RESUMO
Candida albicans is an opportunistic human pathogen that is capable of causing superficial and systemic infections in immunocompromised patients. Extracts of Sapindus saponaria have been used as antimicrobial agents against various organisms. In the present study, we used a combination of two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to identify the changes in protein abundance of C. albicans after exposure to the minimal inhibitory concentration (MIC) and sub-minimal inhibitory concentration (sub-MIC) of the butanolic extract (BUTE) of S. saponaria and also to fluconazole. A total of six different proteins with greater than 1.5 fold induction or repression relative to the untreated control cells were identified among the three treatments. In general, proteins/enzymes involved with the glycolysis (GPM1, ENO1, FBA1), amino acid metabolism (ILV5, PDC11) and protein synthesis (ASC1) pathways were detected. In conclusion, our findings reveal antifungal-induced changes in protein abundance of C. albicans. By using the previously identified components of the BUTE of S. saponaria(e.g., saponins and sesquiterpene oligoglycosides), it will be possible to compare the behavior of compounds with unknown mechanisms of action, and this knowledge will help to focus the subsequent biochemical work aimed at defining the effects of these compounds.
Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Fluconazol/farmacologia , Proteínas Fúngicas/análise , Extratos Vegetais/farmacologia , Sapindus/química , Candida albicans/química , Eletroforese em Gel Bidimensional , Espectrometria de Massas , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de VarreduraRESUMO
The authors report a case of cutaneous tuberculosis in a 63-year-old female patient, who had an infiltrated, erythematous-ferruginous plaque of indurated aspect on her right leg and a nonreactive PPD skin test. Diagnosis was made by tissue culture and PCR of skin biopsy material. The treatment was performed with pyrazinamide, rifampicin, isoniazid and ethambutol, with good response.
Assuntos
Pele/patologia , Tuberculose Cutânea/patologia , Antituberculosos/uso terapêutico , Biópsia , Feminino , Humanos , Pessoa de Meia-Idade , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase , Testes Cutâneos/métodos , Tuberculose Cutânea/tratamento farmacológicoRESUMO
The authors report a case of cutaneous tuberculosis in a 63-year-old female patient, who had an infiltrated, erythematous-ferruginous plaque of indurated aspect on her right leg and a nonreactive PPD skin test. Diagnosis was made by tissue culture and PCR of skin biopsy material. The treatment was performed with pyrazinamide, rifampicin, isoniazid and ethambutol, with good response.
Assuntos
Feminino , Humanos , Pessoa de Meia-Idade , Pele/patologia , Tuberculose Cutânea/patologia , Antituberculosos/uso terapêutico , Biópsia , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase , Testes Cutâneos/métodos , Tuberculose Cutânea/tratamento farmacológicoRESUMO
OBJECTIVES: The resazurin microtitre plate assay (REMA) was evaluated to determine the susceptibility of Mycobacterium tuberculosis to pyrazinamide, and was compared with the broth microdilution method (BMM), the absolute concentration method (ACM) and pyrazinamidase (PZase) determination. METHODS: Thirty-four M. tuberculosis clinical isolates (26 susceptible and 8 resistant to pyrazinamide) and reference strains M. tuberculosis H37Rv ATCC 27294 and Mycobacterium bovis AN5 were tested. RESULTS: REMA and BMM showed 100% specificity and sensitivity when compared with ACM; BMM, however, demanded more reading time. The PZase determination assay showed 87.50% and 100% sensitivity and specificity, respectively. CONCLUSIONS: All tested methods in this preliminary study showed excellent sensitivity and specificity for the determination of pyrazinamide susceptibility of M. tuberculosis, but REMA was faster, low-cost and easy to perform and interpret. Additional studies evaluating REMA for differentiating pyrazinamide-resistant and -susceptible M. tuberculosis should be conducted on an extended panel of clinical isolates.