Biomedical subjects

Ashok Rattan

Publications and source records attributed to Ashok Rattan.

8 recordsLinked to original sources

PubMed2006

Characterisation of laboratory-generated vancomycin intermediate resistant Staphylococcus aureus strains.

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.

Laboratories↗

PubMed2005

Synthesis and SAR of novel oxazolidinones: discovery of ranbezolid.

Novel oxazolidinones were synthesized containing a number of substituted five-membered heterocycles attached to the 'piperazinyl-phenyl-oxazolidinone' core of eperezolid. Further, the piperazine ring of the core was replaced by other diamino-heterocycles. These modifications led to several compounds with potent activity against a spectrum of resistant and susceptible gram-positive organisms, along with the identification of ranbezolid (RBx 7644) as a clinical candidate.

Animals↗

PubMed2005

Activity of RBx 7644 and RBx 8700, new investigational oxazolidinones, against Mycobacterium tuberculosis infected murine macrophages.

The decision to develop a new chemical entity should not only be based on its ability to inhibit multidrug-resistant tuberculosis (MDR-TB) strains but also on its ability to enter macrophages and be active against intracellular bacteria. RBx 7644 and RBx 8700, two novel extended spectrum oxazolidinones, were investigated for their activity against sensitive and MDR isolates of Mycobacterium tuberculosis and for activity against bacteria within a macrophage cell line. RBx 8700 showed excellent in vitro activity against sensitive as well as MDR M. tuberculosis strains with MIC(50) and MIC(90) values of 0.032 and 0.25mg/L (sensitive) and 0.25 and 1.0mg/L (MDR) strains. The corresponding MIC(50) and MIC(90) values of RBx 7644, linezolid, rifampicin and isoniazid were 8 and 16; 32 and 64; 64 and 64; 64 and 64 mg/L, respectively. RBx 8700 and rifampicin were bactericidal at 0.5 and 0.25mg/L when tested intracellularly whereas linezolid reduced the count by 100-fold at a concentration of 8 mg/L. In combination studies with standard antimycobacterial drugs, RBx 8700 did not show any antagonistic effect.

Acetamides↗

PubMed2005

Adverse effect of staphylococci slime on in vitro activity of glycopeptides.

Adhesion to biomaterial is assumed to be a crucial step in the development of staphylococcal foreign body infections. Production of extracellular slime has major implications for the development and implementation of therapeutic strategies. The effect of extracted slime was investigated on the activity of vancomycin, teicoplanin, linezolid, quinupristin/dalfopristin, rifampicin and ranbezolid against 10 clinical and 4 ATCC staphylococcal isolates. The slime extract caused a 2- to 16-fold increase in the MICs of vancomycin and teicoplanin, with a shift in the MIC(90) from 2 to 32 (vancomycin) and 2 to 16 (teicoplanin), whereas the MICs of linezolid and quinupristin/dalfopristin were only moderately affected. In time-kill studies, a significant decrease in bacterial killing (>3 log(10) cfu/ml) was observed with vancomycin and teicoplanin (4 x MIC) after addition of slime (5 and 20 mg/ml), whereas the effect of killing by linezolid and quinupristin/dalfopristin was very modest. The rifampicin and ranbezolid MICs and kill curves were not influenced by the addition of slime. The present study thus indicated that slime interferes with the antimicrobial effect of glycopeptide drugs (vancomycin, teicoplanin), and that for effective prevention and treatment of prosthetic device-related infections, appropriate and newer antibiotics such as ranbezolid should be considered.

Anti-Bacterial Agents↗

PubMed2004

In vitro activity of RBx 7644 (ranbezolid) on biofilm producing bacteria.

Biofilm associated infections are becoming more common. Treatment outcome of device related infections cannot be predicted by the results of a standard susceptibility test such as the MIC. Microorganisms involved in device related infections have a slow growth rate and adhere to surfaces. Activity against glass adherent bacteria has been shown to be correlated with treatment outcome in animal models of catheter related infections. Drug efficacy can be predicted if glass adherent staphylococci are killed by low drug concentration. The eradication of bacteria adhering to glass beads and inhibition of biofilm formation by ranbezolid and three other antibiotics (quinupristin/dalfopristin, vancomycin and linezolid) was studied. The results indicated that ranbezolid required only (2-4 x MIC) for total clearance of glass-adherent MRSA 562 and MRSE 879, compared with vancomycin (8 x), quinupristin/dalfopristin (1-4 x) and linezolid (4-16 x MIC). In addition ranbezolid inhibited biofilm formation to a greater extent at sub MIC and MIC level. In conclusion, this study indicated that ranbezolid had potent activity against adherent staphylococci isolates and may prove useful in the prevention and treatment of device related infections caused by staphylococci.

Anti-Infective Agents↗

PubMed2003

Antianaerobe activity of RBX 7644 (ranbezolid), a new oxazolidinone, compared with those of eight other agents.

The activity of ranbezolid (RBX 7644), a new oxazolidinone, against 306 anaerobes was compared with those of 11 other agents. The MICs at which 50% of the isolates tested are inhibited and those at which 90% of the isolates tested are inhibited (in micrograms per milliliter) were as follows: ranbezolid, 0.03 and 0.5; linezolid, 2 and 4; vancomycin, >16 and >16; teicoplanin, 1 and >16; quinupristin-dalfopristin, 1 and >8; amoxicillin-clavulanate, 0.5 and 2; imipenem, 0.125 and 1; clindamycin, 0.25 and 8; metronidazole, 1 and 4; gatifloxacin, 0.5 and 4; and moxifloxacin, 0.5 and 2, respectively. Ranbezolid had very good in vitro activity against both gram-negative and -positive anaerobes.

Anti-Bacterial Agents↗

PubMed2003

Antipneumococcal and antistaphylococcal activities of ranbezolid (RBX 7644), a new oxazolidinone, compared to those of other agents.

For 260 pneumococcal and 266 staphylococcal strains, ranbezolid MICs ranged from < or =0.06 to 4 micro g/ml. The MICs for pneumococci were similar irrespective of the strains' beta-lactam, macrolide, or quinolone susceptibilities, and ranbezolid MICs for coagulase-negative staphylococci were lower than those for Staphylococcus aureus. Ranbezolid was bacteriostatic against pneumococci. Ranbezolid MICs were similar to or lower than those of linezolid. Vancomycin and quinupristin-dalfopristin were also very active.

Anti-Bacterial Agents↗

PubMed2002

In vitro low-level resistance to azoles in Candida albicans is associated with changes in membrane lipid fluidity and asymmetry.

The present study tracks the development of low-level azole resistance in in vitro fluconazole-adapted strains of Candida albicans, which were obtained by serially passaging a fluconazole-susceptible dose-dependent strain, YO1-16 (fluconazole MIC, 16 microg ml(-1)) in increasing concentrations of fluconazole, resulting in strains YO1-32 (fluconazole MIC, 32 microg ml(-1)) and YO1-64 (MIC, 64 microg ml(-1)). We show that acquired resistance to fluconazole in this series of isolates is not a random process but is a gradually evolved complex phenomenon that involves multiple changes, which included the overexpression of ABC transporter genes, e.g., CDR1 and CDR2, and the azole target enzyme, ERG11. The sequential rise in fluconazole MICs in these isolates was also accompanied by cross-resistance to other azoles and terbinafine. Interestingly, fluorescent polarization measurements performed by using the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene revealed that there was a gradual increase in membrane fluidity of adapted strains. The increase in fluidity was reflected by observed change in membrane order, which was considerably decreased (decrease in fluorescence polarization values, P value) in the adapted strain (P value of 0.1 in YO1-64, compared to 0.19 in the YO1-16 strain). The phospholipid composition of the adapted strain was not significantly altered; however, ergosterol content was reduced in YO1-64 from that in the YO1-16 strain. The asymmetrical distribution of phosphatidylethanolamine (PE) between two monolayers of plasma membrane was also changed, with PE becoming more exposed to the outer monolayer in the YO1-64 strain. The results of the present study suggest for the first time that changes in the status of membrane lipid phase and asymmetry could contribute to azole resistance in C. albicans.

Antifungal Agents↗