Antibiotic resistance pattern of Enterococci isolates from nosocomial infections in a tertiary care hospital in Eastern India.

BACKGROUND: Resistance to commonly used antibiotics by Enterococci causing nosocomial infections is of concern, which necessitates judicious, responsible and evidence-based use of antibiotics. The present study was conducted to review the prevalence and identify therapeutic options for nosocomial Enterococcal infections in our tertiary care hospital. MATERIALS AND METHODS: Isolates identified by morphological and biochemical characteristics were tested for antibiotic susceptibility using Kirby-Bauer method. RESULT: 153 of 2096 culture positive clinical samples comprised of 101 urine, 30 wound swab/pus, 13 blood and 09 high vaginal swab isolates were identified as Enterococcus faecalis (90.85%), Enterococcus faecium (8.50%) and Enterococcus gallinarum (0.65%). Enterococci accounted for 8.45%, 4.53%, 4.23%, 4.43% of urinary, wound swab or pus, blood, high vaginal swab isolates respectively, causing 7.3% of all nosocomial infections. Significant number of Enterococci isolated from nosocomial urinary tract infection (66.01%) and wound infections (19.6%) were multidrug resistant (MDR). Although all isolates were sensitive to vancomycin and linezolid, resistance to erythromycin (71.24%) and ciprofloxacin (49.67%) was frequently observed. High-level gentamicin resistance was observed in 43.88%, and 61.53% of E. faecalis and E. faecium isolates respectively. Minimal inhibitory concentration of vancomycin of all the isolates were ≤1 µg/ml. 7% of the Enterococcal isolates were MDR strains and vancomycin or linezolid were the only effective antibiotics. CONCLUSION: A combination of vancomycin and/or linezolid were effective against Enterococci causing nosocomial infections in our tertiary care facility, nevertheless continuous and frequent surveillance for resistance patterns are necessary for judicious and evidence based use of antibiotics.

Immunomodulation in host-protective immune response against murine tuberculosis through regulation of the T regulatory cell function.

Tuberculosis, caused by the bacteria Mycobacterium tuberculosis, is characterized by an infection in lung and spleen. In the present study, we have elucidated the mechanism by which Mycobacterium indicus pranii renders protection in in vivo Mycobacterium tuberculosis infection. We observed that Mycobacterium indicus pranii treated infected C57BL/6 mice showed a strong host-protective Th1 immune response along with a marked decrease in immunosuppressive cytokines, TGF-ß, and IL-10-secreting CD4(+) T cells. This Mycobacterium indicus pranii mediated decrease in immunosuppressive cytokines was correlated with the reduction in the elevated frequency of CD4(+)CD25(+) T regulatory cells, along with the reduced TGF-ß production from these T regulatory cells in tuberculosis-infected mice. This reduction in the T regulatory cell population was a result of effective modulation of STAT4-STAT5 transcription factor counter-regulation by Mycobacterium indicus pranii, which in turn, reduced the immunosuppressive activity of T regulatory cells. Thus, these findings put forward a detailed mechanistic insight into Mycobacterium indicus pranii mediated regulation of the T regulatory cell functioning during experimental murine tuberculosis, which might be helpful in combating Mycobacterium-induced pathogenesis.

Arabinosylated lipoarabinomannan (Ara-LAM) mediated intracellular mechanisms against tuberculosis infection: involvement of protein kinase C (PKC) mediated signaling.

Tuberculosis causes severe immunosuppression thereby ensuring the loss of the host protective immune responses. During Mycobacterium tuberculosis infection, the pathogen modulates TLR-2 receptor down-stream signaling, indicating the possible involvement of TLR-2 in the regulation of the host immune response. Moreover, different PKC isoforms are also involved in the course of infection. Arabinosylated lipoarabinomannan (Ara-LAM) possesses immuno-modulatory properties which induce the pro-inflammatory responses via induction of TLR-2-mediated signaling. Here, we found that pretreatment of M. tuberculosis-infected macrophages with Ara-LAM caused a significant increase in the conventional PKC expression along with their active association with TLR-2. This association activated the TLR-2 -mediated downstream signaling, facilitating the activation of MAP kinase P38. All these events culminated in the up-regulation of proinflammatory response, which was abrogated by treatment with PKC-α and P38 inhibitors. Moreover, pretreatment of macrophages with Ara-LAM abrogated the IL-10 production while restored MHC-II expression in the infected macrophages. This study demonstrates that Ara-LAM confers protection against tuberculosis via TLR-2/PKC signaling crosstalk which is responsible for the induction of host protective immune response against tuberculosis.

Immune subversion by Mycobacterium tuberculosis through CCR5 mediated signaling: involvement of IL-10.

Tuberculosis is characterized by severe immunosuppression of the host macrophages, resulting in the loss of the host protective immune responses. During Mycobacterium tuberculosis infection, the pathogen modulates C-C Chemokine Receptor 5 (CCR5) to enhance IL-10 production, indicating the possible involvement of CCR5 in regulation of the host immune response. Here, we found that Mycobacterium infection significantly increased CCR5 expression in macrophages there by facilitating the activation of its downstream signaling. These events culminated in up-regulation of the immunosuppressive cytokine IL-10 production, which was further associated with the down-regulation of macrophage MHC-II expression along with the up-regulation of CCR5 expression via engagement of STAT-3 in a positive feedback loop. Treatment of macrophages with CCR5 specific siRNA abrogated the IL-10 production and restored MHCII expression. While, in vivo CCR5 silencing was also effective for the restoration of host immune responses against tuberculosis. This study demonstrated that CCR5 played a very critical role for the immune subversion mechanism employed by the pathogen.

Design, synthesis and docking studies of new quinoline-3-carbohydrazide derivatives as antitubercular agents.

Three new series of 4-hydroxy-8-trifluoromethyl-quinoline derivatives were synthesized through multi step reactions. All the newly synthesized compounds were characterized by spectral and elemental analyses. The structure of 5j was evidenced by X-ray crystallographic study. The newly synthesized title compounds were evaluated for their antimicrobial activities including antimycobacterial activity. Amongst the tested compounds, 5b, 5e, 5h, 5j, 6c and 7c displayed promising antimicrobial activity. The mode of action of these active compounds was carried out by docking of receptor enoyl-ACP reductase with newly synthesized candidate ligands, 5b, 5e, 5h, 5j and 6c.

New quinolin-4-yl-1,2,3-triazoles carrying amides, sulphonamides and amidopiperazines as potential antitubercular agents.

Three new series of quinoline-4-yl-1,2,3-triazoles carrying amides, sulphonamides and amidopiperazines were synthesized through multi-step reactions. The required intermediate, [1-(6-methoxy-2-methylquinolin-4-yl)-1H-1,2,3-triazol-4-yl]methanol (2) was prepared by treating 4-azido-6-methoxy-2-methylquinoline (1) with propargyl alcohol. Three different series of compounds were synthesized from this intermediate. All the newly synthesized compounds were characterized by spectral and elemental analyses. The structure of 2 was confirmed by X-ray crystallographic study. Further, the title compounds were evaluated for their in vitro anti-bacterial activity against five different bacterial strains and antimycobacterial activity against Mycobacterium tuberculosis H37Rv, Mycobacterium smegmatis (ATCC 19420) and Mycobacterium fortuitum (ATCC 19542). Title compounds, 6a, 6d, 6i, 6j, 7e, 10a and 10i were found to be active against Mycobacterium tuberculosis H37Rv strain and could be lead molecules of interest.

New quinoline derivatives: synthesis and investigation of antibacterial and antituberculosis properties.

Four new series of quinoline derivatives were synthesized starting from 2-trifluoromethyl aniline through multi-step reactions. In the reaction sequence, substituted aniline was cyclized to 4-hydroxy quinoline 1, which was then transformed to 4-chloro-2,8-bis(trifluoromethyl)quinoline 2. The key scaffold 4-hydrazinyl-2,8-bis(trifluoromethyl)quinoline 3, obtained from the compound 2, was successfully converted to target quinoline derivatives, viz. hydrazones 4a-t, ureas 5a-e, thioureas 6a-c and pyrazoles 7a-d, in good yields. The newly synthesized title compounds were evaluated for their in vitro antibacterial activity against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Klebsiella pneumoniae (recultured) and antituberculosis activity against Mycobacterium tuberculosis H(37)Rv and MDR-TB. Preliminary results indicated that most of the hydrazone derivatives demonstrated very good antibacterial and antituberculosis activities while other derivatives showed moderate activity.

Design and synthesis of some new quinoline-3-carbohydrazone derivatives as potential antimycobacterial agents.

A series of 26 new quinoline derivatives carrying active pharmacophores has been synthesized and evaluated for their in vitro antituberculosis activity against Mycobacterium tuberculosis H37Rv (MTB), Mycobacterium smegmatis (MC(2)), and Mycobacterium fortuitum following the broth micro dilution assay method. Compounds 13e, 13i, 13k, 14a, 14c, 14i, and 14k exhibited significant minimum inhibition concentrations, when compared with first line drugs isoniazid (INH) and rifampicin (RIF) and could be ideally suited for further modifications to obtain more efficacious compounds in the fight against multi-drug resistant tuberculosis.

A randomized controlled trial to evaluate the efficacy of lactobacillus GG in infantile diarrhea.

In a randomized, double-blind, placebo-controlled trial, 229 infants hospitalized for acute diarrhea in rural India were given a 10-day course of Lactobacillus rhammosus GG (minimum dose, 10 degrees bacteria) or placebo. There was no difference in groups in the duration of diarrhea or numbers of stool on days 3, 6, or 10 of treatment.

Antibacterial activity of ciprofloxacin and trimethoprim, alone and in combinittion, against Vibrio cholerae O1 biotype El Tor serotype Ogawa isolates.

In this communication, the ciprofloxacin-trimethoprim (Cp-Tm) combination showed synergistic (Fractional Inhibitory Concentration, FIC index 0.399) and additive (FIC index 0.665-0.83) effects against Vibrio cholerae O1 biotype El Tor serotype Ogawa isolates having Cp MICs 10 microg/ml and Cp 0.66 microg/ml, respectively, following agar dilution checkerboard method. The time-kill study results demonstrated synergy between Cp and Tm against both groups of isolates providing 2.04 log10 (for strain with Cp MIC 0.66 microg/ml) and 3.12 log10 (for strain with Cp MIC 10 microg/ml) decreases in CFU/ml between the combination and its most active compound. Thus, the findings of the present study suggest an introduction of Cp-Tm combination treatment regimen against drug resistant cholera and this in turn will help in combating the drug resistance of V. cholerae O1 biotype El Tor serotype Ogawa.

Arabinosylated lipoarabinomannan modulates the impaired cell mediated immune response in Mycobacterium tuberculosis H37Rv infected C57BL/6 mice.

Mycobacterium tuberculosis is a facultative intracellular pathogen that flourishes inside the host macrophages. This organism has the ability to deactivate the cell-mediated immune responses involving the down-regulation of pro-inflammatory cytokines, T cell proliferation, apoptosis of CD4+T cells and impairment of the expression of MHC Class II molecules. We observed that Arabinosylated Lipoarabinomannan (Ara-LAM), a glycolipid present in the cell wall of the avirulent Mycobacterium smegmatis, could effectively restrict the growth of tubercle bacilli, induced the transcription of Th1 cytokines in alveolar macrophages (AMs) and splenocytes, enhanced the frequency of CD4+T cells secreting IFN-gamma and induced the expression of MHC Class II molecules on the splenocyte membrane, compared to that of Mycobacterium tuberculosis H37Rv infected C57BL/6 mice. Collectively our findings strongly suggest that Ara-LAM had the potency to restore the impaired cell mediated immune responses in mice infected with Mycobacterium tuberculosis H37Rv, and hence could be utilized as an effective immuno-prophylactic tool in the control of tuberculosis.