RESUMO
Background: Fingerprint drug concentrations can be used as a noninvasive and convenient alternative to evaluate adherence to pharmacotherapy. Methods: Fingerprints were applied over glass slides, extracted and analyzed by ultra-high performance LC-MS/MS. The assay and drug adherence questionnaires were applied to 30 epilepsy patients. Results: The assay had linearity in the range 0.05-10 ng fingerprint-1, with precision of 2.16-7.9% and accuracy of 95.0-102.8%. Carbamazepine (CBZ) levels in fingerprints were stable at 45°C for 15 days. Concentrations in patient samples were 0.06-9.28 ng fingerprint-1. A significant difference (p = 0.003) was found between CBZ concentrations in fingerprints between patient groups divided as low and medium/high adherence. Conclusion: This method can potentially be applied to the identification of epilepsy patients with low adherence to CBZ pharmacotherapy.
[Box: see text].
Assuntos
Carbamazepina , Epilepsia , Estudos de Viabilidade , Adesão à Medicação , Espectrometria de Massas em Tandem , Carbamazepina/uso terapêutico , Humanos , Epilepsia/tratamento farmacológico , Feminino , Masculino , Espectrometria de Massas em Tandem/métodos , Adulto , Anticonvulsivantes/uso terapêutico , Cromatografia Líquida de Alta Pressão/métodos , Dermatoglifia , Pessoa de Meia-Idade , Adulto JovemRESUMO
INTRODUCTION: The rapid global spread of carbapenem-resistant Enterobacteriaceae (CRE) is a threat to the health system. METHODS: We evaluated the antimicrobial susceptibility profiles of 70 CRE isolated in a tertiary hospital in Brazil between August and December 2015, and determined their resistance mechanisms. RESULTS: The most prevalent microorganism was Klebsiella pneumoniae (95.7%); it showed high-level resistance to carbapenems (>98%), with sensitivity to colistin (91.4%) and amikacin (98.6%). The bla KPC gene was detected in 80% of the CRE isolates. CONCLUSIONS: Evaluation of bacterial resistance contributes to an appropriate treatment, and the reduction of morbimortality and dissemination of resistance.
Assuntos
Enterobacteriáceas Resistentes a Carbapenêmicos/isolamento & purificação , Infecções por Enterobacteriaceae/epidemiologia , Centros de Atenção Terciária/estatística & dados numéricos , Adolescente , Adulto , Anti-Infecciosos/farmacologia , Brasil/epidemiologia , Criança , Pré-Escolar , Citrobacter freundii/isolamento & purificação , Infecção Hospitalar/epidemiologia , Enterobacter cloacae/isolamento & purificação , Infecções por Enterobacteriaceae/microbiologia , Escherichia coli/isolamento & purificação , Feminino , Genótipo , Humanos , Lactente , Klebsiella pneumoniae/isolamento & purificação , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Adulto JovemRESUMO
Abstract INTRODUCTION: The rapid global spread of carbapenem-resistant Enterobacteriaceae (CRE) is a threat to the health system. METHODS: We evaluated the antimicrobial susceptibility profiles of 70 CRE isolated in a tertiary hospital in Brazil between August and December 2015, and determined their resistance mechanisms. RESULTS: The most prevalent microorganism was Klebsiella pneumoniae (95.7%); it showed high-level resistance to carbapenems (>98%), with sensitivity to colistin (91.4%) and amikacin (98.6%). The bla KPC gene was detected in 80% of the CRE isolates. CONCLUSIONS: Evaluation of bacterial resistance contributes to an appropriate treatment, and the reduction of morbimortality and dissemination of resistance.
Assuntos
Humanos , Masculino , Feminino , Lactente , Pré-Escolar , Criança , Adolescente , Adulto , Adulto Jovem , Infecções por Enterobacteriaceae/epidemiologia , Centros de Atenção Terciária/estatística & dados numéricos , Enterobacteriáceas Resistentes a Carbapenêmicos/isolamento & purificação , Brasil/epidemiologia , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Infecção Hospitalar/epidemiologia , Enterobacter cloacae/isolamento & purificação , Citrobacter freundii/isolamento & purificação , Infecções por Enterobacteriaceae/microbiologia , Escherichia coli/isolamento & purificação , Genótipo , Klebsiella pneumoniae/isolamento & purificação , Anti-Infecciosos/farmacologia , Pessoa de Meia-IdadeRESUMO
Biofilm resistance mechanisms are multifactorial and vary from one organism to another. The purpose of this study was to investigate the efficacy of linezolid against indwelling device-related meticillin-resistant Staphylococcus epidermidis (MRSE) biofilm, and compare this with other antimicrobials. MICs, minimum biofilm inhibitory concentrations (MBICs) and minimum biofilm eradication concentrations (MBECs) were determined by the microtitre plate method. Fourteen and thirteen isolates from patients with indwelling device-related bacteraemia (IDB) and indwelling device colonization not associated with bacteraemia, respectively, were assessed. High MBIC was associated with a high intensity of biofilm formation (gentamicin r=0.796; linezolid r=0.477; rifampicin r=0.634; tigecycline r=0.410; and vancomycin r=0.771), but this correlation was not observed with MBEC. Linezolid demonstrated better in vitro antimicrobial activity than other antimicrobials (MBIC - gentamicin P<0.001, rifampicin P=0.019, vancomycin P=0.008; MBEC - gentamicin P<0.001, rifampicin P=0.002, vancomycin P<0.001). Biofilm growth inhibition was strongly associated with biofilm formation intensity; however, biofilm eradication was not cell number dependent. MRSE biofilm eradication would represent a huge advance for IDB, although high concentrations of gentamicin, linezolid, rifampicin, tigecycline and vancomycin were required for that. In general, linezolid reached better in vitro concentrations and was demonstrated to be highly active against MRSE biofilms by inhibiting their growth during biofilm formation.
Assuntos
Acetamidas/farmacologia , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Resistência a Meticilina , Oxazolidinonas/farmacologia , Staphylococcus epidermidis/fisiologia , Bacteriemia/microbiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Técnicas Bacteriológicas , Biofilmes/crescimento & desenvolvimento , Relação Dose-Resposta a Droga , Regulação Bacteriana da Expressão Gênica , Linezolida , Meticilina/farmacologia , Testes de Sensibilidade Microbiana , Infecções Relacionadas à Prótese/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus epidermidis/efeitos dos fármacosRESUMO
INTRODUCTION: Antimicrobial activity on biofilms depends on their molecular size, positive charges, permeability coefficient, and bactericidal activity. Vancomycin is the primary choice for methicillin-resistant Staphylococcus aureus (MRSA) infection treatment; rifampicin has interesting antibiofilm properties, but its effectivity remains poorly defined. METHODS: Rifampicin activity alone and in combination with vancomycin against biofilm-forming MRSA was investigated, using a twofold serial broth microtiter method, biofilm challenge, and bacterial count recovery. RESULTS: Minimal inhibitory concentration (MIC) and minimal bactericidal concentration for vancomycin and rifampicin ranged from 0.5 to 1mg/l and 0.008 to 4mg/l, and from 1 to 4mg/l and 0.06 to 32mg/l, respectively. Mature biofilms were submitted to rifampicin and vancomycin exposure, and minimum biofilm eradication concentration ranged from 64 to 32,000 folds and from 32 to 512 folds higher than those for planktonic cells, respectively. Vancomycin (15mg/l) in combination with rifampicin at 6 dilutions higher each isolate MIC did not reach in vitro biofilm eradication but showed biofilm inhibitory capacity (1.43 and 0.56log10 CFU/ml reduction for weak and strong biofilm producers, respectively; p<0.05). CONCLUSIONS: In our setting, rifampicin alone failed to effectively kill biofilm-forming MRSA, demonstrating stronger inability to eradicate mature biofilm compared with vancomycin.
Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/fisiologia , Rifampina/farmacologia , Vancomicina/farmacologia , Biofilmes/crescimento & desenvolvimento , Humanos , Testes de Sensibilidade MicrobianaRESUMO
INTRODUCTION: Antimicrobial activity on biofilms depends on their molecular size, positive charges, permeability coefficient, and bactericidal activity. Vancomycin is the primary choice for methicillin-resistant Staphylococcus aureus (MRSA) infection treatment; rifampicin has interesting antibiofilm properties, but its effectivity remains poorly defined. METHODS: Rifampicin activity alone and in combination with vancomycin against biofilm-forming MRSA was investigated, using a twofold serial broth microtiter method, biofilm challenge, and bacterial count recovery. RESULTS: Minimal inhibitory concentration (MIC) and minimal bactericidal concentration for vancomycin and rifampicin ranged from 0.5 to 1mg/l and 0.008 to 4mg/l, and from 1 to 4mg/l and 0.06 to 32mg/l, respectively. Mature biofilms were submitted to rifampicin and vancomycin exposure, and minimum biofilm eradication concentration ranged from 64 to 32,000 folds and from 32 to 512 folds higher than those for planktonic cells, respectively. Vancomycin (15mg/l) in combination with rifampicin at 6 dilutions higher each isolate MIC did not reach in vitro biofilm eradication but showed biofilm inhibitory capacity (1.43 and 0.56log10 CFU/ml reduction for weak and strong biofilm producers, respectively; p<0.05). CONCLUSIONS: In our setting, rifampicin alone failed to effectively kill biofilm-forming MRSA, demonstrating stronger inability to eradicate mature biofilm compared with vancomycin.
INTRODUÇÃO: A atividade dos antimicrobianos em biofilmes depende do seu peso molecular, de cargas positivas, coeficiente de permeabilidade e atividade bactericida. Vancomicina é a escolha primária para o tratamento de infecções causadas por Staphylococcus aureus resistentes à meticilina (MRSA) e rifampicina possui interessante propriedade antibiofilme, apesar da sua efetividade ainda ser fracamente definida. MÉTODOS: Foi investigada a atividade da rifampicina sozinha e em combinação com vancomicina frente à MRSA formadores de biofilme, utilizando o método das microplacas com diluição seriada e recuperação bacteriana em biofilme após exposição antimicrobiana. RESULTADOS: Concentração inibitória minima (MIC) e concentração bactericida mínima (MBC) para vancomicina e rifampicina foi de 0,5-1mg/l e 0,008-4mg/l; 1-4mg/l e 0,06-32mg/l, respectivamente. Biofilmes maduros foram expostos à vancomicina e rifampicina, e a concentração mínima para erradicar o biofilme (MBEC) foi 64-32.000 e 32-512 vezes maior do que para células planctônicas, respectivamente. A combinação de vancomicina (15mg/l) com rifampicina (6-diluições maior do que o MIC de cada isolado) não atingiu erradicação do biofilme in vitro, porém apresentou capacidade inibitória do biofilme formado (redução de 1,43 e 0,56log10 UFC/ml para produtores fracos e fortes, respectivamente; p<0,05). CONCLUSÕES: Rifampicina sozinha falhou em efetivamente matar MRSA formadores de biofilme, demonstrando fraca habilidade para erradicação de biofilmes maduros comparado com vancomicina.
Assuntos
Humanos , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/fisiologia , Rifampina/farmacologia , Vancomicina/farmacologia , Biofilmes/crescimento & desenvolvimento , Testes de Sensibilidade MicrobianaRESUMO
This study was designed to evaluate antimicrobial activities against methicillin-susceptible Staphylococcus aureus in both sessile and planktonic forms and to detect genes associated with this biofilm phenotype. Minimal biofilm inhibition and eradication concentrations (MBIC and MBEC, respectively) were determined by an in vitro biofilm model, and icaA, atlA, and sasG genes were detected by polymerase chain reaction. Vancomycin and tigecycline presented better biofilm inhibitory activity (MBIC range: 4-8 µg/mL) (P ≤ 0.05) and lower MBEC/MIC ratios (P ≤ 0.001) than other antimicrobials. All isolates harbored icaA and atlA, whereas sasG was present only in strong biofilm formers (P ≤ 0.05). Interestingly, antimicrobial activities against sasG- weak biofilm formers were significantly higher than those against sasG+ strong biofilm formers (P ≤ 0.05), demonstrating that number of cells in a biofilm matrix affected the antimicrobial activity, which was also variable, and might be associated with specific genetic determinants. To our knowledge, this was the first study reporting the presence of sasG in clinical isolates of S. aureus in South America.