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1.
Antimicrob Agents Chemother ; 58(8): 4283-9, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24550341

RESUMEN

We present here the novel ketolide RBx 14255, a semisynthetic macrolide derivative obtained by the derivatization of clarithromycin, for its in vitro and in vivo activities against sensitive and macrolide-resistant Streptococcus pneumoniae. RBx 14255 showed excellent in vitro activity against macrolide-resistant S. pneumoniae, including an in-house-generated telithromycin-resistant strain (S. pneumoniae 3390 NDDR). RBx 14255 also showed potent protein synthesis inhibition against telithromycin-resistant S. pneumoniae 3390 NDDR. The binding affinity of RBx 14255 toward ribosomes was found to be more than that for other tested drugs. The in vivo efficacy of RBx 14255 was determined in murine pulmonary infection induced by intranasal inoculation of S. pneumoniae ATCC 6303 and systemic infection with S. pneumoniae 3390 NDDR strains. The 50% effective dose (ED50) of RBx 14255 against S. pneumoniae ATCC 6303 in a murine pulmonary infection model was 3.12 mg/kg of body weight. In addition, RBx 14255 resulted in 100% survival of mice with systemic infection caused by macrolide-resistant S. pneumoniae 3390 NDDR at 100 mg/kg four times daily (QID) and at 50 mg/kg QID. RBx 14255 showed favorable pharmacokinetic properties that were comparable to those of telithromycin.


Asunto(s)
Antibacterianos/farmacología , Cetólidos/farmacología , Neumonía Bacteriana/tratamiento farmacológico , Inhibidores de la Síntesis de la Proteína/farmacología , Sepsis/tratamiento farmacológico , Streptococcus pneumoniae/efectos de los fármacos , Animales , Antibacterianos/síntesis química , Antibacterianos/farmacocinética , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Farmacorresistencia Bacteriana , Cetólidos/síntesis química , Cetólidos/farmacocinética , Masculino , Ratones , Pruebas de Sensibilidad Microbiana , Neumonía Bacteriana/microbiología , Neumonía Bacteriana/mortalidad , Neumonía Bacteriana/patología , Inhibidores de la Síntesis de la Proteína/síntesis química , Inhibidores de la Síntesis de la Proteína/farmacocinética , Ribosomas/efectos de los fármacos , Ribosomas/metabolismo , Sepsis/microbiología , Sepsis/mortalidad , Sepsis/patología , Streptococcus pneumoniae/patogenicidad , Streptococcus pneumoniae/fisiología , Análisis de Supervivencia
2.
Arch Microbiol ; 192(8): 663-72, 2010 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-20552170

RESUMEN

CSM2, a cold-sensitive mutant of psychrophilic Pseudomonas syringae, grows like wild-type cells when cultured at 22 and 28 degrees C; but at 4 degrees C, the growth is retarded. In CSM2, AAT (coding for aspartate aminotransferase) is identified as the mutated gene. The expression of AAT in Pseudomonas syringae was transiently enhanced when cells were shifted from 22 to 4 degrees C indicating that AAT is cold-inducible. Complementation of the mutated AAT transformed CSM2 from a cold-sensitive phenotype to a cold-resistant phenotype like the wild-type cells, thus providing evidence for the first time that AAT is required for low-temperature growth.


Asunto(s)
Adaptación Fisiológica , Aspartato Aminotransferasas/metabolismo , Proteínas Bacterianas/metabolismo , Frío , Pseudomonas syringae/enzimología , Secuencia de Aminoácidos , Aspartato Aminotransferasas/genética , Proteínas Bacterianas/genética , Secuencia de Bases , Clonación Molecular , ADN Bacteriano/genética , Prueba de Complementación Genética , Datos de Secuencia Molecular , Mutagénesis , Mutación , Filogenia , Pseudomonas syringae/genética , Pseudomonas syringae/crecimiento & desarrollo , Alineación de Secuencia , Sitio de Iniciación de la Transcripción
3.
Appl Environ Microbiol ; 75(13): 4419-26, 2009 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19429554

RESUMEN

Transposon mutagenesis of Pseudomonas syringae Lz4W, a psychrophilic bacterium capable of growing at temperatures between 2 and 30 degrees C, yielded 30 cold-sensitive mutants, and CSM1, one of these cold-sensitive mutants, was characterized. Growth of CSM1 was retarded when it was cultured at 4 degrees C but not when it was cultured at 22 degrees C and 28 degrees C compared to the growth of wild-type cells, indicating that CSM1 is a cold-sensitive mutant of P. syringae Lz4W. The mutated gene in CSM1 was identified as trmE (coding for tRNA modification GTPase), and evidence is provided that this gene is induced at low temperatures. Further, the cold-inducible nature of the trmE promoter was demonstrated. In addition, the transcription start site and the various regulatory elements of the trmE promoter, such as the -10 region, -35 region, UP element, cold box, and DEAD box, were identified, and the importance of these regulatory elements in promoter activity were confirmed. The importance of trmE in rapid adaptation to growth at low temperatures was further highlighted by plasmid-mediated complementation that alleviated the cold-sensitive phenotype of CSM1.


Asunto(s)
Proteínas Bacterianas/fisiología , Frío , GTP Fosfohidrolasas/fisiología , Pseudomonas syringae/crecimiento & desarrollo , Proteínas Bacterianas/genética , Secuencia de Bases , Elementos Transponibles de ADN , GTP Fosfohidrolasas/genética , Perfilación de la Expresión Génica , Prueba de Complementación Genética , Datos de Secuencia Molecular , Mutagénesis Insercional , Regiones Promotoras Genéticas , Pseudomonas syringae/genética , Elementos Reguladores de la Transcripción , Sitio de Iniciación de la Transcripción
4.
Emerg Infect Dis ; 13(10): 1614-6, 2007 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-18258023

RESUMEN

Decreased susceptibility of Neisseria meningitidis isolates to ciprofloxacin emerged from an outbreak in Delhi, India. Results of antimicrobial susceptibility testing of the meningococcal isolates to ciprofloxacin and further sequencing of DNA gyrase A quinolone-resistance-determining region confirmed the emergence of ciprofloxacin resistance in the outbreak.


Asunto(s)
Brotes de Enfermedades , Farmacorresistencia Bacteriana Múltiple , Infecciones Meningocócicas/epidemiología , Neisseria meningitidis Serogrupo A/efectos de los fármacos , Ciprofloxacina/farmacología , Humanos , India/epidemiología , Infecciones Meningocócicas/tratamiento farmacológico , Pruebas de Sensibilidad Microbiana , Neisseria meningitidis Serogrupo A/clasificación , Neisseria meningitidis Serogrupo A/genética , Serotipificación
5.
Int J Antimicrob Agents ; 27(3): 201-11, 2006 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-16417988

RESUMEN

Vancomycin has been the drug of choice for 30 years for the treatment of methicillin-resistant Staphylococcus aureus (MRSA). Emergence of decreased vancomycin susceptibility in MRSA strains presents a significant clinical problem with few therapeutic options. This study was performed to generate and characterise S. aureus strains with reduced susceptibility to vancomycin. Eighteen S. aureus strains were subjected to serial passaging on vancomycin to generate vancomycin intermediate resistant S. aureus (VISA) strains. Minimum inhibitory concentration (MIC) determination was performed for the parent and the passaged cultures with 13 different antibiotics. The strains were tested by the following five methods: simplified population analysis; CDC method; modified vancomycin agar screen; population analysis profile (PAP); and modified population analysis (PAP-area under the curve (AUC) ratio). Phenotypic changes such as doubling time, synergy with beta-lactam antibiotics and effect on norA efflux pumps were also studied for these strains. The result indicated that 8 VISA mutants (vancomycin MICs, 8-16 microg/mL) were generated in vitro from the 18 S. aureus strains. The CDC and modified agar methods proved to be the most sensitive and specific methods for detection of VISA strains. The PAP for all the VISA strains ranged from 12 microg/mL to > 16 microg/mL, with a PAP-AUC ratio of > 1.3. All mutants showed increased doubling time compared with their parent isolate. Synergism of the vancomycin and beta-lactam combinations was observed for all methicillin-resistant mutants. Upon acquisition of vancomycin resistance, a few mutants showed decreased oxacillin resistance. Two VISA strains were chosen for molecular characterisation of the mecA gene and one mutant showed genotypic changes with deletion of mecA. Loss of norA efflux pumps leading to fluoroquinolone sensitivity was also observed in four mutants.


Asunto(s)
Staphylococcus aureus/efectos de los fármacos , Vancomicina/farmacología , Laboratorios , Pruebas de Sensibilidad Microbiana , Staphylococcus aureus/crecimiento & desarrollo , Resistencia a la Vancomicina
6.
Syst Appl Microbiol ; 27(1): 66-71, 2004 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-15053323

RESUMEN

Strain DVS 5a1 was isolated from a moraine sample from the McMurdo Dry Valleys region of Antarctica. The strain is aerobic, Gram-positive, with white aerial mycelia and brown substrate mycelia, sporulating, has meso-diaminopimelic acid, arabinose and galactose in the cell wall, MK-8 (H4) as the major menaquinone and a mol% G+C content of DNA of 71% thus confirming to the description of the genus Pseudonocardia. Phylogenetic analysis based on the 16S rRNA gene sequence analysis further confirms that DVS 5al which forms a robust clade with P. alni, P. compacta, P. autotrophica and P. kongjuensis is closely related to the genus Pseudonocardia and exhibits maximum similarity of 99.7% with Pseudonocardia alni. However, at whole genome level as determined by DNA-DNA hybridisation DVS 5al exhibits a similarity of only 50% with Pseudonocardia alni. Further, DVS 5al differs from Pseudonocardia alni in that it does not produce acid from D-arabinose, meso-erythritol, melizitose, sorbitol, sucrose, D-trehalose; but produces acid from D-mannitol, D-galactose, D-maltose, D-mannose, inulin, D-ribose and D-xylose. Further, compared to Pseudonocardia alni, it has two additional fatty acids namely Me-C(18:0) and Me-C(19:0) and also possesses one additional unidentified lipid. It also shows distinct differences with P. compacta, P. autotrophica and P. kongjuensis and the other species of Pseudonocardia. It is proposed to assign DVS 5a1 the status of a new species for which the name Pseudonocardia antarctica sp. nov. is suggested.


Asunto(s)
Nocardia/aislamiento & purificación , Microbiología del Suelo , Regiones Antárticas , Secuencia de Bases , ADN Bacteriano/química , ADN Bacteriano/genética , Ácidos Grasos/análisis , Microscopía de Contraste de Fase , Datos de Secuencia Molecular , Nocardia/genética , Nocardia/metabolismo , Hibridación de Ácido Nucleico , Peptidoglicano/análisis , Filogenia , ARN Ribosómico 16S/química , ARN Ribosómico 16S/genética , Alineación de Secuencia
7.
Syst Appl Microbiol ; 26(4): 505-10, 2003 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-14666977

RESUMEN

An orange pigmented bacterium, S1, was isolated from a cyanobacterial mat sample collected in the vicinity of Schirmacher Oasis, Maitri, the Indian station, in Antarctica. The bacterium is Gram-positive and possesses all the characteristics of the genus Planococcus. It is non-sporulating, motile and has A4alpha type peptidoglycan, MK-7 and MK-8 as the major menaquinones and anteiso-C(15:0) as the major fatty acid. Based on the phylogenetic characteristics, the bacterium S1 is identified as a close relative of Planococcus citreus with which it shares 98.12% similarity at the 16S rRNA gene level but exhibits a low similarity of 52% at the whole genome level. Apart from the above major differences, S1 also exhibits phenotypic differences with Planococcus citreus and other members of the genus Planococcus. Based on these differences, the bacterium S1 is identified as a new species of the genus Planococcus for which the name Planococcus maitriensis is proposed. The type strain of Planococcus maitriensis is S1(T) (= MTCC 4827; DSM 15305).


Asunto(s)
Cocos Grampositivos/clasificación , Cocos Grampositivos/aislamiento & purificación , Regiones Antárticas , Frío , ADN Bacteriano/química , ADN Bacteriano/aislamiento & purificación , ADN Ribosómico/química , ADN Ribosómico/aislamiento & purificación , Ácidos Grasos/análisis , Ácidos Grasos/química , Cocos Grampositivos/citología , Cocos Grampositivos/fisiología , Datos de Secuencia Molecular , Movimiento , Hibridación de Ácido Nucleico , Peptidoglicano/análisis , Peptidoglicano/química , Filogenia , Pigmentos Biológicos/biosíntesis , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Vitamina K 2/análisis , Vitamina K 2/química
8.
Can J Microbiol ; 52(12): 1238-46, 2006 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-17473893

RESUMEN

An anaerobic, proteolytic bacterium isolated from lake sediments of Leh, India, was characterized with respect to morphology, biochemical characteristics, and 16S rRNA sequence and was identified as Clostridium species, with closest similarity to Clostridium subterminale. Isolate LP3 was psychrophilic, forming maximum cell mass between 10 and 20 degrees C, and produced extracellular protease. Growth was observed in the pH range of 7.0-8.5, with optimum at pH 7.5. Protease was purified 62.4-fold with a total yield of 17.5%. The effects of temperature, pH, and salt concentration on enzyme activity were studied. Protease was found to be a serine-type metallo-enzyme, active in a broad range of pHs. It was thermolabile and resistant to sodium dodecyl sulfate. Enzyme kinetics showed a tendency to increase Km with an increase in temperature for casein substrate.


Asunto(s)
Clostridium/enzimología , Sedimentos Geológicos/microbiología , Péptido Hidrolasas/metabolismo , Clostridium/clasificación , Clostridium/genética , ADN Bacteriano/química , ADN Bacteriano/genética , Electroforesis en Gel de Poliacrilamida , Estabilidad de Enzimas , India , Cinética , Datos de Secuencia Molecular , Péptido Hidrolasas/genética , Péptido Hidrolasas/aislamiento & purificación , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Temperatura
9.
Int J Syst Evol Microbiol ; 55(Pt 3): 1123-1127, 2005 May.
Artículo en Inglés | MEDLINE | ID: mdl-15879243

RESUMEN

Strain Con a/3(T) is a Gram-positive, motile, endospore-forming, rod-shaped and arsenic-resistant bacterium, which was isolated from a concretion of arsenic ore obtained from a bore-hole. The bacterium grew in the presence of 20 mM arsenate and 0.5 mM arsenite. Diaminopimelic acid was present in the cell wall peptidoglycan, MK-7 was the major menaquinone, and iso-C(15 : 0), anteiso-C(15 : 0), iso-C(16 : 0) and C(16 : 1)(delta7cis) were the major fatty acids. Based on its phenotypic, chemotaxonomic and phylogenetic characteristics, strain Con a/3(T) was identified as a member of the genus Bacillus. It exhibited maximum similarity (97 %) at the 16S rRNA gene level with Bacillus barbaricus (DSM 14730(T)); however, the DNA-DNA relatedness value with B. barbaricus was 60 %. Strain Con a/3(T) also exhibited a number of phenotypic differences from B. barbaricus (DSM 14730(T)). Strain Con a/3(T) was therefore identified as representing a novel species of the genus Bacillus, for which the name Bacillus arsenicus sp. nov. is proposed. The type strain is Con a/3(T) (= MTCC 4380(T) = DSM 15822(T) = JCM 12167(T)).


Asunto(s)
Antibacterianos/farmacología , Arsénico/farmacología , Bacillus/clasificación , Bacillus/efectos de los fármacos , Microbiología del Suelo , Arsenitos/farmacología , Bacillus/aislamiento & purificación , Bacillus/fisiología , Técnicas de Tipificación Bacteriana , Carbonatos , Cianoacrilatos/análisis , Cianoacrilatos/aislamiento & purificación , ADN Bacteriano/química , ADN Bacteriano/aislamiento & purificación , ADN Ribosómico/química , ADN Ribosómico/aislamiento & purificación , Ácido Diaminopimélico/análisis , Ácido Diaminopimélico/aislamiento & purificación , Farmacorresistencia Bacteriana , Ácidos Grasos/análisis , Ácidos Grasos/aislamiento & purificación , Compuestos Férricos , Genes de ARNr , India , Datos de Secuencia Molecular , Movimiento , Hibridación de Ácido Nucleico , Filogenia , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Esporas Bacterianas , Vitamina K 2/análisis , Vitamina K 2/aislamiento & purificación
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