative PA116136. Both isolates were examined by PFGE and SDS-PAGE, which led to the identification of a new insertion sequence, ISPa133. This element was shown to have distinct chromosomal locations in each of the original isolates that appeared to explain the differences in imipenem susceptibilty. In strain PA110514, ISPa133 is located 56 nucleotides upstream of the translational start codon, which has no effect on expression of the porin OprD. However, in strain PA116136 ISPa133 it is located in front of nucleotide 696 and, by interrupting the coding region, causes a loss of OprD expression, thus conferring imipenem resistance. In vitro experiments mimicking the natural conditions of selective pressure yielded imipenem-resistant strains in which ISPa133 similarly interrupted oprD. A mechanism is proposed whereby ISPa133 acts as a mobile switch, with its position in oprD depending on the degree of selective pressure exerted by imipenem (AU)
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Assuntos
Humanos , Carbapenêmicos/farmacocinética , Resistência Microbiana a Medicamentos , Pseudomonas aeruginosa/genética , Testes de Sensibilidade Microbiana/métodos , Infecções por Pseudomonas/microbiologia , Elementos de DNA TransponíveisRESUMO
En el presente trabajo se valora la eficacia antibacteriana del colutorio Tantum verde® y la de su, presuntamente, único principio activo, la bencidamina clorhidrato. Para ello, se ensayó la actividad in vitro de la bencidamina HCl y del Tantum verde mediante la obtención de las correspondientes CMI (Concentración Mínima Inhibitoria) siguiendo la técnica de la dilución en medio sólido. Inicialmente, se estudiaron ocho cepas de uso común en el laboratorio y, posteriormente, el estudio fue ampliado a cepas de patógenos bucales aisladas de muestras clínicas. Los estudios realizados, demuestran una eficacia bactericida real frente a patógenos bucales pertenecientes a géneros tales como Actinomyces, Bacillus, Actinobacillus, Veillonella o Streptococcus. Además, el Tantum verde como colutorio posee, en general, una mayor actividad antibacteriana que la demostrada por su principal principio activo, la bencidamina HCl, por lo que cabe pensar que la presencia de etanol a baja concentración, en su composición contribuye de forma notable a la acción antibacteriana (AU)
