The effects of antibiotics combined with natural polyphenols against clinical methicillin-resistant Staphylococcus aureus (MRSA)

Novel therapies are needed to address the public health problem posed by methicillin-resistant STAPHYLOCOCCUS AUREUS (MRSA). In this study, we determined the effects of combinations of antibiotics and plant polyphenols against 20 clinical isolates of MRSA. The IN VITRO activities of 10 antibiotics and 15 natural polyphenols against the isolates were evaluated by determining minimum inhibitory concentrations (MICs). All isolates were susceptible to vancomycin and resistant to rifampicin, while susceptibilities to ciprofloxacin varied. Among the 15 natural polyphenols, kaempferol (3,4',5,7-tetrahydroxyflavone) and quercetin (3,3',4',5,7-pentahydroxyflavone) showed the lowest MICs. In checkerboard assays, combinations of rifampicin and either kaempferol or quercetin acted synergistically or partially synergistically against the clinical MRSA isolates. Rifampicin combined with kaempferol or quercetin exhibited good beta-lactamase inhibitory effects (57.8 % and 75.8 %, respectively) against a representative isolate according to nitrocefin analysis. The study results and ready availability and low toxicity of plant polyphenols warrant further investigations on the therapeutic potential of combination therapies for MRSA infections.     

Epileptogenic activity of methicillin-resistant Staphylococcus aureus (MRSA) antibiotics in rats

The present study was undertaken to clarify the epileptogenic activity induced by intracerebroventricular injection (i.c.v.) of antibiotics effective in methicillin-resistant Staphylococcus aureus (MRSA) in chronically electrode implanted rats. Teicoplanin (10-100 microg, i.c.v.) caused dose-related electroencephalographic (EEG) seizure characterized by an uninterrupted high voltage and wave complex. At the same time, the rats showed forelimb clonus, head nodding, jumping and severe convulsion. At a high dose (100 microg, i.c.v.), the drug caused a severe twisting immediately after the intracerebroventricular injection (i.c.v.) followed by jumping and violent convulsion with a continuous rhythmic spike and wave complex in EEG. On the other hand, vancomycin (30-1000 microg, i.c.v.) caused no or almost no epileptogenic activity in terms of behavior and in EEG. However, at a high dose (1000 microg, i.c.v.), the drug caused an occasional spike from the hippocampus without showing any behavioral changes in the rats. Fosfomycin (30-1000 microg, i.c.v.), cefazolin (10-100 microg, i.c.v.) and penicillin G (30-300 microg, i.c.v.), used as reference drugs, caused dose-dependent epileptogenic activity in both EEG. From these findings, it was found that teicoplanin caused a potent epileptogenic activity, different to vancomycin. Therefore, it can be concluded that vancomycin may be safety on epileptogenic activity used for the clinical purpose of infections caused by MRSA.     

In vitro combination effect of pazufloxacin with various antibiotics against Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus

The in vitro combination effects of pazufloxacin (PZFX) with various antibiotics were investigated by the checkerboard dilution method using piperacillin (PIPC), tazobactam/piperacillin (TAZ/PIPC), ceftazidime (CAZ), cefozoprane (CZOP), imipenem/cilastatin (IPM/CS), meropenem (MEPM), panipenem/betamipron (PAPM/BP), amikacin (AMK) and isepamicin (ISP) for clinical isolates of 27 Pseudomonas aeruginosa strains, vancomycin (VCM), teicoplanin (TEIC) and arbekacin (ABK) for clinical isolates of methicillin-resistant 26 Staphylococcus aureus (MRSA) strains, respectively. The following results were obtained. 1. For 27 P. aeruginosa strains, the synergistic effects were observed with the combination of PZFX and CAZ or MEPM (11.1%: 3 strains), and PZFX and CZOP or PAPM/BP (3.7%: 1 strain), respectively. The additive and synergistic effects of PZFX were observed with the combination in all beta-lactams tested in the strains more than 50%. No antagonistic effect was observed. The additive effects were also observed with the combination of PZFX and AMK or ISP in the strains more than 50% of the test strains and no antagonistic effect was observed. 2. For 26 MRSA strains, no antagonistic effect was observed with the combination of all antibiotics tested. The indifference was observed with the combination of PZFX and VCM or ABK in the strains more than 60%, and the additive effects were observed with the combination of TEIC in the strains more than 80%. In conclusion, no antagonistic effect was observed in PZFX with the combination of beta-lactams and anti-MRSA agents, suggesting that the combination therapy of PZFX with these antibiotics would be possible to use for the infections caused by P. aeruginosa and MRSA.     

Synergistic effect of rosmarinic acid with antibiotics against Staphylococcus aureus and MRSA

OBJECTIVE To evaluate the synergistic effect of rosmarinic acid(RA)with antibiotics against Staphylococcus aureus(S.aureus)and MRSA and to identify the possible mechanism of action responsible for synergism if any.METHODS The antibacterial activity of Rosmarinic acid was studied for its zone of inhibition by agar well diffusion and MIC determination by liquid broth dilution technique against MRSA and VRSA.The synergistic effect of RA with antibiotics like amoxicillin,ofloxacin and vancomycin was evaluated by Broth checker board method and further confirmed with time kill kinetic studies.Microbial Surface Components Recognizing Adhesive Matrix Molecules(MSCRAMMs)were isolated by protein precipitation technique and its expression was studied by SDS PAGE.Further RA was evaluated for its beta lactamase inhibition property.RESULTS Rosmarinic acid exhibited antibacterial activity against S.aureus and MRSAby showing a MIC value of 0.8mg·mL-1 against S.aureus and 10mg·mL-1 against MRSA.Rosmarinic acid at 1/4X MIC value reduced the MIC of vancomycin,amoxicillin and ofloxacin by 1/4 times against S.aureus.But against MRSA,vancomycin was found to be synergistic.All the synergistic combinations have shown FIC value of 0.5.In order to measure the kinetics of the anti-bacterial activity,the bacterial growth rate with RA,antibiotics and synergistic combinations against S.aureus and MRSA was studied.It is observed that the synergistic combinations showed better time kill kinetics as compared to RA and antibiotics.Further RA could able to destruct the cell surface proteins MSCRAMMs which was studied by SDS PAGE.RA was also found to showβlactamase inhibiting property.CONCLUSION It is concluded that RA possess antibacterial activity but at a very higher concentration(in mg/mL)against S.aureus and MRSA.However it shows synergism with antibiotics and could able to reduce the MIC of antibiotics.Thus RA could be developed as an adjuvant for antibiotics against S.aureus and MRSA caused infections.Further studies are needed to identify the mechanism for its synergism with antibiotics.     

Baicalin synergy with beta-lactam antibiotics against methicillin-resistant Staphylococcus aureus and other beta-lactam-resistant strains of S. aureus

Bacterial resistance to antibiotics is a serious global problem and includes strains of beta-lactam-resistant Staphylococcus aureus and methicillin-resistant S. aureus (MRSA). Novel antimicrobials and/or new approaches to combat the problem are urgently needed. The Chinese herb Xi-nan Huangqin (Scutellaria amoena C.H. Wright) has been used in traditional Chinese medicine to treat a wide range of infectious diseases. In this study we have examined the antibacterial action of baicalin, a flavone isolated from the herb. When combined with 16 microg mL(-1) baicalin, minimum inhibitory concentrations (MICs) of benzylpenicillin against MRSA and penicillin-resistant S. aureus were reduced from 125 and 250 microg mL(-1) to 4 and 16 microg mL(-1), respectively. This activity of baicalin was dose-dependent. Viable counts showed that the killing of MRSA and beta-lactam-resistant S. aureus cells by 10 to 50 microg mL(-1) ampicillin, amoxycillin, benzylpenicillin, methicillin and cefotaxime was potentiated by 25 microg mL(-1) baicalin. From the study it was concluded that baicalin has the potential to restore the effectiveness of beta-lactam antibiotics against MRSA and other strains of beta-lactam-resistant S. aureus. In view of its limited toxicity baicalin offers potential for the development of a valuable adjunct to beta-lactam treatments against otherwise resistant strains of microorganisms.     

Synergistic effect of tetrandrine and ethidium bromide against methicillin-resistant Staphylococcus aureus (MRSA)

Methicillin-resistant Staphylococcus aureus (MRSA) along with other resistant bacteria have become a significant social and clinical problem. Therefore, there is an urgent need to develop bioactive compounds from natural products as alternatives to the very few antibiotics that remain effective. Recently, the efflux mechanism has been identified as the main contributor to antibiotic resistance in bacteria. This study therefore aimed to evaluate tetrandrine (TET), an efflux pump inhibitor (EPI), as a potential antibiotic against MRSA. We investigated the antimicrobial activity of TET against 17 MRSA strains, of which 3 selected strains were studied in further detail using a time-kill assay. When these bacterial strains (1 × 10(6) colony-forming units (cfu)/ml) were incubated with TET in a time-kill assay, log-scale bactericidal activity was observed, which lasted for 24 hr. In addition, TET exhibits a synergistic effect when combined with the multi-drug resistance (MDR)-efflux pump substrate ethidium bromide (EtBr). Structure-function studies of the antibiotic activity of TET in combination with EtBr may lead to the discovery of more effective efflux pump inhibitors.     

Antibacterial activities of various aminoglycosides against clinically isolated methicillin-resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) were isolated from samples collected from various patients during 1986, and antibacterial activities of 6 aminoglycosides (AGs) (netilmicin (NTL), gentamicin (GM), sisomicin (SISO), dibekacin (DKB), tobramycin (TOB) and amikacin (AMK] and 4 beta-lactam antibiotics (cefazolin (CEZ), cefmetazole (CMZ), cloxacillin (MCIPC) and methicillin (DMPPC) against these MRSA were evaluated. Among these 6 AGs, NTL was the most potent, and its MIC50 and MIC80 were 1.56 and 3.13 micrograms/ml, respectively. Antibacterial activities of GM, SISO, DKB and TOB were weak, and MIC50's of GM and DKB were both 100 micrograms/ml, while those of SISO and TOB were 50 and greater than 100 micrograms/ml, respectively. Frequency of highly resistant specimens to AMK was rather low and its MIC50 and MIC80 were 12.5 and 25 micrograms/ml, respectively. As for antibacterial activities of the above 4 beta-lactam antibiotics, the MIC50 and MIC80 of CMZ were 6.25 and 12.5 micrograms/ml, respectively, and therefore, its antibacterial activity to MRSA is relatively good. However, MIC50's of CEZ, MCIPC and DMPPC were all greater than 100 micrograms/ml, showing poor antibacterial activities. Recently, MRSA became a problem in various fields of clinical practice, and a number of literatures reporting refractory infections caused by MRSA have been published. Since MRSA is featured as multiply resistant bacteria, it is known that MRSA is resistant to the majority of existing antibiotics (penicillins, cephems, macrolides, AGs, etc.). In 1985, we reported results of our study concerning the antibacterial activities of a number of CEPs and some of AGs against multiply resistant S. aureus including MRSA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Surface-engineered liposomes for dual-drug delivery targeting strategy against methicillin-resistant Staphylococcus aureus (MRSA)

This study focused on the encapsulation of vancomycin(VAN) into liposomes coated with a red blood cell membrane with a targeting ligand, daptomycin–polyethylene glycol–1,2-distearoyl-sn-glycero-3-phosphoethanolamine, formed by conjugation of DAPT and Nhydroxysuccinimidyl-polyethylene glycol-1,2-distearoyl-sn-glycero-3-phosphoethanolamine.This formulation is capable of providing controlled and targeted drug delivery to the bacterial cytoplasm. We performed MALDI-TOF, NMR and FTIR analyses to conf...     

Phytochemical constituents from Cassia alata with inhibition against methicillin-resistant Staphylococcus aureus (MRSA)

The methanolic extract of the leaves of CASSIA ALATA was sequentially partitioned in increasing polarity to afford the hexane, chloroform, butanol and residual extract. Crude extracts were evaluated against MRSA using the agar well diffusion assay. The butanol and chloroform extracts both exhibited inhibition against MRSA with inhibition indexes of 1.03 +/- 0.16 and 0.78 +/- 0.07 at the concentration of 50 mg/mL. The butanol extracts were further purified using silica gel and reverse phase chromatography to afford kaempferol ( 1), kaempferol 3- O-beta-glucopyranoside ( 2), kaempferol 3- O-gentiobioside ( 3) and aloe emodin ( 4). The four constituents showed varying degrees of inhibition against MRSA. Both 1 and 4 exhibited MIC (50) values of 13.0 +/- 1.5 microg/mL and 12.0 +/- 1.5 microg/mL, respectively. The kaempferol glycosides 2 and 3 were less active with MIC (50) values of 83.0 +/- 0.9 microg/mL and 560.0 +/- 1.2 microg/mL, respectively. A free hydroxyl group at C-3 of the flavonol structure is a structural requirement for the inhibition of MRSA.     

Novel two-chain fatty acid-based lipids for development of vancomycin pH-responsive liposomes against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA)

Abstract

The development of bacterial resistance against antibiotics is attributed to poor localisation of lethal antibiotic dose at the infection site. This study reports on the synthesis and use of novel two-chain fatty acid-based lipids (FAL) containing amino acid head groups in the formulation of pH-responsive liposomes for the targeted delivery of vancomycin (VAN). The formulated liposomes were characterised for their size, polydispersity index (PDI), surface charge and morphology. The drug-loading capacity, drug release, cell viability, and in vitro and in vivo efficacy of the formulations were investigated. A sustained VAN release profile was observed and in vitro antibacterial studies against S. aureus and MRSA showed superior and prolonged activity over 72?h at both pH 7.4 and 6.0. Enhanced antibacterial activity at pH 6.0 was observed for the DOAPA-VAN-Lipo and DLAPA-VAN-Lipo formulations. Flow cytometry studies indicated a high killing rate of MRSA cells using DOAPA-VN-Lipo (71.98%) and DLAPA-VN-Lipo (73.32%). In vivo studies showed reduced MRSA recovered from mice treated with formulations by four- and two-folds lower than bare VN treated mice, respectively. The targeted delivery of VAN can be improved by novel pH-responsive liposomes from the two-chain (FAL) designed in this study.     

Infective endocarditis caused by USA300 methicillin-resistant Staphylococcus aureus (MRSA)

We report seven cases of infective endocarditis caused by USA300 methicillin-resistant Staphylococcus aureus (MRSA) at an urban, tertiary care, academic institution. Five strains were community associated and two were healthcare associated. All patients were injection drug users. Staphylococcus aureus isolates were characterised as USA300-type MRSA using pulsed-field gel electrophoresis. Five cases were right-sided endocarditis and two cases were left-sided. The mean length of in-hospital antimicrobial therapy was 23 days and the mean length of total antibiotic therapy was 55 days. Complications included heart failure resulting in valve replacement in one patient as well as death in that patient. As USA300 strains of MRSA continue to increase in prevalence, clinicians must be aware of the increasing spectrum of illness in considering management and prevention strategies.     

Efficacy of newly generated short antimicrobial cationic lipopeptides against methicillin-resistant Staphylococcus aureus (MRSA)

IntroductionInfection caused by methicillin-resistant Staphylococcus aureus (S. aureus) (MRSA) is a serious clinical challenge and research to develop new antimicrobials is imperative.MethodsThis study investigated the in vitro and in vivo efficacy of the short cationic dialkyl lipopeptides (C₁₀)₂-KKKK-NH₂ and (C₁₂)₂-KKKK-NH₂. The antibacterial efficacy of (C₁₀)₂-KKKK-NH₂ and (C₁₂)₂-KKKK-NH₂ was evaluated in representative clinical methicillin-susceptible S. aureus and MRSA strains by both in vitro (MIC, time-kill curve) and in vivo (wax worms model) approaches.ResultsThese studies revealed that both (C₁₀)₂-KKKK-NH₂ and (C₁₂)₂-KKKK-NH₂ have rapid bactericidal activity, with a decrease of > 3 log₁₀ colony forming units (CFU)/mL achieved in the first 6 hours of treatment. Furthermore, (C₁₀)₂-KKKK-NH₂ performed similarly to daptomycin, with a sustained bacterial killing after 24 hours. Wax worms infected and treated with these lipopeptides showed a decreased survival rate of 90% to 50% within the first day of treatment. Scanning electron microscopy determined that the effect of the short lipopeptides in S. aureus was associated with important morphological structural changes that may suggest cell membrane perturbation.ConclusionThese findings suggest that the short lipopeptides (C₁₀)₂-KKKK-NH₂ and (C₁₂)₂-KKKK-NH₂ may be potential new options for treating MRSA infections.     

Exploration of the activity of 7-pyrrolidino-8-methoxyisothiazoloquinolones against methicillin-resistant Staphylococcus aureus (MRSA)

A series of 7-(3'-substituted)pyrrolidino-8-methoxyisothiazoloquinolone (ITQ) analogues were prepared, and their antibacterial potency against methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), and Escherichia coli were compared. Many of these analogues had MIC ≤ 0.25 μg/mL against quinolone-resistant MRSA strains. The stereochemical preference was explored for a series of 1'-methyl-3'-aminomethylpyrrolidine analogues. Antibacterial activity was generally more favorable with 3'-R, 1'-S configuration. Substitution on the 3'-aminomethyl nitrogen tended to decrease activity, while potency was maintained with disubstitution or aryl substitution at the 1'-carbon. The 7-[(R)-3-((S)-1-aminoethyl)pyrrolidin-1-yl] analogue (6a(R,S)) and the (R)-7-[3-(2-aminopropan-2-yl)pyrrolidin-1-yl] analogue (7a(R)) were found to be the ITQs with the most promising antibacterial profiles. The MICs of these select ITQs versus a panel of clinical MRSA strains were determined, and the ITQs were found to have 8- to 16-fold greater potency than linezolid. These analogues were also evaluated for inhibition of the target enzymes, topoisomerase IV and DNA gyrase, from both wild-type and multidrug resistant strains. The ITQs were up to >30 times more inhibitory against these targets than the fluoroquinolone moxifloxacin.     

Directed alteration of a novel bovine beta-defensin to improve antimicrobial efficacy against methicillin-resistant Staphylococcus aureus (MRSA)

beta-Defensin antimicrobial peptides are critical components of the innate immune response in many species and may be useful against pathogens that have acquired resistance to standard antibiotic therapies. We analysed a panel of recently discovered bovine beta-defensins in order to identify sites that may have particular functional importance against antibiotic-resistant bacteria. Modifications were introduced to increase charge at positively selected (PS) sites, to make charge-neutral changes at PS sites, to increase hydrophobicity and to confer a hydrophilic C-terminal. Whilst all four peptide modifications increased antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) compared with the native form of bovine beta-defensin 123 (P=0.02), conferring the hydrophilic tail caused the most significant increase, with a 50% lethal dose (LD(50)) of 3.91 microg/mL. The peptide with increased charge at PS sites showed the most significant increase in antimicrobial activity against a non-resistant strain of S. aureus (P=0.02). Therefore, informed modifications of the amino acid sequence can significantly affect the specificity and antimicrobial activity of a peptide.     

Synergistic effects of berberines with antibiotics on clinical multi-drug resistant isolates of methicillin-resistant Staphylococcus aureus (MRSA)

N-Methyl-dihydroberberine (M-Ber) was synthesized, and antibacterial activities of Berberine (Ber) and M-Ber alone and combined with antibiotics were studied against ten clinical MRSA isolates. MICs/MBCs (μg/ml, alone) ranges were 32–128/64–256 (Ber) and 64–128/256–1,024 (M-Ber) by a broth microdilution method. Significant synergies of Ber (M-Ber)/Azithromycin and Ber (M-Ber)/Levofloxacin combinations were observed by the chequerboard test. The Ber (M-Ber)/Ampicillin and Ber (M-Ber)/Cefazolin combinations showed indifference. These results demonstrated that Ber and M-Ber enhanced the in vitro inhibitory efficacy of Azithromycin and Levofloxacin, which had potential for combinatory therapy of patients infected with MRSA.     

Costs of hospital-acquired methicillin-resistant Staphylococcus aureus (MRSA) and its control

For most countries badly affected by methicillin-resistant Staphylococcus aureus (MRSA) there have been many years of debate about its relative virulence compared with methicillin-susceptible S. aureus (MSSA) and whether it could be controlled. Now that it is endemic in the majority of hospitals around the world, it is clear that it is at least as virulent as MSSA and is an additional burden of healthcare-acquired infection. There is increasing evidence that, despite this endemicity, control efforts can be successful, although they are often perceived as expensive. In reality, there is a large body of consistent evidence that control is highly cost effective, particularly in the context of the huge societal costs of MRSA and the future ever-greater threats that it poses.     

Antibacterial activity of hydroxychalcone against methicillin-resistant Staphylococcus aureus

Anti-candidal hydroxychalcone, 2,4,2′-trihydroxy-5′-methylchalcone (THMC), was investigated for its antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). THMC showed the minimum inhibitory concentrations of 25.0–50.0 μg/ml against tested 20 strains, at which the effect was based on a bacteriostatic action. THMC of 25.0 μg/ml completely inhibited the incorporation of radio-labelled thymidine and uridine into MRSA cells. In combination with antibiotics, the fractional inhibitory concentration indices were 0.47 for gentamicin and 0.79 for vancomycin, indicating that THMC acts synergistically with these agents. THMC would be a potent therapeutic agent for MRSA infections.     

Susceptibility and resistance genes to fluoroquinolones in methicillin-resistant Staphylococcus aureus isolated in 2002

The activity of six fluoroquinolones (FQs) was determined against 100 methicillin-resistant Staphylococcus aureus (MRSA) isolated in 2002 along with mutations in the grlA and gyrA genes and in the norA promoter of these isolates. Of the isolates tested, 97% had mutations in grlA and gyrA. A single mutation in grlA and gyrA resulted in a decrease of susceptibility to old generation FQs (norfloxacin, enoxacin, ciprofloxacin, fleroxacin, sparfloxacin and levofloxacin) but not to new generation FQs (gatifloxacin and moxifloxacin). Double mutations of both grlA and gyrA resulted in high-level resistance to all FQs tested. All norA mutants (15%) contained double mutations in grlA and gyrA and showed no decrease of MIC in the presence of reserpine, which is known to inhibit the drug-efflux pump. Our results showed that double mutations in grlA and gyrA were necessary for the expression of high-level resistance to new generation FQs. As different FQ-resistant mutants occur in the same PFGE type, FQ-resistant MRSA may well develop individually.     

Meningitis due to methicillin-resistant Staphylococcus aureus (MRSA): review of 10 cases

We evaluated retrospectively, 10 MRSA meningitis cases in our hospital that occurred between January 1999 and June 2004. All were post-neurosurgical and were considered to have hospital-acquired meningitis. Fever, leukocytosis, variable conscious levels were the most common findings. Six patients were treated with regimens including teicoplanin, and four with vancomycin. Mean duration of treatment was 23.5+/-18.8 days (range, 3-60 days). One patient died. In cases of MRSA meningitis, intravenous vancomycin is the mainstay of therapy. However, six of these 10 patients were successfully treated with regimens including teicoplanin, suggesting that this agent may be an alternative to vancomycin in the therapy of these cases.