Spectrophotometric determination of rifampicin S in the presence of rifampicin O]

The limits of the use of the methods of direct photometric and differential spectrophotometric determination of rifamycin S containing rifamycin O as an admixture were discussed. The method of direct photometry did not provide determination of rifamycin S in the presence of rifamycin O since instability of the latter under the analysis conditions. A possibility of using the differential-spectrophotometric method for determination of rifamycin S in the presence of not more than 5 per cent of rifamycin O was shown.     

Effect of rifampicin and doxycycline on the chemiluminescence of leukocytes]

The effect of rifampicin and doxycycline on spontaneous and zymosan-induced chemiluminescence of polymorphonuclear leukocytes was studied on guinea pigs. The cells were incubated in the presence of the antibiotics, washed and stimulated by zymosan. Under such conditions rifampicin in therapeutic doses of 0.1 to 10 micrograms/kg and doxycycline in a dose of 100 micrograms/kg potentiated the leukocyte chemiluminescence. Investigation of the antibiotics effect on the cells without washing failed because of the direct interference of rifampicin and doxycycline with the cell-independent stage of the chemiluminescent reaction.     

Pharmacokinetic study of rifampicin in the body of pregnant animals]

The study on distribution of 14C-rifampicin administered intramuscularly to pregnent animals showed that its concentrations in the blood, liver, kidneys, lungs and other organs did not practically change from those in nonpregnant animals. The concentration of 14C-rifampicin in the fetus organs was much lower than that in the organs of the adult animals. The liver and kidneys of the pregnant animals, as well as the fetus though to a less extent had a capacity for metabolism of 14C-rifampicin. The following products of biotransformation were detected: N-oxide of rifampicin, 25-deacetylrifampicin, 3-formylrifamycin SV and rifamycin SV.     

Characteristics of pharmacokinetics of liposome-incorporated rifampicin in rats after intratracheal administration]

Pharmacokinetics of rifampicin after its single intratracheal administration in the form of the liposome-encapsulated drug and its aqueous solution was studied on rats. It was shown that after the exposure to the liposome-incorporated rifampicin (10 mg/kg) the concentration-time curve in the blood and lungs was sigmoid with the retarded decrease in the blood drug concentration within 9 hours. The plateau segment of the curve provided at least a 4-fold longer maintenance of the rifampicin concentration in the blood and lungs at 3 to 4 micrograms/ml. The use of the liposome-incorporated antibiotic induced 2- and 1.5-fold increases in the AUC in regard to the lungs and blood, respectively.     

Effects of rifampicin and doxycycline on the production of hydrogen peroxide by macrophages]

The influence of rifampicin and doxycycline on oxidative metabolism of macrophages was estimated in vitro by production of hydrogen peroxide. It was shown that low concentrations of rifampicin and doxycycline stimulated production of hydrogen peroxide by macrophages of guinea pigs. In concentrations of 1 to 10 micrograms/ml corresponding to the mean therapeutic ones doxycycline increased both the spontaneous and zymosan-induced production of hydrogen peroxide by the macrophages. The potentiating activity of doxycycline on the cells activated by opsonized zymosan was higher. The maximum increase in the induced production of hydrogen peroxide (by 40 per cent) was observed when the antibiotic concentration was 1 microgram/ml. Rifampicin in concentrations of 0.1 to 1 microgram/ml corresponding to the mean therapeutic ones stimulated the zymosan-induced production of hydrogen peroxide by the macrophages. The maximum increase in the production of hydrogen peroxide (by 22 per cent) was noted at the rifampicin concentration of 1 microgram/ml.     

Immunomodulating, antioxidant and hepatoprotective effects of immobilized dosage forms of rifampicin and cephalexin]

The aim of the study was to lower the immunosuppressory, prooxidant and hepatotoxic effects of rifampicin and cephalexin by their immobilization in erythrocyte vehicles. The experiments were performed on Wistar rats with the use of rifampicin, cephalexin and lysozyme (ZAO Ferane) and hemodes (6% aqueous saline solution of low molecular polyvinylpyrrolidone, mol. wt. 12600+/-2700). Rifampicin- and cephalexin-entrapped erythrocytes were prepared. Spectrophotometric procedures for quantitative assay of the immobilized antibiotics were developed. The impact of the solution concentration and incubation time on the level of the antibiotic entrapping was studied. The erythrocyte vehicles were shown to be able to entrap the antibiotics for 9 days and to preserve their stability for 24 hours. It was observed that the increase of the immunosuppressory, prooxidant and hepatotoxic effects of the antibiotics administered without the vehicles to the laboratory animals infected by staphylococci was dose-dependent. The use of the antibiotics entrapped in the erythrocyte vehicles stimulated the immune reactivity of the animals and normalized the indices of lipid peroxidation, the antioxidant system, cytolysis and cholestasis.     

Evaluation of the sanative action of rifampicin on the meningococcal carrier state]

The results of the epidemiological control experiment on the efficacy of rifampicin in sanation of meningococci carriers are presented. The preliminary study of rifampicin sensitivity of 41 freshly isolated nasopharyngeal meningococcal strains showed that the MIC of the drug for 63 per cent of the isolates was 0.04--0.1 gamma/ml. Sanation was performed for 2 days; 1.2 g of the drug was used during the treatment course. The results of examination of 91 meningococci carriers showed that 4 days after the sanation the specific weight of the persons isolating no meningococci was reliably higher in the experimental group than that in the control group. The coefficient of rifampicin efficiency was 70.8 per cent. 10 days after sanation the difference in the level of the carriers isolating no meningococci in the experimental and the control groups was statistically insignificant. Therefore, the carriers treated with the drug received temporary protection from the causative agent at an average for 1 week. Later on they could become carriers again. As a result of sanation no changes in the meningococcal sensitivity to rifampicin was observed.     

Multi-factor analysis of combined effects of rifampicin and peptidoglycan of microbial origin on immune response]

Multifactorial analysis of the combined action of rifampicin and a microbial peptidoglycan on the immune response to antigens of the vaccine EV fraction 1 was performed. The results were computer-processed and the second order equations describing delayed hypersensitivity (DH) and antibody titers were derived. Nomographs or equal level curves showing interrelations of the investigated factors were plotted. The character of the combined action on DH and antibody titers was heterogeneous. The peptidoglycan had a pronounced immunostimulant action on DH and, to a lesser extent, influenced the humoral response. Conditions for the peptidoglycan use aimed at immunostimulation were optimized with application of multifactorial analysis.     

Increasing the effectiveness of immunotherapy of rabies by using rifampicin in experimental studies]

The experiments showed that in a dose of 35-70 mg/kg rifampicin inhibited reproduction of the fixed rabies virus in the brain of infected animals. The drug had no inhibitory effect on synthesis of the virus-neutralizing antibodies after vaccination. Combination of rifampicin with antirabies gamma-globulin had a marked synergistic effect. The animal survival after the combination use amounted to 75-100 per cent and depended on the infective dose of the virus and the scheme of the drug administration. It was concluded that rifampicin might be used in complex therapy of rabies during the incubation period (along with gamma-globulin and the vaccine) for inhibiting virus reproduction at early infection stages.     

Antibacterial activity of rifampicin against the causative agents of suppurative, septic diseases]

The antibacterial activity of rifampicin was studied in comparison with other antibiotics with respect to clinical strains isolated from cases with various purulent inflammatory processes caused by Staphylococcus, E. coli, Ps. aeruginose, Proteus. The aim of the study was to define the role of rifampicin in the treatment of the above infections. No rifampicin resistant strains were found among staphylococci belonging to the phenotype carrying the determinants of resistance to 2-8 antibiotics. Rifampicin was less active against gramnegative organisms. High heterogeneity of the microbial population of rifampicin was shown with respect to all microbial strains tested. The rate of the spontaneous mutants was high. The average rate of the mutants was 1-7.7-10-8. The studies on the dynamics of the rifampicin resistance increase in the strains of Staphylococci, E. Coli, Ps. aeruginosa and Proteus showed that the resistance increased after 1-2 passages, which means that one-stage mutation was characteristic rifampicin.     

Selective inhibition of ribosomal RNA synthesis by rifampicin in E. coli cells]

Under partial inhibition of total RNA synthesis by rifampicin the formation of beta- and beta'-subunits of RNA polymerase is stimulated and the rRNA synthesis is selectively repressed. The differential rate of synthesis of the beta- and beta'-subunits increases from 1,15% up to 2,88% in the presence of 30 micrograms rifampicin per ml. Simultaneously the differential rate of rRNA synthesis decreases from 41% down to 10%. The degree of inhibition of rRNA synthesis by rifampicin depends on the cell growth rate.     

Effect of rifampicin on the synthesis of antibodies to fraction I of vaccine EV]

Multifactorial analysis was used to study the influence of rifampicin on the dynamics of synthesis of antibodies belonging to IgM and IgG classes in mice immunized by fraction I of the vaccinal strain EV. Equations and quadric surfaces describing individual dynamics of antibody formation within a wide range of antibiotic doses and time of antibody content determination were developed by the experimental data. It was shown that within the range of the doses corresponding to the therapeutic ones in man rifampicin stimulated antibody formation. It had an inhibitory effect on antibody genesis only when used in the doses many times higher than the therapeutic ones.     

基于PARMS技术的结核分枝杆菌利福平耐药基因rpoB分子标记的开发

文中旨在建立结核分枝杆菌利福平耐药基因rpoB的荧光分子标记,为分子药物敏感性试验提供简便、可靠的基因分型检测方法。对比分析利福平耐药菌株rpoB基因531、526、516、511、513等氨基酸位点的基因突变与敏感菌株中等位基因的序列差异,结合PARMS技术 (Penta-primer amplification refractory mutation system),建立rpoB基因的荧光分子标记。利用其对104例结核分枝杆菌临床分离株进行检测,经Sanger测序验证,正确率100%。并采用比例法药敏试验对这104份样本进行了利福平耐药性鉴定,与分子标记结果相符率为94.23%。结果表明,分子标记有较强可靠性,能检出表型药敏无法测出的低浓度耐药样本 (511/533单位点突变)。建立的11组荧光分子标记能覆盖92%–96%的利福平耐药菌株rpoB基因突变类型,为快速检测结核分枝杆菌利福平耐药提供新思路。     

Development of highly organized lymphoid structures in Buruli ulcer lesions after treatment with rifampicin and streptomycin

Background

Buruli ulcer caused by Mycobacterium ulcerans is an infection of the subcutaneous tissue leading to chronic necrotising skin ulcers. The pathogenesis is associated with the cytocidal and immunosuppressive activities of a macrolide toxin. Histopathological hallmark of progressing disease is a poor inflammatory response despite of clusters of extracellular bacilli. While traditionally wide excision of the infected tissue was the standard treatment, provisional WHO guidelines now recommend an eight week pre-treatment with streptomycin and rifampicin.

Methodology/Principal Findings

We conducted a detailed immunohistochemical analysis of tissue samples from Buruli patients who received antibiotic treatment. Cellular immune response along with bacterial load and distribution were monitored. We demonstrate that this treatment leads to the development of highly organized cellular infiltration surrounding areas of coagulative necrosis. Diffuse infiltrates, granulomas and dense lymphocyte aggregation close to vessels were observed. Mycobacterial material was primarily located inside mononuclear phagocytes and microcolonies consisting of extracellular rod-shaped mycobacteria were no longer found. In observational studies some patients showed no clinical response to antibiotic treatment. Corresponding to that, one of five lesions analysed presented with huge clusters of rod-shaped bacilli but no signs of infiltration.

Conclusions/Significance

Results signify that eight weeks of antibiotic treatment reverses local immunosuppression and leads to an active inflammatory process in different compartments of the skin. Structured leukocyte infiltrates with unique signatures indicative for healing processes developed at the margins of the lesions. It remains to be analysed whether antibiotic resistance of certain strains of M. ulcerans, lacking patient compliance or poor drug quality are responsible for the absent clinical responses in some patients. In future, analysis of local immune responses could serve as a suitable surrogate marker for the efficacy of alternative treatment strategies.     

Enhanced activity of rifampicin loaded with polybutyl cyanoacrylate nanoparticles in relation to intracellularly localized bacteria]

Association of rifampicin with polybutylcyanoacrylate nanoparticles provided considerable enhancement of drug antibacterial activity. In vitro nanoparticle-loaded rifampicin was more active against Staphylococcus aureus and Mycobacterium avium, localized in isolated alveolar macrophages. Level of rifampicin in macrophages increased 2-3-fold after incubation with rifampicinloaded nanoparticles comparing to the free drug. High therapeutic efficacy of colloidal rifampicin was demonstrated in vivo. Use of nanoparticles provided 2-fold increase in rifampicin efficacy, comparing with the free drug at treatment of staphylococcus sepsis in mice. Single administration of nanoparticulate rifampicin in the dose 25 mg/kg resulted in 80% survival of mice with salmonellosis, while 50 mg/kg of free rifampicin could provide only 10% survival. It may be considered that high antibacterial efficacy of rifampicin bound to nanoparticles is due to its effective delivery to macrophages.