Clinical efficacy of the combined use of levofloxacin and different courses of isoniazid and rifampicin in the treatment of mild spinal tuberculosis

Purpose: To investigate the clinical effectiveness of the combined use of levofloxacin and different courses of isoniazid and rifampicin in the treatment of mild spinal tuberculosis (TB). Methods: The clinic data of 100 patients with light spinal TB were retrospectively reviewed. A double-blind technique was used to divide the patients into 6-month treatment group (M6 group, n = 32), 12-month treatment group (M12 group, n = 34) and 18-month treatment group (M18 group, n = 34). All patients were given isoniazid and rifampicin, in combination with levofloxacin. The effects of the different treatment courses on mild spinal TB were determined. Results: There were significantly higher post-treatment levels of inflammatory factors in M6 group than in M12 and M18 groups (p < 0.001). Moreover, there were significantly higher Visual Analogue Scale (VAS) score and erythrocyte sedimentation rate (ESR), and larger focus size in M6 group than in M12 and M18 groups (p < 0.05). However, after treatment, M18 group had significantly higher total incidence of adverse reactions than M6 and M12 groups (p < 0.05). Conclusion: Compared with the short-course treatment, long-course treatment with isoniazid and rifampicin in combination with levofloxacin is more effective in reducing the levels of inflammatory factors and decreasing focus size in patients with mild spinal TB. However, patients given the 18-month treatment tend to develop more adverse reactions. Therefore, 12-month treatment with the combined therapy is a better therapeutic option

An atypical GdpP enzyme linking cyclic nucleotide metabolism to osmotic tolerance and gene regulation in Mycoplasma bovis

Nucleotide second messengers play an important role in bacterial adaptation to environmental changes. Recent evidence suggests that some of these regulatory molecular pathways were conserved upon the degenerative evolution of the wall-less mycoplasmas. We have recently reported the occurrence of a phosphodiesterase (PDE) in the ruminant pathogen Mycoplasma bovis, which was involved in c-di-AMP metabolism. In the present study, we demonstrate that the genome of this mycoplasma species encodes a PDE of the GdpP family with atypical DHH domains. Characterization of M. bovis GdpP (MbovGdpP) revealed a multifunctional PDE with unusual nanoRNase and single-stranded DNase activities. The alarmone ppGpp was found unable to inhibit c-di-NMP degradation by MbovGdpP but efficiently blocked its nanoRNase activity. Remarkably, MbovGdpP was found critical for the osmotic tolerance of M. bovis under K+ and Na+ conditions. Transcriptomic analyses further revealed the biological importance of MbovGdpP in tRNA biosynthesis, pyruvate metabolism, and several steps in genetic information processing. This study is an important step in understanding the role of PDE and nucleotide second messengers in the biology of a minimal bacterial pathogen

Thermal Decomposition Kinetics and Structure of Novel Polystyrene Clusters with MTEMPO as a Branching Agent

ABSTRACT: Polystyrene clusters were prepared by using a trace amount of 4-methacryloyloxy-2,2,6,6-tetramethylpiperidine-1-oxy (MTEMPO) as a branching agent. Such clusters can undergo a thermal decomposition into linear chains at temperatures higher than 100°C. The thermal decomposition was studied by a combination of static and dynamic laser light scattering (LLS). The time dependence of the weight-average molar mass (M w), the root-mean-square z-average radius of gyration (〈R g 2 〉 z 1/2 ), and the average hydrodynamic radius (〈Rh〉) was used to monitor the decomposition kinetics and cluster structure. It has been found that Mw ∝ t -R , and the decomposition can be roughly divided into three stages; namely, from large clusters to smaller ones; from smaller clusters to less-branched ones; and finally to short linear chains. The scaling of 〈Rg〉 ∝ Mw 0.33 ( 0.01 in the first stage indicates that these clusters are uniform in density, which is rare and much different from conventional polymer clusters whose density decreases from center to periphery. Moreover, we observed, for the first time, that 〈Rg〉/〈Rh〉 ∝ Mw -0.20 ( 0.01 , revealing that even for a uniform cluster swollen in a good solvent, its periphery is still more hydrodynamically draining

Identification of Atypical Enteropathogenic Escherichia coli O98 from Golden Snub-Nosed Monkeys with Diarrhea in China

Fecal samples (n = 76) were collected from 38 snub-nosed monkeys (Rhinopithecus roxellana) in Shennongjia National Nature Reserve (China) and examined for the presence of enteropathogenic Escherichia coli (EPEC). The 56 samples originated from 30 free-ranging monkeys on the reserve and 20 samples from 8 captive monkeys that were previously rescued and kept at the research center. Eight diarrhea samples were collected from four of the eight captive monkeys (two samples from each monkey), and two EPEC strains (2.6%) (95% confidence interval 0.3–9.2%) were isolated from two fecal samples from two diarrheic monkeys. Both strains belonged to serotype O98 and phylogenetic group D (TspE4C2+, ChuA+). The virulence gene detection identified these strains as an atypical EPEC (aEPEC) (bfpB–, stx1–, and stx2–) with the subtype eae+, escV+, and intiminβ+. These strains were highly sensitive to all the antibiotics tested. The lethal dose 50% of the two isolates in Kunming mice was 7.40 × 108 CFU/0.2 mL and 2.40 × 108 CFU/0.2 mL, respectively, indicating low virulence. Based on the report that this serotype had been isolated from some other non-human animals and humans with diarrhea, the first identification of aEPEC O98 strains and their drug resistance profile in R. roxellana is of ecological significance for disease control in this endangered species

Mycoplasma bovis MBOV_RS02825 Encodes a Secretory Nuclease Associated with Cytotoxicity

This study aimed to determine the activity of one Mycoplasma bovis nuclease encoded by MBOV_RS02825 and its association with cytotoxicity. The bioinformatics analysis predicted that it encodes a Ca2+-dependent nuclease based on existence of enzymatic sites in a TNASE_3 domain derived from a Staphylococcus aureus thermonuclease (SNc). We cloned and purified the recombinant MbovNase (rMbovNase), and demonstrated its nuclease activity by digesting bovine macrophage linear DNA and RNA, and closed circular plasmid DNA in the presence of 10 mM Ca2+ at 22–65 °C. In addition, this MbovNase was localized in membrane and rMbovNase able to degrade DNA matrix of neutrophil extracellular traps (NETs). When incubated with macrophages, rMbovNase bound to and invaded the cells localizing to both the cytoplasm and nuclei. These cells experienced apoptosis and the viability was significantly reduced. The apoptosis was confirmed by activated expression of phosphorylated NF-κB p65 and Bax, and inhibition of Iκβα and Bcl-2. In contrast, rMbovNaseΔ181–342 without TNASE_3 domain exhibited deficiency in all the biological functions. Furthermore, rMbovNase was also demonstrated to be secreted. In conclusion, it is a first report that MbovNase is an active nuclease, both secretory and membrane protein with ability to degrade NETs and induce apoptosis

Comparative Genomics of Mycoplasma bovis Strains Reveals That Decreased Virulence with Increasing Passages Might Correlate with Potential Virulence-Related Factors

Mycoplasma bovis is an important cause of bovine respiratory disease worldwide. To understand its virulence mechanisms, we sequenced three attenuated M. bovis strains, P115, P150, and P180, which were passaged in vitro 115, 150, and 180 times, respectively, and exhibited progressively decreasing virulence. Comparative genomics was performed among the wild-type M. bovis HB0801 (P1) strain and the P115, P150, and P180 strains, and one 14.2-kb deleted region covering 14 genes was detected in the passaged strains. Additionally, 46 non-sense single-nucleotide polymorphisms and indels were detected, which confirmed that more passages result in more mutations. A subsequent collective bioinformatics analysis of paralogs, metabolic pathways, protein-protein interactions, secretory proteins, functionally conserved domains, and virulence-related factors identified 11 genes that likely contributed to the increased attenuation in the passaged strains. These genes encode ascorbate-specific phosphotransferase system enzyme IIB and IIA components, enolase, L-lactate dehydrogenase, pyruvate kinase, glycerol, and multiple sugar ATP-binding cassette transporters, ATP binding proteins, NADH dehydrogenase, phosphate acetyltransferase, transketolase, and a variable surface protein. Fifteen genes were shown to be enriched in 15 metabolic pathways, and they included the aforementioned genes encoding pyruvate kinase, transketolase, enolase, and L-lactate dehydrogenase. Hydrogen peroxide (H2O2) production in M. bovis strains representing seven passages from P1 to P180 decreased progressively with increasing numbers of passages and increased attenuation. However, eight mutants specific to eight individual genes within the 14.2-kb deleted region did not exhibit altered H2O2 production. These results enrich the M. bovis genomics database, and they increase our understanding of the mechanisms underlying M. bovis virulence