Bacteria can proliferate perpetually without ageing, but they also face conditions where they must persist. Mycobacteria can survive for a long period. This state appears during mycobacterial diseases such as tuberculosis and leprosy, which are chronic and develop after long-term persistent infections. However, the fundamental mechanisms of the long-term living of mycobacteria are unknown. Every Mycobacterium species expresses Mycobacterial DNA-binding protein 1 (MDP1), a histone-like nucleoid associated protein. Mycobacterium smegmatis is a saprophytic fast grower and used as a model of mycobacterial persistence, since it shares the characteristics of the long-term survival observed in pathogenic mycobacteria. Here we show that MDP1-deficient M. smegmatis dies more rapidly than the parental strain after entering stationary phase. Proteomic analyses revealed 21 upregulated proteins with more than 3-fold in MDP1-deficient strain, including DnaA, a replication initiator, NDH, a NADH dehydrogenase that catalyzes downhill electron transfer, Fas1, a critical fatty acid synthase, and antioxidants such as AhpC and KatG. Biochemical analyses showed elevated levels of DNA and ATP syntheses, a decreased NADH/NAD+ ratio, and a loss of resistance to oxidative stress in the MDP1-knockout strain. This study suggests the importance of MDP1-dependent simultaneous control of the cellular functions in the long-term survival of mycobacteria.

This study reexamined the kinetics of the condensation reactions of hydroxymethylphenols with the purpose of elucidating the reaction mechanisms. This report discusses experimental results on the self-condensations of 2,4-dihydroxymethylphenols (2,4-DHMP) and 2,4,6trihydroxymethylphenol (THMP), focusing on the order of reaction. The relations between the initial rates of reaction and the initial concentrations of reactants were investigated. Results quite different from those of previous reports were obtained. The order of reaction of the selfcondensation of 2,4-DHMP was found to be 1.1, which did not change with the alkali/2,4-DHMP molar ratio. The order of reaction of the self-condensation of THMP was found to vary with both the concentration of THMP and the alkali/THMP molar ratio. In the region of THMP concentrations above 1.5 mol/1, the order of reaction was confirmed to be 2.0, which did not change with the alkali/THMP molar ratio. In the region of THMP concentrations below 1.0 mol/1, the order of reaction varied with the alkali/THMP molar ratio, showing fractional numbers of 1.2-1.6. These results indicate that unimolecular reaction(s) and bimolecular reaction(s) take place simultaneously as the ratedetermining step in the condensation reactions of 2,4-DHMP and THMP and that the reaction mechanism changes with the species of reactant and, in some cases, with the reaction conditions.

Tuberculosis causes the highest mortality among all single infections. Asymptomatic tuberculosis, afflicting one third of the global human population, is the major source as 5–10% of asymptomatic cases develop active tuberculosis during their lifetime. Thus it is one of important issues to develop diagnostic tools for accurately detecting asymptomatic infection. Mycobacterial DNA-binding protein 1 (MDP1) is a major protein in persistent Mycobacterium tuberculosis and has potential for diagnostic use in detecting asymptomatic infection. However, a previous ELISA-based study revealed a specificity problem; IgGs against MDP1 were detected in both M. tuberculosis-infected and uninfected individuals. Although the tertiary structures of an antigen are known to influence antibody recognition, the MDP1 structural details have not yet been investigated. The N-terminal half of MDP1, homologous to bacterial histone-like protein HU, is predicted to be responsible for DNA-binding, while the C-terminal half is assumed as totally intrinsically disordered regions. To clarify the relationship between the MDP1 tertiary structure and IgG recognition, we refined the purification method, which allow us to obtain a recombinant protein with the predicted structure. Furthermore, we showed that an IgG-ELISA using MDP1 purified by our refined method is indeed useful in the detection of asymptomatic tuberculosis.

Three patients presented with severe spontaneous pain, allodynia and numbness on the lateral side of the left heal, foot and/or toe due to L5 and/or S1 root injury, as a result of repeated failed back surgeries including Love's surgery and laminaectomy (failed back surgery syndrome). The neuropathic pain in the lower extremities did not respond to somatic nerve block, lumbar-sympathetic ganglion block, spinal cord stimulation, and/or medications. At the spots in the foot showing the most severe allodynia, bones were drilled with fl uoroscopic assistance. Spontaneous pain diminished immediately and allodynia was completely resolved. Visual analogue scale score decreased immediately after bone drilling. The analgesic effect was maintained for 30-45 weeks. In three patients, drilling until the marrow cavity of the bones at painful sites effectively relieved chronic neuropathic pain with lasting analgesic effect.

No abstract