RESUMO
Pneumonia is one of the most common infections caused by the bacterium Klebsiella pneumoniae. During the initiation of an infection, the immune system recognizes the pathogen through the release of high mobility group box 1 (HMGB1), thereby triggering the inflammation process. Miana has demonstrated potent inhibitory effects on the inflammatory process during infection in animal models. The aim of this study was to determine the effect of Miana leaf extract on mRNA HMGB1 expression in Balb/c mice infected with K. pneumoniae. Methods: This study comprised a cohort experiment using 20 Balb/c mice divided into four groups. Balb/c mice in each group were intraperitoneally injected with K. pneumoniae. Group 1 was given a placebo; Group 2 was given Miana; Group 3 was given levofloxacin; and Group 4 was given both levofloxacin and Miana. The levels of mRNA HMGB1 expression were measured using real-time PCR before, during, and after the infection as well as after the treatments. Results: The initial examination results showed that the average level of mRNA HMGB1 expression was 5.51 fc. The mRNA HMGB1 expression in mice after being challenged with K. pneumoniae was 9.64 fc. Group 1 that was given a placebo had a mean mRNA HMGB1 expression level of 14.99 fc. Group 2 that was given Miana had a mean mRNA HMGB1 expression level of 13.95 fc. Group 3 that was given levofloxacin had an average mRNA HMGB1 expression level of 6.45 fc, and Group 4 that was given levofloxacin and Miana together had an average mRNA HMGB1 expression level of 5.59 fc. Conclusion: Miana (Coleus scutellarioides (L.) Benth) increased mRNA HMGB1 expression at the initial administration via regulation of the immune system. Administration of Miana following K. pneumoniae infection inhibited the increase in mRNA HMGB1 expression. Treatment with levofloxacin reduced the level of mRNA HMGB1 expression, and the effect was optimized by the administration of Miana leaf extract as a supplement.
RESUMO
OBJECTIVES: This study aimed to determine molecular characteristics of rpoB, katG, rrs, and gyrA genes in Mycobacterium tuberculosis isolated from a cohort of Indonesian patients with tuberculosis. METHODS: Fifty isolates of M. tuberculosis were analysed by testing (DST) for susceptibility to first- and second-line drugs using the proportional method in a liquid medium. The genomic material was extracted to perform multiplex polymerase chain reaction (PCR) for identification and gene sequencing of rpoB, katG, rrs, and gyrA. RESULTS: Approximately 80% (40/50) of the rpoB mutations that were detected outside the hot-spot region (S450L, H445D, D435V, S441L, I491F, and Q432P) conferred rifampicin-resistance on M. tuberculosis. Approximately 11.42% (4/35) of isolates with S315T mutation in katG led to rifampicin-resistance instead of isoniazid-resistance. The mutation in katG gene was found at various locations (P280P, G279R, E340Q, T271I, E340*stop codon, R373G, and S315N). Streptomycin-resistance was detected in 42% (21/50) of the strains, but only two strains had rrs gene mutations (G878A and/or S514R). Approximately 14% (7/50) of M. tuberculosis isolates were kanamycin- and capreomycin-resistant but did not harbour mutations in the rrs gene, while 80% (40/50) of the strains had mutations in the quinolone-resistance determining region (QRDR) of the gyrA gene (S95T, D94V, A90V, and S91P) including the pan-susceptible strain. CONCLUSIONS: Of the 50 strains analysed, most of the mutations in the rpoB gene associated with rifampicin-resistance were also detected in the katG and gyrA genes. Molecular characterisation using DNA sequencing techniques is a highly sensitive approach for detecting mutations.
