Eleven strains of Pseudomonas were isolated by selective enrichment on 4-nitrotoluene (4NT). They all utilized 4NT, 4-nitrobenzyl alcohol (4NBA) or 4-nitrobenzoate (4NBZate) as sole sources of carbon and nitrogen, One strain, TW3, was used for more detailed studies. 4NT-grown cells of TW3 take up 0, when incubated in the presence of 4NBA, 4-nitrobenzaldehyde (4NBZ) and 4NBZate. HPLC analysis of culture supernatants showed that 4NBZ and 4NBZate were formed when 4NT-grown cells were incubated with 4NBA, whereas only 4NBZate was found when they were incubated with 4NBZ. Two dehydrogenases were detected in extracts of 4NT-grown cells. 4NBA dehydrogenase could be assayed by a dye-linked assay whereas 4NBZ dehydrogenase activity was linked to NAD+ reduction. No nitrite was detected in supernatants of 4NBZate-grown cells incubated with 4NBZate but the nitrogen appeared as ammonium. The only aromatic ring-cleavage dioxygenase that was induced during growth on the nitroaromatics was protocatechuate 3,4-dioxygenase. It is proposed that the pathway for 4NT catabolism proceeds via 4NBA, 4NBZ and 4NBZate and ultimately to protocatechuate with release of the nitro group as ammonium.

XF-73 is a dicationic porphyrin drug with rapid Gram-positive antibacterial activity currently undergoing clinical trials for the nasal decolonization of Staphylococcus aureus, including methicillin-resistant Staphylococcus aureus (MRSA). In multistep (55-passage) resistance selection studies in the presence of subinhibitory concentrations of XF-73, retapamulin, mupirocin, fusidic acid, and vancomycin against four Network on Antimicrobial Resistance in Staphylococcus aureus MRSA strains, there was no >4-fold increase in the MIC for XF-73 after 55 passages. In contrast, there was an increase in the MICs for retapamulin (from 0.25 g/ml to 4 to 8 g/ml), for mupirocin (from 0.12 g/ml to 16 to 512 g/ml), for fusidic acid (from 0.12 g/ml to 256 g/ml), and for vancomycin (from 1 g/ml to 8 g/ml in two of the four strains tested). Further investigations using S. aureus NRS384 (USA300) and daptomycin demonstrated a 64-fold increase in the MIC after 55 passages (from 0.5 g/ml to 32 g/ml) with a >4-fold increase in the MIC obtained after only five passages. Sequencing analysis of selected isolates confirmed previously reported point mutations associated with daptomycin resistance. No cross-resistance to XF-73 was observed with the daptomycin-resistant strains, suggesting that whereas the two drugs act on the bacterial cell membrane, their specific site of action differs. XF-73 thus represents the first in a new class of antibacterial drugs, which (unlike the comparator antibiotics) after 55 passages exhibited a <4-fold increase in MIC against the strains tested. Antibacterial drugs with a low propensity for inducing bacterial resistance are much needed for the prevention and treatment of multidrugresistant bacteria both within and outside the hospital setting.

International audienceLittle is known about the senescent phenotype of human vascular smooth muscle cells (VSMCs) and the potential involvement of senescent VSMCs in age-related vascular disease, such as atherosclerosis. As such, VSMCs were grown and characterised to generate senescent VSMCs needed for microarray analysis (Affymetrix). Comparative analysis of the transcriptome profiles of early (14 CPD) and late (39-42 CPD) passage VSMCs found a total of 327 probesets called as differentially expressed: 149 are up-regulated in senescence and 178 repressed (-value < 0.5%, minimum effect size of at least 2-fold differential regulation, explore data at www.madras.cf.ac.uk/vsmc). Data mining shows a differential regulation of genes at senescence associated with the development of atherosclerosis and vascular calcification. These included genes with roles in inflammation (IL1β, IL8, ICAM1, TNFAP3, ESM1 and CCL2), tissue remodelling (VEGF, VEGFβ, ADM and MMP14) and vascular calcification (MGP, BMP2, SPP1, OPG and DCN). The microarray data for IL1β, IL8 and MGP were validated by either, ELISA, Western blot analysis or RT-PCR. These data thus provide the first evidence for a role of VSMC senescence in the development of vascular calcification and provides further support for the involvement of senescent VSMCs in the progression of atherosclerosis

XF-70 and XF-73 remained highly active against various forms of slow-growing or non-dividing S. aureus. The results support the hypothesis that membrane-active agents may be particularly effective in eradicating slow- or non-growing bacteria and suggest that XF-70 and XF-73 could be utilized to treat staphylococcal infections where the organisms are only dividing slowly, such as biofilm-associated infections of prosthetic devices.

XF-73 exhibited rapid membrane-perturbing activity, which is likely to be responsible for inhibition of macromolecular synthesis and the death of staphylococci exposed to the drug.